Journal Cover International Journal of Molecular Sciences
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   ISSN (Print) 1661-6596 - ISSN (Online) 1422-0067
   Published by MDPI Homepage  [202 journals]
  • IJMS, Vol. 19, Pages 14: Generation and Characterization of Antibodies
           against Opioid Receptors from Zebrafish

    • Authors: Juan Arévalo, Enrique Hernández-Jiménez, Ada Jiménez-González, María Torres-Valle, Roman Iwasaki, Roger López-Bellido, Cristina Vicente-García, Raquel Rodríguez
      First page: 14
      Abstract: The opioid system is well conserved among species and plays a critical role in pain and addiction systems. The use of zebrafish as an experimental model to study development and genetics is extraordinary and has been proven to be relevant for the study of different diseases. The main drawback to its use for the analysis of different pathologies is the lack of protein tools. Antibodies that work in other models are not suitable for zebrafish due to the low degree of homology that exists among the opioid receptor protein sequences in different species. Here we report the successful generation and characterization of antibodies against the mu, delta 1 and delta 2 opioid receptors in zebrafish. The antibodies obtained, which are specific for each receptor due to the use of the C-terminus as antigens, work for Western blotting and immunohistochemistry. In addition, the antibodies against mu and delta 1 opioid receptors, but not those against delta 2, are able to immunoprecipitate the corresponding receptor from zebrafish lysates. The development of opioid receptor antibodies is an asset to the further study of the endogenous opioid system in zebrafish.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010014
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 33: Chemical Composition and Bioactivity of Essential
           Oil from Blepharocalyx salicifolius

    • Authors: Fabiana Furtado, Bruna Borges, Thaise Teixeira, Hans Garces, Luiz Almeida Junior, Fernanda Alves, Claudio Silva, Ary Fernandes Junior
      First page: 33
      Abstract: Natural products represent a source of biologically active molecules that have an important role in drug discovery. The aromatic plant Blepharocalyx salicifolius has a diverse chemical constitution but the biological activities of its essential oils have not been thoroughly investigated. The aims of this paper were to evaluate in vitro cytotoxic, antifungal and antibacterial activities of an essential oil from leaves of B. salicifolius and to identify its main chemical constituents. The essential oil was extracted by steam distillation, chemical composition was determined by gas chromatography/mass spectrometry, and biological activities were performed by a microdilution broth method. The yield of essential oil was 0.86% (w/w), and the main constituents identified were bicyclogermacrene (17.50%), globulol (14.13%), viridiflorol (8.83%), γ-eudesmol (7.89%) and α-eudesmol (6.88%). The essential oil was cytotoxic against the MDA-MB-231 (46.60 μg·mL−1) breast cancer cell line, being more selective for this cell type compared to the normal breast cell line MCF-10A (314.44 μg·mL−1). Flow cytometry and cytotoxicity results showed that this oil does not act by inducing cell death, but rather by impairment of cellular metabolism specifically of the cancer cells. Furthermore, it presented antifungal activity against Paracoccidioides brasiliensis (156.25 μg·mL−1) but was inactive against other fungi and bacteria. Essential oil from B. salicifolius showed promising biological activities and is therefore a source of molecules to be exploited in medicine or by the pharmaceutical industry.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010033
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 88: Differences in the Plasma Proteome of Patients
           with Hypothyroidism before and after Thyroid Hormone Replacement: A
           Proteomic Analysis

    • Authors: Assim Alfadda, Hicham Benabdelkamel, Afshan Masood, Anwar Jammah, Aishah Ekhzaimy
      First page: 88
      Abstract: Thyroid hormone is a potent stimulator of metabolism, playing a critical role in regulating energy expenditure and in key physiological mechanisms, such as growth and development. Although administration of thyroid hormone in the form of levo thyroxine (l-thyroxine) has been used to treat hypothyroidism for many years, the precise molecular basis of its physiological actions remains uncertain. Our objective was to define the changes in circulating protein levels that characterize alterations in thyroid hormone status. To do this, an integrated untargeted proteomic approach with network analysis was used. This study included 10 age-matched subjects with newly diagnosed overt hypothyroidism. Blood was collected from subjects at baseline and at intervals post-treatment with l-thyroxine until they reached to euthyroid levels. Plasma protein levels were compared by two-dimensional difference in gel electrophoresis (2D-DIGE) pre- and post-treatment. Twenty differentially expressed protein spots were detected. Thirteen were identified, and were found to be unique protein sequences by MALDI-TOF mass spectrometry. Ten proteins were more abundant in the hypothyroid vs. euthyroid state: complement C2, serotransferrin, complement C3, Ig κ chain C region, α-1-antichymotrypsin, complement C4-A, haptoglobin, fibrinogen α chain, apolipoprotein A-I, and Ig α-1 chain C region. Three proteins were decreased in abundance in the hypothyroid vs. euthyroid state: complement factor H, paraneoplastic antigen-like protein 6A, and α-2-macroglobulin. The differentially abundant proteins were investigated by Ingenuity Pathway Analysis (IPA) to reveal their associations with known biological functions. Their connectivity map included interleukin-6 (IL-6) and tumour necrosis factor α (TNF-α) as central nodes and the pathway identified with the highest score was involved in neurological disease, psychological disorders, and cellular movement. The comparison of the plasma proteome between the hypothyroid vs euthyroid states revealed differences in the abundance of proteins involved in regulating the acute phase response.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010088
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 102: Alpha Lipoic Acid: A Therapeutic Strategy that
           Tend to Limit the Action of Free Radicals in Transplantation

    • Authors: Nella Ambrosi, Diego Guerrieri, Fiorella Caro, Francisco Sanchez, Geraldine Haeublein, Domingo Casadei, Claudio Incardona, Eduardo Chuluyan
      First page: 102
      Abstract: Organ replacement is an option to mitigate irreversible organ damage. This procedure has achieved a considerable degree of acceptance. However, several factors significantly limit its effectiveness. Among them, the initial inflammatory graft reaction due to ischemia-reperfusion injury (IRI) has a fundamental influence on the short and long term organ function. The reactive oxygen species (ROS) produced during the IRI actively participates in these adverse events. Therapeutic strategies that tend to limit the action of free radicals could result in beneficial effects in transplantation outcome. Accordingly, the anti-oxidant α-lipoic acid (ALA) have been proved to be protective in several animal experimental models and humans. In a clinical trial, ALA was found to decrease hepatic IRI after hepatic occlusion and resection. Furthermore, the treatment of cadaveric donor and recipient with ALA had a protective effect in the short-term outcome in simultaneous kidney and pancreas transplanted patients. These studies support ALA as a drug to mitigate the damage caused by IRI and reinforce the knowledge about the deleterious consequences of ROS on graft injury in transplantation. The goal of this review is to overview the current knowledge about ROS in transplantation and the use of ALA to mitigate it.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010102
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 110: The Role of Microglia in Diabetic Retinopathy:
           Inflammation, Microvasculature Defects and Neurodegeneration

    • Authors: Christine Altmann, Mirko Schmidt
      First page: 110
      Abstract: Diabetic retinopathy is a common complication of diabetes mellitus, which appears in one third of all diabetic patients and is a prominent cause of vision loss. First discovered as a microvascular disease, intensive research in the field identified inflammation and neurodegeneration to be part of diabetic retinopathy. Microglia, the resident monocytes of the retina, are activated due to a complex interplay between the different cell types of the retina and diverse pathological pathways. The trigger for developing diabetic retinopathy is diabetes-induced hyperglycemia, accompanied by leukostasis and vascular leakages. Transcriptional changes in activated microglia, mediated via the nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB) and extracellular signal–regulated kinase (ERK) signaling pathways, results in release of various pro-inflammatory mediators, including cytokines, chemokines, caspases and glutamate. Activated microglia additionally increased proliferation and migration. Among other consequences, these changes in microglia severely affected retinal neurons, causing increased apoptosis and subsequent thinning of the nerve fiber layer, resulting in visual loss. New potential therapeutics need to interfere with these diabetic complications even before changes in the retina are diagnosed, to prevent neuronal apoptosis and blindness in patients.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010110
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 111: Insights into the Impact of Linker Flexibility
           and Fragment Ionization on the Design of CK2 Allosteric Inhibitors:
           Comparative Molecular Dynamics Simulation Studies

    • Authors: Yue Zhou, Na Zhang, Xiaoqian Qi, Shan Tang, Guohui Sun, Lijiao Zhao, Rugang Zhong, Yongzhen Peng
      First page: 111
      Abstract: Protein kinase is a novel therapeutic target for human diseases. The off-target and side effects of ATP-competitive inhibitors preclude them from the clinically relevant drugs. The compounds targeting the druggable allosteric sites outside the highly conversed ATP binding pocket have been identified as promising alternatives to overcome current barriers of ATP-competitive inhibitors. By simultaneously interacting with the αD region (new allosteric site) and sub-ATP binding pocket, the attractive compound CAM4066 was named as allosteric inhibitor of CK2α. It has been demonstrated that the rigid linker and non-ionizable substituted fragment resulted in significant decreased inhibitory activities of compounds. The molecular dynamics simulations and energy analysis revealed that the appropriate coupling between the linker and pharmacophore fragments were essential for binding of CAM4066 with CK2α. The lower flexible linker of compound 21 lost the capability of coupling fragments A and B to αD region and positive area, respectively, whereas the methyl benzoate of fragment B induced the re-orientated Pre-CAM4066 with the inappropriate polar interactions. Most importantly, the match between the optimized linker and pharmacophore fragments is the challenging work of fragment-linking based drug design. These results provide rational clues to further structural modification and development of highly potent allosteric inhibitors of CK2.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010111
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 113: Rosiglitazone as a Modulator of TLR4 and TLR3
           Signaling Pathways in Rat Primary Neurons and Astrocytes

    • Authors: Dmitry Chistyakov, Nadezda Azbukina, Alexandr Lopachev, Ksenia Kulichenkova, Alina Astakhova, Marina Sergeeva
      First page: 113
      Abstract: An antidiabetic drug of the thiazolidinedione class, rosiglitazone (RG) demonstrates anti-inflammatory properties in various brain pathologies. The mechanism of RG action in brain cells is not fully known. To unravel mechanisms of RG modulation of toll-like receptor (TLR) signaling pathways, we compare primary rat neuron and astrocyte cultures stimulated with the TLR4 agonist lipopolysaccharide (LPS) and the TLR3 agonist poly I:C (PIC). Both TLR agonists induced tumor necrosis factor (TNFα) release in astrocytes, but not in neurons. Neurons and astrocytes released interleukin-10 (IL-10) and prostaglandin E2 (PGE2) in response to LPS and PIC. RG decreased TLR-stimulated TNFα release in astrocytes as well as potentiated IL-10 and PGE2 release in both astrocytes and neurons. RG induced phosphorylation of p38 and JNK MAPK (mitogen-activated protein kinase) in neurons. The results reveal new role of RG as a modulator of resolution of neuroinflammation.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010113
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 114: Sphingolipids in Ventilator Induced Lung Injury:
           Role of Sphingosine-1-Phosphate Lyase

    • Authors: Vidyani Suryadevara, Panfeng Fu, David Ebenezer, Evgeny Berdyshev, Irina Bronova, Long Huang, Anantha Harijith, Viswanathan Natarajan
      First page: 114
      Abstract: Mechanical ventilation (MV) performed in respiratory failure patients to maintain lung function leads to ventilator-induced lung injury (VILI). This study investigates the role of sphingolipids and sphingolipid metabolizing enzymes in VILI using a rodent model of VILI and alveolar epithelial cells subjected to cyclic stretch (CS). MV (0 PEEP (Positive End Expiratory Pressure), 30 mL/kg, 4 h) in mice enhanced sphingosine-1-phosphate lyase (S1PL) expression, and ceramide levels, and decreased S1P levels in lung tissue, thereby leading to lung inflammation, injury and apoptosis. Accumulation of S1P in cells is a balance between its synthesis catalyzed by sphingosine kinase (SphK) 1 and 2 and catabolism mediated by S1P phosphatases and S1PL. Thus, the role of S1PL and SphK1 in VILI was investigated using Sgpl1+/− and Sphk1−/− mice. Partial genetic deletion of Sgpl1 protected mice against VILI, whereas deletion of SphK1 accentuated VILI in mice. Alveolar epithelial MLE-12 cells subjected to pathophysiological 18% cyclic stretch (CS) exhibited increased S1PL protein expression and dysregulation of sphingoid bases levels as compared to physiological 5% CS. Pre-treatment of MLE-12 cells with S1PL inhibitor, 4-deoxypyridoxine, attenuated 18% CS-induced barrier dysfunction, minimized cell apoptosis and cytokine secretion. These results suggest that inhibition of S1PL that increases S1P levels may offer protection against VILI.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010114
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 115: Transient Expression and Purification of
           Horseradish Peroxidase C in Nicotiana benthamiana

    • Authors: Suzanne Huddy, Inga Hitzeroth, Ann Meyers, Brandon Weber, Edward Rybicki
      First page: 115
      Abstract: Horseradish peroxidase (HRP) is a commercially important reagent enzyme used in molecular biology and in the diagnostic product industry. It is typically purified from the roots of the horseradish (Armoracia rusticana); however, this crop is only available seasonally, yields are variable and often low, and the product is a mixture of isoenzymes. Engineering high-level expression in transiently transformed tobacco may offer a solution to these problems. In this study, a synthetic Nicotiana benthamiana codon-adapted full-length HRP isoenzyme gene as well as C-terminally truncated and both N- and C-terminally truncated versions of the HRP C gene were synthesized, and their expression in N. benthamiana was evaluated using an Agrobacterium tumefaciens-mediated transient expression system. The influence on HRP C expression levels of co-infiltration with a silencing suppressor (NSs) construct was also evaluated. Highest HRP C levels were consistently obtained using either the full length or C-terminally truncated HRP C constructs. HRP C purification by ion exchange chromatography gave an overall yield of 54% with a Reinheitszahl value of >3 and a specific activity of 458 U/mg. The high level of HRP C production in N. benthamiana in just five days offers an alternative, viable, and scalable system for production of this commercially significant enzyme.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010115
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 116: Candidate Genes and Molecular Markers Correlated
           to Physiological Traits for Heat Tolerance in Fine Fescue Cultivars

    • Authors: Yi Xu, Jinyu Wang, Stacy Bonos, William Meyer, Bingru Huang
      First page: 116
      Abstract: Heat stress is one of the major abiotic factors limiting the growth of cool-season grass species during summer season. The objectives of this study were to assess genetic variations in the transcript levels of selected genes in fine fescue cultivars differing in heat tolerance, and to identify single nucleotide polymorphism (SNP) markers associated with candidate genes related to heat tolerance. Plants of 26 cultivars of five fine fescue species (Festuca spp.) were subjected to heat stress (38/33 °C, day/night temperature) in controlled environmental growth chambers. Physiological analysis including leaf chlorophyll content, photochemical efficiency, and electrolyte leakage demonstrated significant genetic variations in heat tolerance among fine fescue cultivars. The transcript levels of selected genes involved in photosynthesis (RuBisCO activase, Photosystem II CP47 reaction center protein), carbohydrate metabolism (Sucrose synthase), energy production (ATP synthase), growth regulation (Actin), oxidative response (Catalase), and stress protection (Heat shock protein 90) were positively correlated with the physiological traits for heat tolerance. SNP markers for those candidate genes exhibited heterozygosity, which could also separate heat-sensitive and heat-tolerant cultivars into clusters. The development of SNP markers for candidate genes in heat tolerance may allow marker-assisted breeding for the development of new heat-tolerant cultivars in fine fescue and other cool-season grass species.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-01
      DOI: 10.3390/ijms19010116
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 117: Theaflavin-3,3′-Digallate Enhances the
           Inhibitory Effect of Cisplatin by Regulating the Copper Transporter 1 and
           Glutathione in Human Ovarian Cancer Cells

    • Authors: Haibo Pan, Eunhye Kim, Gary Rankin, Yon Rojanasakul, Youying Tu, Yi Chen
      First page: 117
      Abstract: Ovarian cancer has the highest fatality rate among the gynecologic cancers. The side effects, high relapse rate, and drug resistance lead to low long-term survival rate (less than 40%) of patients with advanced ovarian cancer. Theaflavin-3,3′-digallate (TF3), a black tea polyphenol, showed less cytotoxicity to normal ovarian cells than ovarian cancer cells. We aimed to investigate whether TF3 could potentiate the inhibitory effect of cisplatin against human ovarian cancer cell lines. In the present study, combined treatment with TF3 and cisplatin showed a synergistic cytotoxicity against A2780/CP70 and OVCAR3 cells. Treatment with TF3 could increase the intracellular accumulation of platinum (Pt) and DNA-Pt adducts and enhanced DNA damage induced by cisplatin in both cells. Treatment with TF3 decreased the glutathione (GSH) levels and upregulated the protein levels of the copper transporter 1 (CTR1) in both cells, which led to the enhanced sensitivity of both ovarian cancer cells to cisplatin. The results imply that TF3 might be used as an adjuvant to potentiate the inhibitory effect of cisplatin against advanced ovarian cancer.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010117
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 118: Seed Transcriptomics Analysis in Camellia
           oleifera Uncovers Genes Associated with Oil Content and Fatty Acid
           Composition

    • Authors: Ping Lin, Kailiang Wang, Changfu Zhou, Yunhai Xie, Xiaohua Yao, Hengfu Yin
      First page: 118
      Abstract: Camellia oleifera is a major tree species for producing edible oil. Its seed oil is well known for the high level of oleic acids; however, little is known regarding the molecular mechanism of lipid biosynthesis in C. oleifera. Here, we measured the oil contents and fatty acid (FA) compositions at four developmental stages and investigated the global gene expression profiles through transcriptomics sequencing. We identified differentially-expressed genes (DEGs) among the developmental stages and found that the distribution of numbers of DEGs was associated with the accumulation pattern of seed oil. Gene Ontology (GO) enrichment analysis revealed some critical biological processes related to oil accumulation, including lipid metabolism and phosphatidylcholine metabolism. Furthermore, we investigated the expression patterns of lipid biosynthesis genes. We showed that most of the genes were identified with single or multiple copies, and some had correlated profiles along oil accumulation. We proposed that the higher levels of stearoyl-ACP desaturases (SADs) coupled with lower activities of fatty acid desaturase 2 (FAD2) might be responsive to the boost of oleic acid at the late stage of C. oleifera seeds’ development. This work presents a comprehensive transcriptomics study of C. oleifera seeds and uncovers valuable DEGs that are associated with the seed oil accumulation.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010118
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 119: Vitamin D Status and the Relationship with Bone
           Fragility Fractures in HIV-Infected Patients: A Case Control Study

    • Authors: Marco Atteritano, Luigi Mirarchi, Emmanuele Venanzi-Rullo, Domenico Santoro, Chiara Iaria, Antonino Catalano, Antonino Lasco, Vincenzo Arcoraci, Alberto Lo Gullo, Alessandra Bitto, Francesco Squadrito, Antonio Cascio
      First page: 119
      Abstract: HIV-infected patients show high risk of fracture. The aims of our study were to determine the prevalence of vertebral fractures (VFs) and their associations with vitamin D in HIV patients. 100 patients with HIV infection and 100 healthy age- and sex-matched controls were studied. Bone mineral density was measured by quantitative ultrasound at the non-dominant heel. Serum osteocalcin and C-terminal telopeptide of collagen type 1 served as bone turnover markers. Bone ultrasound measurements were significantly lower in patients compared with controls (Stiffness Index (SI): 80.58 ± 19.95% vs. 93.80 ± 7.10%, respectively, p < 0.001). VFs were found in 16 patients and in 2 controls. HIV patients with vertebral fractures showed lower stiffness index (SI) (70.75 ± 10.63 vs. 83.36 ± 16.19, respectively, p = 0.045) and lower vitamin D levels (16.20 ± 5.62 vs. 28.14 ± 11.94, respectively, p < 0.02). The majority of VFs (87.5%) were observed in HIV-infected patients with vitamin D insufficiency, and regression analysis showed that vitamin D insufficiency was significantly associated with vertebral fractures (OR 9.15; 95% CI 0.18–0.52, p < 0.04). VFs and are a frequent occurrence in HIV-infected patients and may be associated with vitamin D insufficiency.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010119
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 120: Overview of the Microenvironment of Vasculature
           in Vascular Tone Regulation

    • Authors: Yean Loh, Chu Tan, Yung Ch’ng, Zhao Yeap, Chiew Ng, Mun Yam
      First page: 120
      Abstract: Hypertension is asymptomatic and a well-known “silent killer”, which can cause various concomitant diseases in human population after years of adherence. Although there are varieties of synthetic antihypertensive drugs available in current market, their relatively low efficacies and major application in only single drug therapy, as well as the undesired chronic adverse effects associated, has drawn the attention of worldwide scientists. According to the trend of antihypertensive drug evolution, the antihypertensive drugs used as primary treatment often change from time-to-time with the purpose of achieving the targeted blood pressure range. One of the major concerns that need to be accounted for here is that the signaling mechanism pathways involved in the vasculature during the vascular tone regulation should be clearly understood during the pharmacological research of antihypertensive drugs, either in vitro or in vivo. There are plenty of articles that discussed the signaling mechanism pathways mediated in vascular tone in isolated fragments instead of a whole comprehensive image. Therefore, the present review aims to summarize previous published vasculature-related studies and provide an overall depiction of each pathway including endothelium-derived relaxing factors, G-protein-coupled, enzyme-linked, and channel-linked receptors that occurred in the microenvironment of vasculature with a full schematic diagram on the ways their signals interact. Furthermore, the crucial vasodilative receptors that should be included in the mechanisms of actions study on vasodilatory effects of test compounds were suggested in the present review as well.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010120
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 121: Expression and Activity of COX-1 and COX-2 in
           Acanthamoeba sp.-Infected Lungs According to the Host Immunological Status
           

    • Authors: Natalia Łanocha-Arendarczyk, Irena Baranowska-Bosiacka, Karolina Kot, Izabela Gutowska, Agnieszka Kolasa-Wołosiuk, Dariusz Chlubek, Danuta Kosik-Bogacka
      First page: 121
      Abstract: Little is known about the pathomechanism of pulmonary infections caused by Acanthamoeba sp. Therefore, the aim of this study was to determine whether Acanthamoeba sp. may affect the expression and activity of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2), resulting in the altered levels of their main products, prostaglandins (PGE2) and thromboxane B2 (TXB2), in lungs of immunocompetent or immunosuppressed hosts. Acanthamoeba sp. induced a strong expression of COX-1 and COX-2 proteins in the lungs of immunocompetent mice, which, however, did not result in significant differences in the expression of PGE2 and TXB2. Our immunohistochemical analysis showed that immunosuppression induced by glucocorticoids in Acanthamoeba sp.-infected mice caused a decrease in COX-1 and COX-2 (not at the beginning of infection) in lung tissue. These results suggest that similar to COX-2, COX-1 is an important mediator of the pathophysiology in experimental pulmonary acanthamoebiasis. We suggest that the signaling pathways important for Acanthamoeba sp. induction of lung infection might interact with each other and depend on the host immune status.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010121
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 122: Individualizing Treatment Approaches for
           Epileptic Patients with Glucose Transporter Type1 (GLUT-1) Deficiency

    • Authors: Armond Daci, Adnan Bozalija, Fisnik Jashari, Shaip Krasniqi
      First page: 122
      Abstract: Monogenic and polygenic mutations are important contributors in patients suffering from epilepsy, including metabolic epilepsies which are inborn errors of metabolism with a good respond to specific dietetic treatments. Heterozygous variation in solute carrier family 2, facilitated glucose transporter member 1 (SLC2A1) and mutations of the GLUT1/SLC2A2 gene results in the failure of glucose transport, which is related with a glucose type-1 transporter (GLUT1) deficiency syndrome (GLUT1DS). GLUT1 deficiency syndrome is a treatable disorder of glucose transport into the brain caused by a variety of mutations in the SLC2A1 gene which are the cause of different neurological disorders also with different types of epilepsy and related clinical phenotypes. Since patients continue to experience seizures due to a pharmacoresistance, an early clinical diagnosis associated with specific genetic testing in SLC2A1 pathogenic variants in clinical phenotypes could predict pure drug response and might improve safety and efficacy of treatment with the initiation of an alternative energy source including ketogenic or analog diets in such patients providing individualized strategy approaches.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010122
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 123: Natural Antisense Transcripts: Molecular
           Mechanisms and Implications in Breast Cancers

    • Authors: Guillaume Latgé, Christophe Poulet, Vincent Bours, Claire Josse, Guy Jerusalem
      First page: 123
      Abstract: Natural antisense transcripts are RNA sequences that can be transcribed from both DNA strands at the same locus but in the opposite direction from the gene transcript. Because strand-specific high-throughput sequencing of the antisense transcriptome has only been available for less than a decade, many natural antisense transcripts were first described as long non-coding RNAs. Although the precise biological roles of natural antisense transcripts are not known yet, an increasing number of studies report their implication in gene expression regulation. Their expression levels are altered in many physiological and pathological conditions, including breast cancers. Among the potential clinical utilities of the natural antisense transcripts, the non-coding coding transcript pairs are of high interest for treatment. Indeed, these pairs can be targeted by antisense oligonucleotides to specifically tune the expression of the coding-gene. Here, we describe the current knowledge about natural antisense transcripts, their varying molecular mechanisms as gene expression regulators, and their potential as prognostic or predictive biomarkers in breast cancers.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010123
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 124: Extracorporeal Shock Wave Therapy Alters the
           Expression of Fibrosis-Related Molecules in Fibroblast Derived from Human
           Hypertrophic Scar

    • Authors: Hui Cui, A Ram Hong, June-Bum Kim, Joo Yu, Yoon Cho, So Joo, Cheong Seo
      First page: 124
      Abstract: Extracorporeal shock wave therapy (ESWT) considerably improves the appearance and symptoms of post-burn hypertrophic scars (HTS). However, the mechanism underlying the observed beneficial effects is not well understood. The objective of this study was to elucidate the mechanism underlying changes in cellular and molecular biology that is induced by ESWT of fibroblasts derived from scar tissue (HTSFs). We cultured primary dermal fibroblasts derived from human HTS and exposed these cells to 1000 impulses of 0.03, 0.1, and 0.3 mJ/mm2. At 24 h and 72 h after treatment, real-time PCR and western blotting were used to detect mRNA and protein expression, respectively, and cell viability and mobility were assessed. While HTSF viability was not affected, migration was decreased by ESWT. Transforming growth factor beta 1 (TGF-β1) expression was reduced and alpha smooth muscle actin (α-SMA), collagen-I, fibronectin, and twist-1 were reduced significantly after ESWT. Expression of E-cadherin was increased, while that of N-cadherin was reduced. Expression of inhibitor of DNA binding 1 and 2 was increased. In conclusion, suppressed epithelial-mesenchymal transition might be responsible for the anti-scarring effect of ESWT, and has potential as a therapeutic target in the management of post-burn scars.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010124
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 125: The Effects of Kaempferol-Inhibited Autophagy on
           Osteoclast Formation

    • Authors: Chang-Ju Kim, Sang-Hun Shin, Bok-Joo Kim, Chul-Hoon Kim, Jung-Han Kim, Hae-Mi Kang, Bong-Soo Park, In-Ryoung Kim
      First page: 125
      Abstract: Kaempferol, a flavonoid compound, is derived from the rhizome of Kaempferia galanga L., which is used in traditional medicine in Asia. Autophagy has pleiotropic functions that are involved in cell growth, survival, nutrient supply under starvation, defense against pathogens, and antigen presentation. There are many studies dealing with the inhibitory effects of natural flavonoids in bone resorption. However, no studies have explained the relationship between the autophagic and inhibitory processes of osteoclastogenesis by natural flavonoids. The present study was undertaken to investigate the inhibitory effects of osteoclastogenesis through the autophagy inhibition process stimulated by kaempferol in murin macrophage (RAW 264.7) cells. The cytotoxic effect of Kaempferol was investigated by MTT assay. The osteoclast differentiation and autophagic process were confirmed via tartrate-resistant acid phosphatase (TRAP) staining, pit formation assay, western blot, and real-time PCR. Kaempferol controlled the expression of autophagy-related factors and in particular, it strongly inhibited the expression of p62/SQSTM1. In the western blot and real time-PCR analysis, when autophagy was suppressed with the application of 3-Methyladenine (3-MA) only, osteoclast and apoptosis related factors were not significantly affected. However, we found that after cells were treated with kaempferol, these factors inhibited autophagy and activated apoptosis. Therefore, we presume that kaempferol-inhibited autophagy activated apoptosis by degradation of p62/SQSTM1. Further study of the p62/SQSTM1 gene as a target in the autophagy mechanism, may help to delineate the potential role of kaempferol in the treatment of bone metabolism disorders.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010125
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 126: The Potential Roles of the Apoptosis-Related
           Protein PDRG1 in Diapause Embryo Restarting of Artemia sinica

    • Authors: Wan Zhang, Feng Yao, Hong Zhang, Na Li, Xiangyang Zou, Linlin Sui, Lin Hou
      First page: 126
      Abstract: High salinity and low temperatures can induce Artemia sinica to enter the diapause stage during embryonic development. Diapause embryos stop at the gastrula stage, allowing them to resist apoptosis and regulate cell cycle activity to guarantee normal development after diapause termination. P53 and DNA damage-regulated gene 1 (pdrg1) is involved in cellular physiological activities, such as apoptosis, DNA damage repair, cell cycle regulation, and promotion of programmed cell death. However, the role of pdrg1 in diapause and diapause termination in A. sinica remains unknown. Here, the full-length A. sinica pdrg1 cDNA (As-pdrg1) was obtained and found to contain 1119 nucleotides, including a 228 bp open reading frame (ORF), a 233 bp 5′-untranslated region (UTR), and a 658-bp 3′-UTR, which encodes a 75 amino acid protein. In situ hybridization showed no tissue specific expression of As-pdrg1. Quantitative real-time PCR and western blotting analyses of As-pdrg1 gene and protein expression showed high levels at 15–20 h of embryo development and a subsequent downward trend. Low temperatures upregulated As-pdrg1 expression. RNA interference for the pdrg1 gene in Artemia embryos caused significant developmental hysteresis. Thus, PDRG1 plays an important role in diapause termination and cell cycle regulation in early embryonic development of A. sinica.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010126
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 127: Differential Characterization of
           Temozolomide-Resistant Human Glioma Cells

    • Authors: Sheng-Wei Lai, Bor-Ren Huang, Yu-Shu Liu, Hsiao-Yun Lin, Chun-Chuan Chen, Cheng-Fang Tsai, Dah-Yuu Lu, Chingju Lin
      First page: 127
      Abstract: Glioblastoma multiforme (GBM) is the most common type of primary and malignant tumor occurring in the adult central nervous system. Temozolomide (TMZ) has been considered to be one of the most effective chemotherapeutic agents to prolong the survival of patients with glioblastoma. Many glioma cells develop drug-resistance against TMZ that is mediated by increasing O-6-methylguanine-DNA methyltransferase (MGMT) levels. The expression of connexin 43 was increased in the resistant U251 subline compared with the parental U251 cells. The expression of epithelial–mesenchymal transition (EMT)-associated regulators, including vimentin, N-cadherin, and β-catenin, was reduced in the resistant U251 subline. In addition, the resistant U251 subline exhibited decreased cell migratory activity and monocyte adhesion ability compared to the parental U251 cells. Furthermore, the resistant U251 subline also expressed lower levels of vascular cell adhesion molecule (VCAM)-1 after treatment with recombinant tumor necrosis factor (TNF)-α. These findings suggest differential characteristics in the drug-resistant GBM from the parental glioma cells.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010127
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 128: Metal Ion Effects on Aβ and Tau Aggregation

    • Authors: Anne Claire Kim, Sungsu Lim, Yun Kyung Kim
      First page: 128
      Abstract: Amyloid and tau aggregation are implicated in manifold neurodegenerative diseases and serve as two signature pathological hallmarks in Alzheimer’s disease (AD). Though aging is considered as a prominent risk factor for AD pathogenesis, substantial evidence suggests that an imbalance of essential biometal ions in the body and exposure to certain metal ions in the environment can potentially induce alterations to AD pathology. Despite their physiological importance in various intracellular processes, biometal ions, when present in excessive or deficient amounts, can serve as a mediating factor for neurotoxicity. Recent studies have also demonstrated the contribution of metal ions found in the environment on mediating AD pathogenesis. In this regard, the neuropathological features associated with biometal ion dyshomeostasis and environmental metal ion exposure have prompted widespread interest by multiple research groups. In this review, we discuss and elaborate on findings from previous studies detailing the possible role of both endogenous and exogenous metal ions specifically on amyloid and tau pathology in AD.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-02
      DOI: 10.3390/ijms19010128
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 129: Small Myristoylated Protein-3, Identified as a
           Potential Virulence Factor in Leishmania amazonensis, Proves to be a
           Protective Antigen against Visceral Leishmaniasis

    • Authors: Marcelo Oliveira, Vívian Martins, Thaís Santos, Daniela Lage, Fernanda Ramos, Beatriz Salles, Lourena Costa, Daniel Dias, Patrícia Ribeiro, Mônica Schneider, Ricardo Machado-de-Ávila, Antônio Teixeira, Eduardo Coelho, Miguel Chávez-Fumagalli
      First page: 129
      Abstract: In a proteomics approach conducted with Leishmania amazonensis, parasite proteins showed either an increase or a decrease in their expression content during extensive in vitro cultivation, and were related to the survival and the infectivity of the parasites, respectively. In the current study, a computational screening was performed to predict virulence factors among these molecules. Three proteins were selected, one of which presented no homology to human proteins. This candidate, namely small myristoylated protein-3 (SMP-3), was cloned, and its recombinant version (rSMP-3) was used to stimulate peripheral blood mononuclear cells (PBMCs) from healthy subjects living in an endemic area of leishmaniasis and from visceral leishmaniasis patients. Results showed high interferon-γ (IFN-γ) production and low levels of interleukin 10 (IL-10) in the cell supernatants. An in vivo experiment was then conducted on BALB/c mice, which were immunized with rSMP-3/saponin and later challenged with Leishmania infantum promastigotes. The rSMP-3/saponin combination induced high production of protein-specific IFN-γ, IL-12, and granulocyte-macrophage colony-stimulating factor (GM-CSF) by the spleen cells of the immunized mice. This pattern was associated with protection, which was characterized by a significant reduction in the parasite load in distinct organs of the animals. Altogether, these results have revealed that this new virulence factor is immunogenic in both mice and humans, and have proven its protective efficacy against visceral leishmaniasis in a murine model.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010129
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 130: Endothelial Aquaporins and Hypomethylation:
           Potential Implications for Atherosclerosis and Cardiovascular Disease

    • Authors: Inês da Silva, Madalena Barroso, Teresa Moura, Rita Castro, Graça Soveral
      First page: 130
      Abstract: Aquaporins (AQPs) are transmembrane channels that facilitate water and glycerol permeation through cell membranes. Recently, the water channel AQP1 was suggested to contribute to endothelial homeostasis and cardiovascular health. Less is known about endothelial aquaglyceroporins expression and its implication in cardiovascular disease (CVD). We have previously used cultured human endothelial cells under a hypomethylating environment to study endothelial dysfunction and activation, a phenotype implicated in the establishment of atherosclerosis and CVD. Here, we used the same cell model to investigate aquaporin’s expression and function in healthy or pro-atherogenic phenotype. We first confirmed key features of endothelium dysfunction and activation in our cell model, including an augmented endothelial transmigration under hypomethylation. Subsequently, we found AQP1 and AQP3 to be the most predominant AQPs accounting for water and glycerol fluxes, respectively, in the healthy endothelium. Moreover, endothelial hypomethylation led to decreased levels of AQP1 and impaired water permeability without affecting AQP3 and glycerol permeability. Furthermore, TNF-α treatment-induced AQP1 downregulation suggesting that the inflammatory NF-κB signaling pathway mediates AQP1 transcriptional repression in a pro-atherogenic endothelium, a possibility that warrants further investigation. In conclusion, our results add further support to AQP1 as a candidate player in the setting of endothelial dysfunction and CVD.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010130
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 131: Development of Burdock Root Inulin/Chitosan
           Blend Films Containing Oregano and Thyme Essential Oils

    • Authors: Thi Cao, So-Young Yang, Kyung Song
      First page: 131
      Abstract: In this study, inulin (INU) extracted from burdock root was utilized as a new film base material and combined with chitosan (CHI) to prepare composite films. Oregano and thyme essential oils (OT) were incorporated into the INU-CHI film to confer the films with bioactivities. The physical and optical properties as well as antioxidant and antimicrobial activities of the films were evaluated. INU film alone showed poor physical properties. In contrast, the compatibility of INU and CHI demonstrated by the changes in attenuated total reflectance-Fourier transformation infrared spectrum of the INU-CHI film increased tensile strength and elongation at break of the INU film by 8.2- and 3.9-fold, respectively. In addition, water vapor permeability, water solubility, and moisture content of the films decreased proportionally with increasing OT concentration in the INU-CHI film. Incorporation of OT also increased the opacity of a and b values and decreased the L value of the INU-CHI films. All INU-CHI films containing OT exhibited antioxidant and antimicrobial properties. Particularly, the INU-CHI film with 2.0% OT exhibited the highest 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), 2,2-diphenyl-1-picrylhydrazyl radical scavenging, and antimicrobial activities against four pathogens. Thus, the INU-CHI film containing OT developed in this study might be utilized as an active packaging material in the food industry.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010131
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 132: Why Differentiation Therapy Sometimes Fails:
           Molecular Mechanisms of Resistance to Retinoids

    • Authors: Petr Chlapek, Viera Slavikova, Pavel Mazanek, Jaroslav Sterba, Renata Veselska
      First page: 132
      Abstract: Retinoids represent a popular group of differentiation inducers that are successfully used in oncology for treatment of acute promyelocytic leukemia in adults and of neuroblastoma in children. The therapeutic potential of retinoids is based on their key role in the regulation of cell differentiation, growth, and apoptosis, which provides a basis for their use both in cancer therapy and chemoprevention. Nevertheless, patients treated with retinoids often exhibit or develop resistance to this therapy. Although resistance to retinoids is commonly categorized as either acquired or intrinsic, resistance as a single phenotypic feature is usually based on the same mechanisms that are closely related or combined in both of these types. In this review, we summarize the most common changes in retinoid metabolism and action that may affect the sensitivity of a tumor cell to treatment with retinoids. The availability of retinoids can be regulated by alterations in retinol metabolism or in retinoid intracellular transport, by degradation of retinoids or by their efflux from the cell. Retinoid effects on gene expression can be regulated via retinoid receptors or via other molecules in the transcriptional complex. Finally, the role of small-molecular-weight inhibitors of altered cell signaling pathways in overcoming the resistance to retinoids is also suggested.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010132
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 133: Computational Modeling of the Staphylococcal
           Enterotoxins and Their Interaction with Natural Antitoxin Compounds

    • Authors: Mahantesh Kurjogi, Praveen Satapute, Sudisha Jogaiah, Mostafa Abdelrahman, Jayasimha Daddam, Venkatesh Ramu, Lam-Son Tran
      First page: 133
      Abstract: Staphylococcus aureus is an opportunistic bacterium that produces various types of toxins, resulting in serious food poisoning. Staphylococcal enterotoxins (SEs) are heat-stable and resistant to hydrolysis by digestive enzymes, representing a potential hazard for consumers worldwide. In the present study, we used amino-acid sequences encoding SEA and SEB-like to identify their respective template structure and build the three-dimensional (3-D) models using homology modeling method. Two natural compounds, Betulin and 28-Norolean-12-en-3-one, were selected for docking study on the basis of the criteria that they satisfied the Lipinski’s Rule-of-Five. A total of 14 and 13 amino-acid residues were present in the best binding site predicted in the SEA and SEB-like, respectively, using the Computer Atlas of Surface Topology of Proteins (CASTp). Among these residues, the docking study with natural compounds Betulin and 28-Norolean-12-en-3-one revealed that GLN43 and GLY227 in the binding site of the SEA, each formed a hydrogen-bond interaction with 28-Norolean-12-en-3-one; while GLY227 residue established a hydrogen bond with Betulin. In the case of SEB-like, the docking study demonstrated that ASN87 and TYR88 residues in its binding site formed hydrogen bonds with Betulin; whereas HIS59 in the binding site formed a hydrogen-bond interaction with 28-Norolean-12-en-3-one. Our results demonstrate that the toxic effects of these two SEs can be effectively treated with antitoxins like Betulin and 28-Norolean-12-en-3-one, which could provide an effective drug therapy for this pathogen.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010133
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 134: Structure and Chromosomal Organization of Yeast
           Genes Regulated by Topoisomerase II

    • Authors: Ricky Joshi, Christoforos Nikolaou, Joaquim Roca
      First page: 134
      Abstract: Cellular DNA topoisomerases (topo I and topo II) are highly conserved enzymes that regulate the topology of DNA during normal genome transactions, such as DNA transcription and replication. In budding yeast, topo I is dispensable whereas topo II is essential, suggesting fundamental and exclusive roles for topo II, which might include the functions of the topo IIa and topo IIb isoforms found in mammalian cells. In this review, we discuss major findings of the structure and chromosomal organization of genes regulated by topo II in budding yeast. Experimental data was derived from short (10 min) and long term (120 min) responses to topo II inactivation in top-2 ts mutants. First, we discuss how short term responses reveal a subset of yeast genes that are regulated by topo II depending on their promoter architecture. These short term responses also uncovered topo II regulation of transcription across multi-gene clusters, plausibly by common DNA topology management. Finally, we examine the effects of deactivated topo II on the elongation of RNA transcripts. Each study provides an insight into the particular chromatin structure that interacts with the activity of topo II. These findings are of notable clinical interest as numerous anti-cancer therapies interfere with topo II activity.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010134
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 135: Identification of Conserved and Diverse
           Metabolic Shift of the Stylar, Intermediate and Peduncular Segments of
           Cucumber Fruit during Development

    • Authors: Chaoyang Hu, Huiyu Zhao, Wen Wang, Mingfei Xu, Jianxin Shi, Xiangbo Nie, Guiling Yang
      First page: 135
      Abstract: Cucumber (Cucumis sativus L.) is one of the most important vegetables and contains a high content of nutritionally beneficial metabolites. However, little is known about the metabolic variations among different parts of cucumber fruit and their kinetics during growth. In this study, the dynamic metabolic profiles in the stylar end, the intermediate segment and the peduncular end of cucumber fruit during the development were investigated by employing a non-targeted metabolomics approach, where 238 metabolites were identified. Statistical analyses revealed that both development time and tissue type influenced metabolic changes, while development time seemed to exert more effects than tissue type on the cucumber fruit metabolome. The levels of the most of the detected metabolites decreased gradually, while those of some amino acids, carbohydrates and flavonoids increased across development. The metabolomes of the stylar end and the intermediate segment were similar, although all three parts of the cucumber fruit were separated from each other in orthogonal partial least squares projection to latent structures-discriminant analysis (OPLS-DA) plots. Metabolites association analysis revealed that sn-1 and sn-2 lysophospholipids are synthesized via independent pathways in cucumber fruit. In sum, this study demonstrated both conserved and diverse metabolic kinetics of three parts of cucumber fruit, which will facilitate further study of the regulation of cucumber fruit development as well as their potential applications in nutritious quality improvement of cucumber fruit.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010135
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 136: How Sweet Are Our Gut Beneficial Bacteria' A
           Focus on Protein Glycosylation in Lactobacillus

    • Authors: Dimitrios Latousakis, Nathalie Juge
      First page: 136
      Abstract: Protein glycosylation is emerging as an important feature in bacteria. Protein glycosylation systems have been reported and studied in many pathogenic bacteria, revealing an important diversity of glycan structures and pathways within and between bacterial species. These systems play key roles in virulence and pathogenicity. More recently, a large number of bacterial proteins have been found to be glycosylated in gut commensal bacteria. We present an overview of bacterial protein glycosylation systems (O- and N-glycosylation) in bacteria, with a focus on glycoproteins from gut commensal bacteria, particularly Lactobacilli. These emerging studies underscore the importance of bacterial protein glycosylation in the interaction of the gut microbiota with the host.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010136
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 137: The Sodium-Glucose Cotransporter 2 Inhibitor
           Dapagliflozin Prevents Renal and Liver Disease in Western Diet Induced
           Obesity Mice

    • Authors: Dong Wang, Yuhuan Luo, Xiaoxin Wang, David Orlicky, Komuraiah Myakala, Pengyuan Yang, Moshe Levi
      First page: 137
      Abstract: Obesity and obesity related kidney and liver disease have become more prevalent over the past few decades, especially in the western world. Sodium-glucose cotransporter 2 (SGLT2) inhibitors are a new class of antidiabetic agents with promising effects on cardiovascular and renal function. Given SGLT2 inhibitors exert both anti-diabetic and anti-obesity effects by promoting urinary excretion of glucose and subsequent caloric loss, we investigated the effect of the highly selective renal SGLT2 inhibitor dapagliflozin in mice with Western diet (WD) induced obesity. Low fat (LF) diet or WD-fed male C57BL/6J mice were treated with dapagliflozin for 26 weeks. Dapagliflozin attenuated the WD-mediated increases in body weight, plasma glucose and plasma triglycerides. Treatment with dapagliflozin prevented podocyte injury, glomerular pathology and renal fibrosis determined by second harmonic generation (SHG), nephrin, synaptopodin, collagen IV, and fibronectin immunofluorescence microscopy. Oil Red O staining showed dapagliflozin also decreased renal lipid accumulation associated with decreased SREBP-1c mRNA abundance. Moreover, renal inflammation and oxidative stress were lower in the dapagliflozin-treated WD-fed mice than in the untreated WD-fed mice. In addition, dapagliflozin decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), hepatic lipid accumulation as determined by H&E and Oil Red O staining, and Coherent Anti-Stokes Raman Scattering (CARS) microscopy, and hepatic fibrosis as determined by picrosirius red (PSR) staining and TPE-SHG microscopy in WD-fed mice. Thus, our study demonstrated that the co-administration of the SGLT2 inhibitor dapagliflozin attenuates renal and liver disease during WD feeding of mice.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010137
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 138: Identification and Antithrombotic Activity of
           Peptides from Blue Mussel (Mytilus edulis) Protein

    • Authors: Meiling Qiao, Maolin Tu, Zhenyu Wang, Fengjiao Mao, Hui Chen, Lei Qin, Ming Du
      First page: 138
      Abstract: The blue mussel (Mytilus edulis) reportedly contains many bioactive components of nutritional value. Water-, salt- and acid-soluble M. edulis protein fractions were obtained and the proteins were trypsinized. The resultant peptides were analyzed by ultra-performance liquid chromatography quadrupole time of flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). 387 unique peptides were identified that matched 81 precursor proteins. Molecular mass distributions of the proteins and peptides were analyzed by sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE). The differences between the three protein samples were studied by Venn diagram of peptide and protein compositions. Toxicity, allergic and antithrombotic activity of peptides was predicted using database website and molecular docking respectively. The antithrombotic activity of enzymatic hydrolysate from water-, salt- and acid-soluble M. edulis protein were 40.17%, 85.74%, 82.00% at 5 mg/mL, respectively. Active mechanism of antithrombotic peptide (ELEDSLDSER) was also research about amino acid binding sites and interaction, simultaneously.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010138
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 139: Lysophosphatidic Acid Signaling Axis Mediates
           Ceramide 1-Phosphate-Induced Proliferation of C2C12 Myoblasts

    • Authors: Caterina Bernacchioni, Francesca Cencetti, Alberto Ouro, Marina Bruno, Antonio Gomez-Muñoz, Chiara Donati, Paola Bruni
      First page: 139
      Abstract: Sphingolipids are not only crucial for membrane architecture but act as critical regulators of cell functions. The bioactive sphingolipid ceramide 1-phosphate (C1P), generated by the action of ceramide kinase, has been reported to stimulate cell proliferation, cell migration and to regulate inflammatory responses via activation of different signaling pathways. We have previously shown that skeletal muscle is a tissue target for C1P since the phosphosphingolipid plays a positive role in myoblast proliferation implying a role in muscle regeneration. Skeletal muscle displays strong capacity of regeneration thanks to the presence of quiescent adult stem cells called satellite cells that upon trauma enter into the cell cycle and start proliferating. However, at present, the exact molecular mechanism by which C1P triggers its mitogenic effect in myoblasts is lacking. Here, we report for the first time that C1P stimulates C2C12 myoblast proliferation via lysophosphatidic acid (LPA) signaling axis. Indeed, C1P subsequently to phospholipase A2 activation leads to LPA1 and LPA3 engagement, which in turn drive Akt (protein kinase B) and ERK1/2 (extracellular signal-regulated kinases 1/2) activation, thus stimulating DNA synthesis. The present findings shed new light on the key role of bioactive sphingolipids in skeletal muscle and provide further support to the notion that these pleiotropic molecules might be useful therapeutic targets for skeletal muscle regeneration.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010139
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 140: Identification and Comparative Analysis of
           Premature Senescence Leaf Mutants in Rice (Oryza sativa L.)

    • Authors: Yan He, Liangjian Li, Zhihong Zhang, Jian-Li Wu
      First page: 140
      Abstract: Premature leaf senescence negatively impacts the grain yield in the important monocot rice (Oryza sativa L.); to understand the molecular mechanism we carried out a screen for mutants with premature senescence leaves in a mutant bank generated by ethyl methane sulfonate (EMS) mutagenesis of elite indica rice ZhongJian100. Five premature senescence leaf (psl15, psl50, psl89, psl117 and psl270) mutants were identified with distinct yellowish phenotypes on leaves starting from the tillering stage to final maturation. Moreover, these mutants exhibited significantly increased malonaldehyde content, decreased chlorophyll content, reduced numbers of chloroplast and grana thylakoid, altered photosynthetic ability and expression of photosynthesis-related genes. Furthermore, the expression of senescence-related indicator OsI57 was significantly up-regulated in four mutants. Histochemical analysis indicated that cell death and reactive oxygen species (ROS) accumulation occurred in the mutants with altered activities of ROS scavenging enzymes. Both darkness and abscisic acid (ABA) treatments could induce leaf senescence and resulted in up- or down-regulation of ABA metabolism-related genes in the mutants. Genetic analysis indicated that all the premature senescence leaf mutants were controlled by single non-allelic recessive genes. The data suggested that mechanisms underlying premature leaf senescence are likely different among the mutants. The present study would facilitate us to further fine mapping, cloning and functional characterization of the corresponding genes mediating the premature leaf senescence in rice.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-03
      DOI: 10.3390/ijms19010140
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 141: Computational Investigation of the Missense
           Mutations in DHCR7 Gene Associated with Smith-Lemli-Opitz Syndrome

    • Authors: Yunhui Peng, Rebecca Myers, Wenxing Zhang, Emil Alexov
      First page: 141
      Abstract: Smith-Lemli-Opitz syndrome (SLOS) is a cholesterol synthesis disorder characterized by physical, mental, and behavioral symptoms. It is caused by mutations in 7-dehydroxycholesterolreductase gene (DHCR7) encoding DHCR7 protein, which is the rate-limiting enzyme in the cholesterol synthesis pathway. Here we demonstrate that pathogenic mutations in DHCR7 protein are located either within the transmembrane region or are near the ligand-binding site, and are highly conserved among species. In contrast, non-pathogenic mutations observed in the general population are located outside the transmembrane region and have different effects on the conformational dynamics of DHCR7. All together, these observations suggest that the non-classified mutation R228Q is pathogenic. Our analyses indicate that pathogenic effects may affect protein stability and dynamics and alter the binding affinity and flexibility of the binding site.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010141
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 142: Fungi in Bronchiectasis: A Concise Review

    • Authors: Luis Máiz, Rosa Nieto, Rafael Cantón, Elia Gómez G. de la Pedrosa, Miguel Martinez-García
      First page: 142
      Abstract: Although the spectrum of fungal pathology has been studied extensively in immunosuppressed patients, little is known about the epidemiology, risk factors, and management of fungal infections in chronic pulmonary diseases like bronchiectasis. In bronchiectasis patients, deteriorated mucociliary clearance—generally due to prior colonization by bacterial pathogens—and thick mucosity propitiate, the persistence of fungal spores in the respiratory tract. The most prevalent fungi in these patients are Candida albicans and Aspergillus fumigatus; these are almost always isolated with bacterial pathogens like Haemophillus influenzae and Pseudomonas aeruginosa, making very difficult to define their clinical significance. Analysis of the mycobiome enables us to detect a greater diversity of microorganisms than with conventional cultures. The results have shown a reduced fungal diversity in most chronic respiratory diseases, and that this finding correlates with poorer lung function. Increased knowledge of both the mycobiome and the complex interactions between the fungal, viral, and bacterial microbiota, including mycobacteria, will further our understanding of the mycobiome’s relationship with the pathogeny of bronchiectasis and the development of innovative therapies to combat it.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010142
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 143: Characterization of a Potential Probiotic
           Lactobacillus brevis RK03 and Efficient Production of γ-Aminobutyric Acid
           in Batch Fermentation

    • Authors: Chien-Hui Wu, Yi-Huang Hsueh, Jen-Min Kuo, Si-Jia Liu
      First page: 143
      Abstract: Lactic acid bacteria were isolated from fish and evaluated for their γ-aminobutyric acid (GABA)-producing abilities. Out of thirty-two isolates, Lactobacillus brevis RK03 showed the highest GABA production ability. The effects of various fermentation parameters including initial glutamic acid level, culture temperature, initial pH, and incubation time on GABA production were investigated via a singleparameter optimization strategy. For industrial large-scale production, a low-cost GABA producing medium (GM) broth was developed for fermentation with L. brevis RK03. We found that an optimized GM broth recipe of 1% glucose; 2.5% yeast extract; 2 ppm each of CaCO3, MnSO4, and Tween 80; and 10 μM pyridoxal phosphate (PLP) resulted in a maximum GABA yield of 62,523 mg/L after 88 h following the addition of 650 mM monosodium glutamate (MSG), for a conversion rate of 93.28%. Our data provide a practical approach for the highly efficient and economic production of GABA. In addition, L. brevis RK03 is highly resistant to gastric acid and bovine bile salt. Thus, the discovery of Lactobacillus strains with the ability to synthesize GABA may offer new opportunities in the design of improved health-promoting functional foods.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010143
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 144: Correction: Wentao Wu, et al. Evolution Analysis
           of the Aux/IAA Gene Family in Plants Shows Dual Origins and Variable
           Nuclear Localization Signals. Int. J. Mol. Sci. 2017, 18, 2107

    • Authors: Wentao Wu, Yaxue Liu, Yuqian Wang, Huimin Li, Jiaxi Liu, Jiaxin Tan, Jiadai He, Jingwen Bai, Haoli Ma
      First page: 144
      Abstract: The authors would like to insert some websites and citations in the following sentence, “First, the complete proteomes of these species were downloaded from the Phytozome website (Version 11; Available online: www.phytozome.org)” in the “Materials and Methods” section on page 13, paragraph 3.1 of their paper published in the International Journal of Molecular Sciences [1].[...]
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010144
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 145: Allyl Isothiocyanate Exhibits No Anticancer
           Activity in MDA-MB-231 Breast Cancer Cells

    • Authors: Md. Sayeed, Massimo Bracci, Veronica Ciarapica, Marco Malavolta, Mauro Provinciali, Ernesta Pieragostini, Simona Gaetani, Federica Monaco, Guendalina Lucarini, Venerando Rapisarda, Roberto Di Primio, Lory Santarelli
      First page: 145
      Abstract: It was reported recently that allyl isothiocyanate (AITC) could inhibit various types of cancer cell growth. In the present study, we further investigated whether AITC could inhibit the growth of human breast cancer cells. Unexpectedly, we found that AITC did not inhibit, rather slightly promoted, the proliferation of MDA-MB-231 breast cancer cells, although it did have inhibitory effect on MCF-7 breast cancer cells. Cytofluorimetric analysis revealed that AITC (10 µM) did not induce apoptosis and cell cycle arrest in MDA-MB-231 cells. In addition, AITC significantly (p < 0.05) increased the expression of BCL-2 and mTOR genes and Beclin-1 protein in MDA-MB-231 cells. No significant changes in expression of PRKAA1 and PER2 genes, Caspase-8, Caspase-9, PARP, p-mTOR, and NF-κB p65 proteins were observed in these AITC-treated cells. Importantly, AITC displayed cytotoxic effect on MCF-10A human breast epithelial cell line. These observations suggest that AITC may not have inhibitory activity in MDA-MB-231 breast cancer cells. This in vitro study warrants more preclinical and clinical studies on the beneficial and harmful effects of AITC in healthy and cancer cells.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010145
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 146: Splice Variants of the Castor WRI1 Gene
           Upregulate Fatty Acid and Oil Biosynthesis When Expressed in Tobacco
           Leaves

    • Authors: Xia-Jie Ji, Xue Mao, Qing-Ting Hao, Bao-Ling Liu, Jin-Ai Xue, Run-Zhi Li
      First page: 146
      Abstract: The plant-specific WRINKLED1 (WRI1) is a member of the AP2/EREBP class of transcription factors that positively regulate oil biosynthesis in plant tissues. Limited information is available for the role of WRI1 in oil biosynthesis in castor bean (Ricinus connunis L.), an important industrial oil crop. Here, we report the identification of two alternatively spliced transcripts of RcWRI1, designated as RcWRI1-A and RcWRI1-B. The open reading frames of RcWRI1-A (1341 bp) and RcWRI1-B (1332 bp) differ by a stretch of 9 bp, such that the predicted RcWRI1-B lacks the three amino acid residues “VYL” that are present in RcWRI1-A. The RcWRI1-A transcript is present in flowers, leaves, pericarps and developing seeds, while the RcWRI1-B mRNA is only detectable in developing seeds. When the two isoforms were individually introduced into an Arabidopsis wri1-1 loss-of-function mutant, total fatty acid content was almost restored to the wild-type level, and the percentage of the wrinkled seeds was largely reduced in the transgenic lines relative to the wri1-1 mutant line. Transient expression of each RcWRI1 splice isoform in N. benthamiana leaves upregulated the expression of the WRI1 target genes, and consequently increased the oil content by 4.3–4.9 fold when compared with the controls, and RcWRI1-B appeared to be more active than RcWRI1-A. Both RcWRI1-A and RcWRI1-B can be used as a key transcriptional regulator to enhance fatty acid and oil biosynthesis in leafy biomass.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010146
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 147: Role of Microenvironment in Glioma Invasion:
           What We Learned from In Vitro Models

    • Authors: Ivana Manini, Federica Caponnetto, Anna Bartolini, Tamara Ius, Laura Mariuzzi, Carla Di Loreto, Antonio Beltrami, Daniela Cesselli
      First page: 147
      Abstract: The invasion properties of glioblastoma hamper a radical surgery and are responsible for its recurrence. Understanding the invasion mechanisms is thus critical to devise new therapeutic strategies. Therefore, the creation of in vitro models that enable these mechanisms to be studied represents a crucial step. Since in vitro models represent an over-simplification of the in vivo system, in these years it has been attempted to increase the level of complexity of in vitro assays to create models that could better mimic the behaviour of the cells in vivo. These levels of complexity involved: 1. The dimension of the system, moving from two-dimensional to three-dimensional models; 2. The use of microfluidic systems; 3. The use of mixed cultures of tumour cells and cells of the tumour micro-environment in order to mimic the complex cross-talk between tumour cells and their micro-environment; 4. And the source of cells used in an attempt to move from commercial lines to patient-based models. In this review, we will summarize the evidence obtained exploring these different levels of complexity and highlighting advantages and limitations of each system used.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010147
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 148: Understanding the Progression of Bone Metastases
           to Identify Novel Therapeutic Targets

    • Authors: Annie Schmid-Alliana, Heidy Schmid-Antomarchi, Rasha Al-Sahlanee, Patricia Lagadec, Jean-Claude Scimeca, Elise Verron
      First page: 148
      Abstract: Bone is one of the most preferential target site for cancer metastases, particularly for prostate, breast, kidney, lung and thyroid primary tumours. Indeed, numerous chemical signals and growth factors produced by the bone microenvironment constitute factors promoting cancer cell invasion and aggression. After reviewing the different theories proposed to provide mechanism for metastatic progression, we report on the gene expression profile of bone-seeking cancer cells. We also discuss the cross-talk between the bone microenvironment and invading cells, which impacts on the tumour actions on surrounding bone tissue. Lastly, we detail therapies for bone metastases. Due to poor prognosis for patients, the strategies mainly aim at reducing the impact of skeletal-related events on patients’ quality of life. However, recent advances have led to a better understanding of molecular mechanisms underlying bone metastases progression, and therefore of novel therapeutic targets.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010148
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 149: ФC31 Integrase-Mediated Isolation and
           Characterization of Novel Safe Harbors for Transgene Expression in the Pig
           Genome

    • Authors: Yanzhen Bi, Zaidong Hua, Hongyan Ren, Liping Zhang, Hongwei Xiao, Ximei Liu, Wenjun Hua, Shuqi Mei, Adrian Molenaar, Götz Laible, Xinmin Zheng
      First page: 149
      Abstract: Programmable nucleases have allowed the rapid development of gene editing and transgenics, but the technology still suffers from the lack of predefined genetic loci for reliable transgene expression and maintenance. To address this issue, we used ФC31 integrase to navigate the porcine genome and identify the pseudo attP sites suitable as safe harbors for sustained transgene expression. The combined ФC31 integrase mRNA and an enhanced green fluorescence protein (EGFP) reporter donor were microinjected into one-cell zygotes for transgene integration. Among the resulting seven EGFP-positive piglets, two had transgene integrations at pseudo attP sites, located in an intergenic region of chromosome 1 (chr1-attP) and the 6th intron of the TRABD2A gene on chromosome 3 (chr3-attP), respectively. The integration structure was determined by TAIL-PCR and Southern blotting. Primary fibroblast cells were isolated from the two piglets and examined using fluorescence-activated cell sorting (FACS) and enzyme-linked immunosorbent assay (ELISA), which demonstrated that the chr1-attP site was more potent than chr3-attP site in supporting the EGFP expression. Both piglets had green feet under the emission of UV light, and pelleted primary fibroblast cells were green-colored under natural light, corroborating that the two pseudo attP sites are beneficial to transgene expression. The discovery of these two novel safe harbors for robust and durable transgene expression will greatly facilitate the use of transgenic pigs for basic, biomedical and agricultural studies and applications.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010149
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 150: Probiotic Incorporation in Edible Films and
           Coatings: Bioactive Solution for Functional Foods

    • Authors: Foteini Pavli, Chrysoula Tassou, George-John Nychas, Nikos Chorianopoulos
      First page: 150
      Abstract: Nowadays, the consumption of food products containing probiotics, has increased worldwide due to concerns regarding healthy diet and wellbeing. This trend has received a lot of attention from the food industries, aiming to produce novel probiotic foods, and from researchers, to improve the existing methodologies for probiotic delivery or to develop and investigate new possible applications. In this sense, edible films and coatings are being studied as probiotic carriers with many applications. There is a wide variety of materials with film-forming ability, possessing different characteristics and subsequently affecting the final product. This manuscript aims to provide significant information regarding probiotics and active/bioactive packaging, to review applications of probiotic edible films and coatings, and to discuss certain limitations of their use as well as the current legislation and future trends.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010150
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 151: Computational Insights into the Interactions
           between Calmodulin and the c/nSH2 Domains of p85α Regulatory Subunit of
           PI3Kα: Implication for PI3Kα Activation by Calmodulin

    • Authors: Duan Ni, Dingyu Liu, Jian Zhang, Shaoyong Lu
      First page: 151
      Abstract: Calmodulin (CaM) and phosphatidylinositide-3 kinase (PI3Kα) are well known for their multiple roles in a series of intracellular signaling pathways and in the progression of several human cancers. Crosstalk between CaM and PI3Kα has been an area of intensive research. Recent experiments have shown that in adenocarcinoma, K-Ras4B is involved in the CaM-PI3Kα crosstalk. Based on experimental results, we have recently put forward a hypothesis that the coordination of CaM and PI3Kα with K-Ras4B forms a CaM-PI3Kα-K-Ras4B ternary complex, which leads to the formation of pancreatic ductal adenocarcinoma. However, the mechanism for the CaM-PI3Kα crosstalk is unresolved. Based on molecular modeling and molecular dynamics simulations, here we explored the potential interactions between CaM and the c/nSH2 domains of p85α subunit of PI3Kα. We demonstrated that CaM can interact with the c/nSH2 domains and the interaction details were unraveled. Moreover, the possible modes for the CaM-cSH2 and CaM-nSH2 interactions were uncovered and we used them to construct a complete CaM-PI3Kα complex model. The structural model of CaM-PI3Kα interaction not only offers a support for our previous ternary complex hypothesis, but also is useful for drug design targeted at CaM-PI3Kα protein-protein interactions.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010151
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 152: Estimation of Whole Plant Photosynthetic Rate of
           Irwin Mango under Artificial and Natural Lights Using a Three-Dimensional
           Plant Model and Ray-Tracing

    • Authors: Dae Jung, Joon Lee, Woo Kang, In Hwang, Jung Son
      First page: 152
      Abstract: Photosynthesis is an important physiological response for determination of CO2 fertilization in greenhouses and estimation of crop growth. In order to estimate the whole plant photosynthetic rate, it is necessary to investigate how light interception by crops changes with environmental and morphological factors. The objectives of this study were to analyze plant light interception using a three-dimensional (3D) plant model and ray-tracing, determine the spatial distribution of the photosynthetic rate, and estimate the whole plant photosynthetic rate of Irwin mango (Mangifera indica L. cv. Irwin) grown in greenhouses. In the case of mangoes, it is difficult to measure actual light interception at the canopy level due to their vase shape. A two-year-old Irwin mango tree was used to measure the whole plant photosynthetic rate. Light interception and whole plant photosynthetic rate were measured under artificial and natural light conditions using a closed chamber (1 × 1 × 2 m). A 3D plant model was constructed and ray-tracing simulation was conducted for calculating the photosynthetic rate with a two-variable leaf photosynthetic rate model of the plant. Under artificial light, the estimated photosynthetic rate increased from 2.0 to 2.9 μmolCO2·m−2·s−1 with increasing CO2 concentration. On the other hand, under natural light, the photosynthetic rate increased from 0.2 μmolCO2·m−2·s−1 at 06:00 to a maximum of 7.3 μmolCO2·m−2·s−1 at 09:00, then gradually decreased to −1.0 μmolCO2·m−2·s−1 at 18:00. In validation, simulation results showed good agreement with measured results with R2 = 0.79 and RMSE = 0.263. The results suggest that this method could accurately estimate the whole plant photosynthetic rate and be useful for pruning and adequate CO2 fertilization.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010152
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 153: The Osteogenic Differentiation Effect of the FN
           Type 10-Peptide Amphiphile on PCL Fiber

    • Authors: Ye-Rang Yun, Hae-Won Kim, Jun-Hyeog Jang
      First page: 153
      Abstract: The fibronectin type 10-peptide amphiphile (FNIII10-PA) was previously genetically engineered and showed osteogenic differentiation activity on rat bone marrow stem cells (rBMSCs). In this study, we investigated whether FNIII10-PA demonstrated cellular activity on polycaprolactone (PCL) fibers. FNIII10-PA significantly increased protein production and cell adhesion activity on PCL fibers in a dose-dependent manner. In cell proliferation results, there was no effect on cell proliferation activity by FNIII10-PA; however, FNIII10-PA induced the osteogenic differentiation of MC3T3-E1 cells via upregulation of bone sialoprotein (BSP), collagen type I (Col I), osteocalcin (OC), osteopontin (OPN), and runt-related transcription factor 2 (Runx2) mitochondrial RNA (mRNA) levels; it did not increase the alkaline phosphatase (ALP) mRNA level. These results indicate that FNIII10-PA has potential as a new biomaterial for bone tissue engineering applications.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010153
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 154: Implications of Aquaglyceroporin 7 in Energy
           Metabolism

    • Authors: Francesco Iena, Janne Lebeck
      First page: 154
      Abstract: The aquaglyceroporin AQP7 is a pore-forming transmembrane protein that facilitates the transport of glycerol across cell membranes. Glycerol is utilized both in carbohydrate and lipid metabolism. It is primarily stored in white adipose tissue as part of the triglyceride molecules. During states with increased lipolysis, such as fasting and diabetes, glycerol is released from adipose tissue and metabolized in other tissues. AQP7 is expressed in adipose tissue where it facilitates the efflux of glycerol, and AQP7 deficiency has been linked to increased glycerol kinase activity and triglyceride accumulation in adipose tissue, leading to obesity and secondary development of insulin resistance. However, AQP7 is also expressed in a wide range of other tissues, including kidney, muscle, pancreatic β-cells and liver, where AQP7 also holds the potential to influence whole body energy metabolism. The aim of the review is to summarize the current knowledge on AQP7 in adipose tissue, as well as AQP7 expressed in other tissues where AQP7 might play a significant role in modulating whole body energy metabolism.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-04
      DOI: 10.3390/ijms19010154
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 155: LHCSR Expression under HSP70/RBCS2 Promoter as a
           Strategy to Increase Productivity in Microalgae

    • Authors: Federico Perozeni, Giulio Stella, Matteo Ballottari
      First page: 155
      Abstract: Microalgae are unicellular photosynthetic organisms considered as potential alternative sources for biomass, biofuels or high value products. However, limited biomass productivity is commonly experienced in their cultivating system despite their high potential. One of the reasons for this limitation is the high thermal dissipation of the light absorbed by the outer layers of the cultures exposed to high light caused by the activation of a photoprotective mechanism called non-photochemical quenching (NPQ). In the model organism for green algae Chlamydomonas reinhardtii, NPQ is triggered by pigment binding proteins called light-harvesting-complexes-stress-related (LHCSRs), which are over-accumulated in high light. It was recently reported that biomass productivity can be increased both in microalgae and higher plants by properly tuning NPQ induction. In this work increased light use efficiency is reported by introducing in C. reinhardtii a LHCSR3 gene under the control of Heat Shock Protein 70/RUBISCO small chain 2 promoter in a npq4 lhcsr1 background, a mutant strain knockout for all LHCSR genes. This complementation strategy leads to a low expression of LHCSR3, causing a strong reduction of NPQ induction but is still capable of protecting from photodamage at high irradiance, resulting in an improved photosynthetic efficiency and higher biomass accumulation.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010155
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 156: Molecular Epidemiology and Evolution of European
           Bat Lyssavirus 2

    • Authors: Lorraine McElhinney, Denise Marston, Emma Wise, Conrad Freuling, Hervé Bourhy, Reto Zanoni, Torfinn Moldal, Engbert Kooi, Antonie Neubauer-Juric, Tiina Nokireki, Thomas Müller, Anthony Fooks
      First page: 156
      Abstract: Bat rabies cases in Europe are mainly attributed to two lyssaviruses, namely European Bat Lyssavirus 1 (EBLV-1) and European Bat Lyssavirus 2 (EBLV-2). Prior to the death of a bat worker in Finland in 1985, very few bat rabies cases were reported. Enhanced surveillance in the two subsequent years (1986–1987) identified 263 cases (more than a fifth of all reported cases to date). Between 1977 and 2016, 1183 cases of bat rabies were reported, with the vast majority (>97%) being attributed to EBLV-1. In contrast, there have been only 39 suspected cases of EBLV-2, of which 34 have been confirmed by virus typing and presently restricted to just two bat species; Myotis daubentonii and Myotis dasycneme. The limited number of EBLV-2 cases in Europe prompted the establishment of a network of European reference laboratories to collate all available viruses and data. Despite the relatively low number of EBLV-2 cases, a large amount of anomalous data has been published in the scientific literature, which we have here reviewed and clarified. In this review, 29 EBLV-2 full genome sequences have been analysed to further our understanding of the diversity and molecular evolution of EBLV-2 in Europe. Analysis of the 29 complete EBLV-2 genome sequences clearly corroborated geographical relationships with all EBLV-2 sequences clustering at the country level irrespective of the gene studied. Further geographical clustering was also observed at a local level. There are high levels of homogeneity within the EBLV-2 species with nucleotide identities ranging from 95.5–100% and amino acid identities between 98.7% and 100%, despite the widespread distribution of the isolates both geographically and chronologically. The mean substitution rate for EBLV-2 across the five concatenated genes was 1.65 × 10−5, and evolutionary clock analysis confirms the slow evolution of EBLV-2 both between and within countries in Europe. This is further supported by the first detailed EBLV-2 intra-roost genomic analysis whereby a relatively high sequence homogeneity was found across the genomes of three EBLV-2 isolates obtained several years apart (2007, 2008, and 2014) from M. daubentonii at the same site (Stokesay Castle, Shropshire, UK).
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010156
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 157: Telomere Homeostasis: Interplay with Magnesium

    • Authors: Donogh Maguire, Ognian Neytchev, Dinesh Talwar, Donald McMillan, Paul Shiels
      First page: 157
      Abstract: Telomere biology, a key component of the hallmarks of ageing, offers insight into dysregulation of normative ageing processes that accompany age-related diseases such as cancer. Telomere homeostasis is tightly linked to cellular metabolism, and in particular with mitochondrial physiology, which is also diminished during cellular senescence and normative physiological ageing. Inherent in the biochemistry of these processes is the role of magnesium, one of the main cellular ions and an essential cofactor in all reactions that use ATP. Magnesium plays an important role in many of the processes involved in regulating telomere structure, integrity and function. This review explores the mechanisms that maintain telomere structure and function, their influence on circadian rhythms and their impact on health and age-related disease. The pervasive role of magnesium in telomere homeostasis is also highlighted.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010157
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 158: A High-Resolution Proteomic Landscaping of
           Primary Human Dental Stem Cells: Identification of SHED- and
           PDLSC-Specific Biomarkers

    • Authors: Vasiliki Taraslia, Stefania Lymperi, Vasiliki Pantazopoulou, Athanasios Anagnostopoulos, Issidora Papassideri, Efthimia Basdra, Marianna Bei, Evangelos Kontakiotis, George Tsangaris, Dimitrios Stravopodis, Ema Anastasiadou
      First page: 158
      Abstract: Dental stem cells (DSCs) have emerged as a promising tool for basic research and clinical practice. A variety of adult stem cell (ASC) populations can be isolated from different areas within the dental tissue, which, due to their cellular and molecular characteristics, could give rise to different outcomes when used in potential applications. In this study, we performed a high-throughput molecular comparison of two primary human adult dental stem cell (hADSC) sub-populations: Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs) and Periodontal Ligament Stem Cells (PDLSCs). A detailed proteomic mapping of SHEDs and PDLSCs, via employment of nano-LC tandem-mass spectrometry (MS/MS) revealed 2032 identified proteins in SHEDs and 3235 in PDLSCs. In total, 1516 proteins were expressed in both populations, while 517 were unique for SHEDs and 1721 were exclusively expressed in PDLSCs. Further analysis of the recorded proteins suggested that SHEDs predominantly expressed molecules that are involved in organizing the cytoskeletal network, cellular migration and adhesion, whereas PDLSCs are highly energy-producing cells, vastly expressing proteins that are implicated in various aspects of cell metabolism and proliferation. Applying the Rho-GDI signaling pathway as a paradigm, we propose potential biomarkers for SHEDs and for PDLSCs, reflecting their unique features, properties and engaged molecular pathways.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010158
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 160: Transcriptome Analysis of Flounder (Paralichthys
           olivaceus) Gill in Response to Lymphocystis Disease Virus (LCDV)
           Infection: Novel Insights into Fish Defense Mechanisms

    • Authors: Ronghua Wu, Xiuzhen Sheng, Xiaoqian Tang, Jing Xing, Wenbin Zhan
      First page: 160
      Abstract: Lymphocystis disease virus (LCDV) infection may induce a variety of host gene expression changes associated with disease development; however, our understanding of the molecular mechanisms underlying host-virus interactions is limited. In this study, RNA sequencing (RNA-seq) was employed to investigate differentially expressed genes (DEGs) in the gill of the flounder (Paralichthys olivaceus) at one week post LCDV infection. Transcriptome sequencing of the gill with and without LCDV infection was performed using the Illumina HiSeq 2500 platform. In total, RNA-seq analysis generated 193,225,170 clean reads aligned with 106,293 unigenes. Among them, 1812 genes were up-regulated and 1626 genes were down-regulated after LCDV infection. The DEGs related to cellular process and metabolism occupied the dominant position involved in the LCDV infection. A further function analysis demonstrated that the genes related to inflammation, the ubiquitin-proteasome pathway, cell proliferation, apoptosis, tumor formation, and anti-viral defense showed a differential expression. Several DEGs including β actin, toll-like receptors, cytokine-related genes, antiviral related genes, and apoptosis related genes were involved in LCDV entry and immune response. In addition, RNA-seq data was validated by quantitative real-time PCR. For the first time, the comprehensive gene expression study provided valuable insights into the host-pathogen interaction between flounder and LCDV.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010160
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 161: Characterization of Different Types of
           Excitability in Large Somatosensory Neurons and Its Plastic Changes in
           Pathological Pain States

    • Authors: Rou-Gang Xie, Wen-Guang Chu, San-Jue Hu, Ceng Luo
      First page: 161
      Abstract: Sensory neuron types have been distinguished by distinct morphological and transcriptional characteristics. Excitability is the most fundamental functional feature of neurons. Mathematical models described by Hodgkin have revealed three types of neuronal excitability based on the relationship between firing frequency and applied current intensity. However, whether natural sensory neurons display different functional characteristics in terms of excitability and whether this excitability type undergoes plastic changes under pathological pain states have remained elusive. Here, by utilizing whole-cell patch clamp recordings, behavioral and pharmacological assays, we demonstrated that large dorsal root ganglion (DRG) neurons can be classified into three classes and four subclasses based on their excitability patterns, which is similar to mathematical models raised by Hodgkin. Analysis of hyperpolarization-activated cation current (Ih) revealed different magnitude of Ih in different excitability types of large DRG neurons, with higher Ih in Class 2-1 than that in Class 1, 2-2 and 3. This indicates a crucial role of Ih in the determination of excitability type of large DRG neurons. More importantly, this pattern of excitability displays plastic changes and transition under pathological pain states caused by peripheral nerve injury. This study sheds new light on the functional characteristics of large DRG neurons and extends functional classification of large DRG neurons by integration of transcriptomic and morphological characteristics.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010161
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 162: Effects of Fluoxetine on Hippocampal
           Neurogenesis and Neuroprotection in the Model of Global Cerebral Ischemia
           in Rats

    • Authors: Marina Khodanovich, Alena Kisel, Marina Kudabaeva, Galina Chernysheva, Vera Smolyakova, Elena Krutenkova, Irina Wasserlauf, Mark Plotnikov, Vasily Yarnykh
      First page: 162
      Abstract: A selective serotonin reuptake inhibitor, fluoxetine, has recently attracted a significant interest as a neuroprotective therapeutic agent. There is substantial evidence of improved neurogenesis under fluoxetine treatment of brain ischemia in animal stroke models. We studied long-term effects of fluoxetine treatment on hippocampal neurogenesis, neuronal loss, inflammation, and functional recovery in a new model of global cerebral ischemia (GCI). Brain ischemia was induced in adult Wistar male rats by transient occlusion of three main vessels originating from the aortic arch and providing brain blood supply. Fluoxetine was injected intraperitoneally in a dose of 20 mg/kg for 10 days after surgery. To evaluate hippocampal neurogenesis at time points 10 and 30 days, 5-Bromo-2′-deoxyuridine was injected at days 8–10 after GCI. According to our results, 10-day fluoxetine injections decreased neuronal loss and inflammation, improved survival and functional recovery of animals, enhanced neurogenesis, and prevented an early pathological increase in neural stem cell recruitment in the subgranular zone (SGZ) of the hippocampus without reducing the number of mature neurons at day 30 after GCI. In summary, this study suggests that fluoxetine may provide a promising therapy in cerebral ischemia due to its neuroprotective, anti-inflammatory, and neurorestorative effect.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010162
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 164: The Histone Deacetylase Inhibitor Valproic Acid
           Exerts a Synergistic Cytotoxicity with the DNA-Damaging Drug Ellipticine
           in Neuroblastoma Cells

    • Authors: Tereza Cerna, Jan Hrabeta, Tomas Eckschlager, Eva Frei, Heinz Schmeiser, Volker Arlt, Marie Stiborová
      First page: 164
      Abstract: Neuroblastoma (NBL) originates from undifferentiated cells of the sympathetic nervous system. Chemotherapy is judged to be suitable for successful treatment of this disease. Here, the influence of histone deacetylase (HDAC) inhibitor valproate (VPA) combined with DNA-damaging chemotherapeutic, ellipticine, on UKF-NB-4 and SH-SY5Y neuroblastoma cells was investigated. Treatment of these cells with ellipticine in combination with VPA led to the synergism of their anticancer efficacy. The effect is more pronounced in the UKF-NB-4 cell line, the line with N-myc amplification, than in SH-SY5Y cells. This was associated with caspase-3-dependent induction of apoptosis in UKF-NB-4 cells. The increase in cytotoxicity of ellipticine in UKF-NB-4 by VPA is dictated by the sequence of drug administration; the increased cytotoxicity was seen only after either simultaneous exposure to these drugs or after pretreatment of cells with ellipticine before their treatment with VPA. The synergism of treatment of cells with VPA and ellipticine seems to be connected with increased acetylation of histones H3 and H4. Further, co-treatment of cells with ellipticine and VPA increased the formation of ellipticine-derived DNA adducts, which indicates an easier accessibility of ellipticine to DNA in cells by its co-treatment with VPA and also resulted in higher ellipticine cytotoxicity. The results are promising for in vivo studies and perhaps later for clinical studies of combined treatment of children suffering from high-risk NBL.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-05
      DOI: 10.3390/ijms19010164
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 165: Excessive Endoplasmic Reticulum Stress
           Correlates with Impaired Mitochondrial Dynamics, Mitophagy and Apoptosis,
           in Liver and Adipose Tissue, but Not in Muscles in EMS Horses

    • Authors: Krzysztof Marycz, Katarzyna Kornicka, Jolanta Szlapka-Kosarzewska, Christine Weiss
      First page: 165
      Abstract: Nowadays, endocrine disorders have become more frequent in both human and veterinary medicine. In horses, reduced physical activity combined with carbohydrate and sugar overload may result in the development of the so-called equine metabolic syndrome (EMS). EMS is characterized by insulin resistance, hyperinsulinemia, elevated blood triglyceride concentrations and usually obesity. Although the phenotypic features of EMS individuals are well known, the molecular mechanism underlying disease development remains elusive. Therefore, in the present study, we analyzed insulin-sensitive tissues, i.e., muscles, liver and adipose tissue in order to evaluate insulin resistance and apoptosis. Furthermore, we assessed mitochondrial dynamics and mitophagy in those tissues, because mitochondrial dysfunction is linked to the development of metabolic syndrome. We established the expression of genes related to insulin resistance, endoplasmic reticulum (ER) stress and mitochondria clearance by mitophagy using RT-PCR and Western blot. Cell ultrastructure was visualized using electron transmission microscopy. The results indicated that adipose tissue and liver of EMS horses were characterized by increased mitochondrial damage and mitophagy followed by triggering of apoptosis as mitophagy fails to restore cellular homeostasis. However, in muscles, apoptosis was reduced, suggesting the existence of a protective mechanism allowing that tissue to maintain homeostasis.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010165
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 166: Could Aspirin and Diets High in Fiber Act
           Synergistically to Reduce the Risk of Colon Cancer in Humans'

    • Authors: Pan Pan, Yi-Wen Huang, Kiyoko Oshima, Martha Yearsley, Jianying Zhang, Jianhua Yu, Mark Arnold, Li-Shu Wang
      First page: 166
      Abstract: Early inhibition of inflammation suppresses the carcinogenic process. Aspirin is the most commonly used non-steroid anti-inflammatory drugs (NSAIDs), and it irreversibly inhibits cyclooxygenase-1 and -2 (COX1, COX2). Multiple randomized clinical trials have demonstrated that aspirin offers substantial protection from colon cancer mortality. The lower aspirin doses causing only minimal gastrointestinal disturbance, ideal for long-term use, can achieve only partial and transitory inhibition of COX2. Aspirin’s principal metabolite, salicylic acid, is also found in fruits and vegetables that inhibit COX2. Other phytochemicals such as curcumin, resveratrol, and anthocyanins also inhibit COX2. Such dietary components are good candidates for combination with aspirin because they have little or no toxicity. However, obstacles to using phytochemicals for chemoprevention, including bioavailability and translational potential, must be resolved. The bell/U-shaped dose–response curves seen with vitamin D and resveratrol might apply to other phytochemicals, shedding doubt on ‘more is better’. Solutions include: (1) using special delivery systems (e.g., nanoparticles) to retain phytochemicals; (2) developing robust pharmacodynamic biomarkers to determine efficacy in humans; and (3) selecting pharmacokinetic doses relevant to humans when performing preclinical experiments. The combination of aspirin and phytochemicals is an attractive low-cost and low-toxicity approach to colon cancer prevention that warrants testing, particularly in high-risk individuals.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010166
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 167: Investigation of New Morpholino Oligomers to
           Increase Survival Motor Neuron Protein Levels in Spinal Muscular Atrophy

    • Authors: Agnese Ramirez, Sebastiano Crisafulli, Mafalda Rizzuti, Nereo Bresolin, Giacomo Comi, Stefania Corti, Monica Nizzardo
      First page: 167
      Abstract: Spinal muscular atrophy (SMA) is an autosomal-recessive childhood motor neuron disease and the main genetic cause of infant mortality. SMA is caused by deletions or mutations in the survival motor neuron 1 (SMN1) gene, which results in SMN protein deficiency. Only one approved drug has recently become available and allows for the correction of aberrant splicing of the paralogous SMN2 gene by antisense oligonucleotides (ASOs), leading to production of full-length SMN protein. We have already demonstrated that a sequence of an ASO variant, Morpholino (MO), is particularly suitable because of its safety and efficacy profile and is both able to increase SMN levels and rescue the murine SMA phenotype. Here, we optimized this strategy by testing the efficacy of four new MO sequences targeting SMN2. Two out of the four new MO sequences showed better efficacy in terms of SMN protein production both in SMA induced pluripotent stem cells (iPSCs) and SMAΔ7 mice. Further, the effect was enhanced when different MO sequences were administered in combination. Our data provide an important insight for MO-based treatment for SMA. Optimization of the target sequence and validation of a treatment based on a combination of different MO sequences could support further pre-clinical studies and the progression toward future clinical trials.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010167
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 168: Nfatc1 Is a Functional Transcriptional Factor
           Mediating Nell-1-Induced Runx3 Upregulation in Chondrocytes

    • Authors: Chenshuang Li, Zhong Zheng, Xinli Zhang, Greg Asatrian, Eric Chen, Richard Song, Cymbeline Culiat, Kang Ting, Chia Soo
      First page: 168
      Abstract: Neural EGFL like 1 (Nell-1) is essential for chondrogenic differentiation, maturation, and regeneration. Our previous studies have demonstrated that Nell-1’s pro-chondrogenic activities are predominantly reliant upon runt-related transcription factor 3 (Runx3)-mediated Indian hedgehog (Ihh) signaling. Here, we identify the nuclear factor of activated T-cells 1 (Nfatc1) as the key transcriptional factor mediating the Nell-1 → Runx3 signal transduction in chondrocytes. Using chromatin immunoprecipitation assay, we were able to determine that Nfatc1 binds to the −833–−810 region of the Runx3-promoter in response to Nell-1 treatment. By revealing the Nell-1 → Nfatc1 → Runx3 → Ihh cascade, we demonstrate the involvement of Nfatc1, a nuclear factor of activated T-cells, in chondrogenesis, while providing innovative insights into developing a novel therapeutic strategy for cartilage regeneration and other chondrogenesis-related conditions.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010168
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 169: Radical Scavenging and Anti-Inflammatory
           Activities of Representative Anthocyanin Groupings from Pigment-Rich
           Fruits and Vegetables

    • Authors: Federica Blando, Nadia Calabriso, Helge Berland, Gabriele Maiorano, Carmela Gerardi, Maria Carluccio, Øyvind Andersen
      First page: 169
      Abstract: Anthocyanins, the naturally occurring pigments responsible for most red to blue colours of flowers, fruits and vegetables, have also attracted interest because of their potential health effects. With the aim of contributing to major insights into their structure–activity relationship (SAR), we have evaluated the radical scavenging and biological activities of selected purified anthocyanin samples (PASs) from various anthocyanin-rich plant materials: two fruits (mahaleb cherry and blackcurrant) and two vegetables (black carrot and “Sun Black” tomato), differing in anthocyanin content (ranging from 4.9 to 38.5 mg/g DW) and molecular structure of the predominant anthocyanins. PASs from the abovementioned plant materials have been evaluated for their antioxidant capacity using Trolox Equivalent Antioxidant Capacity (TEAC) and Oxygen Radical Absorbance Capacity (ORAC) assays. In human endothelial cells, we analysed the anti-inflammatory activity of different PASs by measuring their effects on the expression of endothelial adhesion molecules VCAM-1 and ICAM-1. We demonstrated that all the different PASs showed biological activity. They exhibited antioxidant capacity of different magnitude, higher for samples containing non-acylated anthocyanins (typical for fruits) compared to samples containing more complex anthocyanins acylated with cinnamic acid derivatives (typical for vegetables), even though this order was slightly reversed when ORAC assay values were expressed on a molar basis. Concordantly, PASs containing non-acylated anthocyanins reduced the expression of endothelial inflammatory antigens more than samples with aromatic acylated anthocyanins, suggesting the potential beneficial effect of structurally diverse anthocyanins in cardiovascular protection.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010169
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 170: CPT-11-Induced Delayed Diarrhea Develops via
           Reduced Aquaporin-3 Expression in the Colon

    • Authors: Risako Kon, Yuika Tsubota, Moe Minami, Saki Kato, Yukari Matsunaga, Hiroshi Kimura, Yuta Murakami, Tetsuya Fujikawa, Ryoya Sakurai, Rei Tomimoto, Yoshiaki Machida, Nobutomo Ikarashi, Kiyoshi Sugiyama
      First page: 170
      Abstract: While irinotecan (CPT-11) has a potent anti-cancer effect, it also causes serious diarrhea as an adverse reaction. In this study, we analyzed the pathogenic mechanism of CPT-11-induced delayed diarrhea by focusing on water channel aquaporin-3 (AQP3) in the colon. When rats received CPT-11, the expression level of AQP3 was reduced during severe diarrhea. It was found that the expression levels of inflammatory cytokines and the loss of crypt cells were increased in the colon when CPT-11 was administered. When celecoxib, an anti-inflammatory drug, was concomitantly administered, both the diarrhea and the reduced expression of AQP3 induced by CPT-11 were suppressed. The inflammation in the rat colon during diarrhea was caused via activated macrophage by CPT-11. These results showed that when CPT-11 is administered, the expression level of AQP3 in the colon is reduced, resulting in delayed diarrhea by preventing water transport from the intestinal tract. It was also suggested that the reduced expression of AQP3 might be due to the inflammation that occurs following the loss of colonic crypt cells and to the damage caused by the direct activation of macrophages by CPT-11. Therefore, it was considered that anti-inflammatory drugs that suppress the reduction of AQP3 expression could prevent CPT-11-induced delayed diarrhea.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010170
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 171: Psoriasis: A STAT3-Centric View

    • Authors: Enzo Calautti, Lidia Avalle, Valeria Poli
      First page: 171
      Abstract: Signal Transducer and Activator of Transcription (STAT)3 has recently emerged as a key player in the development and pathogenesis of psoriasis and psoriatic-like inflammatory conditions. Indeed, STAT3 hyperactivation has been reported in virtually every cell type involved in disease initiation and maintenance, and this factor mediates the signal of most cytokines that are involved in disease pathogenesis, including the central Interleukin (IL)-23/IL-17/IL-22 axis. Despite the recent availability of effective biological agents (monoclonal antibodies) against IL-17 and IL-23, which have radically changed the current standard of disease management, the possibility of targeting either STAT3 itself or, even better, the family of upstream activators Janus kinases (JAK1, 2, 3, and TYK2) offers additional therapeutic options. Due to the oral/topical administration modality of these small molecule drugs, their lower cost, and the reduced risk of eliciting adverse immune responses, these compounds are being actively scrutinized in clinical settings. Here, we summarize the main pathological features of psoriatic conditions that provide the rationale for targeting the JAK/STAT3 axis in disease treatment.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010171
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 172: Synergistic Association of Valproate and
           Resveratrol Reduces Brain Injury in Ischemic Stroke

    • Authors: Lara Faggi, Giuseppe Pignataro, Edoardo Parrella, Vanessa Porrini, Antonio Vinciguerra, Pasquale Cepparulo, Ornella Cuomo, Annamaria Lanzillotta, Mariana Mota, Marina Benarese, Paolo Tonin, Lucio Annunziato, PierFranco Spano, Marina Pizzi
      First page: 172
      Abstract: Histone deacetylation, together with altered acetylation of NF-κB/RelA, encompassing the K310 residue acetylation, occur during brain ischemia. By restoring the normal acetylation condition, we previously reported that sub-threshold doses of resveratrol and entinostat (MS-275), respectively, an activator of the AMP-activated kinase (AMPK)-sirtuin 1 pathway and an inhibitor of class I histone deacetylases (HDACs), synergistically elicited neuroprotection in a mouse model of ischemic stroke. To improve the translational power of this approach, we investigated the efficacy of MS-275 replacement with valproate, the antiepileptic drug also reported to be a class I HDAC blocker. In cortical neurons previously exposed to oxygen glucose deprivation (OGD), valproate elicited neuroprotection at 100 nmol/mL concentration when used alone and at 1 nmol/mL concentration when associated with resveratrol (3 nmol/mL). Resveratrol and valproate restored the acetylation of histone H3 (K9/18), and they reduced the RelA(K310) acetylation and the Bim level in neurons exposed to OGD. Chromatin immunoprecipitation analysis showed that the synergistic drug association impaired the RelA binding to the Bim promoter, as well as the promoter-specific H3 (K9/18) acetylation. In mice subjected to 60 min of middle cerebral artery occlusion (MCAO), the association of resveratrol 680 µg/kg and valproate 200 µg/kg significantly reduced the infarct volume as well as the neurological deficits. The present study suggests that valproate and resveratrol may represent a promising ready-to-use strategy to treat post-ischemic brain damage.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010172
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 173: Skin Protective Effect of Epigallocatechin
           Gallate

    • Authors: Eunji Kim, Kyeonghwan Hwang, Jongsung Lee, Sang Yun Han, Eun-Mi Kim, Junseong Park, Jae Youl Cho
      First page: 173
      Abstract: Epigallocatechin gallate (EGCG) is a catechin and an abundant polyphenol in green tea. Although several papers have evaluated EGCG as a cosmetic constituent, the skin hydration effect of EGCG is poorly understood. We aimed to investigate the mechanism by which EGCG promotes skin hydration by measuring hyaluronic acid synthase (HAS) and hyaluronidase (HYAL) gene expression and antioxidant and anti-pigmentation properties using cell proliferation assay, Western blotting analysis, luciferase assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, and reverse transcription polymerase chain reaction (RT-PCR) analysis. RT-PCR showed that EGCG increased the expression of natural moisturizing factor-related genes filaggrin (FLG), transglutaminase-1, HAS-1, and HAS-2. Under UVB irradiation conditions, the expression level of HYAL was decreased in HaCaT cells. Furthermore, we confirmed the antioxidant activity of EGCG and also showed a preventive effect against radical-evoked apoptosis by downregulation of caspase-8 and -3 in HaCaT cells. EGCG reduced melanin secretion and production in melanoma cells. Together, these results suggest that EGCG might be used as a cosmetic ingredient with positive effects on skin hydration, moisture retention, and wrinkle formation, in addition to radical scavenging activity and reduction of melanin generation.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-06
      DOI: 10.3390/ijms19010173
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 174: Chemical Characterization of a Renoprotective
           Metabolite from Termite-Associated Streptomyces sp. RB1 against
           Cisplatin-Induced Cytotoxicity

    • Authors: Dahae Lee, Ki Kang, Hae-Jeung Lee, Ki Kim
      First page: 174
      Abstract: Platinum-based anticancer drug therapies can cause renal damage and apoptotic kidney cell damage. The development of reno- and kidney-protective molecules is therefore urgently required. To address this challenge, we explored secondary metabolites of termite-associated Streptomyces sp. RB1 isolated from the cuticle of the South African termite, Macrotermes natalensis for their renoprotective ability using bioassay-guided fractionation and LLC-PK1 cells. Chemical investigation of the MeOH extract of Streptomyces sp. RB1 resulted in the isolation and identification of a renoprotective metabolite, 1-O-(2-aminobenzoyl)-α-l-rhamnopyranoside (ABR) (1) from the active fraction, which ameliorated cisplatin-induced cytotoxicity to 80% of the control value at 25 μM. Upregulated phosphorylation of c-Jun N-terminal kinases (JNK) and p38 following cisplatin treatment was markedly decreased after pre-treatment of cells with ABR. In addition, levels of cleaved caspase-3 and the percentage of apoptotic cells were also significantly reduced after pre-treatment with ABR. These findings provide experimental evidence that blocking the MAPK signaling cascade plays a critical role in mediating the renoprotective effect of ABR, which may inspire the development of novel therapeutic substances to prevent anticancer drug-induced nephrotoxicity.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-07
      DOI: 10.3390/ijms19010174
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 175: Transcriptome Analysis in Haematococcus
           pluvialis: Astaxanthin Induction by High Light with Acetate and Fe2+

    • Authors: Bangxiang He, Lulu Hou, Manman Dong, Jiawei Shi, Xiaoyun Huang, Yating Ding, Xiaomei Cong, Feng Zhang, Xuecheng Zhang, Xiaonan Zang
      First page: 175
      Abstract: Haematococcus pluvialis is a commercial microalga, that produces abundant levels of astaxanthin under stress conditions. Acetate and Fe2+ are reported to be important for astaxanthin accumulation in H. pluvialis. In order to study the synergistic effects of high light stress and these two factors, we obtained transcriptomes for four groups: high light irradiation (HL), addition of 25 mM acetate under high light (HA), addition of 20 μM Fe2+ under high light (HF) and normal green growing cells (HG). Among the total clean reads of the four groups, 156,992 unigenes were found, of which 48.88% were annotated in at least one database (Nr, Nt, Pfam, KOG/COG, SwissProt, KEGG, GO). The statistics for DEGs (differentially expressed genes) showed that there were more than 10 thousand DEGs caused by high light and 1800–1900 DEGs caused by acetate or Fe2+. The results of DEG analysis by GO and KEGG enrichments showed that, under the high light condition, the expression of genes related to many pathways had changed, such as the pathway for carotenoid biosynthesis, fatty acid elongation, photosynthesis-antenna proteins, carbon fixation in photosynthetic organisms and so on. Addition of acetate under high light significantly promoted the expression of key genes related to the pathways for carotenoid biosynthesis and fatty acid elongation. Furthermore, acetate could obviously inhibit the expression of genes related to the pathway for photosynthesis-antenna proteins. For addition of Fe2+, the genes related to photosynthesis-antenna proteins were promoted significantly and there was no obvious change in the gene expressions related to carotenoid and fatty acid synthesis.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-07
      DOI: 10.3390/ijms19010175
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 176: Galectin-12 in Cellular Differentiation,
           Apoptosis and Polarization

    • Authors: Lei Wan, Ri-Yao Yang, Fu-Tong Liu
      First page: 176
      Abstract: Galectin-12 is a member of a family of mammalian lectins characterized by their affinity for β-galactosides and consensus amino acid sequences. The protein structure consists of a single polypeptide chain containing two carbohydrate-recognition domains joined by a linker region. Galectin-12 is predominantly expressed in adipose tissue, but is also detected in macrophages and other leukocytes. Downregulation of galectin-12 in mouse 3T3-L1 cells impairs their differentiation into adipocytes. Conversely, overexpression of galectin-12 in vitro induces cell cycle arrest in G1 and apoptosis. Upregulation of galectin-12 and initiation of G1 cell cycle arrest are associated with driving pre-adipocytes toward terminal differentiation. Galectin-12 deficiency increases insulin sensitivity and glucose tolerance in obese animals. Galectin-12 inhibits macrophage polarization to the M2 population, enhancing inflammation and decreasing insulin sensitivity in adipocytes. Galectin-12 also affects myeloid differentiation, which is associated with chemotherapy resistance. In addition to highlighting the above-mentioned aspects, this review also discusses the potential clinical applications of modulating the function of galectin-12.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-07
      DOI: 10.3390/ijms19010176
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 177: Fusion Proteins of NKG2D/NKG2DL in Cancer
           Immunotherapy

    • Authors: Hui Ding, Xi Yang, Yanzhang Wei
      First page: 177
      Abstract: NKG2D (natural killer group 2, member D) is an important activating receptor in natural killer (NK) cells and some T cells. NKG2D ligands (NKG2DLs) are specifically expressed on most tumor cells. The engagement of these ligands on tumor cells to NKG2D on NK cells will induce cell-mediated cytotoxicity and have target cells destroyed. This gives NKG2D/NKG2DLs great potential in cancer therapeutic application. The creation of NKG2D/NKG2DL-based multi-functional fusion proteins is becoming one of the most promising strategies in immunotherapy for cancer. Antibodies, cytokines, and death receptors have been fused with NKG2D or its ligands to produce many powerful fusion proteins, including NKG2D-based chimeric antigen receptors (CARs). In this article, we review the recent developments of the fusion proteins with NKG2D/NKG2DL ligands in cancer immunotherapy.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-07
      DOI: 10.3390/ijms19010177
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 178: Modulation of Protein Quality Control and
           Proteasome to Autophagy Switch in Immortalized Myoblasts from Duchenne
           Muscular Dystrophy Patients

    • Authors: Marion Wattin, Loïc Gaweda, Pascale Muller, Mathieu Baritaud, Charlotte Scholtes, Chloé Lozano, Kathrin Gieseler, Carole Kretz-Remy
      First page: 178
      Abstract: The maintenance of proteome integrity is of primary importance in post-mitotic tissues such as muscle cells; thus, protein quality control mechanisms must be carefully regulated to ensure their optimal efficiency, a failure of these processes being associated with various muscular disorders. Duchenne muscular dystrophy (DMD) is one of the most common and severe forms of muscular dystrophies and is caused by mutations in the dystrophin gene. Protein quality control modulations have been diversely observed in degenerating muscles of patients suffering from DMD or in animal models of the disease. In this study, we investigated whether modulations of protein quality control mechanisms already pre-exist in undifferentiated myoblasts originating from DMD patients. We report for the first time that the absence of dystrophin in human myoblasts is associated with protein aggregation stress characterized by an increase of protein aggregates. This stress is combined with BAG1 to BAG3 switch, NFκB activation and up-regulation of BAG3/HSPB8 complexes that ensure preferential routing of misfolded/aggregated proteins to autophagy rather than to deficient 26S proteasome. In this context, restoration of pre-existing alterations of protein quality control processes might represent an alternative strategy for DMD therapies.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-07
      DOI: 10.3390/ijms19010178
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 179: Scanning the Immunopathogenesis of Psoriasis

    • Authors: Andrea Chiricozzi, Paolo Romanelli, Elisabetta Volpe, Giovanna Borsellino, Marco Romanelli
      First page: 179
      Abstract: Psoriasis is a chronic inflammatory skin disease, the immunologic model of which has been profoundly revised following recent advances in the understanding of its pathophysiology. In the current model, a crosstalk between keratinocytes, neutrophils, mast cells, T cells, and dendritic cells is thought to create inflammatory and pro-proliferative circuits mediated by chemokines and cytokines. Various triggers, including recently identified autoantigens, Toll-like receptor agonists, chemerin, and thymic stromal lymphopoietin may activate the pathogenic cascade resulting in enhanced production of pro-inflammatory and proliferation-inducing mediators such as interleukin (IL)-17, tumor necrosis factor (TNF)-α, IL-23, IL-22, interferon (IFN)-α, and IFN-γ by immune cells. Among these key cytokines lie therapeutic targets for currently approved antipsoriatic therapies. This review aims to provide a comprehensive overview on the immune-mediated mechanisms characterizing the current pathogenic model of psoriasis.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010179
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 180: Effect of Omega-3 Fatty Acid Supplementation on
           Oxylipins in a Routine Clinical Setting

    • Authors: Christoph Schmöcker, Ingrid Zhang, Stefanie Kiesler, Ursula Kassner, Annika Ostermann, Elisabeth Steinhagen-Thiessen, Nils Schebb, Karsten-H. Weylandt
      First page: 180
      Abstract: Omega-6 polyunsaturated fatty acid (n-6 PUFA) is the predominant polyunsaturated fatty acid (PUFA), especially in Western diet. A high omega-6/omega-3 ratio in Western diets is implicated in the development of cardiovascular diseases and inflammatory processes. Studies in animal models and in humans have demonstrated beneficial effects of omega-3 PUFA (n-3 PUFA) in a variety of diseases, including cardiac arrhythmias and inflammatory diseases, as well as breast and colon cancer. The molecular mechanisms underlying the effects of n-3 PUFA are still not well understood. Possible mechanisms include competition between n-3 and n-6 PUFAs at the cyclooxygenase (COX) and lipoxygenase (LOX) and cytochrome P450 levels, and subsequent formation of oxylipins with specific anti-inflammatory or anti-arrhythmic effects. In this study, we report the impact of routine long-term treatment with prescription-grade n-3 PUFA (either 840 mg or 1680 mg per day) on blood cell membrane fatty acid composition, as well as plasma oxylipin patterns, in a patient population with severe hyperlipidemia and cardiovascular disease who are on standard lipid-lowering and cardioprotective medications. Lipidomics analyses were performed by LC/ESI-MS/MS. Supplementation led to a dose-dependent increase in n-3 PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the blood cell fraction. We also observed a dose-dependent increase in EPA- and DHA-derived epoxy metabolites, whereas the effect of n-3 PUFA supplementation on LOX-dependent EPA- and DHA-derived hydroxy metabolites was less pronounced, with a tendency towards lower metabolites in subjects with higher n-3 PUFA levels. These data thus generally confirm effects of n-3 PUFA supplementation observed previously in healthy individuals. Additionally, they indicate a suppressive effect of high n-3 PUFA supplementation on the formation of LOX metabolites in the context of concomitant aspirin medication.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010180
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 182: Non-Transfusion-Dependent Thalassemia: An Update
           on Complications and Management

    • Authors: Joseph Sleiman, Ali Tarhini, Rayan Bou-Fakhredin, Antoine Saliba, Maria Cappellini, Ali Taher
      First page: 182
      Abstract: Patients with non-transfusion-dependent thalassemia (NTDT) experience many clinical complications despite their independence from frequent transfusions. Morbidities in NTDT stem from the interaction of multiple pathophysiological factors: ineffective erythropoiesis, iron overload (IOL), and hypercoagulability. Ineffective erythropoiesis and hemolysis are associated with chronic hypoxia and a hypercoagulable state. The latter are linked to a high prevalence of thromboembolic and cerebrovascular events, as well as leg ulcers and pulmonary hypertension. IOL in NTDT patients is a cumulative process that can lead to several iron-related morbidities in the liver (liver fibrosis), kidneys, endocrine glands (endocrinopathies), and vascular system (vascular disease). This review sheds light on the pathophysiology underlying morbidities associated with NTDT and summarizes the mainstays of treatment and some of the possible future therapeutic interventions.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010182
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 183: Assessing the Performances of Protein Function
           Prediction Algorithms from the Perspectives of Identification Accuracy and
           False Discovery Rate

    • Authors: Chun Yu, Xiao Li, Hong Yang, Ying Li, Wei Xue, Yu Chen, Lin Tao, Feng Zhu
      First page: 183
      Abstract: The function of a protein is of great interest in the cutting-edge research of biological mechanisms, disease development and drug/target discovery. Besides experimental explorations, a variety of computational methods have been designed to predict protein function. Among these in silico methods, the prediction of BLAST is based on protein sequence similarity, while that of machine learning is also based on the sequence, but without the consideration of their similarity. This unique characteristic of machine learning makes it a good complement to BLAST and many other approaches in predicting the function of remotely relevant proteins and the homologous proteins of distinct function. However, the identification accuracies of these in silico methods and their false discovery rate have not yet been assessed so far, which greatly limits the usage of these algorithms. Herein, a comprehensive comparison of the performances among four popular prediction algorithms (BLAST, SVM, PNN and KNN) was conducted. In particular, the performance of these methods was systematically assessed by four standard statistical indexes based on the independent test datasets of 93 functional protein families defined by UniProtKB keywords. Moreover, the false discovery rates of these algorithms were evaluated by scanning the genomes of four representative model organisms (Homo sapiens, Arabidopsis thaliana, Saccharomyces cerevisiae and Mycobacterium tuberculosis). As a result, the substantially higher sensitivity of SVM and BLAST was observed compared with that of PNN and KNN. However, the machine learning algorithms (PNN, KNN and SVM) were found capable of substantially reducing the false discovery rate (SVM < PNN < KNN). In sum, this study comprehensively assessed the performance of four popular algorithms applied to protein function prediction, which could facilitate the selection of the most appropriate method in the related biomedical research.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010183
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 184: Fungicidal PMMA-Undecylenic Acid Composites

    • Authors: Milica Petrović, Debora Bonvin, Heinrich Hofmann, Marijana Mionić Ebersold
      First page: 184
      Abstract: Undecylenic acid (UA), known as antifungal agent, still cannot be used to efficiently modify commercial dental materials in such a way that this affects Candida. Actually, issues with Candida infections and fungal resistance compromise the use of Poly(methyl-methacrylate) (PMMA) as dental material. The challenge remains to turn PMMA into an antifugal material, which can ideally affect both sessile (attached) and planktonic (free-floating) Candida cells. We aimed to tackle this challenge by designing PMMA-UA composites with different UA concentrations (3–12%). We studied their physico-chemical properties, the antifungal effect on Candida and the cytotoxicity toward human cells. We found that UA changes the PMMA surface into a more hydrophilic one. Mainly, as-preparation composites with ≥6% UA reduced sessile Candida for >90%. After six days, the composites were still efficiently reducing the sessile Candida cells (for ~70% for composites with ≥6% UA). Similar results were recorded for planktonic Candida. Moreover, the inhibition zone increased along with the UA concentration. The antifungal effect of UA was also examined at the surface of an UA-loaded agar and the minimal inhibitory concentration (MIC90) was below the lowest-studied 0.0125% UA. Furthermore, the embedded filamentation test after 24 h and 48 h showed complete inhibition of the Candida growth at 0.4% UA.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010184
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 185: Impaired Skin Barrier Due to Sebaceous Gland
           Atrophy in the Latent Stage of Radiation-Induced Skin Injury: Application
           of Non-Invasive Diagnostic Methods

    • Authors: Hyosun Jang, Hyunwook Myung, Janet Lee, Jae Myung, Won-Suk Jang, Sun-Joo Lee, Chang-Hwan Bae, Hyewon Kim, Sunhoo Park, Sehwan Shim
      First page: 185
      Abstract: Radiation-induced skin injury can take the form of serious cutaneous damage and have specific characteristics. Asymptomatic periods are classified as the latent stage. The skin barrier plays a critical role in the modulation of skin permeability and hydration and protects the body against a harsh external environment. However, an analysis on skin barrier dysfunction against radiation exposure in the latent stage has not been conducted. Thus, we investigated whether the skin barrier is impaired by irradiation in the latent stage and aimed to identify the molecules involved in skin barrier dysfunction. We analyzed skin barrier function and its components in SKH1 mice that received 20 and 40 Gy local irradiation. Increased transepidermal water loss and skin pH were observed in the latent stage of the irradiated skin. Skin barrier components, such as structural proteins and lipid synthesis enzymes in keratinocyte, increased in the irradiated group. Interestingly, we noted sebaceous gland atrophy and increased serine protease and inflammatory cytokines in the irradiated skin during the latent period. This finding indicates that the main factor of skin barrier dysfunction in the latent stage of radiation-induced skin injury is sebaceous gland deficiency, which could be an intervention target for skin barrier impairment.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010185
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 186: A Comparative Study on Phytochemical Profiles
           and Biological Activities of Sclerocarya birrea (A.Rich.) Hochst Leaf and
           Bark Extracts

    • Authors: Daniela Russo, Rocchina Miglionico, Monica Carmosino, Faustino Bisaccia, Paula Andrade, Patrícia Valentão, Luigi Milella, Maria Armentano
      First page: 186
      Abstract: Sclerocarya birrea (A.Rich.) Hochst (Anacardiaceae) is a savannah tree that has long been used in sub-Saharan Africa as a medicinal remedy for numerous ailments. The purpose of this study was to increase the scientific knowledge about this plant by evaluating the total content of polyphenols, flavonoids, and tannins in the methanol extracts of the leaves and bark (MLE and MBE, respectively), as well as the in vitro antioxidant activity and biological activities of these extracts. Reported results show that MLE is rich in flavonoids (132.7 ± 10.4 mg of quercetin equivalents/g), whereas MBE has the highest content of tannins (949.5 ± 29.7 mg of tannic acid equivalents/g). The antioxidant activity was measured using four different in vitro tests: β-carotene bleaching (BCB), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), O2−•, and nitric oxide (NO•) assays. In all cases, MBE was the most active compared to MLE and the standards used (Trolox and ascorbic acid). Furthermore, MBE and MLE were tested to evaluate their activity in HepG2 and fibroblast cell lines. A higher cytotoxic activity of MBE was evidenced and confirmed by more pronounced alterations in cell morphology. MBE induced cell death, triggering the intrinsic apoptotic pathway by reactive oxygen species (ROS) generation, which led to a loss of mitochondrial membrane potential with subsequent cytochrome c release from the mitochondria into the cytosol. Moreover, MBE showed lower cytotoxicity in normal human dermal fibroblasts, suggesting its potential as a selective anticancer agent.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010186
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 187: Protective Effects of Protocatechuic Acid on
           Seizure-Induced Neuronal Death

    • Authors: Song Lee, Bo Choi, A Kho, Jeong Jeong, Dae Hong, Sang Lee, Sang Lee, Min Lee, Hong Song, Hui Choi, Sang Suh
      First page: 187
      Abstract: Protocatechuic acid (PCA) is a type of phenolic acid found in green tea and has been shown to have potent antioxidant and anti-inflammatory properties. However, the effect of PCA on pilocarpine seizure-induced neuronal death in the hippocampus has not been evaluated. In the present study, we investigated the potential therapeutic effects of PCA on seizure-induced brain injury. Epileptic seizure was induced by intraperitoneal (i.p.) injection of pilocarpine (25 mg/kg) in adult male rats, and PCA (30 mg/kg) was injected into the intraperitoneal space for three consecutive days after the seizure. Neuronal injury and oxidative stress were evaluated three days after a seizure. To confirm whether PCA increases neuronal survival and reduced oxidative injury in the hippocampus, we performed Fluoro-Jade-B (FJB) staining to detect neuronal death and 4-hydroxynonenal (4HNE) staining to detect oxidative stress after the seizure. In the present study, we found that, compared to the seizure vehicle-treated group, PCA administration reduced neuronal death and oxidative stress in the hippocampus. To verify whether a decrease of neuronal death by PCA treatment was due to reduced glutathione (GSH) concentration, we measured glutathione with N-ethylmaleimide (GS-NEM) levels in hippocampal neurons. A seizure-induced reduction in the hippocampal neuronal GSH concentration was preserved by PCA treatment. We also examined whether microglia activation was affected by the PCA treatment after a seizure, using CD11b staining. Here, we found that seizure-induced microglia activation was significantly reduced by the PCA treatment. Therefore, the present study demonstrates that PCA deserves further investigation as a therapeutic agent for reducing hippocampal neuronal death after epileptic seizures.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010187
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 188: Carbon Monoxide Potentiates High
           Temperature-Induced Nicotine Biosynthesis in Tobacco

    • Authors: Tielong Cheng, Liwei Hu, Pengkai Wang, Xiuyan Yang, Ye Peng, Ye Lu, Jinhui Chen, Jisen Shi
      First page: 188
      Abstract: Carbon monoxide (CO) acts as an important signal in many physiological responses in plants, but its role in plant secondary metabolism is still unknown. Nicotine is the main alkaloid generated in tobacco and the plant hormone jasmonic acid (JA) has previously been reported to efficiently induce its biosynthesis. Whether and how CO interacts with JA to regulate nicotine biosynthesis in tobacco remains elusive. In this study, we demonstrate that high temperature (HT) induces quick accumulation of nicotine in tobacco roots, combined with an increase in CO and JA concentration. Suppressing CO generation reduced both JA and nicotine biosynthesis, whereas exogenous application of CO increased JA and nicotine content. CO causes an increased expression of NtPMT1 (a key nicotine biosynthesis enzyme), via promoting NtMYC2a binding to the G-box region of its promoter, leading to heightened nicotine levels under HT conditions. These data suggest a novel function for CO in stimulating nicotine biosynthesis in tobacco under HT stress, through a JA signal.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010188
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 189: 3,3′-Diindolylmethane Suppressed
           Cyprodinil-Induced Epithelial-Mesenchymal Transition and
           Metastatic-Related Behaviors of Human Endometrial Ishikawa Cells via an
           Estrogen Receptor-Dependent Pathway

    • Authors: Bo-Gyoung Kim, Jin-Wook Kim, Soo-Min Kim, Ryeo-Eun Go, Kyung-A Hwang, Kyung-Chul Choi
      First page: 189
      Abstract: Cyprodinil (CYP) is a pyrimidine amine fungicide that has been extensively used in agricultural areas. 3,3′-Diindolylmethane (DIM) is a derivative of the dietary phytoestrogen, indole-3-carbinol (I3C), which is derived from cruciferous vegetables and considered to be a cancer-preventive phytonutrient agent. In this study, the effects of CYP and DIM were examined on the cell viability, invasion, and metastasis of human endometrial cancer cells, Ishikawa, via epithelial mesenchymal transition (EMT). CYP increased the level of cell viability of Ishikawa cells compared to DMSO as a control, as did E2. Ishikawa cells lost cell-to-cell contact and obtained a spindle-shaped or fibroblast-like morphology in response to the application of E2 or CYP by the cell morphology assay. In the cell migration and invasion assay, CYP enhanced the ability of migration and invasion of Ishikawa cells, as did E2. E2 and CYP increased the expressions of N-cadherin and Snail proteins, while decreasing the expression of E-cadherin protein as EMT-related markers. In addition, E2 and CYP increased the protein expressions of cathepsin D and MMP-9, metastasis-related markers. Conversely, CYP-induced EMT, cell migration, and invasion were reversed by fulvestrant (ICI 182,780) as an estrogen receptor (ER) antagonist, indicating that CYP exerts estrogenic activity by mediating these processes via an ER-dependent pathway. Similar to ICI 182,780, DIM significantly suppressed E2 and CYP-induced proliferation, EMT, migration, and invasion of Ishikawa cancer cells. Overall, the present study revealed that DIM has an antiestrogenic chemopreventive effect to withdraw the cancer-enhancing effect of E2 and CYP, while CYP has the capacity to enhance the metastatic potential of estrogen-responsive endometrial cancer.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010189
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 190: Visfatin Promotes IL-6 and TNF-α Production in
           Human Synovial Fibroblasts by Repressing miR-199a-5p through ERK, p38 and
           JNK Signaling Pathways

    • Authors: Min-Huan Wu, Chun-Hao Tsai, Yuan-Li Huang, Yi-Chin Fong, Chih-Hsin Tang
      First page: 190
      Abstract: Osteoarthritis (OA), an inflammatory form of arthritis, is characterized by synovial inflammation and cartilage destruction largely influenced by two key proinflammatory cytokines—interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α). Notably, levels of visfatin (a proinflammatory adipokine) are elevated in patients with OA, although the relationship of visfatin to IL-6 and TNF-α expression in OA pathogenesis has been unclear. In this study, visfatin enhanced the expression of IL-6 and TNF-α in human OA synovial fibroblasts (OASFs) in a concentration-dependent manner and stimulation of OASFs with visfatin promoted phosphorylation of extracellular-signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK), while ERK, p38, and JNK inhibitors or siRNAs all abolished visfatin-induced increases in IL-6 and TNF-α production. Moreover, transfection with miR-199a-5p mimics reversed visfatin-induced increases in IL-6 and TNF-α production. Furthermore, we also found that visfatin-promoted IL-6 and TNF-α production is mediated via the inhibition of miR-199a-5p expression through the ERK, p38, and JNK signaling pathways. Visfatin may therefore be an appropriate target for drug intervention in OA treatment.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010190
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 191: Cloning and Functional Analysis of
           Phosphoethanolamine Methyltransferase Promoter from Maize (Zea mays L.)

    • Authors: Gai-Li Niu, Wei Gou, Xiang-Long Han, Cheng Qin, Li-Xin Zhang, Abd Abomohra, Muhammad Ashraf
      First page: 191
      Abstract: Betaine, a non-toxic osmoprotectant, is believed to accumulate considerably in plants under stress conditions to maintain the osmotic pressure and promote a variety of processes involved in growth and development. Phosphoethanolamine N-methyltransferase (PEAMT), a key enzyme for betaine synthesis, is reported to be regulated by its upstream promoter. In the present investigation, by using the transgenic approach, a 1048 bp long promoter region of ZmPEAMT gene from Zea mays was cloned and functionally characterized in tobacco. Computational analysis affirmed the existence of abiotic stress responsive cis-elements like ABRE, MYC, HST, LST etc., as well as pathogen, wound and phytohormone responsive motifs. For transformation in tobacco, four 5′-deletion constructs of 826 bp (P2), 642 bp (P3), 428 bp (P4) and 245 bp (P5) were constructed from the 1048 bp (P1) promoter fragment. The transgenic plants generated through a single event exhibited a promising expression of GUS reporter protein in the leaf tissues of treated with salt, drought, oxidative and cold stress as well as control plants. The GUS expression level progressively reduced from P1 to P5 in the leaf tissues, whereas a maximal expression was observed with the P3 construct in the leaves of control plants. The expression of GUS was noted to be higher in the leaves of osmotically- or salt-treated transgenic plants than that in the untreated (control) plants. An effective expression of GUS in the transgenic plants manifests that this promoter can be employed for both stress-inducible and constitutive expression of gene(s). Due to this characteristic, this potential promoter can be effectively used for genetic engineering of several crops.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010191
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 192: Clinical Consequences and Molecular Bases of Low
           Fibrinogen Levels

    • Authors: Marguerite Neerman-Arbez, Alessandro Casini
      First page: 192
      Abstract: The study of inherited fibrinogen disorders, characterized by extensive allelic heterogeneity, allows the association of defined mutations with specific defects providing significant insight into the location of functionally important sites in fibrinogen and fibrin. Since the identification of the first causative mutation for congenital afibrinogenemia, studies have elucidated the underlying molecular pathophysiology of numerous causative mutations leading to fibrinogen deficiency, developed cell-based and animal models to study human fibrinogen disorders, and further explored the clinical consequences of absent, low, or dysfunctional fibrinogen. Since qualitative disorders are addressed by another review in this special issue, this review will focus on quantitative disorders and will discuss their diagnosis, clinical features, molecular bases, and introduce new models to study the phenotypic consequences of fibrinogen deficiency.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010192
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 193: Twelve Weeks of Medium-Intensity Exercise
           Therapy Affects the Lipoprotein Profile of Multiple Sclerosis Patients

    • Authors: Winde Jorissen, Tim Vanmierlo, Inez Wens, Veerle Somers, Bart Van Wijmeersch, Jeroen Bogie, Alan Remaley, Bert Eijnde, Jerome Hendriks
      First page: 193
      Abstract: Multiple sclerosis (MS) is an inflammatory auto-immune disease of the central nervous system (CNS). Serum glucose alterations and impaired glucose tolerance (IGT) are reported in MS patients, and are commonly associated with the development of cardio-metabolic co-morbidities. We previously found that a subgroup of MS patients shows alterations in their lipoprotein profile that are similar to a pre-cardiovascular risk profile. In addition, we showed that a high-intensity exercise training has a positive effect on IGT in MS patients. In this study, we hypothesize that exercise training positively influences the lipoprotein profile of MS patients. To this end, we performed a pilot study and determined the lipoprotein profile before (controls, n = 40; MS patients, n = 41) and after (n = 41 MS only) 12 weeks of medium-intensity continuous training (MIT, n = 21, ~60% of VO2max) or high-intensity interval training (HIT, n = 20, ~100–200% of VO2max) using nuclear magnetic resonance spectroscopy (NMR). Twelve weeks of MIT reduced intermediate-density lipoprotein particle count ((nmol/L); −43.4%; p < 0.01), low-density lipoprotein cholesterol (LDL-c (mg/dL); −7.6%; p < 0.05) and VLDL size ((nm); −6.6%; p < 0.05), whereas HIT did not influence the lipoprotein profile. These results show that MIT partially normalizes lipoprotein alterations in MS patients. Future studies including larger patient and control groups should determine whether MIT can reverse other lipoprotein levels and function and if these alterations are related to MS disease progression and the development of co-morbidities.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010193
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 194: Calbindin D28k-Immunoreactivity in Human Enteric
           Neurons

    • Authors: Katharina Zetzmann, Johanna Strehl, Carol Geppert, Stefanie Kuerten, Samir Jabari, Axel Brehmer
      First page: 194
      Abstract: Calbindin (CALB) is well established as immunohistochemical marker for intrinsic primary afferent neurons in the guinea pig gut. Its expression by numerous human enteric neurons has been demonstrated but little is known about particular types of neurons immunoreactive for CALB. Here we investigated small and large intestinal wholemount sets of 26 tumor patients in order to evaluate (1) the proportion of CALB+ neurons in the total neuron population, (2) the colocalization of CALB with calretinin (CALR), somatostatin (SOM) and vasoactive intestinal peptide (VIP) and (3) the morphology of CALB+ neurons. CALB+ neurons represented a minority of myenteric neurons (small intestine: 31%; large intestine: 25%) and the majority of submucosal neurons (between 72 and 95%). In the submucosa, most CALB+ neurons co-stained for CALR and VIP (between 69 and 80%) or for SOM (between 20 and 3%). In the myenteric plexus, 85% of CALB+ neurons did not co-stain with the other markers investigated. An unequivocal correlation between CALB reactivity and neuronal morphology was found for myenteric type III neurons in the small intestine: uniaxonal neurons with long, slender and branched dendrites were generally positive for CALB. Since also other neurons displayed occasional CALB reactivity, this protein is not suited as an exclusive marker for type III neurons.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-08
      DOI: 10.3390/ijms19010194
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 195: Surface Functionalization and Targeting
           Strategies of Liposomes in Solid Tumor Therapy: A Review

    • Authors: Muhammad Riaz, Muhammad Riaz, Xue Zhang, Congcong Lin, Ka Wong, Xiaoyu Chen, Ge Zhang, Aiping Lu, Zhijun Yang
      First page: 195
      Abstract: Surface functionalization of liposomes can play a key role in overcoming the current limitations of nanocarriers to treat solid tumors, i.e., biological barriers and physiological factors. The phospholipid vesicles (liposomes) containing anticancer agents produce fewer side effects than non-liposomal anticancer formulations, and can effectively target the solid tumors. This article reviews information about the strategies for targeting of liposomes to solid tumors along with the possible targets in cancer cells, i.e., extracellular and intracellular targets and targets in tumor microenvironment or vasculature. Targeting ligands for functionalization of liposomes with relevant surface engineering techniques have been described. Stimuli strategies for enhanced delivery of anticancer agents at requisite location using stimuli-responsive functionalized liposomes have been discussed. Recent approaches for enhanced delivery of anticancer agents at tumor site with relevant surface functionalization techniques have been reviewed. Finally, current challenges of functionalized liposomes and future perspective of smart functionalized liposomes have been discussed.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010195
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 196: Dissecting Time- from Tumor-Related Gene
           Expression Variability in Bilateral Breast Cancer

    • Authors: Maurizio Callari, Matteo Dugo, Patrizia Miodini, Silvia Veneroni, Giampaolo Bianchini, Maria Daidone, Vera Cappelletti
      First page: 196
      Abstract: Metachronous (MBC) and synchronous bilateral breast tumors (SBC) are mostly distinct primaries, whereas paired primaries and their local recurrences (LRC) share a common origin. Intra-pair gene expression variability in MBC, SBC, and LRC derives from time/tumor microenvironment-related and tumor genetic background-related factors and pairs represents an ideal model for trying to dissect tumor-related from microenvironment-related variability. Pairs of tumors derived from women with SBC (n = 18), MBC (n = 11), and LRC (n = 10) undergoing local-regional treatment were profiled for gene expression; similarity between pairs was measured using an intraclass correlation coefficient (ICC) computed for each gene and compared using analysis of variance (ANOVA). When considering biologically unselected genes, the highest correlations were found for primaries and paired LRC, and the lowest for MBC pairs. By instead limiting the analysis to the breast cancer intrinsic genes, correlations between primaries and paired LRC were enhanced, while lower similarities were observed for SBC and MBC. Focusing on stromal-related genes, the ICC values decreased for MBC and were significantly different from SBC. These findings indicate that it is possible to dissect intra-pair gene expression variability into components that are associated with genetic origin or with time and microenvironment by using specific gene subsets.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010196
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 197: Introduction of Exogenous HSV-TK Suicide Gene
           Increases Safety of Keratinocyte-Derived Induced Pluripotent Stem Cells by
           Providing Genetic “Emergency Exit” Switch

    • Authors: Maciej Sułkowski, Paweł Konieczny, Paula Chlebanowska, Marcin Majka
      First page: 197
      Abstract: Since their invention in 2006, induced Pluripotent Stem (iPS) cells remain a great promise for regenerative medicine circumventing the ethical issues linked to Embryonic Stem (ES) cell research. iPS cells can be generated in a patient-specific manner as an unlimited source of various cell types for in vitro drug screening, developmental biology studies and regenerative use. Having the capacity of differentiating into the cells of all three primary germ layers, iPS cells have high potential to form teratoma tumors. This remains their main disadvantage and hazard which, until resolved, prevents utilization of iPS cells in clinic. Here, we present an approach for increasing iPS cells safety by introducing genetic modification—exogenous suicide gene Herpes Simplex Virus Thymidine Kinase (HSV-TK). Its expression results in specific vulnerability of genetically modified cells to prodrug—ganciclovir (GCV). We show that HSV-TK expressing cells can be eradicated both in vitro and in vivo with high specificity and efficiency with low doses of GCV. Described strategy increases iPS cells safety for future clinical applications by generating “emergency exit” switch allowing eradication of transplanted cells in case of their malfunction.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010197
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 198: Understanding the Role of Intrinsic Disorder of
           Viral Proteins in the Oncogenicity of Different Types of HPV

    • Authors: Elvira Tamarozzi, Silvana Giuliatti
      First page: 198
      Abstract: Intrinsic disorder is very important in the biological function of several proteins, and is directly linked to their foldability during interaction with their targets. There is a close relationship between the intrinsically disordered proteins and the process of carcinogenesis involving viral pathogens. Among these pathogens, we have highlighted the human papillomavirus (HPV) in this study. HPV is currently among the most common sexually transmitted infections, besides being the cause of several types of cancer. HPVs are divided into two groups, called high- and low-risk, based on their oncogenic potential. The high-risk HPV E6 protein has been the target of much research, in seeking treatments against HPV, due to its direct involvement in the process of cell cycle control. To understand the role of intrinsic disorder of the viral proteins in the oncogenic potential of different HPV types, the structural characteristics of intrinsically disordered regions of high and low-risk HPV E6 proteins were analyzed. In silico analyses of primary sequences, prediction of tertiary structures, and analyses of molecular dynamics allowed the observation of the behavior of such disordered regions in these proteins, thereby proving a direct relationship of structural variation with the degree of oncogenicity of HPVs. The results obtained may contribute to the development of new therapies, targeting the E6 oncoprotein, for the treatment of HPV-associated diseases.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010198
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 199: Unfolding Role of a Danger Molecule Adenosine
           Signaling in Modulation of Microbial Infection and Host Cell Response

    • Authors: Jaden Lee, Özlem Yilmaz
      First page: 199
      Abstract: Ectonucleotidases CD39 and CD73, specific nucleotide metabolizing enzymes located on the surface of the host, can convert a pro-inflammatory environment driven by a danger molecule extracellular-ATP to an adenosine-mediated anti-inflammatory milieu. Accordingly, CD39/CD73 signaling have has strongly implicated in modulating the intensity, duration, and composition of purinergic danger signals delivered to host. Recent studies have eluted potential roles for CD39 and CD73 in selective triggering of a variety of host immune cells and molecules in the presence of pathogenic microorganisms or microbial virulence molecules. Growing evidence also suggests that CD39 and CD73 present complimentary, but likely differential, actions against pathogens to shape the course and severity of microbial infection as well as the associated immune response. Similarly, adenosine receptors A2A and A2B have been proposed to be major immunomodulators of adenosine signaling during chronic inflammatory conditions induced by opportunistic pathogens, such as oral colonizer Porphyromonas gingivalis. Therefore, we here review the recent studies that demonstrate how complex network of molecules in the extracellular adenosine signaling machinery and their interactions can reshape immune responses and may also be targeted by opportunistic pathogens to establish successful colonization in human mucosal tissues and modulate the host immune response.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010199
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 200: FM19G11 and Ependymal Progenitor/Stem Cell
           Combinatory Treatment Enhances Neuronal Preservation and
           Oligodendrogenesis after Severe Spinal Cord Injury

    • Authors: Ana Alastrue-Agudo, Francisco Rodriguez-Jimenez, Eric Mocholi, Francesca De Giorgio, Slaven Erceg, Victoria Moreno-Manzano
      First page: 200
      Abstract: Spinal cord injury (SCI) suffers from a lack of effective therapeutic strategies. We have previously shown that individual therapeutic strategies, transplantation of ependymal stem/progenitor cells of the spinal cord after injury (epSPCi) or FM19G11 pharmacological treatment, induce moderate functional recovery after SCI. Here, the combination of treatments has been assayed for functional and histological analysis. Immediately after severe SCI, one million epSPCi were intramedullary injected, and the FM19G11 compound or dimethyl sulfoxide (DMSO) (as the vehicle control) was administrated via intrathecal catheterization. The combination of treatments, epSPCi and FM19G11, improves locomotor tasks compared to the control group, but did not significantly improve the Basso, Beattie, Bresnahan (BBB) scores for locomotor analysis in comparison with the individual treatments. However, the histological analysis of the spinal cord tissues, two months after SCI and treatments, demonstrated that when we treat the animals with both epSPCi and FM19G11, an improved environment for neuronal preservation was generated by reduction of the glial scar extension. The combinatorial treatment also contributes to enhancing the oligodendrocyte precursor cells by inducing the expression of Olig1 in vivo. These results suggest that a combination of therapies may be an exciting new therapeutic treatment for more efficient neuronal activity recovery after severe SCI.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010200
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 201: Mangiferin Accelerates Glycolysis and Enhances
           Mitochondrial Bioenergetics

    • Authors: Zhongbo Liu, Pasha Apontes, Ekaterina Fomenko, Nan Chi, Victor Schuster, Irwin Kurland, Jeffrey Pessin, Yuling Chi
      First page: 201
      Abstract: One of the main causes of hyperglycemia is inefficient or impaired glucose utilization by skeletal muscle, which can be exacerbated by chronic high caloric intake. Previously, we identified a natural compound, mangiferin (MGF) that improved glucose utilization in high fat diet (HFD)-induced insulin resistant mice. To further identify the molecular mechanisms of MGF action on glucose metabolism, we conducted targeted metabolomics and transcriptomics studies of glycolyic and mitochondrial bioenergetics pathways in skeletal muscle. These data revealed that MGF increased glycolytic metabolites that were further augmented as glycolysis proceeded from the early to the late steps. Consistent with an MGF-stimulation of glycolytic flux there was a concomitant increase in the expression of enzymes catalyzing glycolysis. MGF also increased important metabolites in the tricarboxylic acid (TCA) cycle, such as α-ketoglutarate and fumarate. Interestingly however, there was a reduction in succinate, a metabolite that also feeds into the electron transport chain to produce energy. MGF increased succinate clearance by enhancing the expression and activity of succinate dehydrogenase, leading to increased ATP production. At the transcriptional level, MGF induced mRNAs of mitochondrial genes and their transcriptional factors. Together, these data suggest that MGF upregulates mitochondrial oxidative capacity that likely drives the acceleration of glycolysis flux.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010201
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 202: Conditions Inducing Excessive O-GlcNAcylation
           Inhibit BMP2-Induced Osteogenic Differentiation of C2C12 Cells

    • Authors: Hanna Gu, Mina Song, Kanitsak Boonanantanasarn, Kyunghwa Baek, Kyung Woo, Hyun-Mo Ryoo, Jeong-Hwa Baek
      First page: 202
      Abstract: Hyperglycemic conditions in diabetic patients can affect various cellular functions, including the modulation of osteogenic differentiation. However, the molecular mechanisms by which hyperglycemia affects osteogenic differentiation are yet to be clarified. This study aimed to investigate whether the aberrant increase in protein O-linked-β-N-acetylglucosamine glycosylation (O-GlcNAcylation) contributes to the suppression of osteogenic differentiation due to hyperglycemia. To induce osteogenic differentiation, C2C12 cells were cultured in the presence of recombinant human bone morphogenetic protein 2 (BMP2). Excessive protein O-GlcNAcylation was induced by treating C2C12 cells with high glucose, glucosamine, or N-acetylglucosamine concentrations or by O-GlcNAc transferase (OGT) overexpression. The effect of O-GlcNAcylation on osteoblast differentiation was then confirmed by examining the expression levels of osteogenic marker gene mRNAs, activity of alkaline phosphatase, and transcriptional activity of Runx2, a critical transcription factor for osteoblast differentiation and bone formation. Cell treatment with high glucose, glucosamine or N-acetylglucosamine increased O-GlcNAcylation of Runx2 and the total levels of O-GlcNAcylated proteins, which led to a decrease in the transcriptional activity of Runx2, expression levels of osteogenic marker genes (Runx2, osterix, alkaline phosphatase, and type I collagen), and activity of alkaline phosphatase. These inhibitory effects were rescued by lowering protein O-GlcNAcylation levels by adding STO45849, an OGT inhibitor, or by overexpressing β-N-acetylglucosaminidase. Our findings suggest that excessive protein O-GlcNAcylation contributes to high glucose-suppressed osteogenic differentiation.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010202
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 203: Intraarticularly-Injected Mesenchymal Stem Cells
           Stimulate Anti-Inflammatory Molecules and Inhibit Pain Related Protein and
           Chondrolytic Enzymes in a Monoiodoacetate-Induced Rat Arthritis Model

    • Authors: Toru Ichiseki, Miyako Shimazaki, Yoshimichi Ueda, Shusuke Ueda, Masanobu Tsuchiya, Daisuke Souma, Ayumi Kaneuji, Norio Kawahara
      First page: 203
      Abstract: Persistent inflammation is well known to promote the progression of arthropathy. mesenchymal stem cells (MSCs) have been shown to possess anti-inflammatory properties and tissue differentiation potency. Although the experience so far with the intraarticular administration of mesenchymal stem cell (MSC) to induce cartilage regeneration has been disappointing, MSC implantation is now being attempted using various surgical techniques. Meanwhile, prevention of osteoarthritis (OA) progression and pain control remain important components of the treatment of early-stage OA. We prepared a shoulder arthritis model by injecting monoiodoacetate (MIA) into a rat shoulder, and then investigated the intraarticular administration of MSC from the aspects of the cartilage protective effect associated with their anti-inflammatory property and inhibitory effect on central sensitization of pain. When MIA was administered in this rat shoulder arthritis model, anti-Calcitonin Gene Related Peptide (CGRP) was expressed in the joint and C5 spinal dorsal horn. Moreover, expression of A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5), a marker of joint cartilage injury, was similarly elevated following MIA administration. When MSC were injected intraarticularly after MIA, the expression of CGRP in the spinal dorsal horn was significantly deceased, indicating suppression of the central sensitization of pain. The expression of ADAMTS 5 in joint cartilage was also significantly inhibited by MSC administration. In contrast, a significant increase in the expression of TNF-α stimulated gene/protein 6 (TSG-6), an anti-inflammatory and cartilage protective factor shown to be produced and secreted by MSC intraarticularly, was found to extend to the cartilage tissue following MSC administration. In this way, the intraarticular injection of MSC inhibited the central sensitization of pain and increased the expression of the anti-inflammatory and cartilage protective factor TSG-6. As the least invasive conservative strategies possible are desirable in the actual clinical setting, the intraarticular administration of MSC, which appears to be effective for the treatment of pain and cartilage protection in early-stage arthritis, may achieve these aims.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010203
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 207: Synthesized Heparan Sulfate Competitors
           Attenuate Pseudomonas aeruginosa Lung Infection

    • Authors: Nicola Lorè, Noemi Veraldi, Camilla Riva, Barbara Sipione, Lorenza Spagnuolo, Ida De Fino, Medede Melessike, Elisa Calzi, Alessandra Bragonzi, Annamaria Naggi, Cristina Cigana
      First page: 207
      Abstract: Several chronic respiratory diseases are characterized by recurrent and/or persistent infections, chronic inflammatory responses and tissue remodeling, including increased levels of glycosaminoglycans which are known structural components of the airways. Among glycosaminoglycans, heparan sulfate (HS) has been suggested to contribute to excessive inflammatory responses. Here, we aim at (i) investigating whether long-term infection by Pseudomonas aeruginosa, one of the most worrisome threat in chronic respiratory diseases, may impact HS levels, and (ii) exploring HS competitors as potential anti-inflammatory drugs during P. aeruginosa pneumonia. P. aeruginosa clinical strains and ad-hoc synthesized HS competitors were used in vitro and in murine models of lung infection. During long-term chronic P. aeruginosa colonization, infected mice showed higher heparin/HS levels, evaluated by high performance liquid chromatography-mass spectrometry after selective enzymatic digestion, compared to uninfected mice. Among HS competitors, an N-acetyl heparin and a glycol-split heparin dampened leukocyte recruitment and cytokine/chemokine production induced by acute and chronic P. aeruginosa pneumonia in mice. Furthermore, treatment with HS competitors reduced bacterial burden during chronic murine lung infection. In vitro, P. aeruginosa biofilm formation decreased upon treatment with HS competitors. Overall, these findings support further evaluation of HS competitors as a novel therapy to counteract inflammation and infection during P. aeruginosa pneumonia.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010207
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 208: Aeginetia indica Decoction Inhibits Hepatitis C
           Virus Life Cycle

    • Authors: Cheng-Wei Lin, Chieh-Wen Lo, Chia-Ni Tsai, Ting-Chun Pan, Pin-Yin Chen, Ming-Jiun Yu
      First page: 208
      Abstract: Chronic hepatitis C virus (HCV) infection is still a global epidemic despite the introduction of several highly effective direct-acting antivirals that are tagged with sky-high prices. The present study aimed to identify an herbal decoction that ameliorates HCV infection. Among six herbal decoctions tested, the Aeginetia indica decoction had the most profound effect on the HCV reporter activity in infected Huh7.5.1 liver cells in a dose- and time-dependent manner. The Aeginetia indica decoction exerted multiple inhibitory effects on the HCV life cycle. Pretreatment of the cells with the Aeginetia indica decoction prior to HCV infection reduced the HCV RNA and non-structural protein 3 (NS3) protein levels in the infected cells. The Aeginetia indica decoction reduced HCV internal ribosome entry site-mediated protein translation activity. It also reduced the HCV RNA level in the infected cells in association with reduced NS5A phosphorylation at serine 235, a predominant phosphorylation event indispensable to HCV replication. Thus, the Aeginetia indica decoction inhibits HCV infection, translation, and replication. Mechanistically, the Aeginetia indica decoction probably reduced HCV replication via reducing NS5A phosphorylation at serine 235.
      Citation: International Journal of Molecular Sciences
      PubDate: 2018-01-09
      DOI: 10.3390/ijms19010208
      Issue No: Vol. 19, No. 1 (2018)
       
  • IJMS, Vol. 19, Pages 104: Effects of Plant Oil Interesterified
           Triacylglycerols on Lipemia and Human Health

    • Authors: Andreina Alfieri, Esther Imperlini, Ersilia Nigro, Daniela Vitucci, Stefania Orrù, Aurora Daniele, Pasqualina Buono, Annamaria Mancini
      First page: 104
      Abstract: The position of the fatty acids (sn-1, sn-2 and sn-3) (stereospecific numbering (sn)) in triacylglycerol (TAG) molecules produces a characteristic stereospecificity that defines the physical properties of the fats and influences their absorption, metabolism and uptake into tissues. Fat interesterification is a process that implies a positional distribution of fatty acids (FAs) within the TAG molecules, generating new TAG species, without affecting the FA cis-trans natural balance. The interesterified (IE) fats, frequently used in the food industry comprise fats that are rich in long-chain saturated FAs, such as palmitic acid (16:0) and stearic acid (18:0). Within the interesterified fats, a critical role is played by FA occupying the sn-2 position; in fact, the presence of an unsaturated FA in this specific position influences early metabolic processing and postprandial clearance that in turn could induce atherogenesis and thrombogenesis events. Here, we provide an overview on the role of TAG structures and interesterified palmitic and stearic acid-rich fats on fasting and postprandial lipemia, focusing our attention on their physical properties and their effects on human health.
      Citation: International Journal of Molecular Sciences
      PubDate: 2017-12-30
      DOI: 10.3390/ijms19010104
      Issue No: Vol. 19, No. 1 (2017)
       
  • IJMS, Vol. 19, Pages 107: The Role of Compounds Derived from Natural
           Supplement as Anticancer Agents in Renal Cell Carcinoma: A Review

    • Authors: Inamul Haque, Arvind Subramanian, Chao Huang, Andrew Godwin, Peter Van Veldhuizen, Snigdha Banerjee, Sushanta Banerjee
      First page: 107
      Abstract: Renal Cell Carcinoma (RCC) is the most prominent kidney cancer derived from renal tubules and accounts for roughly 85% of all malignant kidney cancer. Every year, over 60,000 new cases are registered, and about 14,000 people die from RCC. The incidence of this has been increasing significantly in the U.S. and other countries. An increased understanding of molecular biology and the genomics of RCC has uncovered several signaling pathways involved in the progression of this cancer. Significant advances in the treatment of RCC have been reported from agents approved by the Food and Drug Administration (FDA) that target these pathways. These agents have become drugs of choice because they demonstrate clinical benefit and increased survival in patients with metastatic disease. However, the patients eventually relapse and develop resistance to these drugs. To improve outcomes and seek approaches for producing long-term durable remission, the search for more effective therapies and preventative strategies are warranted. Treatment of RCC using natural products is one of these strategies to reduce the incidence. However, recent studies have focused on these chemoprevention agents as anti-cancer therapies given they can inhibit tumor cell grow and lack the severe side effects common to synthetic compounds. This review elaborates on the current understanding of natural products and their mechanisms of action as anti-cancer agents. The present review will provide information for possible use of these products alone or in combination with chemotherapy for the prevention and treatment of RCC.
      Citation: International Journal of Molecular Sciences
      PubDate: 2017-12-31
      DOI: 10.3390/ijms19010107
      Issue No: Vol. 19, No. 1 (2017)
       
  • IJMS, Vol. 19, Pages 109: Liver Graft Susceptibility during Static Cold
           Storage and Dynamic Machine Perfusion: DCD versus Fatty Livers

    • Authors: Andrea Ferrigno, Laura Di Pasqua, Clarissa Berardo, Veronica Siciliano, Vittoria Rizzo, Barbara Mannucci, Plinio Richelmi, Anna Croce, Mariapia Vairetti
      First page: 109
      Abstract: We compared static preservation (cold storage, CS, 4 °C) with dynamic preservation (machine perfusion, MP, 20 °C) followed by reperfusion using marginal livers: a model of donation after cardiac death (DCD) livers and two models of fatty livers, the methionine-choline deficient (MCD) diet model, and obese Zucker (fa/fa) rats. CS injury in DCD livers was reversed by an oxygenated washout (OW): hepatic damage, bile flow, and the ATP/ADP ratio in the OW + CS group was comparable with the ratio obtained with MP. Using fatty livers, CS preservation induced a marked release in hepatic and biliary enzymes in obese Zucker rats when compared with the MCD group. The same trend occurred for bile flow. No difference was found when comparing MP in MCD and obese Zucker rats. Fatty acid analysis demonstrated that the total saturated (SFA)/polyunsaturated fatty acid (PUFA) ratio was, respectively, 1.5 and 0.71 in obese Zucker and MCD rats. While preservation damage in DCD livers is associated with the ATP/ADP recovered with OW, injury in fatty livers is linked to fatty acid constituents: livers from obese. Zucker rats, with greater content in saturated FA, might be more prone to CS injury. On the contrary, MCD livers with elevated PUFA content might be less susceptible to hypothermia.
      Citation: International Journal of Molecular Sciences
      PubDate: 2017-12-31
      DOI: 10.3390/ijms19010109
      Issue No: Vol. 19, No. 1 (2017)
       
  • IJMS, Vol. 19, Pages 112: Apolipoprotein M Inhibits Angiogenic and
           Inflammatory Response by Sphingosine 1-Phosphate on Retinal Pigment
           Epithelium Cells

    • Authors: Ryo Terao, Megumi Honjo, Makoto Aihara
      First page: 112
      Abstract: Sphingosine 1-phosphate (S1P) is a potent lipid mediator that modulates inflammatory responses and proangiogenic factors. It has been suggested that S1P upregulates choroidal neovascularization (CNV) and may be deeply involved in the pathogenesis of exudative age-related macular degeneration (AMD). Recent studies have suggested that apolipoprotein M (ApoM), a carrier protein for S1P, modulates the biological properties of S1P in the pathogenesis of atherosclerosis. However, the role of ApoM/S1P in AMD has not been explored. We investigated the effect of S1P on proangiogenic factors in human retinal pigment epithelium (RPE) cell lines in vitro. S1P promoted the expression of vascular endothelial growth factor in RPE cells. Hypoxia inducible factor-1α expression was also upregulated. These S1P-induced enhancements in growth factors and chemotactic cytokines in RPE cells were significantly inhibited by ApoM treatment. Additionally, in vivo experiments using a laser-induced CNV murine model demonstrated that intravitreal ApoM injection significantly reduced the progression of CNV formation. Although the detailed mechanisms remain to be elucidated, the present results provide a novel potential therapeutic target for AMD, and demonstrate a suppressive role for ApoM and S1P in the pathology of CNV progression.
      Citation: International Journal of Molecular Sciences
      PubDate: 2017-12-31
      DOI: 10.3390/ijms19010112
      Issue No: Vol. 19, No. 1 (2017)
       
 
 
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