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European Journal of Biology
Number of Followers: 1  

  This is an Open Access Journal Open Access journal
ISSN (Print) 2602-2575 - ISSN (Online) 2618-6144
Published by İstanbul Üniversitesi Homepage  [18 journals]
  • Induction of Apoptosis in BCR-ABL Fusion Associated Chronic Myeloid
           Leukemia Cells by Camellia kissiWall. (Theaceae) Extract

    • Authors: Nguyen Anh Xuan; Nguyen Trung Quan, Bui Thi Kim Ly, Hoang Thanh Chi
      Abstract: Objective: Camellia kissi, a prominent tea, lacks academic works. In a previous report, this plant substantially affected chronic myeloid leukemia cells. Understanding the mechanism of action of this tea species on leukemia cells will contribute to researching alternative treatment methods in the context of drug resistance in chronic myeloid leukemia, which is constantly increasing.Materials and Methods: C. kissi comes from Lam Dong, Vietnam. The crude tea extract in methanol was obtained. The flow cytometry method with Annexin V and PI staining and the DNA fragmentation assays were used to indicate the apoptosis cells. The reversed transcription real-time PCR reactions were conducted to measure the mRNA level under the treatment.Results: The results showed the apoptosis-inducing capacity of the C. kissi extract on K562 cells, and the impact was suggested to be through the induction of intracellular apoptosis and cell cycle arrest.Conclusion: The apoptosis induction on K562 caused by C. kissi was reported for the first time. Initial recognition of the signaling pathway of inhibition is through the BCR-ABL/PTEN.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Bacterial Diversity of the Corpses

    • Authors: Ahmet Asan
      Abstract: The study presents the importance of forensic bacteriology, its use in forensic cases, the methods for bacteriological sampling from corpses, the types and species of bacteria isolated from human and pig corpses, which are most commonly used in forensic biology. The microbial changes that occur after death remain unclear. Postmortem microbiology is a relatively new field of research. After death, the failure of the immune system and other physical barriers leads to the proliferation and spread of microbes. In order for bacteriological information to be accepted within the scope of forensic bacteriology, the court must find suspicion to be present about whether the bacteria seen on the body will contribute to solving the case. Experts must be appointed to examine the issue in line with this suspicion, and these experts must prepare and submit their reports to the court at the requested time. When considering the literature studies, forensic bacteriology has been suggested to be a scientific discipline in the developmental stage and to only be able to provide circumstantial evidence in forensic cases as opposed to primary evidence. According to the literature review, most bacterial studies isolated from corpses were conducted in Romania. Although bacterial samples were isolated from various parts of the corpses, bacteria were mostly isolated from their blood samples. According to literature searches from various scientific journal databases, no study has occurred with a list of the bacteria isolated from corpses. This study is thought to be able to fill this important gap.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • From Pond Scum to Miracle Molecules: Cyanobacterial Compounds New
           Frontiers

    • Authors: Arbab Husain; Md Nematullah, Hamda Khan, Ravi Shekher, Alvina Farooqui, Archana Sahu, Afreen Khanam
      Abstract: Cyanobacteria are a diverse group of photosynthetic microorganisms known for their production of bioactive compounds with various biological activities. This review explores cyanobacterial bioactive compounds’ current and future prospects and their roles in different fields. These compounds have great potential for pharmaceuticals, agriculture, and environmental remediation applications. Cyanobacterial bioactive compounds, such as cyanotoxins, peptides, polyketides, alkaloids, and terpenoids, exhibit remarkable properties, including antimicrobial, antifungal, antiviral, antioxidant, anti-inflammatory, and anticancer activities. Advances in genomics, metabolomics, synthetic biology, screening techniques, and bioinformatics have facilitated the identification, characterization, and manipulation of cyanobacterial compounds. The future prospects involve exploring untapped cyanobacterial diversity, integrating advanced technologies like machine learning and high-throughput screening, and sustainable production through biotechnological approaches. These efforts hold promise for discovering new bioactive compounds with unique properties and applications, contributing to the development of innovative pharmaceuticals, agricultural solutions, and environmental remedies.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Changes in Histological Features, Apoptosis and Necroptosis, and
           Inflammatory Status in the Livers and Kidneys of Young and Adult Rats

    • Authors: Emine Rümeysa Hekimoğlu; Mukaddes Eşrefoğlu, Birsen Elibol, Seda Kırmızıkan
      Abstract: Objective: Aging entails a gradual rise in low-grade inflammation affected by cellular degeneration and death. Inflammaging refers to the chronic, low-grade inflammation that occurs alongside the aging process. This study attempts to evaluate the hepatic and renal histological changes, apoptosis and necroptosis rates, and inflammaging status of 6-week-old and 10-month-old rats.Materials and Methods: This study uses 12 male rats separated into two groups: Young Group (6-week-old rats; n = 6), and Adult Group (10-month-old rats; n = 6). Animals were sacrificed under anesthesia. The rats’ livers and kidneys were removed, and each organ tissue was divided into two parts: one for the microscopic examination (H&E and TUNEL immunohistochemistry) and the other for biochemical determination (tumor necrosis factor-alpha [TNF-α], nuclear factor-kappa beta [NF-𝜅B], interleukin1-Beta [IL-1β], IL6, receptor-interacting serine/threonine protein kinase [RIP], and RIP3).Results: The histological features of the livers and kidneys of the 6-week-old rats were consistent with healthy mammalian organ features, while some histological changes were detected in sections of the 10-month-old rats. The apoptosis rate indicated by TUNEL immunohistochemistry was seen to have increased in the 10-month-old rats, while the necroptosis rate indicated by RIP3 Western-blotting analysis was conversely determined to have decreased. Significant increases in TNF-α and NF-𝜅B levels were consistent with the increased apoptosis rate in the 10-month-old rats compared to the 6-week-old rats.Conclusion: One of the striking results of this study is that the degenerative changes related to aging began to be seen even in 10-month-old rats. The researchers used healthy rats of this age as control subjects as well as to create experimental models.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Radiofrequency Electromagnetic Field Exposure Amplifies the Detrimental
           Effects of Fetal Hyperglycemia in Zebrafish Embryos

    • Authors: Derya Cansız; Merih Beler, Gizem Eğilmezer, Semanur Işıkoğlu, Zülal Mızrak, İsmail Ünal, Selçuk Paker, Ahmet Ata Alturfan, Ebru Emekli Alturfan
      Abstract: Objective: Radiofrequency electromagnetic field (RF-EMF) exposure during the embryonic period can cause defects in the development of the fetus. The study’s aim is to evaluate the effects of RF-EMF on the lipid accumulation, oxidant-antioxidant system parameters, locomotor activities, and gene expressions of insulin and leptin as genes related to insulin resistance in fetal hyperglycemia-induced zebrafish embryos.Materials and Methods: The study exposed zebrafish embryos to RF-EMF (60 min) and glucose (5%) every day until 96 hours post fertilization (hpf). The study measured lipid peroxidation (LPO), superoxide dismutase, nitric oxide (NO), glutathione S-transferase (GST), and glutathione (GSH) levels to observe the oxidative stress status. The study monitored the development of the zebrafish embryos under a microscope, performed a locomotor activity analysis, measured acetylcholinesterase activity, and conducted oil red O staining to determine lipid accumulation. The study used reverse transcription polymerase chain reactions (RT-PCRs) to determine the expressions of ins and lepa by using RT-PCR.Results: Both the glucose and RF-EMF applications decreased locomotor activity and increased the LPO and NO levels as oxidative damage indicators. Applying RF-EMF alone increased GST and GSH levels, while applying RF-EMF and glucose showed a decrease in the antioxidant defense systems. ins expression increased in the glucose and RF-EMF groups, while lepa expression increased in the glucose group and decreased in the RF-EMF group.Conclusion: The harmful effects of hyperglycemia and RF-EMF exposure during the fetal period on embryo development need to be supported by studies to confirm the changes the current study has identified at the gene and protein levels.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Investigating the Antioxidant Capacity of Lunasin Expressed in Aspergillus
           oryzae

    • Authors: Elif Karaman; Cem Albayrak, Serdar Uysal
      Abstract: Objective: Lunasin is a bioactive protein that possesses anti-carcinogenic, anti-inflammatory, and antioxidant properties. Traditional isolation methods are resource-intensive, and chemical synthesis faces cost and environmental issues. This study aims to achieve cost-effective lunasin expression in Aspergillus oryzae with a focus on exploring its antioxidant properties in vitro.Materials and Methods: The expression vector carrying four lunasin sequences fused with amylase and an 8xHis-tag was introduced into pyrG auxotrophic A. oryzae. Subsequently, the recombinant protein was purified using metal affinity chromatography. The study uses sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), western blot analyses, and size-exclusion chromatography to evaluate the composition and purity of the protein, a linoleic acid assay to demonstrate the inhibitory effect on lipid peroxidation, and the 2,2’-azinobis-[3-ethylbenzothiazoline-6-sulfonic acid] ABTS) assay to evaluate the radical scavenging activity.Results: SDS-PAGE and western blot analyses confirmed sustained lunasin expression in A. oryzae, appearing in both fusion and non-fusion forms. Yields were 5.8 mg/L for non-fusion and 4 mg/L for fusion lunasin expression. Moreover, 0.1 μM non-fusion lunasin surpassed α-tocopherol and butylated hydroxyanisole (BHA; p < 0.05) in reducing lipid peroxidation at 4 and 72 h. Unlike the fusion lunasin, the non-fusion lunasin displayed concentration- and time-independent inhibitory effects on linoleic acid peroxidation as well as significant ABTS scavenging activity (p < 0.05).Conclusion: The study has shown for the first time A. oryzae to efficiently express and secrete both fusion and non-fusion lunasin proteins in a soluble form, with the non-fusion lunasin exhibiting superior antioxidant effectiveness compared to the fusion lunasin. The findings underscore A. oryzae’s potential as a promising host for producing functional lunasin with antioxidant properties, opening avenues for broader applications in biotechnology and bioactive peptides.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Investigating the Involvement of Fibroblast Growth Factors in Adipose
           Tissue Thermogenesis

    • Authors: Serkan Kır
      Abstract: Objective: Thermogenesis in white and brown adipose tissues can be induced by various stimuli, including cold exposure, β-adrenergic stimulation, and tumor growth. Fibroblast growth factor (FGF) 21 has emerged as an important mediator of thermogenesis. This study investigated the involvement of other FGF family members in the regulation of adipose tissue thermogenesis.Materials and Methods: Mice were exposed to cold and administrated a β-adrenergic agonist (CL-316,243) to stimulate a thermogenic response in adipose tissues. Stromavascular fractions isolated from white and brown adipose tissues were cultured and differentiated into primary adipocytes. These cells were treated with recombinant FGFs. Changes in the expression levels of thermogenic genes and FGFs were determined by real-time quantitative PCR.Results: Cold exposure stimulated thermogenic gene expression in the adipose tissue, which was accompanied by the upregulation of certain FGFs. Ffg9 and Fgf21 were prominently induced in white and brown adipose tissues. β-adrenergic stimulation also upregulated thermogenic genes in adipocytes. Fgf21 was identified as the main responder to the β-adrenergic pathway. The administration of recombinant FGFs to cultured primary white and brown adipocytes induced thermogenic genes, including uncoupling protein-1 (Ucp1). FGF2, FGF9, and FGF21 triggered the most significant Ucp1-inducing effects in these cells.Conclusion: FGF21 is as a prominent inducer of thermogenesis in adipose tissue and a promising therapeutic target against cardiovascular and metabolic diseases. FGF2 and FGF9 potently promote thermogenic gene expression in adipocytes. Therefore, their therapeutic targeting should be considered to enhance energy metabolism in adipose tissues.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Induction of Apoptosis through Oxidative Stress Caused by Rubus
           tereticaulis Leaves Extracts in A549 Cells

    • Authors: Gamze Nur Öter; Ezgi Durmuş, Ali Şen, Abdurrahim Koçyiğit
      Abstract: Objective: Plants have been used for medicinal purposes since the beginning of human history and form the basis of modern medicine, and they are also the source of most chemotherapeutic drugs for cancer treatment. This study aims to investigate for the first time the cytotoxic and apoptotic effects of the active ethanol (RTE) and chloroform (RTC) extracts of Rubus tereticaulis leaves in the A549 non-small-cell lung cancer cell line.Materials and Methods: A549 cells were treated with RTE and RTC individually. The MTT assay was used to quantitatively detect RTE and RTC’s cytotoxic effects. The fluorescent signal indicator H2DCF-DA was used to detect cellular reactive oxygen species (ROS) production. Apoptosis was evaluated by fluorescence microscope after acridine orange/ethidium bromide fluorescent staining, annexin V-FITC and immunoblotting analyses, immunofluorescence, and imaging.Results: Both RTE and RTC induced cytotoxicity in A549 cells in a dose-dependently, which was accompanied with induced ROS accumulation. Both early and late apoptotic cells detected by flow cytometry were increased in the RTE- and RTC-treated cells. In addition, the results show RTC to have higher cytotoxic and apoptotic effects and increased ROS-generation capacity than RTE. Therefore, the polarity of the solvent used to exert the anticancer effect of R. tereticaulis leaves is crucial.Conclusion: This is the first anti-cancer activity study on R. tereticaulis. The results suggest R. tereticaulis leaves to have an anti-cancer effect on lung cancer cells through ROS-mediated apoptosis and RTC to be an effective therapeutic/adjuvant strategy in cancer treatment.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Resveratrol Dose-Dependently Protects the Antioxidant Mechanism of
           Hydrogen Peroxide-Exposed Healthy Cells and Lung Cancer Cells

    • Authors: Öznur Yurdakul; Aysun Özkan
      Abstract: Objective: The objective of this study was to investigate the protective effects and the underlying mechanisms of resveratrol against hydrogen peroxide (H2O2 )-induced oxidative stress in healthy human and lung cancer cells.Materials and Methods: The cytotoxic doses and IC50 values of resveratrol and hydrogen peroxide for cells were determined by the Cell Titer Blue-Viability Assay kit. The amount of malondialdehyde (MDA) was determined by fluorescence spectrophotometer. The amount of intracellular reduced glutathione level and antioxidant enzyme activities were analyzed by spectrophotometric methods.Results: In both cells, H2O2 treatment alone (IC50 and IC50) increased MDA, glutathione reductase, glutathione S-transferase, selenium-dependent glutathione peroxidase and non-selenium-dependent glutathione peroxidase activities, but glutathione levels decreased compared to the H2O + 2 resveratrol treatment. In addition, high doses of resveratrol alone (IC50 and IC70) induced more oxidative stress in cancer cells than in healthy cells. High doses of resveratrol alone (IC50 and IC70) also showed cytotoxic effects in cells and decreased cell viability. Resveratrol caused more cytotoxic effects in cancer cells compared to healthy cells.Conclusion: The results of this study show that the increase in MDA level and antioxidant enzyme activity caused by highdose resveratrol treatment reveals the prooxidant effect of resveratrol. Our results also showed an antioxidant effect by reducing oxidative stress in cells pre-incubated with low-dose resveratrol and then exposed to H2O2 . Resveratrol has a dose-dependent biphasic (pro/antioxidant) effect on the antioxidant mechanism of cells. However, more research is needed to confirm this
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • In Silico Evaluation of ERQ Bioactive Tripeptide as an Anticancer Agent
           and an Inhibitor of SARS-CoV-2 Enzymes

    • Authors: Gözde Yılmaz; Sefa Çelik, Ayşen Erbölükbaş Özel, Sevim Akyüz
      Abstract: Objective: Short peptides play a significant role in exploring drugs with higher selectivity and fewer side effects in cancer and COVID-19 therapies. This study evaluated the anticancer and anti-COVID-19 activities of Glu-Arg-Gln (ERQ) tripeptide for the first time. To discover the potentiality of the tripeptide as an anticancer and as a SARS-CoV-2 inhibitor, molecular docking analysis of ERQ tripeptide with DNA (PDB ID: 1BNA) and a variety of SARS-CoV-2 enzymes, namely. Main protease (PDB IDs: 6M03, 6LU7) and Spike glycoprotein (PDB ID: 6VXX) were performed.Materials and Methods: To determine the binding efficiency of ERQ to target DNA and proteins, molecular docking processes were carried out using the Autodock Vina program. The sorts of bonds and interacting residues in ERQ/DNA and ERQ/protein complexes were determined.Results: Molecular docking simulations of ERQ tripeptide against 1BNA, 6M03, 6LU7, and 6VXX were performed, and the interactions between the docked ligand and target residues were determined. The binding mechanisms of ERQ with the receptors were clarified. The binding affinities of ERQ towards the targets were predicted to be between -6.3 and -6.7 kcal/mol. ERQ showed the highest binding affinity to Spike glycoprotein (6VXX), with an estimated binding energy of -6.7 kcal/mol.Conclusion: Molecular docking simulations revealed the potential of ERQ tripeptide as an anticancer and anti-COVID-19 agent. High binding affinity against 1BNA (-6.4 kcal/mol), 6M03 (-6.3 kcal/mol), 6LU7 (-6.6 kcal/mol), and 6VXX (-6.7 kcal/mol) indicated that ERQ could be an excellent new natural therapy for the treatment of cancer and COVID-19.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Investigation of the Relationship between the Multidrug Resistance 1 Gene
           Polymorphisms and Bronchodilator Response in COPD

    • Authors: Ersan Atahan; Buket Çalışkaner Öztürk, Suat Sarıbaş, Bülent Tutluoğlu
      Abstract: Objective: Chronic obstructive pulmonary disease (COPD) is described as partially reversible airflow limitation. P-glycoprotein (P-gp/MDR1), encoded by the Multidrug Resistance 1 (MDR1) gene, is regarded as a protective component for the respiratory tract and is present in tracheobronchial epithelium and lung parenchyma, and removes particles from cells and protects against various xenobiotics. Polymorphisms of MDR1 gene and the alteration in the expression of P-gp are considered to have a negative effect on the severity of COPD pathogenesis and treatment efficacy. We aimed to investigate the relationship of the MDR1 gene polymorphisms with reversibility in COPD patients.Materials and Methods: The MDR1 polymorphisms, specifically the 3435C>T and 2677A/G variations, were analyzed in 90 COPD patients.Results: 15 of the 90 COPD patients had positive reversibility tests. 2677TT (p=0.044) and 3435TT (p=0.003) alleles related to positive reversibility tests. There were no significant differences in the distribution of the MDR1 C3435 alleles and the G2677 alleles (p> 0.05).Conclusion: COPD patients with the TT allele have a higher rate of early reversibility positivity; this suggests that those carrying the allele may respond better to bronchodilator therapy. These markers could help to distinguish COPD patients who respond better to β2-agonists or who may not benefit much and, therefore, need different drugs.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Retrospective Analysis of Transcriptomic Differences between
           Triple-Negative Breast Cancer (TNBC) and non-TNBC

    • Authors: Çağlar Berkel
      Abstract: Objective: Triple-negative breast cancer (TNBC), which has no expression of estrogen receptor, progesterone receptor and HER2, is an aggressive subgroup. Molecular differences between TNBC and non-TNBC should be better understood to develop tailored treatment strategies.Materials and Methods: The expression of the most frequently mutated genes, and of genes for which copy number variation events are observed in the highest percentage of breast cancer patients, was compared between TNBC and non-TNBC samples, in R programming environment, using TCGA-BRCA transcriptomics dataset.Results: 70% of the most frequently mutated genes in breast cancer (CDH1, GATA3, MLL3 (KMT2C), MAP3K1, PTEN, NCOR1, FAT3, MAP2K4, NF1, ARID1A, LRP1B, RUNX1, MLL2 (KMT2D) and TBX3) was found to have decreased expression in TNBC compared to non-TNBC. The expression of 40% of the genes with the highest frequency of copy number gain events in breast cancer (SLC45A3, PTPRC, ELF3, FCGR2B, AKT3, FH, TPM3 and SETDB1) was increased in TNBC compared with non-TNBC. The half of the genes with the highest percentage of copy number loss events in breast cancer (CBFA2T3, CDH1, ZFHX3, CDH11, MAP2K4, GAS7, PER1, RABEP1, NCOR1 and PCM1) was observed to have decreased expression in TNBC compared to non-TNBC. Lastly, the expression of BRCA2, but not of BRCA1, was found to be higher in TNBC than in non-TNBC.Conclusion: This study provides further evidence in support of previous research, which show the presence of a large number of molecular differences between TNBC and non-TNBC, pointing to the need for more tailored treatment strategies for patients with TNBC.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Susceptibility of Staphylococcus aureus Isolated from Different Raw Meat
           Products to Disinfectants

    • Authors: Fatma Özdemir; Seza Arslan
      Abstract: Objective: Staphylococcus aureus, a severe public health hazard, causes foodborne diseases from the consumption of contaminated food. Various antimicrobials and disinfectants are used throughout the food chain to reduce microbial contamination or eliminate microorganisms on food contact surfaces. However, little is known about the susceptibility of disinfectants to food pathogens, including S. aureus, which can develop resistance to antimicrobials and cause severe diseases.Materials and Methods: The antimicrobial activity of triclosan, cetyltrimethylammonium bromide (CTAB), acetic acid, citric acid, and lactic acid against 50 S. aureus isolates, including multidrug-resistant (MDR) isolates originating from ground beef, chicken, and fish, was investigated using the broth microdilution method.Results: The minimal inhibitory concentrations (MICs) of triclosan, CTAB, acetic acid, citric acid, and lactic acid against the isolates were 0.125-16 μg/mL, 0.25-32 μg/mL, 102.5-26250 μg/mL, 187.5-12000 μg/mL, and 703-22500 μg/mL, respectively. Almost all MDR isolates showed resistance to triclosan. There was a statistically significant difference in MICs between triclosan and organic acids, as well as between CTAB and organic acids (p < 0.05). However, a statistically significant difference was not observed in triclosan and CTAB, as well as in acetic acid and lactic acid (p > 0.05). Pearson correlation coefficient revealed a strong relationship between triclosan and multidrug resistance. Based on the multiple linear regression analysis, triclosan had a positive effect on multidrug resistance (p < 0.05).Conclusion: This research gives helpful information on the susceptibility of disinfectants to S. aureus, particularly to resistant S. aureus isolates from meats, which may help to recommend proper disinfectant use in food production.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
  • Astragaloside-IV Inhibits Metastasis by Suppressing the SDF-1/CXCR4 Axis
           and Activating Apoptosis in Cisplatin-Resistant Ovarian Cancer Cells

    • Authors: Burçin İrem Abas; Ömer Erdoğan, Özge Çevik
      Abstract: Objective: Ovarian cancer has the highest mortality rate in women and it has a poor response rate to treatment due to its late diagnosis and is frequently resistant to currently used cisplatin-based treatment methods. Astragaloside IV (As-IV), a bioactive compound and natural tripeptide glycoside known as an antioxidant, has drawn attention in Chinese medicine for its healing properties. Many studies have shown that it has anti-inflammatory, antidiabetic, antitumoral, and anti-angiogenic properties.Materials and Methods: In our study, we first rendered ovarian cancer cells (OVCAR-3) resistant to cisplatin and then applied determined doses of As-IV (40 μg/mL) and (70 μg/mL) to OVCAR-3 cells and cisplatin-resistant ovarian cancer cells (OVCAR-3- CisR). The cell viability capacity, variation of BAX/BCL-2 gene expression, and regulation of the SDF-1/CXCR4 chemokine axis protein and their gene expressions were investigated.Results: According to the findings, As-IV administration suppressed metastasis by lowering the colony formation potential of cisplatin-resistant ovarian cancer and down-regulating the SDF-1/CXCR4 axis, and increasing the ratio of BAX/BCL-2 mRNA and protein levels due to BAX up-regulation and BCL-2 down-regulation.Conclusion: As a result, we showed that As-IV, used as an antioxidant, can be used as an effective anticancer agent to improve response to the currently used cisplatin-based treatment in cases of drug resistance in ovarian cancer.
      PubDate: Thu, 30 May 2024 00:00:00 +030
       
 
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