Melatonin Sensitizes OVCAR-3 Cells to Cisplatin through Suppression of PI3K/Akt Pathway.

This study examined the effect of melatonin on oxidative stress, expression of pro-apoptotic protein, anti-apoptotic proteins, and the activity of the PI3K/Akt signaling pathway in the human ovarian cancer cell line (OVCAR3). OVCAR3 cells were treated with cisplatin, melatonin, cisplatin + melatonin, and siRNA Akt. Reactive oxygen species levels were assessed. The expression of the proteins was determined by Western blot. Melatonin administration significantly increased intracellular ROS generation, the cleavage of caspase 3 and decreased phosphorylation of Akt. Combination therapy of cisplatin and melatonin increases apoptosis in the OVCAR-3 cells by inhibiting of PI3K/Akt signaling pathway and exacerbating oxidative stress.

Evaluation of interleukin-17 receptor (IL-17RA) gene expression in PBMCs of patients with premature coronary artery disease.

OBJECTIVES: Present study has been carried out to analyze the IL-17R gene expression in PBMCs of patients with premature coronary artery disease (CAD) in comparison to normal controls. BACKGROUND: Premature CAD results in disability and lack of quality of life over the years and consequent mortality. Cardiovascular disease (CVD) has global distribution. In 2022, CAD is the leading cause of mortality in the United States and Iran. IL-17 cytokine family plays an important role in promoting inflammation and producing pro-inflammatory cytokines, chemokines, and matrix metalloproteinases. METHODS: Entirely, 60 subjects were entered into this examination. The case group consisted of 30 subjects with CAD as well as the control group which consisted of 30 healthy persons. The real-time quantitative reverse transcription PCR assay was used to find out, the relative expression (fold) level of IL-17R gene. RESULTS: Our findings indicated that, the relative expression (fold) level of IL-17R gene in the patients group showed an increased level as compared to the control group. The analysis of findingsobtained in this study showed that the patient group is significantly different from the control group regarding the IL-17R mRNA level (fold) (p = 0.035). CONCLUSION: It has been concluded that IL-17R plays an important role in the pathogenesis of CAD. It follows that superior understanding of IL-17/IL-17R signaling way will be vital for innovating novel therapeutic targets that will facilitate the designing of new drugs for the management of patients (Ref. 40).

Does GD2 synthase (GD2S) detect cancer stem cells in blood samples of breast carcinomas?

INTRODUCTION: Cancer stem cells (CSCs) are a theorized subset of cells within the tumor that is thought to drive disease recurrence and metastatic spread. The aim of this study is to investigate mRNA and protein levels of ganglioside GD2 synthase (GD2S), in breast cancer (BC) patients. METHODS: 65 PBMCs of preoperative BC patients without chemotherapy were compared to PBMCs after chemotherapy and controls. RESULTS: GD2S were significantly higher in BC patients after chemotherapy compared to pre-chemotherapy at both mRNA and protein. GD2S was higher in pre-chemotherapy blood samples compared to control samples. CONCLUSIONS: Higher expression of GD2S in BC samples compared to healthy control indicates the potential utility of GD2S as a marker of malignancy.

Upregulation of Ganglioside GD2 Synthase (GD2S), as a New Putative Cancer Stem Cell Marker in Breast Carcinomas.

Background: GD2 synthase (GD2S) is the key enzyme required for ganglioside GD2 synthesis. It is commonly expressed in normal tissues and various cancers. Ganglioside GD2 is identified as a breast cancer stem cells (BCSCs) marker that promotes tumorigenesis. As GD2S has been found to be a useful molecular marker in neuroblastoma and retinoblastoma tumors, we suggest that it can be considered as a suitable candidate for the detection of CSCs in breast cancer tissues. Methods: Expression of GD2S was examined in 65 breast tumors compared to adjacent normal tissues, applying quantitative real-time PCR (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA). The association between GD2S expression level and patients' clinical characteristics was also assessed. Results: Our findings showed that GD2S mRNA expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissue samples (4.92-fold change, p<0.001) in advanced grades (p<0.001) and stages (p<0.001). It was also shown that GD2S protein expression was significantly higher in breast cancer tissues in comparison to normal adjacent tissues (4.86-fold change, p=0.010) in advanced grades (p=0.010), stages (p=0.005) and larger tumor size (p=0.002). Conclusion: The current study showed that increased expression of GD2S in advanced breast cancer potentiates it as a promising tumor marker in these patients.

High GD2 expression defines breast cancer cells with enhanced invasiveness.

INTRODUCTION: Breast cancer is the most frequently diagnosed cancer among women. Cancer stem cells (CSCs) are suggested to be responsible for tumor initiation, progression, metastasis, recurrence and drug resistance. This study was conducted to evaluate the clinical significance of GD2, a newly suggested CSC marker and two other traditional CSC markers, CD44 and CD24 in breast cancer patients. MATERIAL AND METHODS: A total of 168 primary breast cancer tissues were evaluated in terms of GD2, CD44 and CD24 expression using tissue microarray. Then, the correlation of expression levels of these markers with patients' clinicopathological characteristics was assessed. RESULTS: Higher GD2 expression was mainly found in patients with advanced histological grade (p = 0.02), presence of lymph node invasion (p = 0.04), larger size of tumors (p = 0.04) and older age (p = 0.04). Breast cancer samples with advanced histological grade also showed higher CD44 (p = 0.03) and CD24 expression (p = 0.05). A significant positive association was found between increased CD24 expression and lymph node involvement (p = 0.01). Furthermore, GD2-high/CD44-high/CD24-low phenotype was frequently seen in breast cancer samples with positive lymph node involvement (p = 0.05). CONCLUSION: In summary, increased expression of GD2 may define more aggressive tumor behavior in breast cancer. GD2 can well be considered as a diagnostic and prognostic marker in breast cancer.

Association of MMP-1 (rs1799750)-1607 2G/2G and MMP-3 (rs3025058)-1612 6A/6A Genotypes With Coronary Artery Disease Risk Among Iranian Turks.

The study was conducted to evaluate the association between MMP-1 (rs1799750)-1607 1G/2G and MMP-3 (rs3025058)-1612 5A/6A polymorphisms/haplotypes and coronary artery disease (CAD) risk among Iranian Turks. Totally, 102 patients with CAD and 102 healthy subjects joined the study. Genomic DNA isolation was carried out using "salting out" method from 3 to 4 mL of whole blood samples. The MMP-1 (-1607 2G/1G) and MMP-3 (-1612 5A/6A) promoter gene polymorphisms were detected via polymerase chain reaction restriction fragment length polymorphism. Our results indicated that the frequencies of the MMP-1 (-1607) 2G alleles and 2G/2G genotypes and the MMP-3 (-1612) 6A alleles and 6A/6A genotypes were higher in CAD patients older than 50 years than in healthy controls (P < 0.05). We failed to show statistically significant differences between the CAD patients younger than 50 years and controls concerning MMP-1 -1607 ins/delG (1G > 2 G, rs1799750) and MMP-3 -1612 ins/delA (5A/6A, rs3025058) polymorphisms (P > 0.05). The frequencies of MMP-3/MMP-1 haplotypes were not statistically different among tested groups (P > 0.05). This examination, as the first study of its own kind in Iranian Turks, reported association between MMP-1 (rs1799750) -1607 2G/2G and MMP-3 (rs3025058) -1612 6A/6A genotypes and CAD risk in patients older than 50 years.

PI3K Inhibition Sensitize the Cisplatin-resistant Human Ovarian Cancer Cell OVCAR3 by Induction of Oxidative Stress.

Background: This study evaluates the effect of simultaneous AKT inhibition and cisplatin therapy in changes of Reactive Oxygen Species (ROS) production, apoptosis induction, and cell survival in cisplatin-resistant OVCAR3 cell. Methods: OVCAR3 cancer cells were treated with cisplatin, Ly 294002 (LY), and cisplatin+Ly to investigate the cytotoxicity effect of the mentioned groups via MTT assay. Then, DCFH-DA (2', 7'-dichlorodihydro fluorescein diacetate) assay kit is used to assess the potential of treated groups in intracellular ROS generation. Protein expression levels of caspase-3, cleaved caspase 3, PI3K, Akt, p-Akt, XIAP, and Survivin are estimated through immunoblotting assay in all three experimental groups. Results: The results showed that all three treated groups, including cisplatin and Ly alone and co-administration of cisplatin+Ly, could reduce the cell vitality of OVCAR3 cancer cells, induced intracellular production of ROS and increased the expression level of activated caspase 3 and Akt protein, whereas down-regulated the phosphorylation of Akt protein. However, the effect of combination therapy was more tangible compared to single therapy and control groups. In contrast, the expression amount of XIAP, Survivin, and PI3K did not show detectable changes in comparison with the control group. Conclusion: The results showed that the AKT inhibition by Ly could sensitize the OVCAR3 cancer cells to the cisplatin and lower the effective dose of cisplatin through hyperactivation of oxidative stress.

A novel GTPBP2 splicing mutation in two siblings affected with microcephaly, generalized muscular atrophy, and hypotrichosis.

A novel splice site mutation in the GTPBP2 gene was identified by whole-exome sequencing in two siblings with microcephaly and progressive generalized muscular atrophy associated with hypotrichosis.

A novel biallelic LMNB2 variant in a patient with progressive myoclonus epilepsy and ataxia: A case of laminopathy.

The report of LMNB2-related progressive myoclonus epilepsy and ataxia due to missense homozygous c.473G>T variant.

Association of angiotensin-1-converting enzyme gene variations with silicosis predisposition.

Interactions between several genes and environmental factors occur in silicosis and other dust-mediated pulmonary fibrosis. It has been suggested that several genes play role in individual susceptibility to silicosis. The aim of this study was to determine the angiotensin-1-converting enzyme (ACE) gene (I/D) variations in patients with silicosis and healthy controls in Iranian Kurdish population. Forty-one male workers with silicosis and 41 healthy male controls were entered in the study. Polymerase chain reaction (PCR) method was carried out for allele and genotype typing. Our finding showed that the frequencies of D and I alleles were 0.65 and 0.35 in the silicotic patients compared to 0.52 and 0.48 in the controls. The frequencies of ACE D/D, D/I, and I/I genotypes were 15 (36.59%), 23 (56.1%), and 3 (7.31%) in patients, and 14 (34.15%), 15 (36.59%), and 12 (29.27%) in the controls, respectively. The results indicate that the frequency of ACE D/D+D/I genotypes were higher in the patients group (92.7% vs. 70.73%) (OR: 5.24, 95% CI: 1.35-20.30, P = 0.01). Also, ACE I/I genotype frequency was lower in patients (7.32% vs. 29.27%) (OR: 0.19, 95% CI: 0.04-0.73, P = 0.01). It can be concluded that ACE D/D and I/D genotypes may play a role in the susceptibility to silicosis; on the contrary, ACE I/I genotype may have protective effect.

Zinc Protects against MDMA-Induced Apoptosis of Sertoli Cells in Mouse via Attenuation of Caspase-3.

BACKGROUND: 3,4-Methylenedioxymethamphetamine (MDMA) disrupts function of the endocrine system and different organs such as heart, blood vessels, kidney, liver and nervous systems. This revision was conducted to evaluate impact of MDMA on apoptosis and Zinc in the MDMA-induced apoptosis of cultured Sertoli cells by measuring Caspase-3 gene expression. MATERIALS AND METHODS: In this experimental study, Sertoli cells were incubated with MDMA (0, 0.5, 1, 3 and 5 mM), Zinc (0, 8, 16, 32, 64 µM) and Zinc (8 µM) prior to adding MDMA (5 mM) for 24 and 48 hours. MTT assay was used for evaluating impacts of these conditions on the viability of Sertoli cells. Caspase-3 gene expression level was detected using quantitative reverse transcription PCR (qRT-PCR) in all of the tested groups. RESULTS: Finding showed that cellular viability was decreased and level of Caspase-3 mRNA was increased in MDMA treated cells. Additionally, pre-treatment with Zinc (8 µM) attenuated MDMA-induced apoptosis and down-regulated caspase-3. The mean of caspase-3 mRNA level (fold change ± SE) was 3.98 ± 1.18, 0.31 ± 0.28, and 1.72 ± 0.28 in respectively MDMA (5 mM), Zinc (8 µM), and Zinc+MDMA groups vs. control group. The mean of Caspase-3 mRNA (fold change) was not statistically different in the tested groups (P>0.05), unless MDMA (5 mM) group (P=0.008). CONCLUSION: We suggest that MDMA toxicity could be involved in apoptosis of Sertoli cells. In addition, Zinc could reduce MDMA-induced apoptosis by down-regulation of Caspase-3 mRNA levels.

Identification of Novel Pathogenic PKD2 Variants in Iranian Patients with Autosomal Dominant Polycystic Kidney Disease.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is a delayed-onset renal disorder that results from a mutation in the PKD1 or PKD2 genes. Autosomal dominant polycystic kidney disease results in end-stage renal disease due to renal cystic dysplasia. The aim of this study was to evaluate, by exon sequencing, the disease-causing variants of PKD2 (exons 4, 6, and 8) in Iranian ADPKD patients. METHODS: Genomic DNA was extracted from 3-5 ml of peripheral blood by the salting-out method. PKD2 exons 4, 6, and 8 were PCR-amplified and sequenced. RESULTS: Three disease-causing PKD2 variants were identified; all three were missense mutations in exon 4. The mutations were AGC → ACC (c.893G>C, cDNA.959G>C, S298T), TAC → TTC (c.1043A>T, cDNA.1109 A>T, Y348F), and GAA → GAT (c.1059A>T, cDNA.1125 A>T, E353D. These novel pathogenic variants may cause loss of the normal protein function. CONCLUSION: Our results suggest that AGC → ACC (c.893G>C, cDNA.959G>C, S298T), TAC → TTC (c.1043A>T, cDNA.1109 A>T, Y348F), and GAA → GAT (c.1059A>T, cDNA.1125 A>T, E353D variants are common in Iranian ADPKD patients. These mutations modify the transmembrane domain and likely influence PC2 function.

Report of a Novel Splicing Mutation in the MYO15A Gene in a Patient With Sensorineural Hearing Loss and Spectrum of the MYO15A Mutations.

INTRODUCTION: Autosomal recessive non-syndromic hearing loss (ARNSHL) is a genetically heterogeneous sensorineural disorder with an approximate incidence of 1.4:1000 in neonates. Mutations in more than 60 genes including the MYO15A gene has been reported in patients affected with ARNSHL. In the present study, we report a novel MYO15A mutation identified by clinical exome sequencing and confirmed by Sanger sequencing in a consanguineous Iranian family with ARNSHL. CASE PRESENTATION: A 22-year-old woman with congenital non-syndromic sensorineural hearing loss referred to our medical genetic center. Her parents were consanguineous with F = 1/16 (first cousin), and clinical examination of the patient exclude dysmorphic features. Sanger sequencing of GJB2 and GJB6 genes, which are the most common causes of ARNSHL, was negative. Then she underwent clinical exome sequencing. OUTCOME: We found a novel homozygote variant (c.9611_9612+8delTGGTGAGCAT) in the MYO15A gene which creates a shift in the reading frame starting at codon 3204. This variant was confirmed by Sanger sequencing in the patient and also in her parents who were heterozygous. DISCUSSION: The present results suggest that the homozygous MYO15A (c.9611_9612+8delTGGTGAGCAT) variant is a pathogenic mutation and to the best of our knowledge, this mutation has not been reported in any database.

The use of stromal vascular fraction (SVF), platelet-rich plasma (PRP) and stem cells in the treatment of osteoarthritis: an overview of clinical trials.

Osteoarthritis (OA) is a major cause of disability across the world, which its prevalence is relatively high in elder population. Current accepted therapies such as exercise, anti-inflammatory drugs and intra-articular inoculation of corticosteroids are aimed at controlling symptoms in the affected patients. Surgical options including arthroplasty, osteotomy and joint replacement are other choices of treatment, which are invasive and can be applied in case of failure of conventional therapies. In the last few decades, efforts to treat musculoskeletal diseases are being increasingly focused on regenerative cellular therapies. Stromal vascular fraction (SVF), which obtained from adipose tissue, contains a variety of cells include mesenchymal stem cells (MSCs) and has shown to be effective in cartilage repair. Autologous blood products such as platelet-rich plasma (PRP) act as an adjuvant of surgical treatment and its intra-articular delivery has shown beneficial effects for OA treatment. Given the efficacy of such treatment approaches in OA, this paper discusses both preclinical and clinical evidence with major focus on clinical trials.

Examining the Role of Polymorphisms in Exon 25 of the PKD1 Gene in the Pathogenesis of Autosomal Dominant Polycystic Kidney Disease in ranian Patients.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is a highly prevalent life-threatening monogenic disorder with high morbidity and mortality. Roughly 1:400-1000 individuals are affected with this disease worldwide. The development of ADPKD is largely attributed to mutations in the polycystic kidney disease (PKD)1 and PKD2 genes. However, the pathogenicity of the different polymorphisms in PDK1 in the development of ADPKD remains unclear. The aim of this study was to further elucidate the role of the polymorphisms in exon 25 of the PDK1 gene in relation to the pathogenesis of ADPKD in Iranian patients. METHODS: The genomic DNA of 36 Iranian patients with ADPKD was isolated using the standard salting out method. The PCR products were directly sequenced and analyzed. RESULTS: The frequencies of CAG>GAG, ATG>GTG, GTC>GTA, and GTG>ATG polymorphisms in exon 25 of the PKD1 gene were 34 (94.44%), 33 (91.67%), 26 (72.22%), and 5 (13.89%), respectively. The most frequent polymorphism associated with ADPKD was the homozygous CAG→GAG which causes an amino acid change of Q[Gln] to E[Glu] at codon 3005. CONCLUSION: Our data suggests that there is potentially a common polymorphism of PDK1 among the Iranian population with ADPKD. This may aid in the diagnosis and genetic screening of at-risk patients for ADPKD.

A High Prevalence Rate of Tibia Hemimelia in a Subregion of West Azarbaijan, Iran.

From 1995 to 2017 an abnormally high number of tibia hemimelia (TH) patients from the Maku subregion in the West Azarbaijan province of Iran were referred to our orthopedic department for treatment. Regarding the occurrence of TH in many families in a restricted region and negative results of available genetic tests, we hypothesized that a founder mutation etiology that is different from previous known genetic disorders might produce the trait of TH in our patients. Through a retrospective study, we collected demographic data including date of birth, patients and parents place of birth, sex, type of TH, presence of other musculoskeletal anomalies, and treatment from the patients who were referred to our department. We obtained a blood sample for genetic studies. We carried out genetic studies in cytogenetics and molecular levels on a patient with familial TH. The prevalence of TH in the Maku subregion of West Azarbaijan was 149.5 (95% confidence interval [CI]: 68.4-283.8) per 1 million live birth. The patients did not fit with any known syndromes with TH. Genetic evaluations of a patient with familial history of TH in this case series exhibited no detectable change in both cytogenetic and molecular levels. There was an obvious increased prevalence rate of TH in this province and particularly in the Maku subregion. The cytogenetic locations for known syndromic TH are not responsible for the observed anomalies in our patients. Our next step for detecting possible genetic mutations in our patients would be mutation analysis via very high-resolution whole genomic sequencing in more patients or genetic linkage study.

The prevalence at birth of overt congenital anomalies in Urmia, Northwestern Iran.

BACKGROUND: Congenital anomalies play a significant role in perinatal and infantile morbidity and mortality. There is a variation in the frequency of congenital anomalies in different populations. Determination of the prevalence of different types of congenital anomalies may help plan primary prevention measures for these anomalies. The objective of this study was to determine the prevalence at birth of overt congenital anomalies in Urmia, in the northwest of Iran. METHODS: In a cross-sectional hospital-based study, charts of 14,121 deliveries including live-births and stillbirths during the period from January 2001 through June 2005 were studied. RESULTS: A total of 264 (187 per 10,000 births) anomalies were detected. The anomaly categories with the highest prevalence were nervous system defects (52.65%) followed by musculoskeletal defects (23.86%). The total prevalence at birth of overt congenital anomalies was 1.87%. The rates for live- and stillbirths were 1.17% and 40.7%, respectively. CONCLUSION: Considering the high frequency of central nervous system anomalies recorded in our study, it seems to be reasonable to pay more attention to the role of periconception vitamin supplementation for the primary prevention of congenital anomalies, particularly neural tube defects.

Tumor Necrosis Factor-alpha Gene Expression in PBMCs of Iranian Azeri Turkish Patients with Premature Coronary Artery Disease (Age .50 Years).

BACKGROUND: Coronary artery disease is the leading cause of disability and mortality in Iran and worldwide. Tumor necrosis factor-alpha is a pro-inflammatory cytokine that plays a key role in inflammatory cascades and atherosclerosis. It regulates cytokine networks and adhesion molecule expression, and activates several signal transduction pathways, being also known as transducer of cardiovascular diseases, especially premature coronary artery disease. OBJECTIVE: The aim of the present study was to investigate tumor necrosis factor-alpha gene expression in Iranian Azeri Turkish patients with premature coronary artery disease (age ≤50 years). MATERIAL AND METHODS: Eighty four individuals (42 cases and 42 controls) were enrolled in the study. Total RNA was extracted from patients with premature coronary artery disease using RNX-Plus Solution (Cat. No.: RN7713C) and reverse transcribed into cDNA. The tumor necrosis factor-alpha mRNA expression level was evaluated using Real-Time PCR. RESULTS: The mean ± SE of fold in cases and controls were 1.1±1.08 and 1.6±3.4, respectively. The mean expression of tumor necrosis factor-alpha mRNA (fold) was not statistically different between the tested groups (P value 0.4). OUTCOMES: Our outcome failed to find evidence for any association between tumor necrosis factor-alpha mRNA expression and premature coronary artery disease. Large scale, more detailed studies are further needed to prove our results and to propose other mechanisms in the pathophysiology of premature coronary artery disease.

Analysis of the mutations in exon 10 of MEFV gene in patients with premature coronary heart disease in west Azerbaijan province of Iran.

Introduction: Premature coronary heart disease (PCHD) affects public health and leads to death. PCHD has several genetic and environmental risk factors. The aim of this study was to analysis of the mutations in exon 10 of MEFV gene in patients with PCHD in West Azerbaijan province of Iran. Methods: Totally 41 PCHD patients who were admitted to the cardiology unit of Sayedoshohada hospital (Urmia, Iran) enrolled in the study. Selection of the patients was done based on the strict criteria, that is, who had a minimum of one angiographically documented coronary artery with the stenosis of 50%. Mutations in exon 10 of MEFV gene were found by direct sequencing. Results: V726A, M680I, K695R, and A744S mutations with 2.44%, 1.22%, 1.22%, and 1.22%, allelic frequency were found, respectively. Five patients (12.2%) with PCHD carried at least one mutated MEFV allele. Heterozygote V726A was the most frequent mutation among tested cases (4.88%), followed by heterozygote M680I, heterozygote K695R, and heterozygote A744S. Conclusion: The results of the present study imply that the frequency of the MEFV gene exon 10 is significantly high in PCHD patients. This is the first report in its own kind in clinically diagnosed PCHD pa-tients of Iranian Azeri Turkish population.

Vitamin D Receptor Gene Haplotype and Late-Onset Obesity in Iranian Azeri Turkish Women.

INTRODUCTION: A large body of literature has revealed the association between vitamin D3-VDR complex and obesity. The aim of the present study was to survey the rate of the VDR polymorphisms in obese women and to determine whether there may be an association between VDR BsmI and Tru9I haplotypes and obesity in Iranian Azeri Turkish women. MATERIAL AND METHODS: 65 Iranian Azeri Turkish women were enrolled in the study and PCR amplification and direct sequencing of PCR products were used for genotypings. RESULTS: The findings of this study showed that VDR BsmIG allele, VDR BsmI G/G genotype, VDR BsmI A/A genotype, Tru9IA allele and Tru9I A/A genotype were more frequent in obese women compared to controls. The frequency of VDR BsmIG/Tru9IA (GA), VDR BsmIG/Tru9IG (GG), VDR BsmIA/Tru9IG (AG), and VDR BsmIA/Tru9IA (AA) haplotypes were 19.74%, 42.11%, 38.16% and 0% in cases, and 11.11%, 40.74%, 42.59 and 5.56% in controls. Statistically significant differences were found between cases and controls regarding the VDR AA haplotype (P=0.03). CONCLUSIONS: Our findings demonstrated that the VDR AA haplotype frequency was significantly lower in subjects with obesity compared with normal controls. This study shows that the VDR AA haplotype is significantly associated with a decreased risk of obesity in the tested group. This report is the first of its kind in the West Azerbaijani population.