Investigating the impact of metformin on severity of COVID-19 in patients with Type 2 diabetes mellitus: Focusing on laboratory findings.

BACKGROUND: In the terrifying pandemic caused by SARS-CoV-2, diabetic patients exhibiting more severe outcomes and mortality rate is high among them. Based on recent studies, metformin as the most prescribed drug for T2DM treatment may improve severe outcomes in diabetic patients infected with SARS-CoV-2. On the other hand, abnormal laboratory findings can help to differentiate between the severe and non-severe form of COVID-19. According to the mentioned issues, the effect of metformin on severity of COVID-19 was examined in T2DM patients with SARS-CoV-2 infection. METHODS: The study included 187 individuals diagnosed with COVID-19, 104 patients were diabetic and divided into two groups according to their anti-diabetic drugs: patients who were treated only with metformin and patients who were treated with other anti-diabetic drugs. The other participants were non-diabetic and diagnosed with COVID-19. Biochemical parameters were measured by routine laboratory methods before, during and after SARS-CoV-2 infection. RESULTS: During infection, FBS, creatinine, ALT, AST, Ferritin and LDH were significantly lower in metformin users than non-users (p-value: .02, .01, .03, .04, .0009 and .01, respectively). Also, after recovery, there were statistically significant differences between metformin users and non-users with respect to most of the study parameters, except FBS, BUN and ALP (p-value: .51, .28 and .35, respectively). CONCLUSION: Our result suggested that metformin might be associated with better outcomes in diabetic patients infected with SARS-CoV-2.

The association study between changes in HbA1C with rs2250486 and rs67238751 genetic variants for SLC47A1 in newly diagnosed Iranian patients with type 2 diabetes mellitus: 6 months follow-up study.

OBJECTIVES: One of the most well-known oral medications for the treatment of T2DM is metformin. Variants have been found in studies to be useful in detecting new genes connected to T2DM aetiology and affecting metformin's mechanism of action. In this research, we aimed to study two variations of the SLC47A1 gene; rs2250486 and rs67238751, in T2DM patients who had been taking metformin for the first 6 months after the diagnosis in the Iranian population for the first time. DESIGN AND METHODS: A total of 200 individuals were recruited for the study. According to their glycosylated haemoglobin (HbA1c) levels, the patients were divided into two groups: responders (HbA1c levels were reduced by at least 1% after 6 months of metformin treatment.) and non-responders. DNA was extracted from whole blood and genotyped by Tetra ARMS PCR. High-performance liquid chromatography (HPLC) was used to measure HbA1c levels at the start of the treatment and again 6 months later. RESULTS: rs2250486 variant in the dominant model reduces the HbA1C levels after 6 months of metformin treatment. In fact, when compared to the T/C + C/C genotypes, the T/T genotype improves HbA1C levels (p-value = .014). Furthermore, in the allelic model, the T allele improves HbA1C levels in comparison to the C allele (p-value = .008). After 6 months of metformin treatment, serum levels of HbA1C in responders were reduced significantly in both groups (T/T and T/C + C/C), (p-value = <.0001). However, the rs67238751 variant did not reveal a meaningful relationship with lower HbA1C levels in any of the models. CONCLUSIONS: This study found that the rs2250486 variant could be associated with reducing HbA1C levels while the rs67238751 variant, had no relationship.

Biochemical and molecular biomarkers: unraveling their role in gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) is the most prevalent metabolic disorder during pregnancy, causing short- and long-term complications for both mother and baby. GDM is a multifactorial disease, and it may be affected by interactions between genetic, epigenetic, and environmental factors. However, the exact etiology is poorly understood. Despite the high prevalence of GDM, there is still debate regarding the optimal time for screening, the diagnostic threshold to apply, and the best strategies for treatment. Identifying effective strategies for therapeutic purposes as well as accurate biomarkers for prognostic and diagnostic purposes will reduce the GDM incidence and improve its management. In recent years, new biochemical and molecular biomarkers such as microRNAs, single-nucleotide polymorphisms, and DNA methylation have received great interest in the diagnosis of GDM. In this review, we discuss current and future diagnostic approaches for the detection of GDM and evaluate lifestyle and pharmacological strategies for GDM prevention.

Can circulating miR-7-1-5p, and miR-33a-5p be used as markers of T2D patients?

PURPOSE: Recent evidence has indicated that miRNAs play an important role in both initiation and progression of many pathologic processes such as diabetes and can be used as an important and more sensitive tool to predict the development of the disease than the currently used biomarkers. This research aimed at comparing miR-7-5p and miR-33a-5p expression levels in the diabetics and pre-diabetics with the control group. METHODS: In this study, we compared expression of miR-7-5p and miR-33a-5p in plasma of three groups including pre-diabetic patients (n = 20), T2D patients (n = 20) and control group (n = 20), using RT-qPCR. Biochemical parameters were measured by auto-analyser. In silico analysis was performed to identify potential target genes of these miRNAs. RESULTS: Compared to the controls, miR-7-1-5p expression was down regulated in the pre-diabetics and the T2D patients; whereas, miR-33a-5p was expressed at higher levels in the T2D patients compared to the control group. Both miRs were correlated with glycaemic status such as FBS and HbA1c levels. The ROC analysis indicated a significant ability for miR-33a-5p in discriminating between the diabetics and the healthy individuals. In silico analysis suggests that both miRs affect biological pathways related to T2DM pathogenesis, such as MAPK, and insulin signalling pathway. CONCLUSION: Our results demonstrated that the miR-7-1-5p and miR-33a-5p expression levels are deregulated in the diabetics and pre-diabetics. Furthermore, miR-33a-5p showed significant ability in discriminating between diabetics and healthy individuals, suggesting a potential diagnostic use of miRNAs in type-2 diabetes detection.

Molecular mechanisms of bilirubin induced G1 cell cycle arrest and apoptosis in human breast cancer cell lines: involvement of the intrinsic pathway.

BACKGROUND: Bilirubin, as an essential constituent of cellular signaling pathways, may have a role in cell growth and apoptosis in breast cancer, although the biochemical relevance is still unclear. The purpose of the present study is to recognize the mechanism underlying bilirubin-induced apoptosis in breast cancer cell lines. METHODS AND RESULTS: To detect the cell viability, MTT assay was carried out. Apoptosis was assessed by flow cytometry analysis and caspase activities were determined by colorimetric method. The expression of AhR, cyclin D1, cyclin A, p53, p27, Bcl-2, and Bax were examined using real-time PCR. The cell viability has been reduced by bilirubin in a dose-dependent manner and an intrinsic apoptotic response has been occurred that was evidenced by the elevation of caspase-3 and - 9 activities. Bilirubin induced cell arrest in cell-cycle progression, which was associated with the induction of AhR expression, down-regulation of cyclin D1, cyclin A, and upregulation of p53 and p27 expression. Following bilirubin treatment, Bcl-2 was decreased and Bax protein was increased in both cell lines. CONCLUSIONS: To discuss, bilirubin, as a naturally occurring antiproliferative molecule, mediates growth inhibition by induction of cell cycle arrest and apoptosis in MCF-7 and MDA-MB-468 breast cancer cells. It is associated with the suppression of cyclin A, D1, and Bcl-2; induction of p53, p27, and Bax together with the activation of caspase-3 and - 9.

Circulating miR-148b-3p and miR-27a-3p can be potential biomarkers for diagnosis of pre-diabetes and type 2 diabetes: integrating experimental and in-silico approaches.

BACKGROUND: In view of the growing global prevalence of type 2 diabetes (T2D), detection of prediabetes and type 2 diabetes in the early stages is necessary to reduce the risk of developing diabetes, prevent the progression of the disease, and dysfunction of different organs. Since miRNAs are involved in the initiation and progression of numerous pathogenic processes, including diabetes, in the present study, we aimed to investigate the expression of miR-148b-3p and miR-27a-3p in prediabetic and T2D patients and to evaluate the diagnostic potential of these miRNAs. METHODS: We evaluated the expression of miR-148b-3p and miR-27a-3p in the plasma of three groups: 20 prediabetic patients, 20 T2D patients, and 20 healthy controls. The biochemical parameters were determined by the auto-analyzer. The possible target genes of these miRNAs were identified using an in-silico approach. RESULTS: Our results showed that, as compared to the healthy controls, there was a significant up regulation and down regulation in the expression of miR-148b-3p and miR-27a-3p in the T2D patients, respectively. The results of receiver operating characteristic curve analysis also suggested that miR-148b-3p acted successfully in discriminating the prediabetic and diabetic patients from the control group. According to in-silico analysis, miRs influence biological pathways involved in T2DM development, such as insulin signaling. CONCLUSIONS: The miR148b-3p and miR-27a-3p expression levels were deregulated in diabetes and pre-diabetes. Furthermore, miR-148b-3p showed significant ability in discriminating between diabetic and healthy individuals, suggesting a potential diagnostic use of miR-148b-3p in the detection of T2D.

The influence of SLC22A3 rs543159 and rs1317652 genetic variants on metformin therapeutic efficacy in newly diagnosed patients with type 2 diabetes mellitus: 25 weeks follow-up study.

BACKGROUND AND AIMS: Among anti-diabetic medications, metformin has been proven to be the preferred initial pharmacologic agent for type 2 diabetes mellitus (T2DM) treatment. Despite its safety and efficacy, the response to metformin varies between individuals. Genetic variations, especially within genes involved in pharmacokinetics and pharmacodynamics of metformin (e.g SLC22A3), have been suggested to be responsible for the observed inter-individual differences. By considering the undeniable role of organic cation transporter 3 in hepatic uptake of metformin, this study was aimed to investigate the association of rs543159 and rs1317652 variants in SLC22A3 gene with response to metformin monotherapy in newly diagnosed patients with T2DM. METHODS: The study included 200 T2DM patients who received metformin monotherapy for 25 weeks. The patients were classified into 2 groups according to their HbA1c values: the responders (reduction in HbA1c levels by at least 1% after 25 weeks treatment with metformin) and non-responders (less than 1% reduction in HbA1c levels after 25 weeks treatment with metformin). We used tetra ARMS-PCR method to determine genotypes of the target variants. RESULTS: For the rs543159, CA and AA genotypes were more frequent in responders as compared to non-responders (OR = 2.48; 95% CI = 1.28-4.78, P-value = 0.0057) under the dominant model. In case of rs1317652 CC and CT genotypes were more frequent in metformin responders as compared to non-responder group (OR = 2.49; 95% CI = 1.32-4.70, P-value = 0.0043) under the dominant model. Parameters such as fasting blood sugar (FBS), HbA1c, and total cholesterol (TC) levels were significantly lower in the responder group after 25 weeks of metformin monotherapy. Moreover, according to the result of multiple linear regression rs543159 and base line HbA1c values are significantly associated with response to metformin monotherapy. CONCLUSION: Our results suggested that rs543159 and rs1317652 in SLC22A3 gene might be associated with variability in response to metformin therapy in T2DM patients.

Can miR-145-5p be used as a marker in diabetic patients?

In the light of emerging global epidemics of type 2 diabetes mellitus significant efforts are continuing to discover novel biomarkers for early detection of the disease. Since miRNAs play an important role in both the initiation and progress of many pathologic processes such as diabetes, in this study we aimed to evaluate expression level of plasma miR-145-5p in diabetics and pre-diabetics in comparison to the control group and assess its use as a biomarker in diagnosis of T2D. The plasma level of miR-145-5p was assessed in three groups including 20 prediabetic patients, 20 T2D patients and 20 healthy controls using RT-qPCR. Biochemical parameters were also measured by the auto-analyzer. Expression level of miR-145-5p was down-regulated in the prediabetics and the T2D patients compared to the controls. In the control group miR-145-5p showed a borderline correlation with FBS (p = .06), while in the prediabetic group miR-145 showed a significant negative correlation with FBS and finally in the T2D patients miR-145 was negatively correlated with HbA1c and TC and showed a negative borderline correlation with FBS. The ROC analysis indicated a significant ability for miR-145-5p in discriminating between the diabetics and pre-diabetics from the healthy subjects. Our results demonstrated that the miR-145-5p expression level is deregulated in the diabetics and the prediabetics. Furthermore miR-145-5p displayed a significant ability to discriminate the diabetics from the healthy subjects. These results suggest that miR-145-5p may be a useful biomarker for the diagnosis of T2DM.

Induction of cell apoptosis by biliverdin reductase inhibitor in MCF-7 and MDA-MB-468 breast cancer cell lines: Experimental and in silico studies.

Biliverdin reductase, biliverdin and bilirubin are known as important components of cellular signaling pathways that play major roles in cell proliferation and apoptosis, although their physiological relevance is still under evaluation. This study was designed to investigate the expression and activity of BVR-A and its apoptotic effect in the breast cancer cell lines, MCF-7 and MDA-MB-468. The expression of BVR-A was examined by real-time PCR and western blot analysis. Bilirubin concentration was measured by HPLC and molecular docking was performed to identify an appropriate inhibitor for BVR-A. To detect cell apoptosis, annexin V-PI staining, caspase-3, -8, and -9 activities were evaluated. Cell viability was reduced by biliverdin, in a dose-dependent manner, and an intrinsic apoptotic response occurred which was evidenced by caspase-3 and -9 activities. The intra- and extracellular concentrations of bilirubin were higher in MCF-7 cells than those of MDA-MB-468 cells. The expression of BVR-A, at mRNA and protein levels, in MCF-7 was also higher than that of MDA-MB-468 cells. Treatment of both cell lines with biliverdin plus DTNB, a BVR-A inhibitor, increased the cell death significantly when compared with biliverdin alone. Using annexin V-PI staining and assessment of caspase-3 activity, it was confirmed that biliverdin together with DTNB increases apoptosis in breast cancer cells. In conclusion, biliverdin has an important role in cell apoptosis and inhibition of biliverdin reductase increases the apoptotic effect of biliverdin.

The effect of A1298c polymorphism of the MTHFR gene on anti-Müllerian hormone levels: experimental and Web-based analysis.

BACKGROUND: The 5,10-methylenetetrahydrofolate reductase (MTHFR) is an important enzyme of folate and methionine metabolism, which is expressed in human oocytes and preimplantation. Due to the involvement of MTHFR in female reproduction, we tend to evaluate the influence of MTHFR A1298C polymorphism on ovarian marker reserves such as serum anti-Müllerian hormone (AMH) levels in women after in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). METHODS: A total of 100 women, who underwent ART treatment due to male factor infertility, were recruited into this study. MTHFR A1298C polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, and serum AMH concentrations were measured by an ultrasensitive enzyme-linked immunosorbent assay (ELISA). RESULTS: Women with the CC genotype had higher AMH levels (4.15 ± 1.67 ng/ml), albeit not significant, than carriers with other genotypes after ovarian stimulation. No significant differences existed in terms of miscarriage and live birth rates among different genotype groups. CONCLUSION: The presence of the C mutant allele of the 1298 polymorphism in the MTHFR gene led to an increasing trend in serum AMH concentrations; however, the numbers of oocytes retrieved decreased in women with mutated genotypes. The influence of the MTHFR C677T polymorphism on embryo quality and pregnancy rate after ART cycles remains unclear.

MicroRNAs in diabetic nephropathy: From molecular mechanisms to new therapeutic targets of treatment.

Despite considerable investigation in diabetic nephropathy (DN) pathogenesis and possible treatments, current therapies still do not provide competent prevention from disease progression to end-stage renal disease (ESRD) in most patients. Therefore, investigating exact molecular mechanisms and important mediators underlying DN may help design better therapeutic approaches for proper treatment. MicroRNAs (MiRNAs) are a class of small non-coding RNAs that play a crucial role in post-transcriptional regulation of many gene expression within the cells and present an excellent opportunity for new therapeutic approaches because their profile is often changed during many diseases, including DN. This review discusses the most important signaling pathways involved in DN and changes in miRNAs profile in each signaling pathway. We also suggest possible approaches for miRNA derived interventions for designing better treatment of DN.

Asthenozoospermia: Cellular and molecular contributing factors and treatment strategies.

Semen sample with poor sperm motility, which called asthenozoospermia, is considered as one of the main factors contributing to male infertility. Recognition of the cellular and molecular pathways contributing to sperm motility reduction may lead to applying novel treatment strategies for overcoming low sperm motility in asthenozoospermia individuals. In this review, we intend to discuss the main causes of sperm motility reduction in asthenozoospermia and some treatment strategies used to overcome low sperm motility.

SCSA results correlated with rate of motility reduction after ejaculation in Asthenozoospermia.

Maintaining sperm motility after ejaculation is important for fertilisation. Apoptosis may play an important role to reduce sperm motility after ejaculation. The aim of this study was to perceive whether or not an increase in apoptosis reduces sperm motility in a higher degree after ejaculation and whether it can be predicted by laboratory tests, such as sperm chromatin structure assay (SCSA). Fifty-one Asthenozoospermia and 20 fertile subjects participated in this study. SCSA was applied using flow cytometry. Fluorescein-labelled inhibitors of Caspases (FLICA) method was used for assessment of active Caspase-3. Motility was assessed every 2 hr after ejaculation for 12 hr. Both SCSA and spermatozoa with active Caspase-3 were significantly correlated with the rate of motility reduction after ejaculation. In the subgroups who had SCSA <27% and active Caspase-3 <40%, the sperm motility reduction significantly occurred 6-8 hr after ejaculation compared to the fresh sample. In the cases of SCSA ≥27% and active Caspase-3 ≥ 40%, a significant decrease in motility was observed between 2 and 4 hr after ejaculation. The result demonstrated a significant trend in the rate of sperm motility reduction with SCSA increase, which suggests SCSA may indirectly show a good scheme of apoptosis status and may forecast the rate of motility reduction after ejaculation in Asthenozoospermia.

Anti-Müllerian Hormone: genetic and environmental effects.

Anti-Müllerian hormone (AMH) is a homodimeric glycoprotein produced by granulosa cells of growing ovarian follicles. AMH appears to have an inhibitory effect on both primordial follicle recruitment and responsiveness of growing follicles to follicle-stimulating hormone (FSH). This hormone is considered to be a reliable marker of ovarian reserve; therefore, it is crucial to determine which factors influence AMH levels for prognostic and diagnostic purposes. In this review we intend to discuss the effect of genetic and environmental factors which may lead to AMH interindividual variability.

Role of vitamin D in female reproduction.

Vitamin D is a fat-soluble vitamin that belongs to the family of steroid hormones. The biological actions of vitamin D are exerted through a soluble protein, the vitamin D receptor (VDR). VDR is a transcription factor located in the nuclei of target cells that mediates the genomic action of the active form of vitamin D (1,25(OH)2D3). This transcription factor is distributed in various tissues, including the reproductive system. The presence of VDR in female reproductive tissue suggests that vitamin D is involved in female reproduction. The present article reviews the impact of vitamin D on anti-Müllerian hormone (AMH), as an ovarian reserve marker, and ovarian steroidogenesis. This article also discusses the impact of vitamin D as a factor that influences infertility and the outcome of in vitro fertilization (IVF), insulin resistance (IR), hyperandrogenism, endometriosis and polycystic ovary syndrome (PCOS).