Reports of Biochemistry and Molecular Biology
Volume:12 Issue: 3, Oct 2023

Neuregulin_4 (NRG4) is one of the adipokines members that synthesize adipose tissues. It has an activating effect on epidermal growth factor receptors (ErbB receptors). NRG4 has indirect effects on the hormonal environment through its interaction to ErbB receptors. Increased insulin resistance and chronic low-grade inflammation may be present when NRG4 levels are high in PCOS. Obesity and polycystic ovarian syndrome have recently gained a lot of attention. However, the literature on the connection between NRG4 and the PCOS phenotype is limited. Thus, this research aimed to identify neuregulin_4's function as a biomarker for insulin resistance in PCOS phenotypes.

A case-control study and included 140 female cases effect by different phenotypes of PCOS. Patients samples were collected at the reproductive fertility consultant of the Teaching Hospital for Obstetrics and Gynecology, Kerbala health directorate, Iraq. The outpatient clinic serum hormonal levels and insulin concentration were determined by the electrochemiluminescence immunoassay “ECLIA” system. Elisa system was used for the detection of Neuregulin-4 protein level.

At the early age of participant NRG4 was increased significantly in all phenotypes of PCOS compared to control with a P< 0.05. interestingly, phenotype A was shown high level of NRG4 following phenotype C than phenotype D and phenotype B. Receiver Operator Characteristic Curves (ROC) analysis for NRG4 was performed and showed good diagnostic performers to word phenotype A.

Females with phenotype A have a higher level of NRG4 than other phenotypes, which could be attributable to the more pronounced metabolic abnormalities in this phenotype.

Vitamin D plays crucial roles in immune cell function, including macrophage activation, immune response modulation, and antimicrobial peptide production. Low vitamin D levels can result in reduced immune response, heightened inflammation, and impaired organ function, thereby exacerbating sepsis severity and impacting patient prognosis. This study investigates the influence of vitamin D binding protein expression and vitamin D levels on the mortality of septic patients.

This analytical observational study employs a case-control approach and involves patients at the Critical Care Unit of Dr. M. Djamil General Hospital in Padang, Indonesia. The study comprises 40 patients in the case group and 40 patients in the control group. Vitamin D and vitamin D binding protein levels are assessed using the enzyme-linked immunosorbent assay method.

Vitamin D and vitamin D binding protein levels were observed to be lower in the case group compared to the control group. In the case group, the majority of patients had vitamin D binding protein levels below 200 µg/mL. A significant association was found between vitamin D levels and mortality in sepsis patients (P< 0.05). Patients with vitamin D levels below 20 µg/mL faced a 2.54 times higher risk of mortality than those with levels exceeding 20 µg/mL.

Diminished levels of vitamin D binding protein and vitamin D contribute to an increased risk of mortality in septic patients.

To overcome cisplatin resistance, the cytotoxicity of a novel antitumor agent on two ovarian cancer cell lines sensitive and resistant to cisplatin was investigated.

MTT assay and flow cytometry were performed to assess the cytotoxicity of a novel water-soluble Pd (II) complex, [Pd(bpy)(pyr-dtc)]NO3 (PBPD), on cisplatin-sensitive and cisplatin-resistant ovarian cancer cell lines. Furthermore, variations in the expression of drug resistance gene cluster of differentiation 99 (CD99), signal transducer and activator of transcription 3 (STAT3), octamer-binding transcription factor 4 (OCT4), and multidrug resistance mutation 1 (MDR1) were evaluated using Real-Time PCR.

The IC50 values of PBPD in resistant cells were higher than those in sensitive cells. Furthermore, PBPD has a deadlier effect on sensitive cells compared to resistant cells, and the cell survival rate is reduced over time. Flow cytometry revealed that PBPD enhanced the population of living-resistant cells while driving them to apoptosis. PBPD, on the other hand, has a greater effect on the living cell population and has dramatically shifted the population toward apoptosis and necrosis in the sensitive cells. Furthermore, gene expression analysis showed that when sensitive and resistant cells were treated with cisplatin, all resistance genes increased significantly relative to the control. In contrast to OCT4, MDR1, STAT3, and CD99 resistance genes were not significantly elevated in sensitive cells treated with PBPD compared to the control. Thus, the expression of resistance genes in resistant cells treated with PBPD was lower than cisplatin.

As a result, PBPD is a promising anticancer agent for CDDP-resistant ovarian cancer.

Congenital liver disease refers to a group of heterogeneous diseases from a clinical genetic point of view. The most crucial features are hepatosplenomegaly and elevated liver enzymes. This study aims to identify genetic variants causing the disease in three Iranian families with congenital liver disease using molecular techniques.

Patients were referred to Next Generation Genetic Polyclinic (NGGC) in Mashhad after confirmed congenital liver disease diagnosis by gastroenterologists. Following informed consent signed by participants, DNA was extracted from blood samples. Whole exome sequencing (WES) was performed for three probands. After the analysis of raw data, candidate variants were confirmed in the patients and their parents.

We have found the possible disease-causing variant as the c.1718G>C variant (p. Trp573Ser) in the SMPD1 gene in the F-1 patient and c.1718G>C (p. Trp573Ser) in the SMPD1 gene in the F-3 patient. Moreover, we have found the c.3175C>T variant (p. Arg1059Ter) in the NPC1 gene in the F-2 patient.

In this study, disease-causing variants were identified in three probands suspected of Niemann-Pick disease. Such results show the relatively high power of molecular techniques to assist clinicians with disease management, therapeutic strategies, and preventive options such as preimplantation genetic diagnosis and prenatal diagnosis.

The significance of HTLV-1 proviral load as a prognostic biomarker in HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) has been a subject of controversy. This study aims to assess the impact of HTLV-1 proviral load (PVL) on the clinical outcome in patients with HAM/TSP.

An absolute quantitative HTLV-1 PVL RT-qPCR, TaqMan method was developed with 100% sensitivity and specificity. Then, from 2005-2018, the HTLV-1 PVL of 90 eligible newly diagnosed HAM/TSP patients were assessed for demographic, clinical symptoms and their associations with HTLV-1-PVL.

The quality control of the designed RT-qPCR showed a sensitivity and specificity of 100%. Spasticity in lower limbs in 58.9% and urinary symptoms in 17.8% of HAM/TSPs were observed. Using this designed RT-qPCR, the HTLV-1-PVL strongly affected spasticity and sphincter disturbance (p=0.05). The multivariate logistic test showed that only the beginning of lower limb weakness along with tremor was associated with PVL (OR: 2.78. 95% CI (0.99-1.02) and p=0.05). Urinary incontinence was prevalent among these patients; however, no association was identified with the HTLV-1 proviral load (PVL).

The absolute RT-qPCR developed for measuring HTLV-1 proviral load (PVL) demonstrated reliable results. Despite a high prevalence of urinary incontinence in these patients, no association was observed with the PVL. Consequently, it appears that HTLV-1 proviral load is specifically associated with developing spasticity in HAM/TSP.

Melatonin, the controlling hormone of the sleep–wake cycle, has acquired attention due to its role in immunomodulation, anti-inflammation, as well as its proapoptotic effects. Wnt/β-catenin signaling can modulate cancer progression by promoting cell division and migration, while miR-let-7b may inhibit cell growth, migration, and invasion by affecting the function of adaptive immune cells. This work was designed to detect the effect of using melatonin as an immunomodulating therapeutic approach to control the progression of chemically induced hepatocellular carcinoma (HCC).

Thirty male rats were equally divided into control, HCC, and melatonin-HCC groups. Animals in the HCC and melatonin-HCC groups were injected with diethylnitrosamine (intraperitoneal single dose) followed by repeated carbon-tetrachloride subcutaneous injection once weekly for six weeks. Melatonin was given from the first week of the study and continued during the process of HCC induction.

In the HCC group, the levels of tumor necrosis factor-α (TNF-α), vascular endothelial growth factor (VEGF), and Wnt/β-catenin expression significantly increased, while there was a downregulation of microRNA Let7b. Melatonin administration reversed these changes, along with an increase in hepatic content of interleukin-2 (IL-2) and caspase-3.

Melatonin exerted hepatic immunomodulating changes, in addition to proapoptotic and antiangiogenic effects, illustrated by increased IL-2, caspase-3, and decreased VEGF levels, respectively. Moreover, the use of melatonin during hepatocarcinogenesis positively modulated the disrupted expression of microRNA let7b and Wnt/β-catenin significantly.

Obesity is an abnormal fat accumulation that adversely affects human health. Studies reported several vitamin deficiencies in obese patients. The current study investigates the deficiencies of vitamins D, B6, and B12 among Jordanian adults with hyperlipidemia and demonstrates the association between serum vitamin levels and metabolic and lipid profile parameters.

 Sixty male subjects were divided into 40 hyperlipidemic patients (age: 45.9 yr. ±10.2) and 20 controls (age: 41.2 yr. ±10.7). The blood levels of triglycerides, total cholesterol, high density lipoprotein (HDL)-cholesterol, hemoglobin A1c, and vitamins D, B6, and B12 were measured.

The hyperlipidemic patients showed significantly increased triglycerides, total cholesterol, non-HDL, cholesterol/HDL ratio, low-density lipoprotein (LDL)- cholesterol levels, and decreased HDL-cholesterol levels compared to the controls. No significant differences were found in the blood levels of vitamin D, vitamin B6, or vitamin B12 between groups. However, 50% of the hyperlipidemic patients and 54.5% of the controls exhibited vitamin D deficiency. Only the hyperlipidemic patients exhibited deficiencies of vitamins B6 and B12 in 5.4% and 3.3% of cases, respectively. In the controls, vitamin B12 level was inversely associated with total cholesterol, whereas in the hyperlipidemic patients, vitamin B6 level was inversely correlated with total cholesterol and non-HDL levels.

 The hyperlipidemic patients exhibited vitamins D, B6, and B12 deficiencies. Additionally, vitamins B6 and B12 levels were inversely correlated with total cholesterol and non-HDL levels. Our findings highlight the importance of routine evaluation of vitamin levels in patients with hyperlipidemia.

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has triggered a global health crisis, with genetic mutations and evolution further creating uncertainty about epidemic risk. It is imperative to rapidly determine the nucleic acid sequence of SARS-CoV-2 and its variants to combat the coronavirus pandemic. Our goal was to develop a rapid, room-temperature, point-of-care (POC) detection system to determine the nucleic acid sequences of SARS-CoV-2 isolates, especially omicron variants.

Based on the conserved nucleotide sequence of SARS-CoV-2, bioinformatics software was used to analyze, design, and screen optimal enzymatic isothermal amplification primers and efficient CRISPR RNAs (crRNAs) of CRISPR/Cas13a to the target sequences. Reverse transcription-recombinase polymerase amplification (RT-RPA) was used to amplify the virus, and CRISPR/Cas13a-crRNA was used to cleave the SARS-CoV-2 target sequence. The sensitivity of nucleic acid detection was assessed by serial dilution of plasmid templates. All reactions were performed at room temperature.

RT-RPA, combined with CRISPR/Cas13a, can detect the SARS-CoV-2 with a minimum content of 102 copies/μL, and can effectively distinguish between the original strain and the Omicron variant with a minimum limit of detection (LOD) of 103 copies/μL.

The method developed in this study has potential application in clinical detection of SARS-CoV-2 and its omicron variants.

Ulcerative colitis (UC), a chronic inflammatory bowel disease (IBD), exerts its impact on both rectal and colonic mucosa, with a growing incidence. This study aims to explore the pharmacogenetic influence of thiopurine methyl transferase (TPMT) gene expression and serum tumor necrosis factor (TNF) levels on the response to Imuran in Iraqi patients with UC.

Seventy individuals with chronic UC and 30 healthy controls were enrolled in this investigation. RNA extraction using the triazole method and enzyme-linked immunosorbent assay (ELISA) for TNF measurement were employed. Patients, aged 15-50 years, underwent Imuran treatment.

Diverse responses to Imuran were observed among patients, with TPMT gene expression levels below 1 in 35 patients leading to side effects, while the remaining 35 patients exhibited positive responses with TPMT gene expression exceeding 1. Patients with varying degrees of severe, moderate, and mild UC associated with TNF showed a significant correlation with Imuran non-response.

 A distinct correlation was identified between TPMT gene expression and Imuran therapy outcomes in UC patients. Further investigation is warranted to elucidate the underlying mechanism, positioning the TPMT gene as a potential therapeutic target for mitigating the impact of UC.

Recent studies have implicated dysregulated long non-coding RNA (lncRNA) levels in the pathogenesis of type 2 diabetes (T2D). This study aimed to assess the expression of circulating HOTAIR and uc.48+, examining their correlation with clinical and biochemical variables in T2D patients, pre-diabetic individuals, and healthy controls.

Peripheral blood levels of lncRNAs were quantified using QRT-PCR in 65 T2D patients, 63 pre-diabetic individuals, and 63 healthy subjects. Pathway enrichment analysis was conducted to explore the functional enrichment of lncRNA-miRNA targets.

Analysis revealed a significantly elevated circulating level of HOTAIR in both T2D (P < 0.0001) and pre-diabetic patients (P = 0.04) compared to controls. ROC analysis demonstrated that, at a cutoff value of 9.1, with a sensitivity of 80% and specificity of 62%, HOTAIR could distinguish T2D patients from controls (AUC = 0.723, 95% CI 0.637-0.799, P < 0.0001). Spearman correlation analysis identified a significant positive correlation between HOTAIR expression, HbA1c, and insulin resistance (P < 0.005). MiRNA enrichment analysis indicated significant enrichment of diabetes-related pathways among HOTAIR's miRNA targets. Conversely, no significant difference in uc.48+ circulating levels between groups was observed, but a significant positive correlation emerged between uc.48+ and systolic blood pressure.

This study provides evidence that elevated HOTAIR expression levels are associated with T2D progression, suggesting their potential as biomarkers for early diagnosis and prognosis.

A genetic polymorphism that causes abnormal folate metabolism may lead to genomic instability and increase susceptibility to malignancies such as Acute Lymphoblastic leukemia (ALL). The purpose of this research is to identify methylene tetrahydrofolate reductase (MTHFR C677T) (NCBI ID: 4524) mutation in ALL patients.

The study was a descriptive case-control hospital-based study with one hundred Sudanese participants divided equally into fifty (50) Sudanese ALL diagnosed patients as cases and fifty (50) Sudanese individuals as controls. The MTHFR C677T mutant allele was detected using conventional PCR, with the primer sequence of MTHFR C677T F-TGAAGGAAGGTGTCTGCGGGA R-AGGACGGTGCGGTGAGAGTG. The study was conducted from January to March 2023, and samples were collected from the Radiation and Isotops Center at Khartoum Hospital.

The investigation revealed that 12 of the 50 patients in the case group (24%) had the MTHFR C677T mutant allele, and the study also revealed that there is significant correlation with the control group. There is no significant relationship between socio-demographic variables and MTHFR mutation detection in ALL patients. Also, the sociodemographic variables predictors of MTHFR mutation among ALL patients adjusted for smoking habit revealed no significant relationship.

According to the findings of this study, the mutant allele of the Methylene Tetra Hydro Folate Reductase C677T was detected and demonstrated varying degrees of significance. It was concluded that the MTHFR C677T gene mutation was associated with acute lymphoblastic leukemia in Sudanese patients.

Urinary tract infection (UTI) is one of the common bacterial infections. Escherichia coli is the most common cause of UTI. In this research, the prevalence of several virulence factors and beta-lactam resistance genes was investigated.

One hundred E. coli isolates were collected from patients’ specimens with UTI referred to Allame-Bohlol Gonabadi hospital. Polymerase chain reaction (PCR) was performed to identify five pathogenic genes (fimH, aer, pap, hly, traT) and three antibiotic resistance genes (blaTEM, blaCTX, blaSHV).

The frequencies of blaSHV, blaTEM and blaCTX beta-lactamase genes among extended-spectrum-beta-lactamases (ESBLs) positive isolates were 11.1%, 48.1%, and 93.3%, respectively. A significant number of isolates were resistant to the most commonly used antibiotics.

Pathogenic genes may also increase the severity, progression, and expansion of urinary tract infections. Therefore, identifying these genes as critical controllers of illness can use for better manage the treatment.

Fast diagnosing ischemic stroke (IS) is a critical issue in clinical studies, as it allows more effective therapy and stops the progression of IS. The blood level of circular RNAs (CircRNAs) after stroke may be a rapid diagnostic marker.

In this study, the blood level of circRNAs was evaluated using a real-time polymerase chain reaction (PCR). We used logistic and linear regression analysis to assess the potential of circRNAs levels with the risk of IS.

circRNA DLG associated protein 4 (CircDLGAP4) was decreased in patients compared with controls, and logistic regression showed its expression negatively associated with IS risk. The expression level of human genome version 38_Circular_0008980 (hg38_circ_0008980) was reduced significantly in patients with small vessel disease (SVD), and the linear regression analysis showed a negative relationship between hg38_circ_0008980 expressions with SVD subtype. hg38_circ_0008980 expression relative to controls showed a significant association with IS risk.

Taken together, we found a significant decrease in the level of hg38_circ_0008980 after IS; it may act as a novel circRNA in IS pathophysiology with a positive correlation with stroke severity.

This study explores the association between growth arrest-specific 5 (GAS5) rs145204276, nuclear paraspeckle assembly transcript 1 (NEAT1) rs512715, and Maternally Expressed 3 (MEG3) rs4081134 polymorphisms and their impact on susceptibility to papillary thyroid carcinoma (PTC), considering differential expression of long noncoding RNAs (lncRNAs) in PTC.

A case-control study involving 125 papillary thyroid carcinoma (PTC) patients and 125 controls was conducted. Genotyping of polymorphisms was performed using tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP) methods.

No significant association was found between the two groups regarding genotypes and allelic frequencies of GAS-5 145204276 and MEG3 rs4081134 polymorphisms. Genetic models also showed the same results. Regarding NEAT1 rs512715, The PTC group had more GC genotypes and over-dominant models of NEAT1 rs512715 than controls, while controls showed a higher frequency of recessive models.

GAS5 rs145204276 and MEG3 rs4081134 polymorphisms showed no significant association with papillary thyroid carcinoma (PTC) risk. In contrast, NEAT1 rs512715 exhibited a significant impact on PTC development.

In the current study, the effects of cerium oxide nanoparticles (nanocerium; NC) on doxorubicin (DOX)-induced cardiomyopathy and its possible underlying mechanisms were addressed.

32 adult male rats were allocated into 4 groups; i) control group, ii) NC group; rats received NC (0.2 mg/kg, i.p., daily), iii) DOX group; rats received DOX 4 mg/kg (2 injections with a 14-day interval), and iv) DOX+NC group as DOX but rats received NC. At the end of the experiment, ECG and ECHO recordings and assessments of the levels of cardiac enzymes (CK-MB, LDH), and myocardial oxidative stress (MDA, catalase, and GSH), the expression of LC3 and beclin1 (markers of autophagy), caspase3 (marker of apoptosis) by immunohistochemistry, the expression of acetyl-CoA carboxylase alpha (ACCA) by PCR, and 5’adenosine monophosphate-activated protein kinase (AMPK) levels in the heart tissues were performed.

The DOX group displayed a prolonged corrected QT interval, an increase in cardiac enzymes (CK-MB and LDH), myocardial oxidative stress (high MDA with low catalase and GSH), expression of ACCA, caspase-3, beclin1, and LC3 in myocardial tissues, with reduction in myocardial AMPK levels, and myocardial contractility (low ejection fraction, and fractional shortening). On the other hand, administration of NC with DOX resulted in significant improvement of all studied parameters.

NC offers a cardioprotective effect against DOX-induced cardiomyopathy. This effect might be due to its antioxidant and antiapoptotic effects as well as to the modulation of autophagy and metabolic dysfunctions induced by DOX in the heart tissues.