Reports of Biochemistry and Molecular Biology最新文献

Background: 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).

Methods: 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.

Results: 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.

Conclusions: 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.

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Background: 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.

Methods: 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.

Results: 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.

Conclusions: 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.

Background: 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.

Methods: 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.

Results: 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.

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

Background: 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.

Methods: 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).

Results: 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.

Conclusion: 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.

Background: Red blood cell distribution (RDW), an index of the size variability of erythrocytes, is significantly associated with coronary stenosis and can strongly predict the mortality risk in coronary artery disease (CAD). The biological mechanisms involved are not fully understood but may include oxidative stress. We sought to investigate the relationship between RDW and markers of oxidative stress in patients with CAD.

Methods: Participants were 112 consecutive patients referred to department of cardiac surgery for evaluation of chest pain. 32 patients had stable CAD, 40 patients had unstable CAD and 40 subjects were diagnosed as non-CAD. The levels of lipid peroxidation (TBARS) were measured in plasma and membrane samples by a fluorometric method. The plasma levels of glutathione (GSH) and total antioxidant capacity (TAC) were determined using spectrophotometric methods.

Results: Lipid peroxidation levels were significantly higher in the erythrocyte membrane of stable CAD patients than non-CAD patients. The levels of TAC were significantly lower in both stable and unstable groups when compared to that of the control group (P< 0.019 and P< 0.001, respectively), but did not differ between stable and unstable CAD. In addition, there was no significant difference in the serum GSH levels among the study groups. Membrane TBARS was directly associated with RDW in three groups of study.

Conclusions: We found an independent association between RDW levels and membrane lipid peroxidation in patients with CAD. This finding suggests that oxidative stress may be a potential underlying biological mechanism for increased RDW in CAD patients.

Background: The number of erythromycin-resistant Streptococcus pneumoniae has significantly increased around the world. The present study aimed to determine the serotype distribution and molecular epidemiology of the erythromycin-resistant Streptococcus pneumoniae (ERSP) isolated from patients with invasive disease.

Methods: A total of 44 Streptococcus pneumoniae isolates were tested for susceptibility to several antimicrobial agents. Additionally, the polymerase chain reaction (PCR) was applied to evaluate ERSP isolates in terms of the presence of erythromycin resistance genes (e.g., ermB and mefA). The isolates were serotyped using the sequential multiplex-PCR method, and molecular epidemiology was assessed through the multilocus sequence typing (MLST) analysis.

Results: The results represented multidrug resistance (MDR) in approximately half of the pneumococcal isolates. Among 22 ERSP isolates, 20 (90.9%) and 12 (56%) ones contained ermB and mefA, respectively. Further, 14 (31.8%), 3 (22.7%), and 19A (18.1%) were the common serotypes among the isolates. No significant correlation was observed between serotypes and erythromycin resistance genes. Furthermore, the MLST results revealed 18 different sequence types (STs), the top ones of which were ST3130 (3 isolates) and ST166 (3 isolates). Population genetic analysis disclosed that CC63 (32%), CC156 (18%), and CC320 (18%) were identified as the predominant clonal complexes.

Conclusions: The ERSP isolates exhibited high genetic diversity. The large frequency of MDR isolates suggests the emergence of high resistant strains, as well as the need to implement vaccination in the immunization schedule of Iran. These accumulating evidences indicate that 13-valent pneumococcal conjugate vaccines provided higher serotype coverage in the ERSP isolates.

Background: Preeclampsia (PE) is a multisystem pregnancy disorder that increases maternal-perinatal morbidity and mortality significantly. MicroRNA-155 (miR-155) overexpression in the sera of pregnant women has been linked to preeclampsia. Researchers discovered that miR-155 acts during pregnancy by down-regulating and reducing the cysteine-rich angiogenic inducer 61 (CYR61), which causes local ischemia as well as oxidative stress.

Methods: The level of miR-155 expression in all serum samples was quantified using real-time polymerase chain reaction (RT-PCR), and serum CYR61 was measured using enzyme-linked immunosorbent assays. Together with the Cyr-61/miR-155 ratio, they were evaluated as biomarkers for PE pathogenesis and severity prediction.

Results: MiR-155 expression, serum CYR61 levels, and Cyr-61/miR-155 ratios were all significantly higher in PE patients compared to the control group. Serum CYR61 levels and the Cyr-61/miR-155 ratio differed significantly between mild and severe PE patients.

Conclusions: MiR-155 expression, serum CYR61 levels, and Cyr-61/miR-155 may serve as biomarkers for PE pathogenesis and severity prediction.

Background: Allergic rhinitis (AR) is an inflammatory disorder of the nasal mucosa, caused by exposure to environmental allergens. It is known that 15-lipoxygenase (15-LOX) is involved in the biosynthetic pathways of anti-inflammatory lipid mediators, including resolvins and protectins.

Methods: In this study, which was performed on 130 AR patients and 130 healthy controls, we aimed to investigate the association of susceptibility to AR with two selected single-nucleotide polymorphisms (SNPs), that is, rs2619112:A>G and rs7217186:C>T, in the intron regions of arachidonic acid 15-LOX (ALOX15) gene, using SNPinfo and Regulome DB tools.

Results: The results showed that the CT genotype of rs7217186: C>T was significantly associated with the increased risk of AR compared to the CC genotype (P= 0.037, OR=1.943, CI: 1.038-0.638). However, there was no strong evidence of the association of rs2619112: A>G with susceptibility to AR (P> 0.05).

Conclusions: The present results indicated that rs7217186 polymorphism of ALOX15 gene might be a potential biomarker for susceptibility to AR.

Background: Systemic Lupus Erythematosus (SLE) is a chronic autoimmune condition that affects multiple organs significantly impacts morbidity and mortality. The development of SLE is influenced by genetic predisposition and dysregulated immune response. Our objective was to investigate miR-21, IL-10, and PDCD4 expression in SLE patient plasma and analyze their correlations and potential diagnostic and prognostic values.

Methods: The study included 100 healthy subjects, 50 newly diagnosed (ND), and 50 under-treatment (UT) SLE patients. The patients were observed for 24 weeks to track relapses. miR-21 and PDCD4 gene expression levels were measured using real-time RT-PCR, and IL-10 production was measured using ELISA.

Results: miR-21 and IL-10 expression levels were significantly greater in SLE patients than in healthy subjects, with the highest levels observed in ND patients. PDCD4 expression was also significantly greater in SLE patients than in subjects, with the highest levels observed in UT patients. ROC curve analyses and Cox-Mantel Log-rank tests indicated miR-21, PDCD4, and IL-10 as proper diagnostic and prognostic biomarkers for SLE. The study also revealed a significant positive correlation between miR-21 and PDCD4 and IL-10 levels in SLE patients.

Conclusions: The studies suggest that dysregulation of miR-21, PDCD4, and IL-10 in patients with SLE may contribute to disease development and provides new diagnostic and prognostic markers. Additionally, the observed correlation between miR-21, PDCD4, and IL-10 levels in SLE patients signifies a potential interplay between these molecules.