Reports of Biochemistry and Molecular Biology
Volume:13 Issue: 1, Apr 2024

People with autism frequently exhibit poor social skills, communication difficulties, and repetitive and stereotyped behaviors. MicroRNAs (miRNAs) are potential and promised targets in developing of new treatment strategies for autism.This study aimed to assess the relative expression of miR-124a, miR-34a-3p, miR-545-3p, miR-153, and BDNF in the blood samples of autistic children.

The children autism rating scale (CARS) was used to determine the severity of autism and to confirm the diagnosis. Blood samples were obtained from 50 patients and 40 age-/sex-matched healthy controls. Expressions of miR-545-3p, miR-34a-3p, miR-124a, and BDNF were evaluated using qRT-PCR. Pearson's correlation coefficient and regression analysis were used to check correlations between relative expressions of the miRNAs and BDNF. Biomarker potencies were assessed by ROC curve analysis.

 qRT-PCR analysis showed that the relative expressions of miR-545-3p, miR-34a-3p, miR-124a, and BDNF were significantly higher in the patients' group than the healthy controls. However, the relative expression of miR-153 was significantly lower in the case group than the control group. The relative expression of miR-124a was positively correlated with those of miR-545-3p and BDNF among the patients group. Also, the relative expressions of miR-545-3p and BDNF were positively correlated with each other. The ROC curve data also indicated that miR-124a, miR-34a-3p, miR-545-3p, miR-153, and BDNF could be possible diagnostic biomarker for CARS diagnosis (AUC=0.8328, AUC=0.8354, AUC=0.6727, AUC=0.8518 and AUC=0.8214, respectively).

Deregulation of miR-124a, miR-454-3p and BDNF might be considered as potential biomarkers for severity of autism.

Hypertrophic scar (HS) is a cutaneous condition results from abnormal wound healing following deep tissue injury. To date, there is no optimal treatment for this skin disorder. Catechins possess anti-inflammatory, antioxidant and anti-fibrotic properties. In this study we investigated the effects of catechin hydrate (CH) in rabbit ear model of HS.

A rabbit ear model of hypertrophic scar was set up. Ten New Zealand white rabbit were divided into 5 equal groups: non-treatment group, vehicle control, treated with intralesional injection of dimethyl sulfoxide (DMSO), and test groups, received intralesional injection of CH/DMSO solution at concentration of 0.25, 1.25 and, 2.5 mg/ml, respectively. The treatments were initiated ‏‎35 days after wounding once a ‏week for ‏‎4 ‎weeks‎.‎ ‏ The scar elevation index (SEI) and the epidermal thickness index (ETI) were measured using Hematoxylin and Eosin (H&E) ‏staining and the amount of collagen deposition were determined after Masson‎' ‎trichrome staining‎. ‎In addition‎, the enzyme-linked immunosorbent assay (ELISA) ‎‏method was used to determine the levels of ‎ type І ‏and ІІІ collagen and matrix metalloproteinase ‏‎1 (MMP1) in scar tissues.

CH improved abnormal scarring at concentrations of 1.25 and 2.5 mg/ml and significantly (P<0.001) reduced the SEI and ETI. The levels of collagen type І and type ІІІ, and total collagen deposition were significantly (P<0.05) decreased in scar tissues of CH treated groups and no significant effect on MMP1 levels.

Our findings demonstrated that CH has the potential for the treatment of HSs.

Prostate cancer is a classic public health problem in males and has broadly different levels of mortality and morbidity. As an endocrine gland, adipose tissue synthesizes and secretes a variety of bioactive peptides, such as irisin and omentin-1. Adipokines and oxidative stress potentially contribute to the proliferation of prostatic carcinoma cells. The relationship between irisin, omentin-1, and oxidative stress has not been widely investigated in prostate cancer. Therefore, the present research assessed whether there is a significant correlation between irisin and omentin-1 levels and oxidative status in prostate cancer individuals.

The present research recruited 40 individuals diagnosed with prostate cancer and 40 healthy individuals for comparative purposes. All individuals underwent demographics, biochemicals, and serum adipokines (irisin and omentin-1) data analysis.

The means of total prostate-specific antigen (43.3±20.5 vs. 2.5±1.2) and free prostate-specific antigen (2.1±1.4 vs. 0.08±0.02) were highly significant increases in the prostate cancer patients than in the healthy individuals. Furthermore, the means of omentin-1 (31.6±12.8 vs. 23.5±14.1) and total oxidant stress (22.4±10.6 vs. 9.1±3.6) were highly significant increases in patients with prostate cancer than in healthy individuals. In contrast, the means of irisin (343.5±240.2 vs. 716.4±142.3) and total antioxidant capacity (2.2±1.2 vs. 3.3±1.3) were highly significant decreases in patients with prostate cancer than in healthy individuals. No significant relationship was demonstrated between all parameters in the two groups under study.

The study findings indicate that irisin and omentin-1 could serve as biomarkers for predicting prostate cancer.

The progression of acute limb ischemia (ALI) is being significantly influenced by changes in immune system function. The study aimed to determine the dominant immune cell responses (Th1 or Th2) in ALI patients by measuring serum levels of IL-4, IL-12, and IFN-γ. Previous studies indicate altered cytokine levels in cerebral ischemia, but there is no prior research on these cytokines in ALI patients.

This study involved 34 patients with ALI and 34 healthy controls. Blood samples were analyzed for hematological factors such as erythrocyte sedimentation rate (ESR), white blood cell (WBC) count, red blood cell (RBC) count, platelet (Plt) count, hemoglobin (Hb), and hematocrit (HCT). The levels of serum cytokines IL-4, IL-12, and IFN-γ were measured in both patients and control subjects using enzyme-linked immunosorbent assay (ELISA). The statistical analyses were conducted using SPSS and GraphPad Prism.

The results showed that serum levels of IL-4 in ALI patients did not significantly differ from those in control groups. Acute limb ischemia exhibited significantly elevated levels of IL-12 and IFN-γ compared to healthy individuals. In addition, no correlation between the production of cytokines and the hematological parameters was found.

Th1 responses are believed to play a role in the pathogenesis of ALI, but further research is needed to fully understand their exact role.

Asthma is a common and major allergic disease in the world. We aimed to investigate the impact of supplements with vitamin D, folic acid, selenium, zinc, and copper in patients with moderate to severe asthma.

In this clinical trial study 70 patients above six years old with moderate to severe asthma, were divided into two groups, randomly; one group received daily Asmavit syrup, 10 ml (Asmavit, Vitabiotics Ltd, London, UK), and the other group received daily 1000 IU vitamin D3 drops (Asmavit, Vitabiotics Ltd, London, UK) for two months along with ordinary treatment for asthma. Clinical and physical examinations, immunological and biochemical tests were carried out for each patient before and after the treatment.

The mean age of patients was 39.9± 14.7 years old, and the mean disease duration was 8.8 ± 9.8 years. A significant increase in lung function, asthma control, and quality of life score tests was observed in both groups after the treatment (P< 0.05). There was no significant difference in cytokines expression levels before and after the treatment with vitamin D3 or Asmavit (P> 0.05). Serum levels of selenium and folic acid before treatment were correlated with disease severity, while post-treatment vitamin D levels significantly increased FEV1 (P> 0.05). Oxidative stress levels reduced in both groups, with greater reduction in the vitamin D group (P< 0.05).

Supplements, particularly vitamin D, when combined with standard asthma treatment, may effectively improve clinical symptoms and enhance the quality of life for asthmatic patients.

Individuals experiencing severe cases of Coronavirus Disease 2019 (COVID-19) exhibited elevated fibrinogen levels and decreased albumin levels, potentially linked to the presence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) proteins. Consequently, our study endeavors to examine the impact of SARS-CoV-2 ORF9b on the expression of fibrinogen and albumin genes within the Hep-G2 cell line.

In this study, the Hep-G2 liver cell line was utilized alongside the plasmid pcDNA3.1 hyg+ containing ORF9b from the SARS-CoV-2 strain originating in Wuhan. Transfection procedures were executed, and the transfected cells were selected utilizing hygromycin B. Validation of ORF9b expression was conducted through SYBR green-based real-time PCR, and the expression of the Fibrinogen α (FGA), Fibrinogen β (FGB), Fibrinogen γ (FGG), and Albumin (ALB) genes was quantified using the same method.

The real-time PCR analysis revealed a significant upregulation of fibrinogen genes—α (P=0.03), β (P=0.02), and γ (P=0.029) in Hep-G2 cells containing ORF9b compared to control cells. Furthermore, the findings indicated a markedly lower expression level of albumin in Hep-G2 cells harboring ORF9b compared to the control cells (P=0.028).

The findings suggest that SARS-CoV-2 ORF9b could potentially influence the course of SARS-CoV-2 infection by triggering the expression of α, β, and γ fibrinogen gene chains while suppressing the albumin gene. Further investigations are warranted to validate these observations across various SARS-CoV-2 strains exhibiting differing levels of pathogenicity.

Oral Squamous Cell Carcinoma (OSCC) is a pressing global health challenge. Long non-coding RNAs (lncRNAs) have emerged as pivotal regulators. Among these, the lncRNA ANRIL (antisense non-coding RNA in the INK4 locus) has a role in cancer progression. The aim of this study was to look into possible links between a certain genetic variant of lncRNA ANRIL, rs10757278 A/G, and OSCC risk and tumor features in the Iranian population.

We conducted a case-control study, enrolling 101 OSCC patients and 115 healthy controls. We took out the genomic DNA and used the tetra-primer ARMS-PCR (tetra-primer amplification refractory mutation system-polymerase chain reaction) method to find the rs10757278 genotype. We evaluated the associations between genotypes and both OSCC susceptibility and various tumor characteristics.

Although we did not observe significant differences in allele and genotype frequencies between cases and controls, we revealed compelling associations between genotypes and tumor characteristics. Genotypes AG and GG were linked to smaller tumor sizes, while genotypes with at least one wild-type allele (A) were linked to well differentiated OSCC. Specific genotypes exhibited significant associations with tumor sites, with the tongue demonstrating the strongest correlation.

The rs10757278 A/G variant did not show a direct link with OSCC risk, but its complex effect on tumor behavior suggests that it may play a bigger role in the development of OSCC. These findings open avenues for future investigations to uncover hidden genetic interactions, and potentially inform more targeted therapeutic strategies.

The most common cause of Nephrotic Syndrome (NS) in children is idiopathic NS, also called nephrosis. The most prominent clinical signs are hyperlipidemia, severe proteinuria, edema, swelling of body tissues, and an increased risk of infection. The object of this study was to examine the correlation of the ABCB1 gene (rs10276036, C > T), IL-18, and TNFα to the prevalence of NS among Egyptian children having NS.

This study included 100 participants with NS and 100 healthy controls. To analyze the ABCB1 gene (rs10276036 C >T) variant PCR technique was used. IL-18 and TNF levels were estimated using Enzyme-Linked Immunosorbent Assay (ELISA).

Increased frequency of CT and TT genotypes of the ABCB1 gene (rs10276036 C / T) in NS patients compared to controls, with p-value = 0.001, OR = 2.270, CI = (1.550-3.327) for CT genotype and p-value = 0.001, OR = 5.070, CI = (2.463-10.438) for TT genotype. The frequencies of ABCB1 (rs10276036 C >T) genotypes were statistically significant in the dominant model (OR 2.560; p< 0.001) and in the recessive model OR, 3.231; p= 0.001). Significantly high levels of both IL-18 and TNFα were found in NS patients compared to controls.

The ABCB1gene (rs10276036 C/T), IL-18, and TNFα are associated with the prevalence of NS in Egyptian children and might be considered as independent risk factors for its incidence.

Infertility affects about 15% of couples during reproductive age and male factor infertility accounts for half of these cases. This study aimed to evaluate the expression of leptin, KISS1, leptin receptor, and HOTAIR genes in blood and semen samples of individuals diagnosed with oligospermia in comparison to healthy controls.

In the current investigation, we studied 36 semen and 30 blood samples from fertile oligospermic men as well as the same number of healthy controls. RNA was extracted and cDNA was synthesized. Real-time polymerase chain reaction (PCR) was conducted to assess the gene expression levels. Statistical analysis was performed using Graph Pad Prism software. The results were reported as mean±SEM and any P< 0.05 were considered statistically significant.

A significant increase in the expression of the leptin gene in infertile males, particularly in semen samples (P< 0.001) was found when compared to the healthy controls. On the other hand, significant decrease in the expression of the KISS1 (P< 0.0001) and HOTAIR (P< 0.001) genes, particularly in semen samples were found when compared to the controls. As it was shown, the expression of leptin receptor had no significant effects on male infertility. Additionally, there may be a potential correlation between the expression of HOTAIR and the leptin and KISS1 genes, with a negative correlation observed between HOTAIR and leptin, and a positive correlation between HOTAIR and KISS1 in both blood and semen samples. However, further investigations are necessary to establish the statistical significance of these correlations.

According to the results, leptin, KISS1, and HOTAIR genes seem to be affected in oligospermia, however, further studies with higher sample sizes are necessary.

Amino acid analysis is an important tool for the diagnosis of metabolic disorders in newborns. Today, Liquid Chromatography tandem mass spectrometry (LC-MS/MS) has emerged as a powerful technique for amino acid analysis. We aimed to determine the local normal range of amino acids in dried blood spot (DBS) samples of neonates using LC-MS/MS.

A total of 1005 samples from healthy neonates of northeast and east of Iran aged 2-7 days were utilized for normal range determination. The amino acids were extracted from dried blood spot samples using organic solvent and then analyzed using LC-MS/MS system. The 1%, 2.5%, 97.5%, and 99% percentiles were calculated, and the results were compared to the global cut-off values.

The results showed that glutamic acid has the highest concentration range among amino acids evaluated in this study (178.94 – 421.31mmol/L). Moreover, the plasma concentrations of Glycine (142.65 – 397.06 mmol/L), Alanine (97.00–349.72 mmol/L), Proline (63.77 – 236.53 mmol/L), and Tyrosine (25.79 – 150.58 mmol/L) were in the next ranks. Comparing the obtained results with the global values obtained in the R4S study indicated a slight difference between the obtained local normal values and the global values.

The calculated values were slightly different from global values obtained in the R4S study and regional values calculated in other studies. This further emphasized the importance of the local establishment of reference values, which facilitates the correct interpretation and diagnosis in the Newborn Screening Programs.

Several studies provide evidence for a role of serum cytokines imbalance including IL-10 and IL-27 in immune thrombocytopenia pathogenesis and prognosis. The aim of this study was designed to investigate the role of serum levels of IL-10 and IL-27 in prognosis the efficiency of treatment in thrombocytopenic Iraqi children

This case controls study was carried out at Department of Biochemistry, College of Medicine, University of Baghdad, during the period from October 2023 to March 2024. It included 88 children, 63 children previously diagnosed with immune thrombocytopenia, and 25 apparently healthy children who served as control group. The included immune thrombocytopenic children were sub-grouped according to their treatment into three groups: Romiplostim group (group 1), Prednisolone group (group 2), Prednisolone and intravenous immunoglobulin (IVIG) or Prednisolone and mycophenolate group (group 3). Investigations included serum level measurements of IL-10 and IL-27 by using enzyme linked immunosorbent assay ELISA. Platelet count of each included children was measured by Huma Count 30 TS Human, Germany.

The mean (±SEM) values of serum IL-10 and IL-27 levels of immune thrombocytopenic children were insignificantly lower than that of controls. In addition, there was non- significant differences in serum levels of IL-10 and IL-27 among and between the three groups of patient children. The mean value of platelet count of patient children was significantly increased by all types of treatment in whole immune thrombocytopenic children (117.48±18.15*10^9/L).

Measurement of serum IL-10 and IL-27 are helpful biomarker in prognosis of thrombocytopenia irrespective of type of treatment.

Anti-Müllerian hormone (AMH) has lately been connected to polycystic ovary syndrome (PCOS) in a growing body of research, even though hyperandrogenism is one of the primary symptoms of PCOS.    
Evaluate the association of serum anti-Müllerian hormone and free testosterone with different phenotypes of polycystic ovary syndrome.

This is cross-sectional study was carried out at Department of Biochemistry, College of Medicine, University of Baghdad, during the period from November 2023 to March 2024. It included 111 women, 91 of these women, age range (18-40 year) were diagnosed with polycystic ovary syndrome according to 2003 Rotterdam Consensus criteria, and 20 women were apparently healthy women. The PCOS women were sub-grouped into four phenotype groups (A, B, C and D). Investigations included serum measurements of free testosterone and anti-müllerian hormone by using enzyme linked immunosorbent assay (ELISA) technique in all included women.

 The results revealed that phenotype A is the predominant one of PCOS, while the B phenotype is the rare one. The mean (±SEM) values of free testosterone levels of phenotypes A, B, and C were significantly higher those of phenotype D and controls (p=0.001). The mean (±SEM) value of serum anti-Mullerian hormone levels was significantly increased in phenotypes A (p=0.04) and D (p=0.01) than C phenotype.

Phenotype A is the predominant one of PCOS phenotypes and is associated with highest serum AMH, free testosterone and obesity. Both free testosterone and AMH are helpful in differentiation of different phenotypes of PCOS.

Infection of diabetic foot ulcer is very common and leads in 20% of cases to amputation. Antibiotic-resistant Staphylococcus aureus is the main cause of severe infection. Antibiotic resistance is a major challenge to the global health system. This work aimed to investigate the antibacterial efficacy of some algae extracts against Staphylococcus aureus isolated from diabetic foot ulcers.

freshwater river samples were collected to isolate the algae, and PCR was used for identification. The ethanol, water, and ethyl acetate extract of these algae were prepared and analyzed using high-performance liquid chromatography-mass spectrometry to determine the key components that have antibacterial properties. The antibacterial activity of these extracts against S. aureus was determined by broth dilution and well diffusion methods.

Chlorella vulgaris and Anabaena flos-aquae were isolated from freshwater river and identified by PCR. Anabaena flos-aquae has a greater antibacterial efficacy against Staphylococcus aureus in comparison to Chlorella vulgaris, and the ethanolic extract demonstrated superior outcomes compared to the aqueous and ethyl acetate extracts. The MS spectrum of both algae had a very similar pattern, but the frequency of detected peaks was different

Ethanolic extract of A. flos-aquae and Chlorella vulgaris can be suggested to treat and control diabetic foot ulcer infection caused by S. aureus. Further studies are required to explore the full potential of these algae safely and extensively.

The envelope (E) protein of globally circulating severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) is highly conserved. This study aimed to find the mutation rate of the E genes in COVID-19 patients, and also to evaluate the conformational characteristics of viral E protein.

In this study, 120 patients with SARS-CoV-2 positive test results were selected according to real-time PCR assay. Specific primers for conventional PCR have been used to amplify E gene; furthermore, to identify the E gene mutations, direct sequencing of the E genes was also done. Bioinformatics techniques were used to investigate the possible effects of antigenic changes and 3D characteristics of amino acid substitutions. Also, the immunogenicity of wild-type and mutant E was analyzed utilizing a ClusPro docking server and the IEDB online platform.

A total of 120 COVID-19 patients were included (57.5% were male and 42.5% female), with an overall mean age of 55.70±10.61 years old. Of 10 nucleotide changes, 8 (80%) were silent. Also, 2 (20%) missense mutations (amino acid altering) were found in the E gene (L73F and S68F).

These mutations insert some new helix structures in the E mutants. Also, the results of molecular docking studies indicated that both S68F and L73F mutations could notably enhance the stability and binding affinity of protein E's C-terminal motif to the Protein Associated with LIN7 1, MAGUK P55 Family Member (PALS1) which may probably increase local viral spread, and infiltration of immune cells into lung alveolar spaces.

high body mass index (BMI) is known to be associated with high blood pressure. Levels of matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), and interleukin-6 (IL-6) are also increased in hypertensive patients. The aim of this study was to determine the correlation of BMI with MMP-9, TIMP-1, and IL-6 based on blood pressure.

The study design was cross-sectional with subjects aged >= 36 years, male and female and divided into 3 groups: group with normal blood pressure (NBP), group with controlled hypertension (CHT), and group with uncontrolled hypertension (UcHT). Height, weight, and blood pressure were measured, as well as serum levels of MMP-9, TIMP-1 and IL-6 using the ELISA method. The correlation was considered significant at p-value of < 0.05.

 The BMI in group UcHT was higher than in the other groups. THERE was a positive correlation between BMI and MMP-9; BMI and TIMP-1; and BMI and IL-6 (r=0.480, p=0.007; r=0.620; p=0.000; r=374, p=0.042 respectively) in group UcHT.

An increase in BMI is accompanied by an increase in levels of MMP-9, TIMP-1, and IL-6 in group UcHT, signifying that it is necessary to control BMI to maintain stable levels of MMP-9, TIMP-1, and IL-6, thereby keeping blood pressure under control.