Hemoglobin is a tetrameric oxygen transport protein in animal bodies. However, there is a paucity of information regarding differences between alpha and beta subunits of hemoglobin in terms of oxygen affinity. The sequential model of Koshland, Nemthy and Filmer (KNF model) has attributed similar affinities to both alpha and beta subunits. The main purpose of the present study is to construct a new sequential model for hemoglobin oxygenation based on higher oxygen affinities for alpha subunits. To this end, coordinate files of 19 oxy and 41 deoxy hemoglobin structures were used as starting structures. These files were processed using Microsoft Excel and SPSS software in order to calculate Euclidean distances between each pair of proximal and distal histidine Fe2+ as well as other pairs of atoms of interest. The calculated distances were then compared for either set of hemoglobin conformations, i.e. oxy and deoxy conformations. Our results showed that a2 subunit show higher structural changes that could be related to oxygen affinity. This subunit could be introduced as initiator of hemoglobin oxygenation and cooperativity. Subunit a2 in our sequential model induces relaxed conformation in a1, b2 and b1 respectively. The order of oxygen affinity in our model is as follow: a2>a1>b1>b2.

Activation of chloride gated GABA receptors regulates the excitatory transmission in the epileptic brain. Positive allosteric modulation of these receptors via distinct recognition sites is the therapeutic mechanism of antiepileptic agents which prevents the hyperexcitability associated with epilepsy. These distinct sites are based on subunit composition which determines binding of various drugs like benzodiazepines. The binding of antiepileptic agents to this recognition site increases the affinity of GABA receptor for modulating the inhibitory effects of GABA-induced chloride ion flux. In the pentameric complex structure of these receptors, the a/g interface forms the benzodiazepine (BZD) binding site on extracellular domain. Thus the a subunit is shown as highly required for functional modulation of the receptor channels by benzodiazepines. The extracellular domain of a subunit of human GABAA is modeled and docking studies are performed with clonazepam, clobazam, clorazepate, diazepam, midazolam, lorazepam. In order to accomplish this, the amino acid sequences human gamma-aminobutyric acid receptor subunit alpha-1 precursor was obtained from National Center for Biotechnology Information. Three-dimensional structure of protein sequences were received from phyre2 protein fold recognition server. Molecular and structural properties of drugs were taken by drug bank server. Autodock4.2 software was used for docking purpose. This study shows hydrogen bond interactions of GABRA1 with selected drugs and binding modes and the interacting amino acid residues involved in recognition of the compound. The results obtained from this study would be useful in understanding the modulatory mode of GABRA1 with benodiazipine drugs.

Phycocyanin is a blue pigment in two eukaryote algal genera and in cyanobacteria as Spirulina. This pigment has various biology activities and are utilised in a number of applications in foods, cosmetics and pharmaceuticals. A lot advantage of phycocyanin studied by many researchers but the scale-up of these methods is difficult and expensive while production of recombinant phycocyanin is more convenience and inexpensive to scale up protein desire. The purpose of this study was to isolation and cloning of phycocyanin alpha subunit gene in expression vector and production of recombinant protein in E.coli to provide industrial production of phycocyanin. The genomic DNA of Spirulina platensiswas prepared and used for PCR as template. phycocyanin alpha subunit gene amplified by designed specialize primers was cloned in a pET43.1a+ expression vector, under the control of T7 promoter usingNdeI and NotI restriction enzymes. The cloning of phycocyanin alpha subunit gene is confirmed by colony PCR, digestion and DNA sequencing. The constructs were transformed in to E.coli strain BL21 (DE3). Expression of phycocyanin alpha subunit gene was examined by 12.5 % SDS-PAGE analysis at 8 hrs after induction by IPTG. The SDS-PAGE analysis showed that alpha subunit phycocyanin was produced in E.coli expression system. Our study provided the production of recombinant phycocyanin. Also overexpression of the synthetic alpha subunit phycocyanin in a bacterial system (E.coli BL-21) showed that E.colican be used to produce this desire protein in large quantity.

Introduction: Anterior pituitary glycoprotein hormones include thyroid stimulating hormone, luteinizing hormone, follicle stimulating hormone, and gonadotropin hormone. Each of them contains alpha and beta subunits. The alpha subunit gene is the same in all of these hormones and contains 4 exons and 3 introns.The beta subunit is responsible for specific function of each hormone. The aim of this study was to clone alpha chain cDNA of glycoprotein hormones in a proper vector for eukaryotic system.Material and Methods: To clone cDNA, alpha subunit of glycoprotein hormones was amplified by using one pair primers and T.vector as template and cloned in Not I and Bam HI sites of pcDNA3.1 plasmid. The recombinant plasmid transformed to E.coli Top 10F¢ cell and colonies that contain plasmid were selected by Colony PCR. The accuracy of extracted plasmid of these clones was approved by enzyme digestion and sequencing.Results: Enzyme analysis showed that pcDNA3.1-F351a had correct structure and sequencing confirmed by 100% homology of the gene with reported alpha gene in Gene Bank.Conclusion: Because of its proper structure, this plasmid is able to transform to Eukaryotic system and translation.

Introduction: Tetralogy of Fallot (TOF) is the most common cyanotic form of congenital heart defects. However, there is no effective therapeutic approach and current therapies have limited curative efficacy. Moreover, the exact etiology of TOF has remained largely unknown. Improved understanding of molecular mechanisms can give an insight into TOF pathogenesis and development of therapeutic approaches. Methods: Here, we conducted a systematic study on the right ventricular myocardium of 24 infants (16 ToF/8 control) using weighted gene co-expression network analysis (WGCNA) to identify meaningful modules or candidate biomarkers. Results: Co-expression network analysis by WGCNA suggested that a highly preserved turquoise module with 2, 493 genes and a P-value of 3×10-11 was significantly correlated to TOF. The top 5 hub genes of this module were PSMA2, MYL12A, C11ORF71, COMMD6, and CREG1. The result of turquoise module enrichment showed that the most correlation topic in biological processes and KEGG pathways were positive regulation of cardiac neural crest migration involved in outflow tract morphogenesis and positive regulation of neural crest cell differentiation. Also, we recognized 4 FDA-approved drug candidates for other indications could potentially use for the treatment of TOF patients through regulation of two hub genes of the coexpression network (PSMA2 and NDUFA4). Our findings also showed that the 13 experimentally validated microRNAs regulated the co-expression network through 5 hub genes. Conclusion: We systematically recognized co-expressed gene modules and hub genes associated with TOF progression, which offered insights into the mechanisms underlying TOF progression and some potential drugs for the treatment of TOF.

Background: Autism spectrum disorders (ASDs) (MIM 209850) are a group of distinct neurodevelopmental disorders characterized by impaired social interactions and communication abilities and abnormal repetitive activities. Many genetic variants have been shown to be associated with ASD. Channelopathies are among putative culprits in the pathogenesis of many neurodevelopmental disorders, including autism. The calcium channel, voltage-dependent, L type, alpha 1C subunit gene (CACNA1C) encodes an alpha-1 subunit of a voltage-dependent calcium channel. Genetic variants within this gene have been associated with psychiatric disorders including Autism Spectrum Disorders (ASD). Our aim was to determine whether the SNPs rs1006737, rs4765905, and rs4765913 were associated with ASD in an Iranian population. Methods: In the present case-control study we investigated the associations of rs1006737, rs4765905, and rs4765913 polymorphisms within CACNA1C and the risk of ASD in a population of 529 Iranian ASD patients and 480 age, gender, and ethnicity-matched healthy subjects Results: None of these SNPs were associated with ASD risk in the assessed population. Although previous studies have shown an association between these polymorphisms and psychiatric disorders and an association between rs4765905 and ASD, we did not replicate those results in our study. Conclusions: Our data indicate that these CACNA1C variants are not involved in the pathogenesis of ASD in the Iranian population.

Background: Hepatocellular carcinoma (HCC) cell development was investigated in relation to the regulation of Sec61 translocon alpha 1 subunit (SEC61A1) by microRNA (miR)-18a-5p. Materials and Methods: After collection of clinical samples, the transfection of interfering vectors of miR-18a-5p or SEC61A1 was into HCC cells to figure out their roles in development of HCC. The Pearson test and starBase analyzed the association and target prediction of miR-18a-5p. Subsequently, through the dual luciferase reporter experiment, SEC61A1 can be regulated via miR-18a-5p. The salvage experiment revealed that miR-18a-5p influence the degradation process of Hepatocellular carcinoma through combining SEC61A1. Results: In HCC cells, we found that SEC61A1 was elevated and miR-18a-5p was downregulated. HCC cells deteriorating was considerably slowed down by the enhanced miR-18a-5p level. The outcomes demonstrated that miR-18a-5p can interact and regulate SEC61A1. Additionally, the effects of earlier therapy on HCC cell proliferation can be restored by overexpressing SEC61A1. Conclusion: Overall, in HCC cells and tissues miR-18a-5p was significantly downregulated, and inhibited the proliferative ability of HCC cells by targeting SEC61A1.

Background: Atopy and/or allergy are in the same meaning. Patients with allergy are individuals who have positive family history and the number of eosinophil in their blood test and immunoglobulin E (IgE) level in their skin test are high against allergen, which is demonstrated by allergic rhinitis، eczema، asthma and urticaria. Many genetic factors are involved in disease onset one of which is interleukin-7 receptor subunit alpha (IL7R-α). In this study we aimed to evaluate the frequency of this polymorphism in two groups of case and control.Methods: 101 patients with allergy and 201 controls were selected. After DNA extraction، different genotypes of C/T polymorphisms were studied using high-resolution melt real-time polymerase chain reaction (HRM real-time PCR) technique.Findings: There was a significant association between the frequency of this gene polymorphism and atopy. In fact، individuals with TT genotype were approximately 4 times more at a risk of this disease.Conclusion: The frequency of this polymorphism with same results was also studied in both Germany and India، which shows that this polymorphism is significantly associated with allergy. Studies have also shown that IL-7 plays a pivotal role not only in eosinophil survival and operation but also in immune system homeostasis.

Background and purpose: Although many recent studies have analyzed the validation of integrin subunit alpha 4 (ITGA4) biomarker for cancer detection in patients with various malignancies, the diagnostic value of ITGA4 methylation for malignant tumors remains uncertain. We performed a systematic review and metaanalysis to unravel the relationship between ITGA4 promoter methylation status and malignant tumors. Experimental approach: A meta-analysis was performed using the metaphor package in R 3. 5 and Meta-Disc 1. 4 software. Data were derived from a search of main electronic databases up to January 2022. SROC analysis was used to evaluate the status of ITGA4 promoter methylation in colorectal cancer (CRC) and other cancers. A total of 1232 tumor samples and 649 non-tumor samples from 13 studies were analyzed. Findings/Results: The pooled results including all types of cancer provided evidence that ITGA4 hypermethylation was more frequent in tumor samples than non-tumor samples (OR 13. 32, 95% CI 7. 96-22. 29). Methylation of ITGA4 has a pooled sensitivity of 0. 95 (95% CI: 0. 94-0. 97), a pooled specificity of 0. 57 (95% CI: 0. 54-0. 60), and an area under the curve (AUC) of 0. 94. When the analysis was performed independently for CRC, it revealed a higher association (OR = 20. 77, 95% CI: 9. 15-47. 15). The assessment of ITGA4 methylation of tissue samples resulted in a pooled sensitivity of 0. 99 (95% CI: 0. 97-1. 00) and a pooled specificity of 0. 90 (95% CI: 0. 86-0. 93), and AUC of 0. 94 for the diagnosis of CRC. Conclusion and implications: ITGA4 methylation analysis is a reliable method for CRC screening in tissue samples.