Avicenna Journal of Medical Biotechnology
Year:2019 | Volume:11 | Issue:3|Papers Count:11

Despite the advent of several antidepressant medications, treatment of Major Depressive Disorder (MDD) is still far from optimal 1-3. A large proportion of patients with MDD do not respond to their first medication. To achieve favorable response, these patients are generally treated with either switching to another treatment or augmentation therapy. In the recent decade, several augmentative strategies for treatment of MDD have been developed. Some of these treatment modalities focus on recently developed hypotheses of pathophysiological processes in patients with MDD 1-3. These mainly include immune system dysfunction, hypothalamic-pituitary-adrenal (HPA) axis and metabolic derangements, impaired neuroprotection, or neuroinflammation 1-3. ...

Background: Marine environment is a valuable source of bioactive compounds with variable medicinal properties. Previously, it was shown that Ophiocoma erinaceus ex-tracted polysaccharide has prominent cytotoxic effect on HeLa human cervical cancer cells. In the present study, the anti-cancer properties of polysaccharide extracted from Ophiocoma scolopendrina (O. scolopendrina) were examined in comparison with paclitaxel as a conventional drug against resistant ovarian cancer; also, its related mechanism against A2780cp ovarian cancer cells was investigated. Methods: The A2780cp cancer cells and NIH3T3 normal cells were cultured and treat-ed with different concentrations of polysaccharide extracted from O. scolopendrina for 24 hr and 48 hr. Then, cell toxicity was studied by MTT assay, morphology of cells was observed under inverted microscopy and the type of induced cancer cell death was assessed by annexin V-FITC, propodium iodide and acridine orange staining. Finally, the apoptosis pathway was determined by measurement of caspase-3 and caspase-9 activity and assessment of p53 and Bcl-2. The statistical analysis was performed by SPSS software, one way ANOVA and p<0. 05 was considered significant. Results: Our observations from MTT assay and morphological assessment exhibited that O. scolopendrina isolated polysaccharide inhibited proliferation of ovarian cancer cells with IC50 of 35 μ g/ml, while paclitaxel suppressed tumor cell growth with IC50=10 μ g/ml. In contrast, MTT observations revealed low cytotoxicity of these chemothera-peutic agents against NIH3T3 normal cells. Also, the analysis correlated with induced cell death elucidated that concurrent treatment of polysaccharide plus paclitaxel had a further anti-cancer effect against A2780cp cells mainly through restoration of p53 and mitochondrial apoptosis cell death induction. Conclusion: Taken together, our research supports the finding that application of pol-ysaccharide extracted from O. scolopendrina can be considered a promising marine chemotherapeutic approach for advancing efficacy of paclitaxel in treatment of re-sistant ovarian cancer. Additional in vivo experiments are required to elucidate the role of brittle star polysaccharides in animal and clinical trials.

Background: Although Saccharomyces cerevisiae has several industrial applications, there are still fundamental problems associated with sequential use of carbon sources. As such, glucose repression effect can direct metabolism of yeast to preferably anaer-obic conditions. This leads to higher ethanol production and less efficient production of recombinant products. The general glucose repression system is constituted by MIG1, TUP1 and SSN6 factors. The role of MIG1 is known in glucose repression but the evalu-ation of effects on aerobic/anaerobic metabolism by deletion of MIG1 and constructing an optimal strain brand remains unclear and an objective to be explored. Methods: To find the impact of MIG1 in induction of glucose-repression, the Mig1 dis-ruptant strain (Δ MIG1) was produced for comparing with its congenic wild-type strain (2805). The analysis approached for changes in the rate of glucose consumption, bio-mass yield, cell protein contents, ethanol and intermediate metabolites production. The MIG1 disruptant strain exhibited 25% glucose utilization, 12% biomass growth rate and 22% protein content over the wild type. The shift to respiratory pathway has been demonstrated by 122. 86 and 40% increase of glycerol and pyruvate production, respectively as oxidative metabolites, while the reduction of fermentative metabolites such as acetate 35. 48 and ethanol 24%. Results: Results suggest that Δ MIG1 compared to the wild-type strain can significantly present less effects of glucose repression. Conclusion: The constructed strain has more efficient growth in aerobic cultivations and it can be a potential host for biotechnological recombinant yields and industrial interests.

Background: Intrafamilial spread of Hepatitis B virus (HBV) infection in Iran has only been investigated with serological testing without using molecular studies as the most informative and definitive type of analysis. Methods: In the present study, intrafamilial transmission of HBV among family mem-bers of Iranian index HBsAg carriers was investigated using phylogenetic analysis of the S region of the viral genome. Nested polymerase chain reaction was used for de-tection of HBV DNA in serum samples from 22 index and 43 contact patients with chronic HBV infection. HBV DNA was detected in 37 samples (14 indexes, 23 contacts). The S gene region of the DNA isolates was subjected to direct sequencing and phylo-genetic analysis by Bioedit, Mega and Phylip programs. Results: All isolates (from 26 patients) were clustered with genotype D, of which 24 strains were of subgenotype D1, subtype ayw2, while 2 additional strains were of sub-genotype D2, subtype ayw3. Evidence of intrafamilial transmission of the virus was found in 8 families studied phylogenetically. Overall, 60 changes were detected in the amino acid sequences of the surface antigen protein in 23 patients. Four premature stop codons occurred in 3 isolates at residues 69 and 182. Seven out of 8 families dis-played 25− 100% common amino acid substitutions among their members. Conclusion: Our data corroborated intrafamilial transmission of HBV, as evidenced by concordant HBV genotype among household members, viral sequence homology and close genetic relatedness of the strains on the phylogenetic tree, and horizontal trans-mission of S gene mutations among family members.

Background: Some products of bacteria are reported as an immunomodulator. The Helicobacter pylori (H. pylori) outer membrane proteins play an important role in stimulation of immune system. The present study was performed to determine the in vitro effect of recombinant HopH of H. pylori on Nitric Oxide (NO) production and viability of mouse peritoneal macrophages. Methods: H. pylori recombinant HopH was produced in this study. Mice peritoneal macrophages were purified and cultured. Different concentrations of recombinant HopH were used for stimulation of macrophages in order to evaluate NO production. The cell viability was detected by MTT assay. NO amounts released in to the superna-tants of cultured macrophages and LPS-stimulated macrophages (10 μ g/ml) were de-tected by Griess reagent. Results: Results demonstrated that the suppressive effect of high concentrations of re-combinant HopH on NO release and the stimulation effect of protein was shown in 15 μ g/ml, compared to the control group. NO stimulation was significant in all the con-centrations of LPS stimulated with HopH groups. Conclusion: According to our findings, recombinant HopH has a toxic effect in high concentration on cell. So it can be an anticancer candidate.

Background: Patients with diabetes present with lipid disorders, including hypercho-lesterolemia, which can be a high-risk factor for atherosclerosis. Recently, increasing interest has been focused on anti-lipidemic function of herbal medicines, especially Zingiber officinale (known as ginger), in diabetes. However, the mechanism underly-ing the effect of ginger on some players involved in cholesterol homeostasis of Central Nervous System (CNS) among diabetic patients remains unclear. To our knowledge, this is the first study to investigate the effect of ginger on brain regulation of Hydro-xymethylglutaryl-CoA Reductase (HMG-CoA reductase) and Cholesterol 24-hydroxy-lase (CYP46A1), which provides a rational model for understanding brain dyslipidem-ia mechanisms associated with diabetes. Methods: Brains of rats were isolated from four groups: control, non-treated diabetic, and treated diabetic groups receiving 200 or 400 mg/kg of hydroalcoholic extracts of ginger for eight weeks. HMG-CoA reductase and CYP46A1 levels in brain homoge-nates were determined by western-blot technique. Results: Ginger root extract caused a significant decrease in HMG-CoA reductase and an increase in CYP46A1 levels in treated diabetic groups compared to diabetic con-trol. In comparison to diabetic group, these effects were more remarkable with 400 mg/kg concentration of ginger extract. Conclusion: The findings showed that ginger extract has a regulatory effect on pro-teins involved in cholesterol homeostasis in CNS by a significant down-and up-re-gulation of HMG-CoA reductase and CYP46A1 levels, respectively. It can be suggested that adding ginger to daily diet of diabetic patients has useful effects and may ame-liorate diabetes complications.

Background: Type 2 Diabetes Mellitus (T2DM) is a serious problem in the world. 5-Hydroxytryptamine (5-HT, serotonin) plays an important role in obesity, glucose con-trol and insulin resistance. The polymorphism of the serotonin transporter gene linked promoter region (5-HTTLPR) might influence 5-HTT expression and serotonin uptake. The polymorphism results in two alleles of L (Long) and S (Short). The aim of the pre-sent study was to evaluate the association between 5-HTTLPR genotypes in type 2 diabetes mellitus (T2DM), obesity as well as serum biochemical profiles in Iranian population from 2012 until 2015. Methods: 180 patients with T2DM and 180 controls were selected and the frequency of S and L alleles was determined by PCR. Then, the relationship between genotypes, body mass index (BMI) and serum biochemical variables was investigated. Results: The frequency of S and L alleles in experimental and control groups was the same [for the L allele p=0. 754, OR (95%CI)=1. 103 (0. 597 to 2. 041) and for the S allele p=0. 906, OR (95%CI)=(0. 490 to 1. 676)]. However, the mean triglyceride, cholesterol, LDL-C, systolic and diastolic blood pressure levels in the diabetic subjects with LL gen-otype were significantly higher than LS and SS genotypes (p<0. 001) in this population. Conclusion: The L allele of 5-HTTLPR was related to the increased serum lipids and blood pressure in the diabetic patients. However, there was no relationship between the polymorphism of 5-HTTLPR L/S and T2DM in Iranian population.

Background: In the process of sperm cryopreservation, apart from cryoinjury, the production of Reactive Oxygen Species (ROS) can adversely affect the integrity of chromatin and cellular membranes. Addition of natural antioxidants to freezing me-dium is an approach to reduce the destructive effects of ROS on sperm. Methods: In this study, during 60 min of cooling process, the ejaculates of five stallions were diluted in the following media: INRA 82 medium as Control (C), INRA 82 medi-um supplemented with 0. 25% Sericin (S), INRA 82 medium supplemented with 1. 5 mM Glutathione (G), and INRA 82 medium supplemented with 0. 25% Sericin+1. 5 mM Glutathione (S+G). Results: In the frozen/thawed sericin supplemented group, while the integrity of DNA and the activity of catalase and Glutathione Peroxidase (GPx) were increased, the li-pid peroxidation and midpieceab normality decreased, compared with other groups (p<0. 05). The proportions of sperms with abnormal head in group S and the sperm with distal droplet in G and S+G groups decreased, compared with group C (p<0. 05). In CTC assay, the percentage of capacitated spermatozoa in treatment groups was lower than control (p<0. 01). Conclusion: In conclusion, the presence of sericin in freezing medium of stallion semen could improve sperm DNA integrity and its resistance to ROS and lipid peroxidation.

Wiskott-Aldrich Syndrome (WAS) is a rare X-linked recessive Primary Immunodefi-ciency (PID) caused by mutations in WAS gene which encodes a protein known as WASp. WASp plays important roles in cytoskeletal functions that compromise multi-ple aspects of normal cellular activity including proliferation, phagocytosis, immune synapse formation, adhesion and directed migration. WASp defect particularly causes platelets abnormality which is presented in forms of decrease of Mean Platelet Vol-ume (MPV) and thrombocytopenia in most WAS conditions; nevertheless, some stud-ies reported WAS patients with a normal or large size of platelets in recent years. This phenomenon is unique and the exact mechanism of thrombocytopenia with a normal or large size of platelets is still unknown. In this study, Next Generation Sequencing (NGS) was utilized to discover the causing mutation in WAS gene; furthermore, an at-tempt was made to evaluate the possibility of other mutations or genes especially WASp interacting proteins and inherited platelet disorder genes in patient clinical symptoms for the purpose of understanding the origin of such unique symptom and to perform further analysis if it is required. Therefore, clinical manifestations and im-munologic functions of the patient were checked and Whole Exome Sequencing (WES) was performed to analyze all exonic variations which can be associated with patient phenotypes. Finally, a novel de novo mutation in WAS gene which truncates WASp to half of its normal size was determined as the only cause of clinical manifes-tation.

Background: The CRISPR/Cas9 genome editing system is a powerful and simple gene editing method. The format of the CRISPR components is one of the important factors in targeting efficiency. Compared to plasmid or mRNA (IVTs) format, using the CRISPR/Cas9 system as Cas9– crRNA– tracrRNA RNP format is more efficient and rap-id, especially in minimizing some of the pitfalls of CRISPR-mediated gene editing. In addition to efficient in vivo applications of the CRISPR RNP format in a variety of cell types and organisms, another advantage of this approach is usability for in vitro ap-plications in which the crRNAs in the tracrRNA– crRNA structure guides the Mg2+-dependent RNAdirected DNA endonuclease to introduce double-strand breaks at specific sites in DNA. Methods: Here, Cas9– crRNA– tracrRNA RNP system was used to test the designed crRNAs for in vitro DNA cleavage by Cas9 protein in RAG1, RAG2 and IL2RG genes. Results: The results of cleavage reveal theCas9– crRNA– tracrRNA RNP system is a rapid and efficient way to pre-validate the efficiency of CRISPR cleavage with crR-NAs designed for RAG1, RAG2 and IL2RG genes. Conclusion: one step in vitro cleavage of DNA by CRISPR/Cas9 ribonucleoprotein complex can be used to pre-validate the functionality and relative efficiency of CRISPR system for targeting genes.

Background: Brucellosis is still an important health problem in under developing countries and researches for finding efficient vaccine are going on. Brucella melitensis (B. mellitensis) bp26 gene is a good candidate for brucellosis vaccine and investiga-tions showed that Lactococcus lactis (L. lactis) with several positive characteristic are attractive for protein expression as a live delivery vectors. These fast growing bacteria need no aeration, are easy to handle, have no exotoxin, endotoxin and protease, so the cost of culturing is inexpensive. Methods: B. mellitensis bp26 gene was cloned in food grade pNZ 8149 vector and ex-pressed in L. lactis NZ 3900. Results: Results showed that we can produce a food-grade recombinant L. lactis pro-ducing the B. melitensis BP26 protein. Conclusion: In this study, for Future evaluation about ability of L. lactis as a live deliv-ery vector, a food-grade recombinant L. lactis producing the B. melitensis BP26 pro-tein was produced.