The effect of pentoxifylline drug on bax/bcl2 gene dosage expression changes following ischemic reperfusion injury in kidney

Global cerebral ischemia (GCI) and reperfusion induced apoptosis that lead to cell injury and death. The bax and bcl-2 are pro-apoptotic and anti-apoptotic genes, respectively. These genes belong to The B-cell lymphoma-2 (bcl-2) family.In this study; we assessed the effect of pentoxifylline drug on bax/bcl2 gene dosage expression changes following   ischemic reperfusion injury in kidney. In this experimental study, 20 male wistar rats were accidently divided them on two tenth group of control and treatment groups. In the control group, celiotomy was performed by ventral midline incision. The left kidney was isolated, and then both the renal artery and vein were obstructed. After 60 minutes of warm ischemia, vessel obstruction resolved and the right kidney was removed. 72 hours after reperfusion, tissue samples were taken from left kidney for histopathology. All these steps in treatment group were exactly repeated after administration of 45 mg/kg/PO pentoxifylline (3 hours before operation) and in this group treatment was continued every 12h until 3 days. In this research quantitative real-timePCR is used for the detection expression Bcl2 and  Bax genes in ischemia group and PNT drug group and  compared to  normal sample. The results showed the gene dosage ratio of Bax/bcl2 in PNT group  decline  than to  ischemia group. Therefore, the pentoxyfylline might have a role in control of apoptosis result from Ischemia- reperfusion

The analysis of endothelial nitric oxide synthase gene polymorphism in intron 4 with hypertension disease

          Nitric oxide is an important vasorelaxant factor that it inhibits platelet adhesion and proliferation of smooth muscle cells. NO is synthesized from L-arginine by means of endothelial nitric oxide synthase (eNOS) which is an isoform of  Nitric Oxide Synthase.In the present study, we examined  possible association between the 27 base pair (bp) repeat  polymorphism in intron 4 of the  eNOS3 gene and hypertension in an Iranian  population. 40 patients with hypertension and 40 healthy controls were included in the research.Genotyping was performed by polymorphism chain reaction (PCR). The frequency of 4a4a, 4a4b , 4b4b  genotypes  were  52.6% , 26.3%,  15.7% respectively  in hypertension groups .The frequency of 4a4a, 4a4b , 4b4b  genotypes  were 73.8% , 15.7% , 10.5% respectively  in   control  groups. The hypertensive patient group showed a significant higher frequency of the 4a allele compared to the controls (p<0.01).The present study showed a singificant  association between the eNOS3 gene polymorphism and hypertension in the Iranian population

Cloning of c-Myc gene in embryonic stem cells

Embryonic stem cells (ESCs) are pluripotent, self-renewing cells. These cells can be used in applications such as cell therapy, drug discovery, disease modelling, and the study of cellular differentiation. In this experimental study embryonic stem cells cultured in the laboratory and were amplified. Total RNA was extracted from cells and converted to cDNA by reverse transcription-polymerase chain reaction (RT-PCR). Then c-Myc gene was amplified by (PCR) and inserted into the pTZ57R/T vector. Ligated product was transformed into susceptible bacteria and transformed bacteria were screened on a selective medium. Plasmids extracted from bacteria and enzyme digestion to confirm the sequencing was performed. The cloned c-Myc gene can be used to prepare a gene cassette to produce stem cells from somatic cell.

Study of pentoxifylline drug effect on Bax gene expression changes in kidney after ischemic/reperfusion in rat

Ischemia Reperfusion injury is the tissue damage caused when blood supply returns to the tissue after a period of ischemia or lack of oxygen. Ischemia Reperfusion induces cell death and endemic reaction that is one of the most important clinical problems with acute renal failure and renal transplantation. In this study, the effect of pentoxifylline on rat kidney function and cell injury following Ischemia Reperfusion were evaluated. In this experimental study, 20 male wistar rats with average weight of 250-300g were selected and then were accidently divided them on two tenth group of control and treatment groups. In the control group, celiotomy was performed by ventral midline incision. The left kidney was isolated, and then both the renal artery and vein were obstructed. After 60 minutes of warm ischemia, vessel obstruction resolved and the right kidney was removed. 72 hours after reperfusion, tissue samples were taken from left kidney for histopathology. All these steps in treatment group were exactly repeated after administration of 45 mg/kg/PO pentoxifylline (3 hours before operation) and in this group treatment was continued every 12h until 3 days. In this research quantitative real-time PCR is used for the detection expression Bax gene in ischemia group and PNT drug group and  compared to  normal sample .The results showed the gene dosage ratio of 1.24 for ischemia groups  and 0.64 for drug group. The results showed the expression Bax gene in PNT group  decline  than to  ischemia group. Therefore, quantitative real time PCR could be used as a direct method for detection of Bax gene expression in tested and normal samples.

Association of tRNAThr 15927G→A Mutation with the Incidence of Coronary Artery Disease

One of the leading causes of death in the world are cardiovascular diseases, among which coronary artery disease (CAD) is the most common one. It occurs as a result of narrowing of the arteries which supply blood to the heart due to a theroma plaque formation. This kind of heart disease can be considered as a multifactorial one as genetic and environmental factors are involved in its incidence. The already conducted studies have investigated the relationship between some of polymorphisms in different loci and CAD with the aim of on-time diagnosis of this disease, which may lead to its prevention and appropriate treatment. The aim of this study was to examine the association of tRNA Thr 15927 G → A mutation with the risk of CAD incidence to offer programs for early diagnosis and treatment of this disease in Iran. Fifty patients with CAD were included in the patient group, and fifty healthy participants were selected for the control group. 5mL of peripheral blood samples drawn from subjects in patient and control groups were collected in the tubes containing Ethylenediaminetetraacetic acid (EDTA). After DNA extraction from blood with the employment of DNA extraction kit, its quality and quantity were measured using electrophoresis and NanoDrop devices, respectively. Then, the isolated DNA with the implementation of  amplification-refractory mutation system (ARMS)  and restriction fragment length polymorphism (RFLP) techniques was examined to determine and evaluate the target polymorphism in the intended locus. Sequencing method was also used to confirm the findings. To this end, 4 samples were randomly selected and sequenced.In the control group, 6 screened patients had the mutation while the others did not. Similar result was observed in the patient group. The findings of the present study reveal that there was not any significant relationship between tRNAThr 15927G → A mutation and risk of CAD incidence

Moral challenges of research on embryos in pre-implantation genetic diagnosis and IVF

     Purposes of pre-implantation genetic diagnosis (PGD) include screening for single gene mutations for late onset disorders or susceptibility to cancer. The problematic issue is that PGD is to produce a healthy baby, causing the destruction of some embryos that have been transferred by in vitro fertilization (IVF). Some PGD embryos may be discarded because they are excess to the woman or couples’ goal for family creation, and have been diagnosed as being affected by a particular genetic condition that woman/couple wish to avoid. So, the controversial issue is the destruction of embryos as a consequence of fertility treatment that raises questions on whether the moral status of an embryo of 3 days is the same as that of a born, living adult human being

Molecular focus in p63 and correlated human diseases

The p63 gene, a member of the p53 gene family, is expressed into at least six protein isoforms.The transcription factor p63 is a homologue of the tumor suppressor p53. Unlike p53, which is dispensable for normal development, p63 is critical for the development of stratified epithelial tissues such as epidermis, breast, and prostate. p63, , is transcribed from two different promoters giving rise to two different proteins: p63, a member of the p53 family, is transcribed from two different promoters giving rise to two different proteins: TAp63 that contains the N-terminal transactivation domain and ΔN that lacks this domain. p63 encodes multiple protein isoforms with both transactivating and transcriptional repressor activities that can regulate a wide spectrum of target genes. p63 is also implicated in tumor formation and progression in stratified epithelia, with evidence for both tumor suppressive and oncogenic properties.

Simulating fluid-solid interaction problems using an immersed boundary-SPH method

In this work, the Immersed Boundary Method (IBM) is adapted and implemented in the context of Smoothed Particle Hydrodynamics (SPH) method to study moving solid bodies in an incompressible fluid flow. The proposed computational algorithm is verified by solving a number of benchmark particulate flow problems. The results are also compared with those obtained using the same SPH scheme along with a direct solid boundary imposition technique

Therapeutic Value of CD73 as a Biomarker in Human Cancer

Context: Over the past several years, biomarkers have emerged as a diagnostic and therapeutic tool for cancer therapy. CD73 (ecto-5'-nucleotidase) which is a cell surface ectonucleotidase, mediates the conversion of extracellular Adenosine monophosphate (AMP) into adenosine through the purinergic signaling pathway. In this study the role of CD73 in different human cancers was investigated. Evidence Acquisition: The present study reviewed the articles related to the importance of CD73 as a therapeutic tool in human cancers which were published from 1990 to 2019. The publications were found by searching the valid databases for instance PubMed, Google Scholar, ProQuest, Scopus and Science Direct using keywords CD73, ectonucleotidase, therapy, cancer, etc. During the review process 90 articles were selected. Results: Different studies about the effect of CD73 on human malignancies show that CD73 is overexpressed in various types of cancer. Recent findings demonstrate that the extracellular adenosine can promote tumor growth and invasion. The significant regulatory role of CD73 has made it a suitable biomarker for cancer treatment. Conclusion: This study outlines the impact of CD73 on tumor growth, metastasis and angiogenesis. The findings are promising and can highlight the efficacy of this protein as a therapeutic tool in the context of anti-CD73 cancer therapy

Amidolytic Activity of Factor VII Expressed in Iranian Lizard Leishmania

Over many years, variable gene expression systems have been used obtaining of Factor VII protein, but each one has certain limitations. Therefore, the main goal of this study was to assess the biological activity of purified and recombinant factor VII expressed in Iranian lizard Leishmania. After transferring recombinant construct containing FVII gene to Leishmania, first, the expression of 55 kDa FVII protein in transfected cells was confirmed by analyzing cell lysate using SDS-PAGE and Western Blotting techniques. Then, FVII was purified by NI-NTA-His Tag- resin through an affinity chromatography. The chromogenic activity of human recombinant Factor VII (amidolytic activity) in supernatant and pellet fractions of Leishmania promastigotes was done using ELISA method. The amidolytic activity of rhFVII was about 0.0125 IU/ml in the concentrated cell sediment and 0 IU/ml in the supernatant in the first 60 minutes and after that time, none of the samples showed acceptable activity. HIGHLIGHTS• Factor VII has important role in the treatment of coagulation and hemophilic disease.• Recombinant FVII was expressed in Iranian Lizard Leishmania.• The amidolytic activity of rhFVII was about 0.0125 IU/ml in cell pellet.• No amildolytic activity was observed in the supernatant