فهرست مطالب reza khodarahmi

Caspase-9 is a key enzyme in the intrinsic pathway of apoptosis that its activity is regulated by various mechanisms such as phosphorylation. It has been reported that phosphorylation of serine 310 in murine caspase-9 prevents enzyme processing. The role of this residue in human caspase-9 activity in not clear. In this study we investigated the effect of negative charge on serine 310 in caspase-9 activity.

Considering that phosphorylation leads to negative charge on caspase-9, the codon of serine 310 in human capsase-9 was mutated to aspartate via quick change site-directed mutagenesis. Recombinant wild type and mutant caspase-9 were expressed in BL21(DE3) and purified by affinity chromatography. The temperature profile and activity of the mutant caspase-9 were assessed by chromogenic substrate of Ac-LEHD-pNA in vitro, and compared to those of wild enzyme. Student’s t test was used for data analysis.

The results showed that kinetics parameters of S310D mutant and wild type caspase-9 were similar, but their temperature profiles were different. Comparison of S310D mutant enzyme and wild type caspase-9 showed that S310D mutant enzyme had higher activity at 37 oC and lower activity at 4, 15, 45 and 60 oC.

In our study, the negative charge on serine 310 in caspase-9 led to change in the profile temperature of the enzyme with no effect on kinetic parameters.

COVID-19 is a worldwide health problem. Although the most infected patients experience a mild-to-moderate disease, some patients (especially older people) develop pulmonary distress with fatal lung failure and multi-organ damage. There is currently no known effective treatment for this disease. Sofosbuvir, an FDA-approved drug for the treatment of hepatitis C virus, is also able to inhibit other members of positive strand RNA viruses with conserved polymerase and may be helpful for the treatment of SARS-CoV-2. The goal of the current trial is to determine the usefulness of “standard of care (SOC) plus hydroxychloroquine and lopinavir/ritonavir” vs. “SOC plus a combination of lopinavir/ritonavir hydroxychloroquine and sofosbuvir/velpatasvir” in patients hospitalized with COVID-19.

 In this randomized controlled trial, patients over 18 years who have been diagnosed with COVID-19 by the positive SARS-CoV-2 reverse transcriptase–polymerase chain reaction (RT–PCR) test or compatible chest computed tomography (CT) scan were candidates for the study. Eighty patients from Kermanshah province, West of Iran were allocated to treatment with SOC plus hydroxychloroquine and lopinavir/ritonavir (dual therapy) or SOC plus a combination of hydroxychloroquine and lopinavir/ ritonavir and sofosbuvir/velpatasvir (triple therapy) for 10 days. Allocation was conducted using simple randomization. The primary outcomes were reducing mortality up to 28 days after hospitalization. Adverse events were handled and reported in accordance with the Good Clinical Practice guidelines. Participants: Patients who were hospitalized with COVID-19 (with positive SARS-CoV-2 RT–PCR test and/or compatible chest CT scan) were screened for eligibility at Farabi Hospital, Kermanshah University of Medical Sciences (KUMS), Kermanshah, Iran.

Both arms received active treatment and none was given placebo. The intervention arm received hydroxychloroquine 400 mg single dose and lopinavir–ritonavir (400 and 100 mg) twice daily plus sofosbuvir–velpatasvir (400 and 100mg) once daily orally, plus SOC for 10 days. The comparator arm received hydroxychloroquine 400 mg single dose and lopinavir–ritonavir (400 and 100 mg) twice daily orally, plus SOC for 10 days. SOC includes oxygen therapy, non-invasive and invasive ventilation, antibiotic agents, vasopressor support, renal-replacement therapy, and corticosteroids.

The main outcomes are reducing mortality until 28 days after hospitalization. Other outcomes can be found in full protocol file. Randomization: For the purpose of allocation sequence generation, using an Excel file (randomnumbers table) and simple random allocation, 80 included patients entered to the study, 40 patients in each group (1:1 ratio). In order to maintain the allocation sequence concealment, the details of treatment for each patient were contained in a sealed envelope, labeled by the numbers from 1 to 80. In fact, our study was a randomized open label clinical trial in which all the physicians and nurses plus all patients were aware of the type of treatment. Blinding: Our study was a randomized open label clinical trial in which all the physicians and nurses plus all patients were aware of the type of treatment. Numbers to be Randomized (Sample Size): Eighty included patients entered to the study, 40 patients in each group using simple random allocation. Trial Status: The finalized protocol version 1.5 was used in the trial study and the recruitment/intervention process started on April 11, 2020, finished on May 11, and the related follow-up finished on June 8, 2020.

This clinical trial has been registered on March 30, 2020 under IRCT number 46790, in the Iranian Registry of Clinical Trials (https://www.irct.ir/ trial/46790) and by KUMS under Grant No. 990097. Full Protocol: The full protocol and other details are attached as a Supplementary File (full protocol), accessible from the journal website. Preliminary Data: The sofosbuvir/velpatasvir regimen does not improve survival, clinical improvement, and duration of hospitalization in hospitalized COVID-19 patients.

ancreatic β-cells recognize blood glucose changes and release insulin that is a peptide hormone responsible for stable glycemia. Diabetes, a chronic disorder of insulin insufficiency, leads to disturbed glucose homeostasis and multi-organ problems. Glucose and insulin are key markers in the follow-up and control of this disease. Mitochondrial metabolism of pancreatic beta cells is a crucial part of glucose-stimulated cascade of insulin secretion. Effective factors on β-cells mitochondrial function in production of compounds such as tricarboxylic acid intermediates, glutamate, nicotinamide adenine dinucleotide phosphate, and reactive oxygen species can have great effects on the secretion of insulin under diabetes. This review enhances our knowledge of factors influencing mitochondrial function as a key mediator of glucose-induced insulin release that accordingly will be helpful to further our understanding of the mechanisms implicated in the progressive beta cellfailure that results in diabetes.

Discovering the association between genetic variations of metabolizing enzymes with idiopathic diseases such as ulcerative colitis (UC) may not only be an auxiliary agent in diagnosis but also could be an effective pharmacotherapy for inflammatory bowel disease (IBD). The aim of the present case-control study was to determine the association of cytochrome P450 2D6 (CYP2D6 *4), N-acteyltransferase-2 (NAT2*7) and multidrug resistance 1 (MDR1) 3435 C/T genotypes with UC susceptibility and thiopurine methyltransferase (TPMT) enzyme activity. TPMT activity was measured by high performance liquid chromatography (HPLC) and genotypes for the 3 mentioned polymorphisms were determined in 215 unrelated UC patients and 212 unrelated healthy controls by polymerase chain reactionrestriction fragment length polymorphism (PCR-RFLP) in a Kurdish population from Iran. CYP2D6*4 A allele, NAT2*7 A and MDR1 3435 C/T alleles act synergistically to increase the risk of UC by 3.49 times. The frequency of the A allele of CYP2D6*4 was significantly higher in UC patients (12.6%) compared to control subjects (8.5%, P = 0.046) that significantly increased the risk of UC by 1.56-fold (P = 0.047). The frequencies of NAT2*7 genotypes and alleles were similar in both studied groups. The most important outcome of this study is that for the first time we demonstrated the simultaneous presence of TMDR1, A CYP2D6*4 and A NAT2*7 alleles robustly increased the risk of developing UC by 3.49-fold. The current study suggests that CYP2D6*4 and MDR1 3435 C/T gene polymorphisms may be risk factors for UC susceptibility.

Polymorphism in the genes encoding CYP2C9 enzyme and VKORC1 reductase significantly influence warfarin dose requirement since patients with CYP2C9*2, CYP2C9*3 and VKORC1 mutant alleles require lower warfarin maintenance doses. Studies have reported the ethnic variations in the frequency of these genes within the various populations in Iran and other parts of the world. However, no such study has been done yet on Kurdish population in Kermanshah. From Kurdish population of Kermanshah province in Iran, a total of 110 patients who had heart surgery and taking warfarin, were genotyped for polymorphisms of VKORC1-1639 G>A, CYP2C9*2, and CYP2C9*3. Polymorphism genotyping was performed by sequencing as well as polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using restriction enzymes of MspI, AVAII and KpnI, respectively. The frequencies of VKORC1-1639 GG, GA, and AA genotypes were 42%, 36%, and 22%, respectively and for CYP2C9 1*/1*, 1*/2*, 2*/2*, 1*/3*, 3*/3*, 2*/3* were 71%, 17%, 5.4%, 1.8%, 4.5%, and 0%, respectively. The frequency of VKORC1-1639A allele was 42.3% and the frequencies of CYP2C9*2 and *3 alleles were 14% and 5.4%, respectively. It was indicated that low warfarin dose requirements are strongly associated with the presence of CYP2C9 and VKORC1-1639 variant alleles. Our results confirmed the supply to understand the distribution of genomic biomarkers related to the drugs metabolism for future planning health programs.

CYP2C9 and VKORC1-1639 G>A genes as the genetic factors significantly influence the warfarin dose requirement in individuals. The patients with genetic variations in CYP2C9 and VKORC1 are at increased risk of adverse warfarin-related events. A young patient with atrial septal defect being highly sensitive to normal daily dose of warfarin was subjected to the study. The patient consented to genetic testing. Furthermore, DNA was isolated and PCR-RFLP performed. The patient required low warfarin dose of 11 mg/week to achieve the target international normalized ratio (INR). Genetic testing revealed that the patient carried VKORC1-1639 AA and CYP2C9*1*2 genotypes. Our findings reaffirm the significance of pharmacogenetic analysis prior to the warfarin therapy to achieve an efficient treatment and the least side/adverse drug effects.

Aldehyde oxidase, a molybdenum cofactor-containing cytosolic enzyme, is extensively distributed throughout the animal kingdom. The enzyme is mainly active in liver and other tissues of mammalian species and involved in the metabolism of wide range of aldehydes and nitrogen-containing molecules. A continuous spectrophotometric method for the quantitative determination of aldehyde oxidase, AO, enzyme activity is described in this article. This method is based on the coupling reaction between 3-methyl-2-benzothiazolinone hydrazone (MBTH) and the o-quinone. Dopamine as substrate for AO, is converted/oxidized to o-quinone. The latter react with MBTH to produce intensely colored products that absorb light maximally in the visible region. Then, AO activity has been kinetically characterized; the Km (Michaelis–Menten constant) and Vmax (maximum initial velocity) values for the oxidation of dopamine by AO were evaluated. The existence of MBTH in the reaction medium and production of stable color as well as the high ϵ values at 510 nm of MBTH-Q adduct make this direct technique more sensitive than other continuous methods for AO assay. The optimized MBTH reaction may be useful for biological staining of AO activity isolated from various biological sources in electrophoresis gels.

Omics, referred to a part of biological science that evaluates information, systematically and broadly. Although initially genomics and proteomics have been focused, but along on advances in analytical instruments, potential capabilities of subfields such as Lipidomics recognized, increasingly. Lipidomics studies have been largely able to change the past limited viewpoint to lipids as basic components of cell structure and sources of energy. For instance, to date results of several Lipidomics studies have confirmed the role of lipids as cell messengers, antigenic agents, cell integrator. New insights on function of lipids and potential features of Lipidomics, was followed by using of these studies to information synergy with genomics and proteomics, as well as developments in pharmaceutical-medicine fields. By consider of limited development of this scientific field in Iran, in this article we try to introduce the needed tools and applications of lipidomics.

Allura red (AR) is a widely used colorant in food industry, but there is debate on its potential security risk. In this study, in vitro inhibitory properties of the dye against carbonic anhydrase (CA) were evaluated. The esterase activity of purified CA decreased in the presence of AR, in a dose-dependent manner. Regarding literature review and observed results, this preliminary study may provide new horizons in safety of AR and the other dye additives.

Acute Myelogenous Leukemia (AML) is an aggressive hematologic malignancy that cause by abnormal proliferation and accumulation of hematopoietic progenitor cells. A 37-year-old woman referred to oncologic clinic with a self-detected mass and pain in her left breast. The stage of tumor was ΙΙΙA. She was treated with the combination of anthracycline and cyclophosphamide for four courses, followed by four courses of paclitaxel with trastuzumab for one year. After 18 months of the first treatment for breast cancer, her bone marrow biopsy was compatible with AML-M2.Here, we are reporting a young woman case with breast cancer that developed AML malignancy during short interval of therapy.

The purification of biomolecules is a necessary step in many biochemical researches. In this regard, developments of convenient, specific and low cost methods of purification are of particular interest. Given the human hemoglobin (Hb) affinity toward some charged carbohydrates, interaction of this molecule with human chorionic gonadotropin (hCG) which is a glycoprotein hormone containing sialic acid, was examined. In the current study, we gathered evidence of free hCG and free Hb interaction using spectroscopic and radiometric techniques. Then, based on the affinity of hemoglobin (Hb) toward charged carbohydrates on human chorionic gonadotropin (hCG), a known sialic acid containing glycoprotein hormone, Hb-sepharose as well as native and denatured globin columns for isolation of the hormone were prepared. Sepharose-6B was activated by cyanogens bromide. Native Hb, normal globin and denatured globin were bound to cyanogen bromide–activated sepharose. Then, uptake of hCG by these gels were compared. Among the columns only native hemoglobin-sepharose column was able to catch a limited number of serum proteins such as hCG. Using the above column hCG hormone was purified with fold purification of 34 and efficiency of 80%. The chromatographic behavior of growth hormone (GH) and hCG in binding to the DEAE-Cellulose column were identical but GH showed no binding to Hb-sepharose column, indicating that the retention mechanism of hCG to Hb-sepharose column is not a simple ion exchange mode. Since globin had no property to attach to hCG but native Hb-sepharose was able to catch hCG, the BPG cavity of Hb is suggested as the possible binding site for hCG to Hb.

Chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML) are the most common leukemias of the elderly (>43 year). However, the sequential occurrence of CML followed by CLL in the same patient is extremely rare. In our report, a 52-year-old female was diagnosed with CLL (type of bone marrow (BM) infiltration was nodular and interstitial) and was treated with chlorambucil. 64 months after the diagnosis of CLL, she developed CML. She was treated with imatinib (400mg/day). After a few months, signs of CML were disappeared and CLL became dominant. This is first reported case.

Timolol is a non-selective beta-adrenergic receptor antagonist administered for treating glaucoma, heart attacks and hypertension. In the present study, we set out to determine whether or not timolol can provoke cataract formation, thus the influence of timolol on the amyloid-type aggregation of crystallin was investigated. We then provided experimental evidence of crystallin aggregation and its induction by timolol using different spectroscopic measurements. Turbidimetric measurements as well as ThT fluorescence data indicated that timolol induce extent of crystallin amyloid formation. The kinetic of protein aggregation was also changed in presence of increasing concentrations of the drug suggesting that long-term drug administration may contribute to the development of cataract. Since the consequence of timolol-crystallin interaction has yet to be identified, additional data on it may help us to postpone amyloid cataract formation.

Development of phytotherapies aimed at angiogenesis inhibition, in combination with classical anti-cancer therapies, is among the most intensively studied approaches for treatment of cancer. Epidemiological and animal studies have indicated that consumption of Allium species like shallot is associated with a reduced risk of cancer development. As a continuation of our efforts to study and characterize the effective anti-angiogenic agents from Allium species, here, we investigated the effects of aqueous extract of shallot on critical steps and mediators of in vitro angiogenesis. The anti-proliferation, -migration, and -tubulogenesis properties of the aqueous extract of shallot (at 100 - 1500 µg/ml) were evaluated using three-dimensional capillary tube formation as well as a wound-healing assay in endothelial cell-based experimental systems. In addition, the effect of the extract on vascular endothelial growth factor (VEGF) secretion and matrix metalloproteinase (MMP-2 and -9) expression was assayed using ELISA, gelatin zymography, and RT-PCR techniques. Treatment with the aqueous extract of shallot at ≥ 500 µg/ml concentrations resulted in significant decreases in endothelial cell proliferation, migration, and tubulogenesis. Moreover, the extract caused a dose-related inhibition of VEGF secretion and MMP-2/-9 expression. Taking all the data into account, the current study indicated that shallot – containing potent anti-angiogenic properties - exerts its inhibitory effect mainly through down-regulation of VEGF and MMP-2/-9; essential angiogenic mediators in many malignant and chronic inflammatory diseases.

The DNA molecule has been known to be the cellular target for many cytotoxic anticancer agents for several decades. Understanding how drug molecules interact with DNA has become an active research area in the interface between chemistry, molecular biology and medicine. DNA extraction has been suggested as a main step affecting molecular DNA technology such as PCR and PCR-based methods. Therefore, researchers have used several modified protocols for efficient DNA extraction from whole blood. In this study, we focused on a fast and reliable protocol with inexpensive and non-poisonous reagents for DNA extraction from whole blood. Current method was optimized based on a combination of conventional salting-out and boiling methods. Also the quality and quantity of the extracted DNA were surveyed by gel electrophoresis and Nanodrop spectrophotometry methods, respectively. Results showed that high quantity and quality of isolated DNA by this method is enough to do hundreds of PCR-based reactions and also to be utilized in other DNA manipulation assay such as restriction digestion, drug- DNA interaction and methylation detection survey. In conclusion, we described a fast, low-cost, non-toxic and enzyme free protocol for high yield genomic DNA extraction from whole blood.

Curcumin is a natural polyphenolic compound with anti-cancer, anti-inflammatory, and anti-oxidation properties. Low water solubility and rapid hydrolytic degradation are two challenges limiting use of curcumin as therapeutic agent. In the current study, the role of the Bovine Serum Albumin (BSA), β-lactoglobulin and casein, as food-grade biopolymers and safe drug delivery systems, on the physical activity of curcumin were surveyed. It appears that BSA and casein as protein vehicles are useful tools to increase stability of curcumin, as a health promoting agent.

Polyphenol oxidase (PPO), also known as tyrosinase (EC 1.14.18.1), is a copper-containing enzyme widely distributed in microorganisms, animals, and plants. This enzyme is responsible not only for browning in plants but also for melanization in animals. Thus, tyrosinase inhibitors have a huge impact on industry and the economy. In the current study, we first purified the enzyme, then evaluated inhibitory potency of three benzaldehyde derivatives: 2,4-dihydroxybenzaldehyde, 3,4-dihydroxybenzaldehyde and 4-dimethylaminobenzaldehyde on diphenolase activity of the purified mushroom tyrosinase, compared to kojic acid. Despite their close structural similarity, 2,4-dihydroxybenzaldehyde was found as potent competitive inhibitor while uncompetitive inhibition was observed for 4-dimethylaminobenzaldehyde. Further complementary studies on these types of inhibitors, as potential drugs for treating abnormal melanin pigmentation, are needed.

Molecular Chaperones or Amyloid-binding compounds? Perspective on Their Application as Possible Therapeutic Agents in Reduction of Cytotoxicity of Amyloid Oligomers

The prevalence of obesity as one of the most health concerns has increased globally. This kind of disease has been accounted for several diseases such as type 2 diabetes, different types of cancer, heart disease, and Alzheimer. Obesity is a multifactorial disease that both environmental factors and genetics play important role in its susceptibly. In molecular biology, characterization of the adipocyte secretome is important in signaling to other organs and in regulating energy balance for evaluating underlines mechanism. Since better understanding of this disease lead to both preventive and post treatment of obesity which is achieved by molecular evaluations, this review underlies the importance of some molecular approaches in the field of obesity.

Because of the huge amounts of proteomic data and demand for new methods of laboratory analysis results, proteins collective analysis, in addition to taking less time, biostatistician assist at identification of new patterns in the data set. In this study, rat hippocampus proteome in normal and Alzheimer''s disease (AD) were analyzed by using proteomic techniques and bioinformatics’ analysis. Protein extracts from normal and Alzheimer''s rats were separated by using two-dimensional electrophoresis (2DE). The silver staining method was used for detecting spots. Bioinformatics analysis of proteome were performed by progensis same spots software. Bioinformatics and statistical analysis of 2DE gel techniques obtained 760 protein spots were detected in both normal and AD rats. Comparisons between controls and Alzheimer gel containing 20 common proteins were expressed significantly differences. 16 new proteins were expressed in AD, while 36 proteins were suppressed. Proteins clustering by using correlation analysis evaluated 3 clusters in the proteome; Principal component analysis also confirmed the results of clustering. Finally, we can conclude that a significant expression of Alzheimer changes in the hippocampus proteome which are associated with specific biological processes summarized in 3 main clusters indicated 3 principal biological pathways of AD.

Curcumin is a natural polyphenolic compound with anti-cancer, anti-inflammatory, and anti-oxidation properties. Low water solubility and rapid hydrolytic degradation are two challenges limiting use of curcumin as therapeutic agent. In the current study, the role of the native/modified forms of bovine serum albumin (BSA) and casein, as food-grade biopolymers and safe drug delivery systems, on the physical and biological activity of curcumin were surveyed. Analyses of quenching of proteins fluorescence by curcumin indicated that chemical modification decreased binding affinity of curcumin toward albumin whereas it significantly increased for casein and average number of binding sites also doubled in modified casein. Measurement of cell viability using LDH assay showed that cytotoxicity of protein-bound curcumin is higher than free curcumin. Moreover, in the presence of native proteins, curcumin revealed elevated in vitro anti-cancer activity (against MCF7 and SKNMC) compared to modified forms. It appears that BSA and casein as protein vehicles are useful tools to increase both food quality and the bioavailability of curcumin as health promoting agent. However, results imply that the chemical modification of proteins cannot improve the anti-cancer activity of curcumin despite increasing of their binding affinity.

A number of 6-hydroxy-2-benzylidene-3-coumaranones were synthesized from condensation of 6-hydroxy-3-coumaranone with appropriate aldehydes and were evaluated for their antioxidant activities. The antioxidant activity was assessed using two methods, including, 1,1-biphenyl-2-picrylhydrazyl (DPPH) radical scavenging, and reducing power assays. Some of the benzylidene coumaranones showed antioxidant activity more than Trolox as reference antioxidant.

Angiogenesis is the process in which the new blood vessels are formed from pre-existing ones. The development of the vascular system is one of the earliest events in organogenesis. Since angiogenesis is a key process in the promotion of cancer and its metastasis (most common cause of cancer death in human), inhibition of angiogenesis is one of the promising approaches for treatment of tumor growth and metastasis. In this study, several derivatives of 4-aryl-4H-chromenes bearing methoxy substituent have been synthesized with boric acid as catalyst and investigated with potential anti-angiogenesis effects as well as anti-angiogenic mechanism. Anti-angiogenic and anti-proliferative effects of the prepared compounds checked on three-dimensional culture of human umbilical vein endothelial cells (HUVECs) in collagen matrix and HUVEC proliferation assay as well as matrix metalloproteinase (MMP) gelatinase assay in the endothelial cell-based experimental system. Finally, anti-angiogenic mechanism of the compounds was identified, using gel zymography method. Among the synthesized compounds 4a and 4h were as the most active in these series. The compound 4a was caused angiogenesis inhibition with complete suppression at 3 µg/ml (8.537 µM) and 4h at 1.6 µg/ml (4.196 µM). At end, chromene derivatives can serve as the lead molecules for further development of a new class of anti-angiogenesis agents

Universities as science production centers, have grate roles in the development of the societies. So, the scientific activities of their researchers should be evaluated. The current study aimed to evaluate science production in Kermanshah University of Medical Sciences (KUMS), from 1995 to 2011, based on ISI Web of Science (ISI-WoS) service. This study was performed based on the data retrieved from WoS. The obtained data was analyzed using descriptive statistical analyses. The WoS-derived results showed that KUMS researchers have published 354 scientific papers while total number of citations to these published articles was 1375. Moreover, the highest number of published papers was in 2010. KUMS has experienced triggering growth in scientific productivity (74 papers in 2010) but with slow onset (2 in 1995). Among the factors contributing to this growth are; establishment of efficient motivating policies, development of scientific relationships, accessibility of essential scientific databases and the growth of internet and its various services can be overall considered as possible causes of KUMS improved productivity.