{"title":"Investigation of the association of thee chemokine CCL22 gene polymorphism rs4359426 with multiple sclerosis","authors":"A. Jafarzadeh","doi":"10.22037/amls.v1i2.9179","DOIUrl":"https://doi.org/10.22037/amls.v1i2.9179","url":null,"abstract":"Objective: CCL22 is a chemokine that induces the migration of Th2- and regulatory T cells to the inflammatory sites. The aim of this study was to investigate the association of a single nucleotide polymorphism (SNP), rs4359426, in CCL22 gene, with multiple sclerosis (MS) in patients from southeast of Iran. Methods: The blood samples collected from 150 patients with MS and 150 healthy subjects as a control group. The serum levels of CCL22 measured by ELISA and the DNA analyzed for CCL22 polymorphism using PCR-RFLP method. Results: There were no significant differences in the frequencies of genotypes and alleles at SNP rs4359426 in CCL22 gene between MS patients and controls. No significant differences also observed between controls and patients with RRMS, SPMS, PPMS and PRMS patterns regarding the genetic variation at rs4359426. In both MS and control groups, No significant differences were observed between subjects with CC, CA and AA genotypes or between subjects with C and A alleles at rs4359426 with respect to the serum levels of CCL22. Conclusion: These results do not show any association between the investigated genotypes and alleles at at rs4359426 in CCL22 gene with MS or its patterns in MS patients. The serum levels of chemokine did not also influence by genetic variation at SNP rs4359426.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"100 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78919106","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Cloning and expression of HIV 1 Tat Protein and injection of Tat protein in camel to isolation of nanobody","authors":"A. Abdoli","doi":"10.22037/amls.v2i1.13302","DOIUrl":"https://doi.org/10.22037/amls.v2i1.13302","url":null,"abstract":"Designing a novel therapeutic agent has been a critical a challenge for HIV virus treatment. The aim of the present study was the isolation of VHH (Nanobody) gene from camel antibody library in order to present a therapeutic nanobody against Tat protein of HIV. Human immunodeficiency virus (HIV-1) needs to Tat protein for the replication. The results of recent studies have showed that neutralizing antibodies against Tat protein can inhibit HIV virus replication. At first, a DNA fragment of encoding Tat protein of HIV-1 was synthesized and expressed in E.coli. Next, Tat protein was purified by NTA affinity chromatography. The purified recombinant protein was formulated with Freund's adjuvant and injected to one camel for five times. Then, total RNA was extracted from camel lymphocytes and VHH fragments synthesized and amplified using RT-PCR and Nested- PCR methods.The 17KD Tat protein purification efficiency and its conformation were confirmed by SDS-PAGE and Western blot, respectively. The 600 and 400bp of VHH genes were produced by RT-PCR and Nested- PCR shown by gel electrophoresis, respectively. The nanobody may be a useful a promising drug for treatment of HIV. The small sizes of nanobodies give them the potency of the recognizing the cryptic epitopes of tat and neutralizing the virus.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"43 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79065345","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Evaluation of miR-122 level in chronic HBV infection and liver cirrhosis patients","authors":"A. Moradi, M. Ebrahimifard, M. Zandi","doi":"10.22037/AMLS.V2I3.13675","DOIUrl":"https://doi.org/10.22037/AMLS.V2I3.13675","url":null,"abstract":"MicroRNA is a kind of small RNAs with about 22 nucleotide length; it is expressed in most eukaryotes and acts as a regulating key of gene expression through attaching to the end of 3’mRNA target. Most popular miRNAs are found in liver that are extensively important in determining the biological and clinical functions. MicroRNA, in host cells, may affect on replication of viruses positively or negatively. MicroRNA processing mechanism and inactivating RISC complex could affect on virus replication. In this study, miR-122 expression was examined and compared in three patient groups with chronic hepatitis, hepatitis B virus-associated cirrhosis and healthy group. In this study, 108 samples were taken in 5 ml complete blood with EDTA from each participant. Extracting RNA by RNX-plus was done based on manufacturer protocol; and evaluating miRNA expression was conducted through Relative real time PCR. The results indicated that miR-122 expression increased in patients with chronic hepatitis B and hepatitis B virus-associated cirrhosis about 1.8 times more than control group which is significant statistically (P < 0.05). According to the results, measuring miR-122 expression may estimate the disease progress from chronic hepatitis B to hepatitis B virus-associated cirrhosis ; and it can be used as a biomarker that needs more investigation with more samples.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"101 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73623326","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Samira Rashidian, M. Derakhshan, E. Aryan, Roghayeh Teimourpour, Aida Gholoobi, Z. Meshkat
{"title":"Designing and construction of a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis","authors":"Samira Rashidian, M. Derakhshan, E. Aryan, Roghayeh Teimourpour, Aida Gholoobi, Z. Meshkat","doi":"10.22037/AMLS.V3I1.16982","DOIUrl":"https://doi.org/10.22037/AMLS.V3I1.16982","url":null,"abstract":"Background and Aim : Novel TB vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis . Materials and Methods: Tb10.4 fragment was amplified by PCR and the products were digested with restriction enzymes. Next, it was cloned into the pcDNA3.1 + plasmid. The ag85a gene and pcDNA3.1 + / tb10.4 plasmid were digested by EcoRI and BamH1 restriction enzymes. Eukaryotic cells were transfected with pcDNA3.1 + / tb10.4 - ag85a plasmid for confirming expression of tb10.4 - ag85a in these cells. Results: Using electrophoresis of PCR products, fragments 297 bp for tb10.4 and 1017 bp for ag85a were observed. Eukaryotic cells transfection with pcDNA3.1 + / tb10.4 - ag85a vector was confirmed with cDNA synthesis and existence of tb10.4-ag85a was confirmed with RT-PCR. Conclusion: In this study, we constructed a DNA vaccine encoding tb10.4-ag85a fusion fragment. It can be used for development of more new DNA vaccines in future studies .","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"125 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88628851","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Tacrolimus toxicity in organ transplantation: an overview","authors":"A. Sharifi, Azadeh Aminzadeh","doi":"10.22037/AMLS.V3I3.20832","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.20832","url":null,"abstract":"Tacrolimus is a macrolide lactone antibiotic, and acts as a calcineurin inhibitor. It is widely used to prevent organ transplant rejection. It has been approved as first-line treatment after organ transplantation. Tacrolimus has narrow therapeutic range and wide individual variability in its pharmacokinetics. In organ transplantation, immunosuppression is associated with important risks, in particular, related to infections and cardiovascular diseases, which are the predominant causes of death in those with a functioning graft. This review focuses on toxicity of tacrolimus after transplantation. Tacrolimus toxicity is a major determinant of morbidity and mortality in organ recipients after transplantation. Therefore, reducing toxicities has become a priority. To decrease the incidence of side effects, and expand graft survival, the appropriate initial and maintenance dose of tacrolimus is essential. Clinical conditions that influence tacrolimus pharmacokinetics, such as hemorrhage, systemic inflammation and shock, all result in higher variations of tacrolimus concentrations. In addition, unbound plasma concentration is a major important reasonable parameter for monitoring of receiving optimal tacrolimus dosing in the unstable patient. Therefore, the approach of tacrolimus monitoring is vital and will support to avoid tacrolimus toxicity in the early days after transplantation.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"29 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89507372","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The effects of Quercetin on miRNA-21 expression in MCF-7 cells","authors":"G. Panahi","doi":"10.22037/AMLS.V3I3.21696","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.21696","url":null,"abstract":"Background : Cancer prevention, by the use of natural dietary or secondary metabolites in plants is one of the strategies has been attracting some of the scientific interests. One of these natural agents is Quercetin, which has anti-metastatic and anti-cancer effects. MiR-21 among different miRNAs is one of the most important frequently up-regulated miRs in numerous cancers including breast and increase cell proliferation and decrease apoptosis and therefore leads to increases in cancer incidence. We assessed the effects of Quercetin on cell viability, MiR-21, Maspin and PTEN gene expression in the MCF-7 cell line. Materials & Methods : The human MCF-7 breast cancer cell line was cultured in RPMI1640 and treated with different concentrations of Quercetin (0.01-100 μM) for 24 hours. The cytotoxic effect of Quercetin on MCF-7 viability was determined using Methyl-Thiazolyl-Tetrazolium (MTT) assay by IC50 determination. The relative expression of MiR-21, Maspin, and PTEN gene expression were determined by real-time Polymerase Chain Reaction (PCR). Results : The maximum inhibitory effect of Quercetin on cell viability was observed at 100 μM after 24-hour incubation. The expression of MiR-21 in the treated cells compared to controls was significantly decreased after treatment with three different concentrations of Quercetin. In addition, expression of Maspin and PTEN in the treated cells compared to controls was significantly increased. Conclusions : Quercetin decreases cell viability and miR-21 gene expression in a dose-dependent manner. Also, Quercetin decreases mir-21 gene expression and increases Maspin and PTEN expression in MCF-7 breast cancer cell line. The growth inhibitory properties and therapeutic effect of Quercetin on the breast cancer may be mediated by reduction of miR-21 expression, and for verifying this hypothesis and the possible therapeutic implication of Quercetin in this direction further studies are necessary.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"99 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85755358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Probing the Biological evaluations of a new designed Palladium (II) complex using spectroscopic and theoretical approaches: Human Hemoglobin as a Target","authors":"M. Abbasi, Omid Abazari","doi":"10.22037/AMLS.V3I3.21712","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.21712","url":null,"abstract":"Background : Previous studies reported that Palladium (Pd)(II) drug compounds showed significant anti-tumor activity in comparison with cis-platin. Materials and Methods : In this study, we investigated the biological evaluations of a designed Pd (II) complexes (bi pyridine ethyl dithiocarbamate palladium II nitrate) via its anti-proliferative effects on the alterations in the function and structure of human hemoglobin (Hb) at different temperatures of 25 and 37°. Also for further investigation, multi-spectroscopic methods such as fluorescence and the far-UV circular dichroism (CD) with hemoglobin target were assessed. Results : Fluorescence data showed the pure ability of Pd(II) complex to quench the intrinsic fluorescence of Hb. The binding constant, number of binding sites, and thermodynamic parameters at two temperatures were assessed and the results demonstrated the major possibility of occurring electrostatic and hydrophobic interactions in the Pd (II) complex–Hb interaction. For evaluating the change of secondary structure of Hb upon interaction with various concentrations of complex, far-UV CD spectra was applied and it was observed that in high dose of complex, significant changes occurred which is indicative of some side effects in overdosing of this complex. Conclusion : Our results suggested that using palladium complex as an anticancer agent might cause some disorders in structure and function of Hb as well as improve understanding of the side effects of newly designed metal anticancer drugs.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"92 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78682347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"The effect of drinking cold water on fasting blood glucose, urea, uric acid, creatinine, liver transaminase enzymes activities, lipid profiles and thyroid hormones in ratsfiles and thyroid hormones in rats","authors":"J. Amri","doi":"10.22037/AMLS.V3I3.22355","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.22355","url":null,"abstract":"Background: The current study was designed to investigate the changes of fasting blood glucose (FBG ), urea, uric acid, creatinine, aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), very low-density lipoprotein cholesterol (VLDL-c), high-density lipoprotein cholesterol (HDL-c), thyroid-stimulating hormone (TSH), thyroxin (T4) and triiodothyronine (T3) of rats after long-term drinking of cold water. Materials & Methods: In this experimental study, 12 adult male Wistar rats weighting 220-200 g were used. The rats were divided into 2 equal groups including, control and experimental groups. The control and experimental groups received normal water (20 ° C) and cold water (4 ° C) for 60 days, respectively. At the end of the 60 days, blood was taken from the heart of animals. After separating the serum, concentration of FBG, urea, uric acid and creatinine, ALT, AST, TG, TC, HDL-c were assayed by spectrophotometer and LDL-c, VLDL-c were calculated by the Friedewald formula. The serum concentrations of TSH, T4 and T3 were identified by ELISA. Results: Results showed that cold water significantly increased the levels of ALT,AST,TG, LDL,VLDL,TSH,T4 and T3 (P<0.05) and had no significant effect on urea, uric acid, creatinine, TC and HDL levels in experimental group compared to control group. Conclusion: Cold water can have a devastating effect on the metabolism of the body in the long-term. Although more studies are needed.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76111034","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Real-time PCR and pp65-antigen test for monitoring human cytomegalovirus infection in kidney transplant patients","authors":"Somayeh Shokri","doi":"10.22037/AMLS.V3I3.18853","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.18853","url":null,"abstract":"Background: Human cytomegalovirus (HCMV) is a common virus that infects people of all ages. HCMV infection is an important and common complication in immunocompromised patients, especially transplant recipients. Antigenemia test and real-time PCR are one of the most common assays for diagnosis and monitoring of CMV infections. The aims of this study were to compare two common detection methods in order to identify clinically useful CMV infection in kidney transplant patients. Material and methods : One hundred and fifty peripheral blood samples from kidney transplant patients, including 78 men and 72 women aged from 4 to 73 years; with mean age of 36 years, collected during March 2016 to June 2016. Then samples were investigated for pp65-antigen on polymorphonuclear cells and HCMV DNA viral load on plasma and whole blood. Results: Out of 150 samples analyzed, HCMV DNA was detected in 47(31.33%) cases; with 26 (55.32%) and 21(44.68%) cases in men and women, respectively. The pp65 antigen was detected in 42(28%) casas; with 23 (54.76%) and 19 (45.24%) cases in men and women, respectively. Of the 150 samples, 42 (28%) were positive for both assays and 108 (72%)were negative. Conclusion: Our findings showed both tests were significantly correlated and can be useful for monitoring of CMV infection. Hence, higher viral loads have been found to be associated with increase of disease complication, Real- time PCR is more suitable. The findings merit more investigations involving larger numbers of samples","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84444579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Detection of Efflux MexAB-associated Multidrug Resistant (MDR) Pseudomonas aeruginosa Isolated from Patients in Torbat Heydarie, Northeast Iran","authors":"Mohammad Jafari, S. Dolatabadi, H. Miri","doi":"10.22037/AMLS.V3I3.19458","DOIUrl":"https://doi.org/10.22037/AMLS.V3I3.19458","url":null,"abstract":"Aims of the study : Pseudomonas aeruginosa ( P. aeruginosa ) is one of the leading causes of hospital acquired infections. Infections with P. aeruginosa are often hard to treat because of existence of different mechanisms of antibiotic resistance changes in permeability of drugs and activity multidrug efflux pumps . The aim of current study was to determine the antibiotic resistance pattern of P. aeruginosa and existence of efflux pump MexAB genes using PCR technique. Materials and Methods: 506 isolates cultured from different clinical specimens of patients hospitalized at Nohom Dey and Razi hospitals of Torbat Heydarie (northeast Iran) were collected and used in this study. Isolates were identified using conventional bacteriology and their susceptibility to different antibiotics were assessed using agar disk diffusion method. The PCR assay was used to detect efflux pump MexAB genes. Results: From 506 isolates, 50 were identified as P. aeruginosa and these were isolated from isolated from blood, tracheal, burn, and wound. Incidence of P. aeruginosa was greater in males than females , wound infections had the highest number of occurrence and patients between 30-50 years were the most infected age group. In total, 60.86% of strains were multidrug resistant (MDR). The PCR technique revealed that most of the P. aeruginosa isolates and all the MDR strains contained MexA and MexB genes. Conclusions: The emergence of MDR microorganisms poses serious therapeutic problems for patients. Determining bacterial resistance mechanisms is complex. In this way, efflux systems were responsible for antibiotic resistance and played an important role in the MDR phenotype among P. aeruginosa isolates.","PeriodicalId":18401,"journal":{"name":"Medical laboratory sciences","volume":"115 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2018-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80291849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}