International Journal of Medical Laboratory
Volume:4 Issue: 2, May 2017

Hepatitis B virus (HBV) infection is a serious global health problem affects many people. Recently, because of vaccination, the incidence of HBV infection has been reduced, but in high risk population also health care students (HCS) adequate immunization is serious. The aim of this survey was to evaluate HBV immunization in the population of the paramedical students based on the demographic characteristics in Iran.
Anti-HBsAg titer was prospectively assayed in all vaccinated of the 95 HCS using Enzyme-linked immunosorbent assay.
Results and Anti-HBsAg titer in the studied population was estimated about 92.8±80.5 mIU. Anti-HBsAg titer response was significantly higher in females to males. A low proportion of vaccinated HCS had low titers antibody against HBV infection. Therefore, measuring anti-HBsAg titer may help to drop in HBV incidence in HCS. This reduction can be correlated with the effectiveness of national immunization program.

Six sigma is the latest version of total quality management. It is quantitative goal for process performance. With increasing demands for improved accuracy and reliability of the results, Six Sigma is gaining increased visibility in the clinical laboratory process outcomes. The aim of study was to evaluate the quality of analytical phase performance in a clinical biochemistry laboratory by calculating sigma metrics.
The study was conducted in a hospital laboratory of Bhatia Hospital, Mumbai. Mean, coefficient of variation, bias, and sigma values were calculated for 24 biochemistry parameters. The guidelines used for total error allowable (TEa) values were clinical laboratory improvement amendments (CLIA), RILIBAK and college of american pathologists (CAP). Four months’ internal and external quality control data were extracted for the following parameters-albumin, alkaline phosphatase (ALP), alanine aminotransferase (ALT), amylase, aspartate aminotransferase (AST), bilirubin direct, bilirubin total, Ca, cholesterol, creatine kinase (CK), creatinine, triglycerides (TG), Uric acid (UA), unsaturated iron binding capacity (UIBC), urea, gamma-glutamyl transferase (GGT), glucose, high-density lipoproteins (HDL), iron, lactate dehydrogenase (LDH), lipase, Mg, phosphorus and total protein.
Albumin, ALP, AST, ALT, amylase, bilirubin total, Ca, cholesterol, CK, creatinine, TG, uric acid, GGT, glucose, iron, LDH, lipase, Mg, Phosphorus, total protein showed the performance of more than six Sigma for both level of controls. Bilirubin direct, Urea, for level 1; UIBC, urea, HDL for level 2 showed sigma from 3-6.
Based upon sigma metrics, laboratory quality control strategy can be planned and reevaluated for continuous monitoring and improvement of test methods.

Mucosal-associated invariant T (MAIT) cells are striking lymphocyte population in the blood and their importance in immune responses is growing fast. The current study was conducted to evaluate leptin hormone effects on MAIT cell functions.
Five healthy male donors in ages of 22-30 years were selected and peripheral blood mononuclear cells (PBMCs) were enriched by Ficoll-density gradient. The cells were stimulated by different doses of human recombinant leptin. Using anti-CD3, anti-CD161 and anti-Vα7. 2 antibodies, positive CD3/CD161/ Vα7.2 MAIT cells were selected among stimulated PBMCS and proliferation alterations (after 5 days) and intercellular tumor necrosis factor (TNF)-α production (after 24 hours) were determined by flow cytometer.
Stimulation of MAIT cells in doses of under 800 ng/ml of leptin did not alter the frequency of the cells significantly. However, in 800 ng/ml of leptin the number of the cells declined substantially, but statically analysis did show a significant difference with unstimulated and other leptin concentrations (p=0.12). When the frequency of intracellular TNF-α positive MAIT cells investigated, it revealed that in doses of 250 and 400 ng/ml of leptin, the number of the TNF-α. positive cells significantly increased compared to other concentrations (p=0.002). In high concentration of leptin (800 ng/ml), the frequency of positive TNF-α cell decreased compared to 400 ng/ml of the hormone.
Leptin hormone in doses of 250 and 400 ng/ml has affected MAIT cells’ ability to produce TNF-α cytokine. Therefore, in adipose tissue leptin might be considered as a new source of inflammatory cytokines.

Patients with Hyper-IgE syndrome suffer from fungal and bacterial infections, especially Candida albicans and Staphylococcus aureus. Due to the important role of T helper17 (Th17) lymphocytes in defense against fungal infections, the percentage of Th17 lymphocytes was studied in the patients with autosomal recessive hyper-IgE syndrome (AR-HIES).
In this case-control study, six patients with AR-HIES (with DOCK-8 mutation) and seven healthy age and sex-matched controls were included. Peripheral blood mononuclear cells were isolated from their venous blood and the percentage of Th17 lymphocytes were determined by flow cytometry.
There was no statistical difference between the percentage of Th17 lymphocytes (p=0.15) in the case and control groups. Also in comparison to the control subjects, the numbers of eosinophils were dramatically increased (p=0.000). Also, there was a significant negative correlation between serum IgE levels and Th17 lymphocytes’ percentage (r=-0.927, p=0.006) and a significant positive correlation between eosinophils number and Th17 lymphocytes’ percentage (r=0.557, p=0.01). Serum IgE levels showed a significant positive correlation with the numbers of eosinophils in the patients’ peripheral blood with AR-HIES (r=0.961, p=0.003).
The numbers of Th17 in the patients with AR-HIES may not show statistical differences between the cases and controls. The numbers of eosinophils significantly increased in the patients AR-HIES compare to the controls.

Chondrocytes and their differentiation play a central role in joint diseases. Effect of the transforming growth factor (TGF)-β1 on chondrocyte characteristics and differentiation is not clearly understood. This study was undertaken to investigate the effects of TGF-β1 on tissue characteristics and morphology of chondrocytes against degradation induced by interleukin (IL)-lα in bovine nasal cartilage (BNC) explant culture.
BNC explants were cultured in DMEM and samples were divided into four groups. In group A (control); samples were only cultured in DMEM and group B; was treated with IL-1α (10 ng/ml), group C; treated with TGF-β1 (10 ng/ml) and group D; treated with IL-1α (10 ng/ml) TGF-β1 (10 ng/ml) for 14 days. At days 3, 7 and 14 the media were collected and replaced with fresh media. Then, samples were harvested on days 3, 7 and 14 and chondrocyte morphology was assessed by invert microscopy. We used Masson's Trichrome stain to visualize collagen distribution and synthesis, and Safranin O and Alician blue to highlight the proteoglycan content.
In the presence of IL-lα, most of the chondrocytes were transformed into fibroblast-like morphology with pyknotic nuclei at day 14 and proteoglycan and collagen in extracellular matrix (ECM) were destructed at this time. In the presence of TGF-β1, chondrocytes preserved their ordinary normal features. Also, TGF-β1 inhibited collagen and proteoglycan destruction and cartilage ECM showed normal characteristics.
IL-1α induced significant morphological changes in chondrocytes and increased the destruction of ECM. TGF-β1 could strongly preserves cartilage from IL-1α degradation effects.

Backgroung and Aims: L-Asparaginase II is a cornerstone of treatment protocols for acute lymphoblastic leukemia. Only asparaginase II obtained from E. coli K12 and Erwinia chrysanthemi have been used in human as therapeutic drug. The therapeutic effects of asparaginase II from E. coli K12 and Erwinia chrysanthemi is accompanied by side effects. It is desirable to search for other asparaginase II sources with novel properties that could be therapeutic and produce an enzyme with less adverse effects.
Previously, we performed the in vitro studies, including cloning, sequencing and expression of L-asparaginase II genes (ansB) from Citrobacter freundii 1101, Erwinia chrysanthemi DSM 4610, E. coli BL21 and Klebsiella pneumoniae ATCC 10031. In this article, the obtained results were compared bioinformatically. The nucleotide and amino acid sequence alignments were carried out by ClustalW2. Protein localization and signal peptides were predicted by PSORT and SIG-Pred softwares, respectively. Percentages of hydrophobic and hydrophilic residues were calculated by Genscript software. The physicochemical parameters were computed using Expasy’s ProtParam prediction server. The secondary and 3D structures were predicted by SOPMA and the online server Phyre2, respectively. The antigenicity of the asparaginase IIs was predicted using Semi-empirical method.
E. coli BL21 and Citrobacter freundii 1101 had the most similarity in physicochemical parameters and antigenicity with E. coli K12. Also, Erwinia chrysanthemi DSM 4610 had the most similarity in physicochemical parameters and antigenicity with Erwinia chrysanthemi.
In spite of these similarities with drug types, the potentiality of other low-similar asparaginase IIs should also be determined and compared with drug types.

The aim of this study was profiling glycated hemoglobin (HbA1c) level, lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) activity in obese women with gestational diabetes mellitus (GDM) and evaluating the correlation between them.
Sample size was 90 subjects admitted to the clinical laboratory, who were divided into three groups, in each group (n=30). Subjects glycemic control was checked by HbA1c; ALP, LDH activity and serum glucose were determined with commercial kit. Age and body mass index (BMI) was recorded for each subject. The correlation analysis between blood activity of ALP, LDH activity, HbA1c, glucose, BMI and age in diabetic and normal pregnant women was carried out.
The mean of HbA1c levels was significantly higher in the GDM obese women than in women with normal pregnancy (p=0.01). In contrast, the means of ALP and LDH activity were lower in the GDM obese women than in women with normal pregnancy (p=0.09, and p=0.15, respectively). Also, an increase from the first to the third trimester of pregnancy in Hb-A1c levels was occurred from 3.6±0.016 to 4.0±0.15 mmol/L, from the first to the third trimester of pregnancy an increase in ALP activity was occurred from 174.4±12.2 to 177.5±16.3 U/L and there is an increase towards the third trimester.
Analysis of HbA1c level, LDH and ALP activity provides the evidence about health during the pregnancy. Using the HbA1c, LDH and ALP as a biomarker for monitoring GDM will be useful.

Rotavirus enteritis is an acute viral infectious disease among infants. VP7 protein has a key role in attachment and entry virus into the target cell. The VP7 protein is involved in inducing the production of neutralizing antibodies that protect infants against reinfection of the virus. The aim of this study was to heterologous expression of the VP7 gene of bovine rotavirus as a fusion protein of Trx–VP7 in a genetically engineered bacteria.
Total RNA was extracted from MA104 cells infected with bovine rotavirus strain RF. BRV VP7 gene was amplified using polymerase chain reaction. Another gene was the VP7 synthetic that made in the pBSK plasmid. The pBSK-VP7 plasmid was digested using BamHI and SacI restriction endonucleases, then recombined into the prokaryotic expression vector pET32a. The pET32a-VP7 amplified and pET32a-VP7 synthetic were transformed into BL21 (DE3) competent cells of Escherichia coli, respectively, and induced by different concentrations of Isopropyl β-D-1-thiogalactopyranoside at 30°C and 37°C in luria bertani and terrific broth media, then analyzed using SDS-PAGE and western blotting.
SDS-PAGE analysis showed that the both pET32a-VP7 amplified and pET32a-VP7 synthetic were not expressed in the BL21 (DE3) cells, But the expression of pET32a-VP7 synthetic was weak at 30° C in luria bertani media.
This is the first report of the production of bovine rotavirus (RF strain) Full-Length VP7 in prokaryotic system expression. VP7 is a membrane protein and has toxic domains that show high toxicity in this expression system.