Medical Laboratory Journal
Volume:9 Issue: 5, Nov-Dec-2015

Probiotics are living microorganisms that have beneficial effects on the health of digestive system. The aim of this study was to evaluate the antimicrobial ability of acidic and neutral supernatants (culture supernatant) of lactic acid bacteria against common bacterial pathogens.

Four species of lactic acid bacteria (Lactobacillus plantarum PTCC1745, Lactobacillus PTCC1608, Lactobacillus Saki PTCC1712 and Lactobacillus Lactis PTCC1336) were obtained from the microbial collection of Iranian Research Organization for Science and Technology in Lyophilized form. The antimicrobial activity of neutral and acidic supernatants against bacterial pathogens was investigated using the Disk and Well Diffusion Agar methods.

Lactic acid bacteria showed good antimicrobial ability against six pathogenic bacteria with the highest inhibitory effect observed in Lactococcus lactis against E. coli PTCC1399 through well method with an average diameter of 14 mm inhibition zone. In this study, the well diffusion method was far more sensitive compared to the disk method and acidic supernatants showed higher antimicrobial efficiency compared to neutral types.

the Metabolites produced by lactic acid bacteria are able to inhibit the growth of pathogenic bacteria that can be an important and practical solution for the prevention and treatment of infections and ultimately improve human health.

BabA2 and Hpa genes are involved in adherence of Helicobacter pylori (H.pylori) to gastric mucosal tissue. This study aimed to investigate the frequency of these genes in isolates of H. pylori from gastric biopsies and their relationship with gastritis, peptic ulcer and gastric cancer.
Gastric biopsy samples were obtained from patients with gastritis, peptic ulcer and gastric cancer. A sample was sent to the laboratory for urease test and histopathology study, and another sample for DNA extraction. The frequency of BabA2 and Hpa genes was investigated using their specific primers by PCR.
Among the 80 analyzed biopsy samples, 51 (63%) were BabA2 positive, and the frequency of this gene in the samples of gastric cancer, gastritis and peptic ulcer was 61.1, 58.3 and 73.3%, respectively. In addition, 57 samples (71%) were Hpa positive, and the frequency of this gene in the samples of gastric cancer, gastritis and peptic ulcer was 55.5, 69.4 and 84.6%, respectively. There was no significant correlation between the presence of these genes and the type of H.pylori-related diseases.
Frequency of BabA2 and Hpa genes is higher in the samples of peptic ulcer but there was no significant relationship between these genes and H.pylori-related diseases.

Hirudin is an anticoagulant polypeptide secreted from the salivary glands of leeches. Recombinant hirudin is a strong anticoagulant agent in arterial and venous thrombosis. The aim of this study was to evaluate the effect of inserting protein A signal peptide sequence of pEZZ18 plasmid on expression and secretion of the recombinant hirudin in E.coli.
the synthetic hirudin gene was amplified by PCR using specific primers. First, the gene was purified and cloned into PTG19-T cloning vector, and then it was subcloned into pEZZ18 expression vector by SalI / SacI enzymatic digestion and finally transformed into E.coli JM107. After the expression of recombinant hirudin protein, different cellular fractions were isolated and analyzed on SDS-PAGE and further confirmed by Western blotting.
PCR product (522 bp) was first subcloned into the T-Vector (replicating vector) and then successfully subcloned into the pEZZ18 (expression vector). Cloning and subclonig were confirmed by enzymatic digestion and Colony PCR. After the expression and isolation of fractions, the presence of hirudin (about 29 kDa) in different cell fractions due to the effects of signal peptide was observed in SDS-PAGE and finally confirmed by Western blotting.
The gene of anticoagulant hirudin protein (desirudin) was cloned into the pEZZ18 vector containing Protein A signal peptide sequence and later transformed into E.coli JM107. The recombinant hirudin protein expression in the extracellular space was approved.

Aluminum salts are among the most common useful additive compounds in preparation of human and animal vaccines. Aluminum phosphate and aluminum hydroxide are two additives that show good immunoadjuvant effects with many antigens. Aluminum-containing vaccines lead to a better and longer immune response compared to adjuvant-lacking vaccines. The Chromogenic methods used for determination of aluminum amounts in manufacturing centers are time-consuming and requires some experienced technicians to obtain accurate results. This study aimed to design and validate a simple polarographic method to measure aluminum in recombinant hepatitis B vaccine.
In this study, the effects of temperature, pH, potential range and potential scan rate on the polarographic method of measuring aluminum in hepatitis B vaccine was evaluated and the optimal values for each of these factors were achieved.
In order to measure aluminum, temperature of 60 °C and pH of 4.5 were found as the optimal values. Implementation of polarographic method in the potential range of -0.25 to 0.1 volts had a better signal.
Since the polarography method is more simple, accurate and faster than the chromogenic methods, it is suitable to be used for the measurement of aluminum in hepatitis B vaccine and it is recommended to be used in quality control laboratories for biological products.

Various cellular factors affect the process of HIV activity. One of these cellular factors are structures known as microRN that are expected to be involved in controlling HIV replication and infectivity. The expression of one or a set of them may represent the patient's clinical conditions. In this study, the expression of miR-29a and miR-29b involved in regulating viral genes’ expression was evaluated in three HIV-positive groups and a healthy control group. Later, the expression level of these microRNAs was compared between the cases and controls.
Total RNA extraction was performed on the collected samples using RNx-plus kit and then the microRNA expression levels were evaluated using Relative Real-time PCR. The obtained data was entered into SPSS 22 and Graphpad softwares and analyzed using Kruskal-Wallis and Man-Whitney tests. P-value of less than 0.05 was considered as statistical significance level.
The expression level of miR-29a was reduced in patients under treatment and drug-resistant patients ( P ≤ 0.05) . All three HIV-positive groups including people without drug treatment, patients under treatment and drug-resistant patients showed reduced miR-29b expression level compared to control group (P ≤ 0.05).
the decreased expression of miR-29a and miR-29b in patients under treatment and drug-resistant patients indicates an increased viral replication and reduced CD4 cell count. It may be possible to predict the progression of the disease by miRNA measurement or control viral replication using these mir-RNAs that requires further studies.

Normal hemoglobin (Hb) is formed of a heme group and a protein group known as globin. Globin is made of four polypeptide chains and in hemoglobinopathies, the structure of one of these four polypeptide chain becomes abnormal. Cellulose acetate method is a common way to differentiate haemoglobinopathies. Inability to identify the components of Hb low concentrations and incapability to isolate all Hb types are among the disadvantages of this method. The aim of this study was to report the prevalence of hemoglobinopathies in the North of Iran by capillary electrophoresis method.
All patients with suspected hemoglobinopathies, referred by physicians for electrophoresis, have been studied in a private center in the city of Gorgan, Iran. The level of HbA2, HbA, HbF and other Hb was recorded.
Overall, 725 blood samples were analyzed using the capillary method. HbE was reported in 2 patients, HbH was observed in 2 patients and Hb Barts was reported in 3 patients. Using the capillary method, among patients with the SDG area, only 4 of 38 (10.52%) had HbS and the majority of them (89.48%) had HbD.
HbD is the most common hemoglobinopathy in the North of Iran.

the Formation of urinary infection by uropathogenic E.coli needs numerous virulence factors and biofilm formation is among these factors. Bacteria that form biofilms express phenotype traits that appear according to the bacteria type. Cellulose is an important compound on the outside of E.coli causing bacterial cell-cell reactions and connection to nonliving surfaces. Curli pili cause the reaction between cell-cell and surface-cell in biofilms and lead to bacteria aggregation. Microorganisms’ ability to form biofilm on a surface depends on the surface nature and its conditions. This study aimed at determining the production ability of cellulose polysaccharide and curli pili in UPEC strains, and its correlation with formation and intensity of biofilm.
In this study carried out to compare the ability of cellulose and pili curli production ability in 40 uropathogenic E.coli isolates ,by morphotype method in Congo Red medium (CR), each isolate was incubated at 37 oC, for 24 hours. After 24 hours, all colonies’ morphology characteristics were studied
It was shown that 67.5% of strains produced cellulose and 72.5% produced curli pili. In addition, 92.6% and 89% of isolates that produce cellulose and curli, respectively, had a moderate to strong biofilm. Moreover, it was shown that there is a significant correlation between cellulose and / or curli pili production with biofilm intensity.
About 70% of E.coli isolates from patient's urine are able to produce cellulose or curli pili; therefore, it can be concluded that the production of these two combinations is effective in amount and intensity of biofilm formation.

Paratuberculosis has been repeatedly reported from Iranian ruminant herds. The extrem fastidious nature of Mycobacterium avium subspecies paratuberculsos hinders genomic diversity studies of the pathogen. Short Sequence Repeat analysis is one of the genome-based approches recently developed to overcome this difficulty. In this study we describe the application of SSR genotyping on three Iranian MAP type strains plus the III & V vaccinal strain.
All the bacteria were examined by PCR-F57 and PCR-IS900 experiments in order to authenticate their identity as MAP. SSR genotyping using SSR1 & SSR2 loci was conducted according to the Amonsin method. PCR amplicons were sequenced to guarantee the accuracy of findings.
At SSR1 locus two allels were identified, a larger allel of 770 bp and a smaller allel of 763 bp long. At SSR2 only a single allele, 800 bp long, was detected. Two Iranian bovine and ovine MAP isolates along with the vaccinal III & V strain shared a single SSR1/SSR2 pattern while a different SSR1/SSR2 was represented by the third (caprine) Iranian MAP isolate.
While finding a shared SSR type between the two Iranian MAP isolates and the III & V strain might represent a mutual ancestral background but this has to be assessed through further studies. Detection of two SSR genotypes between three Iranian type strains is likely a reflection of more MAP clones in Iran.

Nonalcoholic fatty liver disease is the most common chronic liver disorder and is also currently considered as the hepatic manifestation of metabolic syndrome. Regular exercise training may decrease fatty liver disease complications, although its impact is not yet clear. This study aimed to evaluate the effects of six weeks of aerobic training on liver enzymes and other factors contributing to metabolic syndrome in young inactive women.
In this quasi-experimental study, 37 inactive overweight women were randomly divided into an experimental and a control group. The experimental one participated in a controlled aerobic training program (5-minute interval walking) at 65-90% maximum heart rate for 6 weeks, 45-90 minutes per session and 4 sessions per week. Blood samples were taken following 12 hours of fasting, both before and after the training program.
The levels of aspartate aminotransferase and alkaline phosphatase decreased in both groups. Alanine aminotransferase level, weight and waist circumference were significantly decreased in the experimental group following the 6-week exercise training (P Six weeks of aerobic training may help prevent hepatic damage through decreasing serum levels of liver enzymes, anthropometric factors and some metabolic syndrome factors associated with nonalcoholic fatty liver disease.

Diabetes mellitus is the most common risk factor for coronary artery disease (CAD). Cholesteryl ester transfer protein (CETP) TaqIB polymorphism is associated with changes in lipid profile and may be a risk factor for CAD in patients with diabetes. This study aimed to evaluate the association of CETP TaqIB polymorphism with CAD in patients with type 2 diabetes.
In this case-control study, 292 diabetic patients were divided into two groups based on angiography reports (150 participants with normal angiogram as the control group and 142 participants with more than 50% stenosis of at least one coronary artery as the case group). The CETP TaqIB genotypes were determined by PCR-RFLP analysis. Fasting blood glucose was measured using glucose oxidase and lipid profile (triglycerides, total cholesterol, high density lipoprotein-cholesterol and low density lipoprotein-cholesterol) by an enzymatic method.
There was no significant difference in the frequency of genotypes and alleles between the case group and controls (the control group: B1B1, 17.3%; B1B2, 63.3%; and B2B2, 19.3%; the case group: B1B1, 18.3%; B1B2, 64.1%; and B2B2, 17.6%) (P=0.92). In the control group, heterozygous participants (genotype B1B2) had higher levels of cholesterol compared with other genotypes (B1B1 and B2B2). Also, the patients with genotype B1B2 had significantly higher weight (P=0.013).
There is no significant correlation between CETP TaqIB polymorphism and the increased risk of coronary artery disease in patients with type 2 diabetes.