Research in Medicine
Year:2013 | Volume:37 | Issue:1|Papers Count:12

Background: Human amniotic epithelial cells (hAECs) have been introduced as a valuable source of stem cells with therapeutic applications. Despite the extensive study and emerging evidence, cell surface marker signature of hAECs remained controversial. The aim of the present study was to establish an efficient and optimized isolation procedure of hAECs and their characterization in terms of proliferation and stem cell markers.Materials and methods: Four human placentas were collected from healthy women undergoing elective caesarian delivery at term labor under sterile condition. Amniotic membranes were carefully isolated and subjected to trypsin digestion. Single cell suspensions were obtained and the epithelial origin was confirmed by assessment of cytokeratin expression. Expression of CD9, CD10, CD29, CD34, CD38, CD44, CD45, CD73, CD105, CD133, MHC-I, HLA-DR, HLA-G, SSEA-4, STRO-1 and OCT-4 was then evaluated by flow cytometry. Results: The overall yield of hAEC ranged from 80-130×106 cells per placenta with more than 98% viability. Microscopic examination revealed that hAECs are large and refractive cells with great capacity to adhere to plastic surfaces. The cells substantially expressed cytokeratin implying their epithelial origin. Flow cytometry data revealed that hAECs are a heterogenic population consisting of immune phenotypically mixed cells. Nearly all purified cells expressed mesenchymal markers, CD9, CD10, CD29, and CD73. A large proportion of cells also expressed HLA-I, HLA-G, STRO-1, OCT-4 and SSEA-4. Isolated cells, however, failed to express CD34, CD38, CD44, CD45, CD105, CD133 and HLA-DR.Conclusion: The procedure presented here is a simple and cost-effective protocol for isolation of a large numbers of viable hAECs. Surface markers and such features as proliferation capacity of hAECs are among the characteristics that are strongly dependent upon isolation and cell culture conditions. Thus such variables should be taken in to consideration when stem cell properties of hAECs are to be interpreted.

Background: Opinions differ about the role of smoking cigarettes on homogeneity of ventricular repolarization. The aim of this study was to investigate the short term effect of cigarette smoking on QT dispersion (QTD) as a measure to evaluate the ventricular repolarization homogeneity in patients with Congestive Heart Failure (CHF) and healthy smokers.Materials and methods: In this experimental study, 40 patients with CHF and 40 healthy individuals, (all smokers), matched for gender and age, were enrolled. The magnitude of QTD before and immediately after smoking a cigarette with 1.7 mg nicotine was measured and compared between them.Results: QTD in patient with CHF was 55.1±13.7 msec before and 64.2±13.7 msec after smoking a cigarette; in the control group, the figures were 42.16±14.2 and 47.3±17.1 respectively. Difference between two groups was statistically significant (p=0.001). Also, the scale of QTD was significantly higher in patients with CHF before and after smoking compared to control group (p=0.032 and 0.001 respectively). Finally we observed that increase in QTD in patients with CHF (9.4±4 msec) was significantly greater than that of healthy group (4.7±2.5). (p=0.001).Conclusion: Cigarette smoking, even for a short period, can increases QTD and may lead to ventricular arrhythmia and sudden cardiac death both in healthy smokers and in patients with CHF; magnitude of this effect is much higher in the latter group. It is crucial that patients with CHF stop smoking and avoid places polluted with cigarette smoke.

Background: Removal of prostate and radiotherapy are used routinely for treatment of prostate cancer; however in advanced stages of the disease these treatments are ineffective. Researchers are exploring strategies for preventing this malignancy. This study aims to design and develop a phage based vaccine for preventing prostate cancer.Materials and methods: After conducting bioinformatic research on designing and evaluating the effect of the vaccine, the vaccine was produced and the immune response tested in rabbit. Phage display technology was used to express the prostate specific membrane antigen (PSMA) on the surface of pVIII of M13 bacteriophage. Recombinant PSMA gene was cloned in the pAK8-GVO77 vector which contained the pVIII sequence. Recombinant construct was transferred into TG1 (an E. coli strain) bacterium and the transformed bacteria were infected with M13k07 helper phage. Recombination phages were evaluated by monoclonal antibodies via ELISA. To assess the immunogenicity, recombinant phages were injected subcutaneously and intramuscularly into white rabbit. The immune response against PSMA was assessed by ELISA method.Results: The developed vaccine was able to raise an immune response in the laboratory animal.Conclusion: This research showed that it is possible to design and develop a phage based vaccine that raises the humoral immunity; assessment of the cancer inhibitory effect of the vaccine requires more experimental studies.

Background: Pepper Spray could be utilized for personal defense against human and animals assailants. The symptoms would be exhibited immediately if this spray comes in contact with eyes, nose, throat and the lungs. No comprehensive information is available on the effect of the period of exposure to this spray on anxiety behavior; this study was done to determine the effect of OS pepper spray on anxiety (fear) behavior in rats.Materials and methods: Adult male Wistar rats, with a mean weight of 210±20 grams were divided into 4 experimental groups and a control group. 6 rats were placed within a chamber that had been sprayed with OS spray for 3 seconds.. The rats remained for 30, 60, 90, and 120 seconds within the chamber, and were then returned to their cages. After 90 minutes, Plus-maze was used to measure animal anxiety levels.Results: There was a significant difference in anxiety behavior between the experimental and control groups; in addition a significant increase in anxiety was observed with increasing periods of exposure to the spray, (p<0.05).Conclusion: Increasing OS exposure time would result in rising anxiety.

Background: Chlorpromazine has been widely used to treat schizophrenia; there are some reports about negative side effects of phenothiazines on the reproductive system. The present study was undertaken to consider the effects of the Chlorpromazine on the reproductive functions in female rats.Materials and methods: This experimental study was conducted on 32 adult female wistar rats divided into four groups. One group served as control was given 5 mL/kg of 0.5% methylcellulose solution; female rats in the experimental groups were administered CPZ (3, 10, and 30 mg/kg, PO) once daily for 28 days. On day 29 all rats were sacrificed by Co2 inhalation. Histopathological analysis of the mammary glands and vagina was carried out. Serum levels of prolactin and potential fertility of animals was evaluated.Results: Female rats exposed to CPZ had significant increase in serum prolactin concentrations at high doses (p<0.01, p<0.05). CPZ also induced histopathological changes in the mammary glands and vagina at high doses (p<0.01, p<0.05). CPZ decreased potential fertility in female rats.Conclusion: CPZ has adverse effect on the reproductive system in the female rat.

Background: Usual diagnostic methods for detection of Cryptosporidium include smear preparation, modified Zeihl-Neelsen staining, microscopic examination and observation of oocysts. No studies have been done to compare PCR and Zeihl-Neelsen staining in Iran. This study was done to determine the positive predictive value, (PPV) and the negative predictive value (NPV) of PCR in comparison to modified Zeihl-Neelsen staining for isolation of Cryptosporidium in children with diarrhea.Materials and methods: Direct smears were prepared from fecal samples of 2500 children with diarrhea. Smears were stained by modified Zeihl-Neelsen method and examined by microscope. All samples positive for cryptosporidia and some randomized negative samples were selected for DNA extraction for PCR assay.Results: From 2500 fecal samples, 30 positive cases were detected by modified Zeihl-Neelsen, (Z-N) staining; these tested positive by PCR method too; however two samples, found to be negative on microscopic examination were positive by PCR technique. PPV and NPV of microscopic examination after staining with Z-N were computed as 94.1% and 100%, PPV and NPV of PCR method were computed as 100%.Conclusion: PCR is a sensitive technique for detection of Cryptosporidium, especially useful in immunocompromised individuals.

Background: Nocturnal hypoxia is an important factor increasing the risk of mortality in patients with chronic heart failure and is associated with atrial and ventricular arrhythmias. QT dispersion, an index of myocardial electrical activity, is used as a prognostic tool in determining future malignant arrhythmias and sudden cardiac death. In the current study we investigated the effect of nocturnal oxygen therapy (NOT) on QTd in patients with CHF.Materials and methods: In this clinical study 54 patients with CHF (EF≤40%) were enrolled. Patients were administered NOT; electrocardiography was taken before and after NOT and QTd measured each time. Paired t-test was used to compare pre- and post-NOT QTd.Results: QTd significantly decreased from 56.3±18.2 msec before NOT to 46.3±17 msec after NOT (p=0.001).Conclusion: In this study NOT decreased QTd in patients with CHF which could consequently decrease risk of malignant arrhythmias and sudden cardiac death. However, the mechanisms by which NOT decreases QTd is not clear and requires further investigation.

Background: Two types of Cyclooxygenase enzymes, which produce prostaglandins, are present in humans. Prostaglandins play a role in the functions of various organs including the immune system, blood circulation, and cell division. Reports indicate that the level of these enzymes is increased in patients with metastases in several malignancies, including larynx, lung, stomach, colon, and prostate. The aim of this study was to investigate the association between -765 G/C promoter polymorphism of cyclooxygenase 2 and initiation and progression of lung cancer.Materials and methods: This is a case-control, retrospective study. Genotyping of cyclooxygenases was carried out by testing blood samples from 120 lung cancer patients and 110 controls using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) technique. The observed numbers of each cyclooxygenase genotype were compared with that expected for a population in Hardy-Weinberg equilibrium by using a χ2 test. The significance of the differences of observed alleles and genotypes between groups was tested using the odds ratio analysis. Results: The genotype GG of polymorphism -765G/C seen in 62.5% of patients did not indicate a significant statistic increase in comparison with the control group (55.45%; P<0.6); however, when the patients were divided into two groups i.e. those with metastases and without metastases, a weak link was identified between genotype GG of polymorphism G/C, with metastasizing cancer, (P<0.04). Dividing patients into sub-groups such as sex and smoking habit showed no difference.Conclusion: It seems that there is no association between -765 G/C polymorphism and developing of lung cancer statistically. But genotype GG of polymorphism -765G/C is linked weakly with lung cancer metastases.