Pathobiology Research
Year:2013 | Volume:16 | Issue:2|Papers Count:10

Objective: The prevalence of respiratory allergies, especially those induced by fungi such as Alternaria alternata, has dramatically increased over the past decade. This increase has caused major health problems worldwide. This study aimed to investigate the role of A. alternatain the etiology of allergic asthma, by using the skin prick test and assessment of IgE specific to the fungus in the patient's sera.Methods: This study enrolled 202 patients with allergic asthma, aged 12 to 83 years.Participants included 40.1% males and 59.9% females who were enrolled after recording demographic information. A skin prick test with the whole cell extract of A. alternata was performed on the epidermis of the patients' forearms. Histamine and normal saline were used as positive and negative controls, respectively. Serum levels of IgE specific for A. alternatawere measured for all patients using the ImmunoCAP Phadiatop method in which the specific A. alternata allergen cocktail that connected to the solid phase reacted to IgE antibodies in each patient's sera. Data were analyzed by analysis of variance and chi-square tests.Results: Among 202 patients with allergic asthma, 14 (6.93%) had mild asthma, 73 (36.10%) were moderate asthmatics and 115 (56.90%) had severe asthma. In total, 14 (6.93%) patients were positive for both the skin test and IgE specific to A. alternata, 35 (17.33%) had negative specific IgE and positive skin test results, and 36 (17.82%) had a positive specific IgE and negative skin test. A total of 117 (57.92%) patients were negative for both tests.Conclusion: The results of this study showed the presence of IgE specific for A. alternata in 50 of 202 (24.75%) patients diagnosed with allergic asthma. The skin prick test was successfully used as a screening test. The results were further confirmed by solid-phase immunoassay of the IgE specific forA. alternata crude allergenic extract.

Objective: Human cytomegalovirus (HCMV) is a beta-herpesvirus that causes persistent infection in humans, as well as severe disease in fetuses and immunocompromised individuals. Although HCMV is not currently causally implicated in human cancer, emerging evidence suggests that HCMV infection may be specifically associated with malignancies such as gliomas. Gliomas are one of the most common brain tumors that affect humans. It is classified into four grades. In this study, we have developed and used a real-time PCR method for the detection and diagnosis of HCMV infection in glioma brain tumor samples.Methods: Paraffin-embedded tumor samples were chosen from patients who referred to Imam Khomeini Hospital Neurosurgery Ward. DNA was extracted from paraffinembedded tissues by a DNA extraction kit. After designing specific primers for the HCMV US28 region, a real-time PCR method was developed for detection of HCMV US28.Results: The results of qualitative real-time PCR on 4/18 patients (22.2%) were positive.Two patients with positive HCMV results died.Conclusion: This is the first study that has monitored HCMV genes in samples from glioma patients in Iran. Considering the results of this study and controversies associated with other studies, a more comprehensive study using this and other diagnostic methods is suggested.

Objective: Diabetic neuropathy leads to axonal transport abnormalities. However its mechanism and the beneficial effects of exercise on these abnormalities are not well documented. The present study aims to investigate KIF1B mRNA in spinal cord sensory neuron tissue of Wistar male rats with diabetic neuropathy following endurance training.Methods: We randomly assigned 12 male Wistar rats into three groups: diabetic trained, diabetic untrained and healthy control. Intraperitoneal injection of a STZ (streptozotocin) solution (45 mg/kg) was used to induce diabetes. At two weeks after STZ injections, the mechanical allodynia and thermal hyperalgesia tests demonstrated the presence of diabetic neuropathy. A moderate endurance training protocol was performed for a sixweek period. At 24 hours after the final training session, the rats were sacrified and the L4-L6 sensory neurons of the spinal cord tissue were removed. KIF1B mRNA expression was performed using real time-PCR.Results: Diabetic neuropathy led to increased KIF1B gene expression in the diabetic untrained group compared with the intact control group (p=0.03). Compared with the diabetic untrained group, training significantly decreased KIF1B gene expression (P<0.05) and blood glucose levels (P=0.0001) in the diabetic trained group.Conclusion: KIF1B mRNA up-regulation in sensory neurons of STZ-diabetic rats is a factor which can be involved in abnormal axonal transport. Endurance training as a nonpharmacotherapy strategy can modulate and return KIF1B to approximate normal levels.

Objective: Approximately three-fourths of women experience an episode of vaginal candidiasis.Candida albicans (C. albicans) is the etiological agent in over 80% of cases.C. albicanshas numerous virulence factors such as the agglutinin-like sequence (ALS) genes which code the large glycoprotein family that has a role in the adherence of Candida. The present study aims to evaluate expressions of the ALS 2, 9 genes in C.albicanswhich have been isolated from vaginal candidiasis.Methods: We collected 150 wet vaginal swabs from patients diagnosed with vaginal candidiasis. Samples were cultured on sabouraud dextrose and CHROMagar for morphological analysis. Then, DNA was extracted by glass bead and lysis buffer. We performed RFLP-PCR to confirm the presence of C. albicans. For investigation of semiquantitative expression of ALS2 and ALS9 genes, we performed RT-PCR by using specific primers.Result: From 55 clinical isolates of C. albicans, 36.36% expressed both the ALS2 and ALS9 genes. There were 23 (41.81%) isolates that expressed only the ALS2 gene and 21 (38.1%) expressed the ALS9 gene.Conclusion: Expressions of the ALS9 (41.8%) and ALS 2 (38.1%) genes in Candida isolates may indicate that these genes play a critical role in adhesion and biofilm formation of vaginal infection. However the presence of both genes in 36.36% of the isolates suggests a positive role for these genes in augmentation of their activities.

Objective: In vitro ovarian follicle culture provides a tool to investigate folliculogenesis and may be used as an assisted reproductive technology (ART). Thisstudy aims to comparesurvival and development rates in mouse ovarian follicles after two and three dimensional in vitro cultures.Methods: Preantral follicles were isolated from the ovaries of 14-day old female mice and cultured inα-MEM medium supplemented with 5% FBS for 12 days in a two dimensional and three dimensional culture with different concentrations of sodium alginate (0.25%, 0.5%, or 1%). The follicle diameter, survival and maturation rate during culture were analyzed and compared with one-way analysis of variance (ANOVA). P<0.05 was considered as statistically significant.Results: The mean diameter of preantral follicles that capsulated with 0.5% alginate was significantly higher than other concentrations in each group on days 6 and 12 (P<0.001).The percentages of follicles which released metaphase II (MII) oocytes in the two dimensional groups and in the three dimensional groups at 0.25%, 0.5% and 1% concentrations of sodium alginate were 29.03%, 33.33%, 44.18% and 35.89% respectively.The percentage of MII oocytes was significantly higher at the 0.5% concentration of sodium alginate (P<0.001).Conclusion: Follicles encapsulated in 0.5% sodium alginate in three dimensional culture displayed the highest survival development and maturation rate.

Objective: Diabetes plays an important role in the progression of tissue damage.Influencing factors of the cellular matrix can lead to changes in the structure and function of tissues or their failure. Herbal consumption can be effective in reducing tissue failure.The aim of the this study is to research the effect Portulaca oleracea seeds on the levels of matrix metalloproteinase 2, 9 and tissue inhibitor matrix metalloproteinase 1 in patients with type 2 diabetes.Methods: This semi-empirical study included 15 women diagnosed with type 2 diabetes in the experimental and control groups. Participants' mean age was 45 years.Portulaca oleracea seeds at a total dose of 7.5 g per day (2.5 mg with lunch and 5 mg with dinner) were consumed for eight weeks. Blood sampling was carried out before and after the eight-week period. Participants fasted for 12 hours prior to blood sampling. The t-test was used to analyze study results. P<0.05 was considered statistically significant.Results: After eight weeks, the levels of matrix metalloproteinase 2, 9 significantly reduced in the experimental group (P<0.05). However, there was no significant difference between groups. Tissue inhibitor matrix metalloproteinase 1 levels increased significantly in the experimental group. There was also a significant difference between the experimental and control groups (P<0.05).Conclusion: The results have shown that Portulaca oleracea seed did not adequately improve matrix metalloproteinases in diabetic patients. Thus, more research is needed to derive accurate conclusions.

Objective: E-cadherin is widely down-regulated and tightly associated with tumor invasion and metastasis in multiple human cancer types. Recent studies have shown that aberrant methylation of the E-cadherin gene promoter contributes to its silencing.However, information regarding epigenetic inactivation of E-cadherin in colorectal cancer is insufficient. Herein, we correlate association of the methylation of the ECadherin promoter with pathological features of colorectal cancer as well as history and demographic data.Methods: We used methylation specific polymerase chain reaction (MSPCR) to examine methylation status of the 5’ CpG island of E-cadherin along with its expression by using RT-PCR following surgical resection of 66 unrelated patients with colorectal cancer.Results: Results showed that 35 out of 66 tumor DNA samples (53%) showed aberrant methylations. In contrast, all normal tissues were unmethylated.Conclusion: The obtained results show a similarity with the Japanese (54.5%) and Greek (55.7%) populations. The results have confirmed methylation of this gene in sporadic colorectal cancer cases (40.8%) in the Iranian population by researchers in Shiraz. These data suggest that epigenetic silencing via aberrant methylation of the E-cadherin promoter plays a critical role in the inactivation of this tumor suppressor gene in colorectal cancer.

Objective: This study presents an efficient, cost-effective method to improve proliferation and colonization of spermatogonial stem cells (SSCs) in vitro.Methods: Isolated SSCs from neonate mice were cultured in DMEM culture medium with 10% fetal bovine serum (FBS). In the first phase of the study, the temperature was controlled by low intensity pulsed ultrasound stimulation (LIPUS) of the plate that contained the culture medium. In the next phase, SSCs were stimulated by LIPUS with 200 mW/cm2 with 20% and 40% duty cycle for five days. Proliferation and colonization of SSCs were on the seventh day.Results: LIPUS treatment of mouse SSCs increased the proliferation rate and colonization of SSCs in the experimental groups compared to the control group. Average proliferation rate in the 20% duty cycle group was 1.46±0.06, in the 40% duty cycle group it was 2.00±0.1 and for the control group, it was 1.26±0.06. The average number of colonies in the 20% duty cycle group was 24±7.7, whereas the 40% duty cycle group had 62±1.4 colonies and the control group had an average of 19±5.5 colonies. Average colony diameters were as follows: 186.6±2.07 mm (20% duty cycle group), 185.3±4.4 mm (40% duty cycle group) and 190.0±2.0 mm (control group). Our results showed a significant increase in proliferation rate and number of colonies in the experimental groups compared to the control group (P<0.05), whereas no significant differences were observed between groups in colony diameters.Conclusion: These results suggested that LIPUS treatment can be an efficient, costeffective method to improve proliferation and colonization of SSCs during in vitro culture.