Cathepsin-D is a lysosomal protease enzyme belonging to the aspartic family. Due to its serious pathological and physiological effects, it has recently gained remarkable importance. This study is intended to isolate cathepsin-D from human LEUKOCYTES in 3 steps. The procedure was done through ionic exchange chromatography and completed with sephadex G-200 following gel chromatography. A single bond on polyacrilamid gel and a single peak on the 280 nm region were observed. Its total inhibition with pepstatin indicated the full purification of cathepsin-D.

Transendothelial migration of LEUKOCYTES (diapedesis), have vital role in both the innate and adaptive immune responses. This process consists of overlapping steps, including: activation of LEUKOCYTES, formation of weak adhesions, and translocation along the endothelium followed by stronger adhesions resulting in transmigration of LEUKOCYTES. Although the surface molecules used by LEUKOCYTES and endothelial cells during diapedesis have been well characterized, the mechanisms by which they regulate this process is poorly understood. In this minireview we introduce some of the involved molecules and mechanisms that play roles in this process.

Infection is one of the important causes of morbidity and mortality in diabetic patients. It has been reported that poorly controlled patients are more susceptible to infection, hence we examined the chemiluminescence of LEUKOCYTES from insulin dependent diabetic patients in response to a soluble (phorbol meristate acetate) and particular stimulus (opsonized zymosan)The patients were divided into two separate groups, only controlled and well controlled, in regard to their blood glucose. Using PMA as a stimulus LEUKOCYTES from both groups, patients showed no significant difference comparing with healthy controls, but that of the two groups of patients were significantly different (P<0.05). When opsonized zymosan was used as the stimulus, no statistically significant difference was observed between all of the coupled groups. However, the chemiluminescence of LEUKOCYTES from poorly controlled patients was lower than the other groups.

Introduction: Chronic obstructive pulmonary disease (COPD) is characterized by airflow limitation that is not completely reversible by administration of inhaled bronchodilators. Many studies propose that telomere length shortening might have occurred in COPD patients. We aimed to determine the telomere length in COPD patients and compare the results of non-smoking and smoking control subjects.Materials and Methods: In our case-control study, 84 clinically stable COPD patients were recruited on admission to Masih Daneshvari Hospital. Eighty-five healthy controls were also selected including 45 non-smokers and 40 smokers admitted for diseases other than COPD. Spirometry was done for all subjects.Telomere length was measured by quantitative real time PCR as described by Cawthon. The telomere repeat copy number (T) to single-gene copy number (S) ratio was calculated using the comparative Ct method.Results: The mean ±SD of age was 64.33±10.04 years in patients and 65.06±10.02 years in controls (P=0.693). The mean ±SD of FEV1 was 1.62±0.75 L in patients, 2.84±0.54 L in smoker controls and 2.83±0.56 L in non-smoker controls; significant differences were detected in this regard between cases and controls (P<0.001). T/S ratio was significantly lower in COPD patients (0.61±0.08) than in the control subjects (0.69±0.09) (P<0.001). However, telomere length was shorter in the patients than in controls in each age group (P<0.001).Additionally, there were no statistically significant differences in telomere length between the smoker and non-smoker control subjects. Regarding the correlation between BMI and telomere length, there were no significant differences among the patients and control groups.Conclusion: In conclusion, we found that telomere length in COPD patients was shorter than that in smoker and non-smoker controls, irrespective of age, sex, spirometric variables, BMI and history of cigarette smoking.

Background: Radioprotective effects of famotidine, an antagonist of H2 receptor clinically used for peptic ulcer treatment, was previously shown on radiation-induced micronuclei and chromosomal aberration in human peripheral blood lymphocytes and mouse bone marrow cells. This study was conducted to investigate radioprotective property of famotidine against radiation induced apoptosis in human peripheral blood LEUKOCYTES. Materials and Methods: Peripheral blood was obtained from 6 healthy volunteers including three males and three females. 12 µL of blood sample diluted in 1 ml RPMI-1640 supplemented with antibiotics and foetal calf serum was irradiated a dose of 8 Gy gamma rays generated from a Co-60 source at a dose rate of 1.27 Gy/min. After 48 h incubation in a 37 oC incubator, cells embedded in low melting point agarose were transferred to a slide precoated with normal agarose. Cells were lysed and subjected to electrophoresis under neutral condition. Slides were then stained with ethidium bromide and analysed under a fluorescence microscope. 500 cells were analysed for each sample for the presence of apoptosis. The data were statistically evaluated using Man-Whitney non-parametric and ANOVA tests.Results: Results show a significant increase in apoptosis induction following 8 Gy γ-irradiation comparing with controls (p<0.001). The presence of famotidine at 50 and 100 µg/ml did not show any protective effect against radiation induced apoptosis; however, the presence of famotidine at higher concentration (200 µg/ml) significantly deceased radiation induced apoptosis (p<0.001).Conclusion: The obtained results suggest that famotidine suppresses radiation-induced apoptosis at 200 µg/ml, probably via OH radical scavenging and an intracellular antioxidation mechanism. Famotidine appears to be a useful candidate for the future development of post-irradiation radioprotectors.

Background and Purpose: The clinical and epidemiologic features of Kawasaki disease (KD) suggest an infectious etiology; however, the agent(s) remain unknown. Our purpose was to isolate the causative bacterial gene from peripheral blood LEUKOCYTES of patients with acute KD, by Universal polymerase chain reaction (UPCR), in Tehran Children's Medical Center.Materials and Methods: Universal polymerase chain reaction (UPCR) assay was used to amplify the bacterial16S ribosomal RNA gene (rDNA).Results: Forty three (28 boys and 15 girls) were diagnosed with acute Kawasaki disease included in this study. The median age at diagnosis was 3.5 years (range: 0.5 -9 years). Twenty Nine (29) cases had typical KD criteria and 14 patients had atypical KD at diagnosis. Two of the 43 KD patients were positive for the Universal PCR assay for 16S rRNA, prior to intravenous g-globulin therapy (IVGT), while all specimens were negative by conventional blood culture. In our study, there was fever in 100%, conjunctivitis in 62.7%, rash in 83.72%, oral mucosal changes in 76.74%, peripheral changes in 37.20%, and cervical lymphadenopathy in 39.53% cases.Conclusion: The 16S rDNA sequence was positive in 4.65% of acute KD patients; this data shows that an infectious KD agent is traced in peripheral LEUKOCYTES. The question remains as to what true frequency ofthe16S rDNA sequence in KD is.

Introduction: An important factor involved in infertility is reactive oxygen species (ROS). ROS can damage sperm DNA, and involve in lipid peroxidatrion. ROS elevation is under the influence of leukocyte activation. The objective of this prospective study was to evaluate the level of ROS as well as leukocyte in normozoospermic (NO) and oligoasthenoteratozoospermic (OAT) ejaculates.Materials and Methods: The study population consisted of 75 individuals who were referred to research and clinical center for infertility in Yazd for semen analysis. 50 out of 75 men were NO, and the rest were OAT. ROS was measured with laminator, while leukocyte concentration was done with ENDTZ test.Results: The results showed that ROS level in OAT was significantly higher compared with NO (1253.49±200.95 vs. 75.64±149.52; p=0.00). Also, men with OAT were divided into 2 groups with sperm morphology and motility >5% and <5%. In group >5%, ROS level was significantly higher than group with <5% (3627.55±407.79 vs. 81.29±100.48; p=0.007). In addition, leukocyte concentration in NO was 0.07±0.22x106; while, it was 0.12±0.20x106 in OAT samples; p=0.35).Conclusion: The results indicate that although ROS is present in normal seminal samples, but it is significantly higher in OAT. This shows the vital role of antioxidants, which may improve the sperm quality. Further clinical studies will pinpoint the antioxidant capacity in improving the seminal contents.