Cell Journal (Yakhteh)
Year:2004 | Volume:6 | Issue:21|Papers Count:9

Introduction: This investigation was performed to study the anatomy and ultrastructure that is interstitial cells of cajal (ICC), of colon in Iranian patients with Hischsprung's disease (HD).   Material and Methods: Samples were obtained from healthy and pathologic segments of colon in 10 childern under 10 years of age in which the distal part of colon was resected for surgical treatment of H.D. The specimens were prepared for electron microscopic studies and the ultrathin sections were obtained. Then the location & morphology of ICC in pathologic segments were compared with healthy ones.   Results: In the affected segment, on ICC was detected among the smooth muscle cells and nerve endings, but the ultrastructural study showed mild differences with healthy parts.   Conclusion: Lack of intermediate position of ICC in pathologic segments, considering it's inhibitory role on smooth muscle cell function, could be the reason for spasm in involved areas.    

Introduction: Considering the role of HLA matching in transplant outcome, the quality of HLA-DR typing is clearly an important issue.In recent years serologic methods have been replaced with DNA based typing methods.The main objective of this study was to compare HLA-DR typing data obtained from existing serologic method with that obtained by the new PCR-SSP method.   Material and Methods: Fifty-five peripheral blood samples were collected from randomly selected individuals who reffered to the transplantation laboratory of Isfahan in Aliasghar Hospital and was typed for HLA-DR antigens by both methods. HLA-DR typing by serologic method was performed using 30 different antisera and for PCR-SSP method specific primers were used for HLA-DRB1*01-10(except DR6, 8, 10) and also for HLA-DR52 and DR53. After DNA extraction 13 pairs of specific primers were used for each sample separately and PCR reaction was performed. In this study the third intron of DR locus was used as internal positive control. After PCR amplification, electrophoresis on 2% agarose gel was performed on reaction products, and after photography of gel ,interpretation and comparison of results was performed.   Results: The results of 31 samples (56.3%) correlated completely with serologic method. 12 samples (22%) had been assigned heterozygous in serology and homozygous in molecular typing, 7 samples (12.7%) were heterozygous in both methods but different in one allele, 2 samples (3.6%) were homozygous in serology and heterozygous in molecular typing, and also one sample (1.8%) was homozygous in both methods; that is serologically DR14 positive and moleculary DR11 positive. And finally 2 samples of 55 (3.6%) were not definable by in serologic method.     Conclusion: The data obtained from the conventional and the new method were compared. Sensitivity,specificity and rate of discrepancy between the two methods were discussed.  

Introduction: It is well known that glycoconjugates on the cell surface of embryonic cells as well as extracellular matrix (ECM) are involved in many developmental phenomena including cell differentiation and maturation. These molecular events have not received proper attention during pituitary cell differentiation. The purpose of this study was to  investigate glycoconjugate distribution on cell surface during adenohypophysis embryogenesis. Material and Methods: Using lectin histochemistry, 5µm normalin fixed, paraffin embedded Rat embryonic sections from day 10 to 20 of gestation (N=90) were incubated with different HRP-lectins from Triticum vulgaris (WGA) Arachis hypogaea (PNA) and Griffonia simplicifolia (GSA1- B4) specific for terminal sugars Sialic acid, N-acetyl galactosamine and α-D- galactose ofcomplex glycoconjugates respectively. On the basis of intensity of staining that was determined by Gong's method, sections were graded and non-parametric statistical test (Kruskal Wallis) was used to compare differences between samples. Results: The results demonstrated that the reaction of adenohypophysis cells with WGA started from gestational day13(E13) and increased with proceeding differentiation during the following days (P<0.05). A few cells reacted with PNA from E13 and increased to E14 (P<0.05) and then identified to E17 and decreased afterward (P<0.05). GSA1-B4 didn't react with any cells during development. Our findings also indicated that glycoconjugates with terminal sugars sialic acid and N-acetylgalactosamine may play a critical role in adenohypophysis development. Conclusion: The appearance and changes of Glycoconjugates on the cell surface with terminal sugars such as acid sialic and N-acetyl galactosamine may play a key role in tissue interactions and lead to  developmental changes in certain  embryoniorgans such as adenohypophysis

Introduction: In this study the immature mouse oocytes (Germinal Vesicles: GV), which were arrested in metaphase (MI), were activated with DC pulses and the effect of DC pulse frequency on immature oocyte activation and their subsequent in vitro development were studied.     Material and Methods: Immature oocytes successfully passed the meiosis processes. That is, germinal vesicle stage of oocytes changed to germinal vesicle breakdown (GVB) and finally the first polar body extruded and reached metaphse II, (MII) and formation of 2-cell, 4-cell, 8-cell embryos. Immature Oocytes were separated from NMARY mice (4-6 week old) ovary in different phases. They were placed in M2 medium droplet and then activated with DC pluse (50V, 30µs).    Results: Immature oocytes (GV) began meiosis resumption and GVs changed to GVB and extruded first polarbody and some of them reached metaphase II.  After 24 hours evaluation by inverted microscope was performed. Ovulated oocytes were inseminated with the capacitded epididymal sperm of the same strain of mice. One to four pulses with a duration of 30مs induced more oocyte activation (67% to 89%), maturation (68 to 77%) and embryo formation (44% to 84%). Embryo formation increased significantly with more than two DC pulses (55-88%) compared with a frequency of less than two (51%) groups.    Conclusion: This study revealed that electroactivation is helpful for in-vitro maturation, fertilization, embryo formation and development in female infertility, especially in those with irrgular secretion of follicle stimulating and luteinizing hormone (FSH, LH)

Introduction: The process of wound healing sets in motion a complex and dynamic series of events, which includes the remodeling of the extracellular matrix degradation of matrix proteins, predominantly collagen fibers, mediated by matrix metalloproteinase (MMPs). Our aim was to demonstrate whether fibroblasts derived from various strains of human fetal foreskin (FS) and adult skin (AS), when cultivated in a 3-D tissue model, exhibit different matrix metalloproteinase activities as compared to plastic environment. Material and Methods: A total of 13 cell strains, 8 and 5 strains of fetal foreskin and adult dermal fibroblasts, respectively, were grown under sterile conditions. Synchronized cultures up to 80-90% confluency of either study groups were used for further analyses. The rates of collagenase activity and gelatinase zymography of conditioned medium derived from adult cells relative to fetal fibroblasts in plastic vs. 3-D gel were determined and compared. Results: Adult fibroblasts showed higher collagenase I activity vs. FS strains both in plastic and in 3-D substrate (14.38±1.33. vs. 9.79±2.27 in plastic, and, 27.05±1.28 vs. 25.2±2.97.± in 3-D gel, (p<0.05). FS strains produced nearly three times more collagenase I in 3-Dgel than in plastic; this increased activity rate was less than two times for AS strains. This finding was further added by enhanced gelatinase activities seen in both strains in 3-D gel.  However, FS strains expressed more gelatinolytic activities in 3-Dgel, indicating different cellular functions derived from varied morphologies. Conclusions: These findings showed physiological differences in matrix components and could be due to phenotypic variations adopted by cells in response to local milieu, e.g. aging process. However, more studies are needed to address the question whether these phenotypic discrepancies could be further modified by the action of either growth modulators or cytokines

Introduction: In the present study the effect of nucleus basalis of Meynert (NBM) destruction on the temporal characteristics of response integration evoked by multiple whisker stimulations in rat barrel cortex was studied. Material and Methods: Wistar male rats weighing 250-350 g were used. The animals were randomly distributed into two groups: In the first group bilateral NBM lesions was induced by 5µg ibotonic/5µl phosphate-buffered saline; rats in the other group were sham perated. Extracellular single-unit recording and controlled whisker stimuli were used to compare response properties of cells in barrel cortex between the two groups. Two neighboring principal and adjacent whisker in the same row were deflected  alone or in paired combination in a condition-test paradigm, to assess excitatory and inhibitory receptive field (RF) characteristics. Results: Results show that NBM lesion significantly decreased response magnitude (P<0.05) but did not  affect response latencies. Units discharge to subsequent deflection of adjacent whisker were reduced in a time dependent fashion in both groups. But the magnitude of responses were significantly reduced at each inter-deflection interval in rats with NBM lesion (P<0.001). The excitatory FR of  barrel units, which are the most tightly focused on the principal whisker, were the most greatly defocused by NBM lesion and  NBM lesion increased surround inhibition. Conclusion: Our results suggest that NBM has an important role in changing the balance of excitation and inhibition in the barrel cortex and can affect on the temporal integration of tactile inputs in the cortex.

Introduction: The aim of this study was to evaluate the effect of sperm chromatin anomalies on fertilization outcome post ICSI.Material and Methods: Therefore along with semen parameters, CMA3 staining for protamine deficiency, aniline blue staining for excessive histones, SDS for sperm chromatin stability and SDS+EDTA for ability of sperm to undergo decondensation were carried out on 55 semen samples from patients referring for ICSI to Isfahan fertility and infertility cente.Result: The results showed that among the aforementioned tests and semen parameters, only CMA3 showed a significant correlation with fertilization outcome post ICSI. Patients were also grouped according to CMA3 level of <30% or >30 % or fertilization rate of <50%, or >50%. The results showed that the mean percentage fertilization and mean percentage of CMA3 positivity is different inboth groups respectively. The area under ROC curve showed that CMA3 is a highly sensitive and specific test for prediction of fertilization outcome post ICSI.Conclusion: Therefore, it can be concluded that sperm protamine deficiency has profound effect on fertilization failure in ICSI.

Introduction: Selected probiotic lactobacilli have inhibitory effect and therefore may be use as biological preservative, so, the aim of this study was to present some data on isolation, growth, antimicrobial activity, effect of pH, heat, and sensitivity to proteolytic enzymes of lactobacillus. Material and Methods: Isolation of lactic acid bacteria from sausage was done by using 1g of samples in MRS medium. Each isolate of lactobacillus species was identified by biochemical tests and comparison of  their sugar fermentation pattern. Antibacterial activities was done by an agar spot, well diffusion and blank disk method. Stability of antibacterial activity was measured in boiling water and autoclaving for 15 min. Enzyme sensitivity of supernatant fluid and concentrated cell free culture after treatment with a-amylase, lysozyme and trypsin was determined.  Results: The isolated bacteria were Lacto. plantarum, Lacto delbruekii, Lacto. acidophilus, Lacto. brevis. The isolated bacteria had strong activity against indicator strains. The antibacterial activity was stable at 100ºC for 10 minutes and at 56ºC for 30 minutes, but activity was lost after autoclaving. The maximum production of bacterocin was obtained at 25-30°C at pH 6.5. Discussion: If Lactobacilli that are used to induce sausage fermentation have antimicrobial activity with heat stable bacteriocin, these bacteria may be considered to be a healthy probiotic diet. Lactobacilli originally isolated from meat products are the best candidates as probiotic bacteria to improve the microbiological safety of these foods.