[Designing of human cartilage tissue, by differentiation of adipose-derived stem cells with BMP-6 in alginate scaffold]

In the present study the effect of BMP-6 was investigated on chondrogenesis of adipose-derived stem cells. Mesenchymal stem cells derived from subcutaneous adipose tissue were cultured on alginate scaffold to induce chondrogenesis in experimental group, with chondrogenic medium having BMP-6 growth factor for 3weeks. In control group medium without BMP-6 was applied. The harvested constructs were examined with immunohistochemical and RT-PCR methods for assessment of cartilage-specific characteristics. The results of immunohistochemical method revealed the presence of typical cartilage extracellular matrix components such as type II collagen and aggrecan in constructs induced by BMP-6 growth factor on alginate scaffold. In addition evaluation of the results of RT-PCR analysis confirmed the expression of cartilage- specific genes, such as type II collagen and aggrecan, in the differentiated cells under the influence of growth factor BMP-6. It can be concluded that BMP-6 promotes chondrogenesis of ADSC in 3-D and adipose-derived stem cells could be used for cartilage tissue engineering

Determining the behavior management technique's acceptance of mothers referred to the department of pediatric dentistry in Qazvin [2007]

For dental treatment of the children who do not cooperate with the dentist, the use of behavior management techniques is inevitable. Behavior management techniques are the means by which the dental health team effectively performs treatment for the child and at the same time, instills a positive dental attitude. Determining the behavior management technique's acceptance of mothers referred to the department of pediatric dentistry in Qazvin [2007]. This was a cross-sectional descriptive study in which a film including 4 different behavior management techniques marked as: general anesthesia [GA], parents' separation [PS], hand-over- mouth [HOM], and voice control [VC] was produced. Before showing each technique, a brief explanation over the technique's characteristics, method, and the objective was given by a pedodontist in the film. A total of 50 mothers who were referred to the pediatric dentistry department of Qazvin University watched the film and later filled out a questionnaire. The data were analyzed using SPSS. Mothers' acceptance of P.S was 100%, V.C 92%, HOM 50%, and G.A 30%. There was a statistically significant relationship between the acceptance of voice control and child's order, child's age, and the number of children. Also, a significant relationship between the acceptance of general anesthesia and the child's order and mother's education was found. According to our data, the parents' separation and general anesthesia are the most and the least accepted technique, respectively

[Congenital adrenal hyperplasia report of a case]

Congenital adrenal hyperplasia [CAH] is an autosomal recessive disorder with an incidence of 1/15000, which is sometimes called the adrenogenital syndrome. This disorder which occurs during fetal development is a consequence of inborn errors in cortisol biosynthesis. Compensatory overproduction of ACTH by the fetus to overcome cortisol deficiency produces hyperplastic but inefficient adrenal cortices. More than 90% of cases result from mutations of CYP21 gene, leading to 21- hydroxyllase deficiency, which is essential not only for cortisol biosynthesis but also for aldosterone formation. Deficient or excessive mineralocorticoid production is the other major clinical problem associated with CAH syndrome. The distinctive clinical features of the CAH syndrome result from inappropriate production of both sex steroids and mineralocorticoids. In its classical form, CAH is severe and consists of the virilizing [increase of androgens] and salt wasting [lack of aldosteron] phenotype. When a probond exists, early prenatal diagnosis for CAH can be performed by direct molecular analysis in the first trimester. In recently reported case, the patient is a 10 year old young boy whose parent's chief complaint were unerupted maxillary right central incisor. After taking a complete history the patient was diagnosed with CAH. His 18 year old sister was affected too. Up to now, there has been no report regarding the oral and dental status of these patients in the literature. It is probable that delayed eruption and root formation of some teeth can be related to either this congenital disorder or drug consumption

[ efficiency of Zeta method in separation of sperm with normal morphology and chromatin structure]

Selection of mature sperm with normal morphology and chromatin structure is needed for assisted reproduction technology [ART] procedures. At present, various sperm separation methods exist to select mature sperm. The selection of mature sperm based on electrical charge of membrane is one of these methods. Therefore, the aim of this study was to evaluate the efficiency of the Zeta potential for selection of normal sperm in term of morphology and chromatin structure integrity. In this descriptive-analytical study, semen samples from 70 infertile couples referring to Isfahan Fertility and Infertility center were evaluated. Semen analysis was carried out according to WHO criteria. The remaining of semen samples were used for Zeta method. Then Sperm morphology, protamine deficiency and DNA fragmentation were assessed using papanicolaou staining, chromomycin A3 [CMA3] staining, and sperm chromatin dispersion [SCD] test, respectively. In addition, we compared the result of Zeta group with control group. Coefficients of correlation and paired-samples t-test were carried out using SPSS 11.5, to compare results among between two groups. The results of this study show that the mean of abnormal morphology, protamine deficiency and DNA fragmentation were 88.19 +/- 7.37, 67.17 +/- 17.34, 32.87 +/- 8.65 in control group, and 80.17 +/- 10.26, 58.40 +/- 18.20, 18.19 +/- 8.64, in the Zeta group, respectively. Zeta group can decrease significantly the percentage of abnormal morphology, protamine deficiency and DNA fragmentation compared to control group [P<0.05]. Zeta method can be used in separation of mature sperm with normal morphology, normal protamine contained and intact DNA

Effects of acrosomal activity and morphology on fertilization rates following ICSI

Fertilization failures after ICSI may be due to different factors related to oocyte, sperm or both of them. Considering the importance of sperm morphology, acrosomal activity in oocyte activation and fertilization rates, this study was done to evaluate the relationship between a series of events occurring during spermiogenesis such as sperm morphology and acrosomal activity as an index for acrosomal integrity and the relationship between sperm ability to induce oocyte activation with fertilization rates following ICSI. Semen samples from 68 infertile couples undergoing ICSI at Isfahan Fertility and Infertility Center were assessed. Some of each semen sample was analyzed for semen parameters according to WHO criteria, most of it was used for ICSI and the rest for Papanicolaou staining and gelatinolysis test to evaluate sperm morphology based on the strict criteria and acrosin activity respectively. The results were analyzed by SPSS software [Version 11.5] and correlation coefficient was determined. P-values less than 0.05 were considered statistically significant. Results from gelatinolysis showed that the mean halo diameter had a significant positive correlation with sperm concentration [r=0.343, p=0.004], motility [r=0.282, p=0.020], sperm morphology according to the WHO criteria [r=-0.314, p=0.009], fertilization rate [r=0.270, p=0.026] and the percentage of halo formation [r=0.853, p=0.001]. The results of the study revealed that during ICSI, spermatozoa with small acrosome, which are likely to have reduced gelatinolysis test parameters [Smaller percentages of halos and smaller mean halo diameters], have lower fertilizing potential. As gelatinolysis test is considered as an index for acrosomal and perinuclear theca integrity, lower fertilizing ability in these spermatozoa could be likely due to their reduced levels of sperm associated oocyte activating factors [SAOAFs]. Therefore, studies for the identification and measurement of SAOAF levels in these kinds of spermatozoa are proposed

[Effect of seminal vesicles and dithiotritol [DTT] on stability of sperm chromatin in IVF and ICSI patients]

Different studies have shown that there is no relation between sperm chromatin stability andfertilization rate in both IVF and ICSI patients. However, the relation between SDS tests, as a detergent, alongwith DTT as reducer of disulphide bridges has not been studied so far in ICSI patients. Since differentconcentrations of DTT can induce different degrees of sperm chromatin decondensation, the aim of this studywas to evaluate the effect of different concentrations of DTT on sperm chromatin decondensation in IVF andICSI cases.During this study, 85 patients were divided into two groups according to their treatment procedure[IVF or ICSI].Semen samples of each patient was evaluated for sperm chromatin tests including SDS,SDS+EDTA and SDS+DTT for assessment of free thiole groups level [-SH], amount of non covalent bondbetween Zn and thioles[-SH Zn SH-] and levels of disulfide bond [-S-S-] in sperm chromatin, respectively. Inthis study, seminal fructose concentration, corrected seminal fructose level and true corrected fructose level asindicators of seminal vesicle function on sperm chromatin stability were assessed.No correlation was observed between any of the above tests and rate of fertilization, both in IVF andICSI cases. However, in IVF patients, a significant correlation was observed between SDS, SDS+DTT test andseminal fructose level, while in ICSI patients, only a significant correlation was observed between SDS+DTTand corrected or true fructose concentration.Since no correlation was observed between sperm chromatin test and fertilization rate, it issuggested that the chromatin status of these samples are adequate for fertilization to take place and extent ofdisulphide bridges has no effect on fertilization rate. However, the amount of disulphide bound present in spermsof ICSI and IVF patients are different, and this difference is related to seminal vesicle performance in thesepatients

Effect of sperm protamine deficiency on fertilization and embryo development post-ICSI

The aim of this study was to evaluate the effect of sperm protamine deficiency on fertilization and embryo development post ICSI. Semen samples from 27 patients candidate for ICSI were assessed for semen parameters. Sperm processing was carried out using pure sperm and percentage of protamine deficient sperm was assessed by Chromomycin A3 post processing. Correlation between fertilization rate, embryo quality and cleavage score with protamine deficiency was assessed using SPSS statistical programme. A significant negative correlation was observed between fertilization rate and embryo quality score on day 3 with percentage of protamine deficient sperms. Semen samples with high percentage of protamine deficient sperms have lower fertilization rate and lower potential to develop good quality embryos

[Effect of human sperm chromatin anomalies on fertilization outcome post ICSI]

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 center

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 in both 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