Effect of follicular fluid and platelet-activating factor on lactate dehydrogenase C expression in human asthenozoospermic samples

Background: Application of follicular fluid [FF] and plateletactivating factor [PAF] in artificial insemination improves sperm motility. Lactate dehydrogenase C [LDH-C] is a key enzyme for sperm motility. In this study, the effects of FF and PAF on the sperm motility index and LDH-C expression were investigated. Moreover, LDH-C expression was compared between asthenozoospermic and normozoospermic samples

Methods: The expression of LDH-C was examined by quantitative real-time polymerase chain reaction [q-RT PCR] and western blotting after it was treated with optimized concentrations of FF and PAF in twenty asthenozoospermic samples. Also, LDH-C expression was evaluated in five normozoospermic samples

Results: Samples with 75% FF and 100 nM of PAF had an increase in their percentages of progressive and slowly motile sperms and a decrease in their percentages of non-progressive and non-motile sperms. Moreover, LDH-C mRNA transcripts were not changed following PAF and FF treatment, and LDH-C protein was detected in highly progressive motile specimens treated with FF in the asthenozoospermic samples. Furthermore, LDH-C expression was more detectable in the normal sperms

Conclusion: Our results indicated that PAF had more beneficial effects than FF on sperm motility in the asthenozoospermic samples [P=0.0001], although the LDH-C expressions of the sperms were not changed significantly in both groups. We found no association between LDH-C expression and sperm motility after FF and PAF actions. This finding, however, requires further investigation. The fact that LDH-C protein was detected in the normozoospermic, but not asthenozoospermic, samples could be cited as a reason for the infertility in these patients

Cardiomyocyte marker expression in mouse embryonic fibroblasts by cell- free cardiomyocyte extract and epigenetic manipulation

Background: The regenerative capacity of the mammalian heart is quite limited. Recent reports have focused on reprogramming mesenchymal stem cells into cardiomyocytes. We investigated whether fibroblasts could transdifferentiate into myocardium

Methods: Mouse embryonic fibroblasts were treated with Trichostatin A [TSA] and 5-Aza-2-Deoxycytidine [5-aza-dC]. The treated cells were permeabilized with streptolysin O and exposed to the mouse cardiomyocyte extract and cultured for 1, 10, and 21 days. Cardiomyocyte markers were detected by immunohistochemistry. Alkaline phosphatase activity and OCT4 were also detected in cells treated by chromatinmodifying agents

Results: The cells exposed to a combination of 5-aza-dC and TSA and permeabilized in the presence of the cardiomyocyte extract showed morphological changes. The cells were unable to express cardiomyocyte markers after 24 h. Immunocytochemical assays showed a notable degree of myosin heavy chain and ?-actinin expressions after 10 days. The expression of the natriuretic factor and troponin T occurred after 21 days in these cells. The cells exposed to chromatin-modifying agents also expressed cardiomyocyte markers; however, the proportion of reprogrammed cells was clearly smaller than that in the cultures exposed to 5-aza-dC, TSA, and extract

Conclusion: It seems that the fibroblasts were able to eliminate the previous epigenetic markers and form new ones according to the factors existing in the extract. Since no beating was observed, at least up to 21 days, the cells may need an appropriate extracellular matrix for their function

use of aloe vera extract as a novel storage media for the avulsed tooth

Background: Tooth avulsion is one of the most severe dental traumas which most often occur in children. When immediate replantation is not possible, storage in a proper media may lead to a prolonged survival rate. Aloe Vera is a cactus like plant with green, tapered leaves that are filled with a transparent viscous gel. This medicinal plant has significant anti-inflammatory, antioxidant, antibacterial and antifungal effects. The purpose of this study was to assess the effectiveness of different concentrations of Aloe Vera extract compared to DMEM [cell culture medium] and egg white

Methods: The periodontal ligament [PDL] cells were cultured and certain number of cells were treated with Aloe Vera extract [in four different concentrations], egg white and culture media for 1, 3, 6, and 9 hours. Cell viability was determined by using the [3-[4, 5-dimethylthiazolyl-2]-2, 5-diphenyltetrazolium bromide] assay. Moreover, One-way ANOVA and post hoc [LSD] test were used for analyzing the study groups

Results: The results indicate that culture media and Aloe Vera extract [10, 30, and 50% concentration] were statistically similar and significantly preserved more PDL cells compared to other experimental storage media

Conclusion: Aloe Vera 10, 30, and 50% may be recommended as a suitable storage media for avulsed teeth

comparison of the multiple oocyte maturation gene expression patterns between the newborn and adult mouse ovary

The interaction between follicular cells and oocyte leads to a change in gene expression involved in oocyte maturation processes. The purpose of this study was to quantify the expression of more common genes involved in follicular growth and oocyte developmental competence. In this experimental study, the expression of genes was evaluated with qRT-PCR assay in female BALB/c mice pups at 3-day of pre-pubertal and 8 week old virgin adult ovaries. The tissue was prepared by H and E staining for normal morphological appearance. The data were calculated with the 2-delta Ct formula and assessed using non-parametric two-tailed Mann-Whitney test. The p<0.05 was considered as significant. The data showed a significant increase in the level of Stra8 and GDF9 in adult compared with newborn mice ovaries [p=0.049]. In contrast, a significant decrease in the level of Mvh, REC8, SCP1, SCP3, and ZP2 was observed in adult mice ovaries compared to those in the newborn mice ovaries [all p=0.049 except SCP1: p=0.046]. There was no significant difference in the level of OCT4 and Cx37 expression between adult and newborn mice ovaries. The modifications in gene expression patterns coordinate the follicular developmental processes. Furthermore, the findings showed higher expression level of premeiotic gene [Stra8] and lower level of meiotic entry markers [SCP1, SCP3, and REC8] in juvenile than newborn mouse ovaries

Stereological study of the effect of ginger's alcoholic extract on the testis in busulphan-induced infertility in rats

In traditional medicine zingiber officinale used to regulate female menstural cycle and treat male infertility. Recent studies have suggested the possible role of ginger extract in improving the testicular damage of busulfan. The aim of this study was to evaluate the effects of zingiber officinale on the sperm parameters, testosterone level and the volume of the testes and seminiferous tubules by stereological methods. Fifty rats were divided into four groups. All the rats were given a single intraperitoneally injection of 5mg/kg busulfan solution. The first group was kept as busulfan control, while the other groups were orally administrated ginger extract in graded doses of 50, 100 and 150mg/kg b.wt, for 48 consecutive days. At the end, all animals were anesthetized and their testes and vas deference were removed, fixed, embedded, and stained. The volume of testes and seminiferous tubules were estimated by cavalieri methods. The result showed, that zingiber officinale increased the volumes of seminiferous tubule in 100mg/kg treated group compared to control group. Sperm count [706x10[5] and 682x10[5]] and the level of testosterone [50.90 ng/mL and 54.10 ng/mL] enhanced in 100 mg/kg and 150 mg/kg treated groups compared to control group [p=0.00]. It seems that zingiber officinale stimulate male reproductive system in induce busulfan infertility

Effects of L-carnitine and pentoxifylline on the activity of lactate dehydrogenase C[4] isozyme and motility of testicular spermatozoa in mice

Extracted sperm from the testis have poor motility. Moreover, their motility changes during their journey through epidydimis. Meanwhile, they face high concentration of L-carnitine. In addition, lactate dehydrogenase C[4] [LDH-C[4]] gene disorders has been shown to cause impaired sperm motility, leading to infertility in male mice. The aim of this study was to evaluate sperm motility and LDH-C[4] enzyme activity upon L-carnitine [LC] and Pentoxifylline [PTX] administrations in mice. We extracted testicular sperm of 48 mice and divided them into three equal parts. One part was incubated with Ham's F10 medium [control], the other parts were treated with Ham's F10 containing LC and PTX with a final concentration of 1.76 mM, for 30 min at room temperature. Sperm motility was assessed according to the World Health Organization [WHO] criteria. Sperm LDH-C[4] enzyme activity was measured by spectrophotometery method. Statistical analyses were performed using ANOVA and Fisher's LSD test, and a p-value less than 0.05 was considered as a statistically significant difference. Sperm motility increased after 30 min of incubation in LC- and PTX-treated group [p<0.001]. LC and PTX administrations showed a significant increase in the LDHC[4] enzyme activity of sperm compared to that of the controls after 30 min [P=0.04 and 0.01, respectively]. The effects of LC and PTX on motility of sperm can be explained by an increase in LDH-C[4] enzyme activity that may influence male fertility status. We suggest that LC as a non-toxic antioxidant is more suitable for use in assisted reproductive technique protocols than PTX