Effect of anti-estrogen and anti-progesterone on spermatogenesis, testosterone production and expression of steroidogenic enzyme genes in adult male rats.

The present study was planned to investigate the anti-spermatogenic and anti-steroidogenic effects of Clomiphene Citrate (CC) an anti-estrogen and Mifepristone (MT) an anti-progesterone in the testis of male rats. Following the oral administration of 1.0 mg and 5.0 mg/kg b.w/day of each for the duration of 30 and 60 days, quantitation of spermatogenesis, RIA for serum and intra-testicular testosterone levels, western blotting and RT-PCR for expression of StAR, 3ß-HSD and P450arom enzymes in the testis was done. Clomiphene Citrate at 5.0 mg/kg b.w/day for 60 days significantly reduced testosterone (T) levels however the effect was not significant with the lower doses. Reproductive parameters in animals treated by Mifepristone remained mostly unaffected, however, a significant decline in testosterone levels and altered expression of selected genes was observed in 5.0 mg for the 30d treatment group. Clomiphene Citrate at higher doses affected the weights of the testis and secondary sex organs. Seminiferous tubules revealed hypo-spermatogenesis with a significant decrease in the number of maturing germ cells and a reduction in tubular diameter. Attenuation in serum testosterone was associated with the downregulation of expression in StAR, 3ß-HSD, and P450arom mRNA and protein levels in the testis even after 30 d of CC administration. The results indicate that the anti-estrogen (Clomiphene Citrate) but not anti-progesterone (Mifepristone) induces hypo-spermatogenesis in rats which are associated with a downregulation of expression of two of the steroidogenic enzymes, 3ß-HSD and P450arom mRNA and StAR protein.

Antifungal Biofilm Strategies: A Less Explored Area in Wound Management.

BACKGROUND: The treatment of wound-associated infections has always remained a challenge for clinicians, with the major deterring factor being microbial biofilms, majorly bacterial or fungal. Biofilm infections are becoming a global concern owing to resistance to antimicrobials. Various fungal pathogens form fungal biofilms, namely Candida sp., Aspergillus fumigates, Trichosporon sp., Saccharomyces cerevisiae, Cryptococcus neoformans, among others. The rising cases of fungal biofilm resistance add to the burden of wound care. Additionally, with an increase in the number of surgical procedures, transplantation, and the exponential use of medical devices, the fungal bioburden is rising. OBJECTIVES: The review discusses the methods of biofilm formation and the resistance mechanisms against conventional treatments. The potential of novel delivery strategies and the mechanisms involved therein are also highlighted. Further, the prospects of nanotechnology-based medical devices to combat fungal biofilm resistance have been explored. Some clinical trials and up-to-date patent technologies to eradicate biofilms are also mentioned. CONCLUSION: Due to the many challenges faced in preventing/eradicating biofilms, only a handful of approaches have made it to the market. Eradication of fungal biofilms are a fragmentary area that needs further exploration.

Quercetin supplementation restores testicular function and augments germ cell survival in the estrogenized rats.

Quercetin, as a flavonoid, has been recognized to possess dual properties of an oxidant and antioxidant as well. The role of quercetin (QC), as an antioxidant in countering estradiol-3-benzoate (EB) induced adverse effects and germ cell apoptosis in adult rat testis was presently investigated. Adult rats received EB (0.075 mg/rat/5th day) alone or EB+QC (15 mg/kg bw/alternate day) simultaneously for 30 days. Revival of spermatogenesis following QC intervention was associated with a significant restoration in serum and intra-testicular levels of testosterone. Decline in lipid peroxidation and simultaneous improvement in the activities of superoxide dismutase, catalase and glutathione s-transferase were very much evident. Identically, total antioxidant capacity and glutathione demonstrated a marked improvement. QC augmented germ cell survival leading to a decrease in cell apoptosis. Expression of downstream apoptotic markers, caspase-3 and poly-ADP-ribose polymerase (PARP) presented a significant reduction. Down regulation with respect to upstream markers, caspase-8 and -9, Fas, FasL, Bax, and p53 was similarly observed. Taken together, the above findings indicate that with the dose presently used quercetin with its antioxidant and antiestrogenic properties restored testicular function leading to revival of spermatogenesis. It also augmented germ cell survival primarily mediated through downregulation in the expressions of upstream, downstream and other markers in the pathways of metazoan apoptosis.

Clomiphene citrate potentiates the adverse effects of estrogen on rat testis and down-regulates the expression of steroidogenic enzyme genes.

OBJECTIVE: To investigate the antiestrogenic effect of clomiphene citrate (CC) in male rats estrogenized with estradiol-3-benzoate (EB). DESIGN: Prospective experimental study. SETTING: Laboratory. ANIMALS: Adult male albino rats (Holtzman strain). INTERVENTION(S): CC was given alone or in combination with EB. MAIN OUTCOME MEASURE(S): Testicular function and steroidogenic enzyme gene expression were evaluated in control versus treated groups. RESULT(S): EB after 30 days of treatment induced a rise in TUNEL-positive germ cells adversely affecting spermatogenesis with complete absence of elongated spermatids or sperms. CC alone had only a moderate effect. In contrast, CC+EB synergistically inflicted more adverse effects as apoptotic germ cells per tubule rose further. Significant down-regulation in expression of testicular steroidogenic enzyme genes StAR, p450scc, 3ß-HSD, and p450c17 was observed. In the EB-alone group, aromatase gene expression in the testis was up-regulated but reversed in brain and liver tissues. CC alone had little modulatory effect on aromatase expression. On the other hand, CC+EB countered the EB-induced rise of aromatase expression in the testis. CONCLUSION(S): The above findings indicate that CC in the presence of estrogen synergistically potentiates more adverse effects in testis, inhibiting expression of upstream steroidogenic enzyme genes and leading to disruption of steroidogenesis.

Role of estrogen in the regulation of spermatogenesis in the Indian wall lizard Hemidactylus flaviviridis.

The role of estrogen in the Indian wall lizard, Hemidactylus flaviviridis during PMSG induced spermatogenesis in the regression phase and during normal spermatogenesis in the active breeding phase was investigated. Blood hormone levels demonstrated a high testosterone to estrogen ratio in the breeding and vice verse during the regressed phase. PMSG treatment (30 IU in 100 µl saline/lizard/alternate day for 30 days) during the regressed phase stimulated spermatogenesis which was associated with a significant (p<0.001) rise in plasma testosterone levels. Complete spermatogenesis with sperms was resolved in many tubular sections. However, co-administration of PMSG plus estrogen in high doses (2 µg of estradiol benzoate/alternate day) for the same period not only curtailed germ cell proliferation significantly but also induced apoptosis in germ cells. There was no significant reduction in testicular weight but sperms were found completely absent in all the tubules. Decline in the plasma testosterone was more pronounced in high compared to low estrogen treated groups. Further, low estrogen administration had little effect either on raising the plasma levels of estrogen or subsequently on spermatogenesis which was identically observed in the breeding phase too. Estrogen intervention (2 µg) in the breeding phase also profoundly suppressed spermatogenesis leading to a severe depletion in germ cells. Simultaneously, there was a significant rise in germ cell apoptosis which was associated with an up-regulation of extrinsic (caspase 8, Fas, FasL) and intrinsic (caspase 9, Bax, Bcl2) markers in these cells. Taken together, the above data indicate that the estrogen plays a key role in regulating spermatogenesis in the wall lizard retarding it during testicular quiescence and eliminating germ cells through apoptosis during the active breeding phase.