Acrosome reaction is crucial to the penetration of spermatozoa through the zona pellucida (ZP). Glycosylation of ZP glycoproteins is important in spe

ObjectiveTo evaluate the anti-diabetic potential of leaf extract from Urena lobata (U. lobata) through dipeptidyl peptidase IV (DPP-IV) inhibitory activity.MethodsU. lobata leaf was extracted in hot water and ethanol. The activity of DPP-IV inhibitor was tested by in vitro study using gly-pro-p-nitroanilide as substrat of DPP-IV and vildagliptin, as standard reference. A product of the reactions between gly-pro-p-nitroanilide and DPP-IV, was observed by microplate readers with λ = 405 nm. All data were expressed as mean ± SD and the IC50 value was determined by non linear regression curve fit. Active substances in leaf extract of U. lobata was analyzed by liquid chromatography-mass spectrometry. DPP-IV inhibitory activity of active compounds was evaluated in silico using docking server.ResultsThe ethanolic extract of U. lobata showed stronger DPP-IV inhibitor activity than water extract with the IC50 values of 1 654.64 and 6 489.88 μg/mL, respectively. Vildagliptin, based on standard reference for DPP-IV inhibitor activity, has IC50 value of 57.44 μg/mL. Based on in silico analysis, mangiferin, stigmasterol and β-sitosterol in U. lobata extract have a strong inhibitory activity on DPP-IV.ConclusionsThe results showed that DPP-IV inhibitory activity of U. lobata is related to its active compounds such as mangiferin, stigmasterol and β-sitosterol

Fertilization starts with the binding of the spermatozoa to the zona pellucida (ZP) of the oocyte. Such binding is a carbohydrate-mediated event and consists of a series of tightly regulated events. Molecular interactions between spermatozoon and ZP in human are not well characterized due to limited availability of oocytes for research. Our current technology cannot generate recombinant human ZP (hZP) glycoproteins with native glycosylation. In this study, hZP glycoproteins, hZP2 (∼120 kDa), hZP3 (∼58 kDa) and hZP4 (∼65 kDa) were purified from ZP (purity >88%) by immunoaffinity columns. The binding sites of the purified native hZP3 and hZP4 were localized to the acrosome region of the capacitated human spermatozoa, and were lost after acrosome reaction. Purified human hZP2 bound to this region only in acrosome-reacted spermatozoa. Differential binding of the three glycoproteins to the post-acrosomal region and the midpiece of the spermatozoa was observed. In addition, hZP3, but not hZP2 and hZP4, induced hyperactivation. The stimulatory activity was dependent partly on N-linked glycosylation of hZP3. This manuscript describes the biological activities of purified hZP glycoproteins from the native source for the first time.

ObjectiveTo observe the effects of Centella asiatica (C. asiatica) methanolic extract on α-synuclein aggregation and its expression in rotenone-exposed zebrafish.MethodsZebrafish (Danio rerio) were exposed to 5 μg/L rotenone for 28 days and co-incubated with 2.5, 5.0 and 10.0 μg/mL of C. asiatica methanolic extract. The medium was changed every 48 h for maintain the concentration of rotenone and extract. After 28 days zebrafish were sacrificed on the ice block and protein was isolated from zebrafish brain for ELISA of dopamine and Western blotting of α-synuclein. Immunohistochemistry was conducted to observe the α-synuclein expressions from histopathological preparation of zebrafish brain. The head were soaked in 10% formaline for less than 24 h and embedded onto paraffin block, then sliced for immunohistochemistry using anti α-synuclein antibody. We also measured zebrafish motility for 5 min in each week.ResultsC. asiatica has important bioactive compounds such as asiaticoside that has anti-inflammatory and antioxidant properties. It may inhibit cascade reaction due to oxidative stress induced by rotenone. Decreasing reactive oxygen species proposed probability of radical attack to α-synuclein protein that caused aggregation and increase of its expression. The motility of zebrafish was also maintained in C. asiatica groups due to the increasing dopamine level in rotenone-induced zebrafish. High level of reactive oxygen species inactivated enzyme for dopamine synthesis such as tyrosine hydroxylase, and oxidized dopamine itself. Oxidized dopamine increased α-synuclein aggregation. Thus, the dopamine level decreased in rotenone-induced zebrafish, but C. asiatica increased dopamine level.ConclusionsC. asiatica has a potential to be developed as an anti-Parkinson's disease treatment due to its capability for minimized the sign of Parkinson's such as α-synuclein aggregation and expression, increasing motility and dopamine as well

Objective: To prove that the decrease in the expression of protamine 2 affects the fertilization ability and the number of child production. Methods: During the development stage of elongating spermatids, human spermatozoa chromatin undergoes a complex transition where histone is extensively replaced by protamine. Histone substitution by protamine 1 and 2 plays a significant role in condensing chromatin required to induce the quality of potent spermatozoa. The substitution also has a key role in protecting spermatozoa from effects of free radicals which can degrade the spermatozoa quality. Results: The results revealed that protamine deficiency instigated a severe disruption of spermatogenesis affecting male infertility. In addition, the protamine expression disorder caused a decrease in number, motility, and morphology of spermatozoa. Conclusion: Our results of a study in mice confirmed that inhibin B injection caused a reduction in the expression of protamine 2 in cauda epididymis. It has implications in the decrease of motility, concentration, and spermatozoa viability so that it affects the fertilization ability and the number of child production