Foad Farrag

Desalted and lyophilized bovine seminal plasma delays induction of the acrosome reaction in frozen-thawed bovine spermatozoa in response to calcium ionophore

Cryopreservation is partially damaging and induces capacitation-like changes in spermatozoa. Seminal plasma (SP) contains a variety of biochemical components, such as protein and lipids, which are specific for the regulation of sperm cell function including those effective for decapacitation of spermatozoa. Therefore, this study tested the hypothesis that desalted and lyophilized SP could prevent premature capacitation (cryocapacitation) of Japanese Black bull spermatozoa. Seminal plasma was desalted by using Sephadex G-25 desalting column and lyophilized before added to semen extender at final concentrations 0, 2.5, 12.5, and 25 mg/mL. Frozen-thawed sperm progressive motility, acrosomal integrity, abnormal morphology, and the calcium ionophore A23187-induced acrosome reaction were assessed. Protein and lipid compositions in SP were analyzed by SDS-PAGE and thin-layer chromatography, respectively. The results revealed that progressive motility, intact acrosome, and abnormal morphology were not substantially modified by addition of SP. Stimulation of spermatozoa with calcium ionophore A23187 resulted in a time-dependent induction of the acrosome reaction, which was delayed by the desalted and lyophilized SP. There was no difference in the protein profile of SP before and after gel filtration. In total, 19 protein bands with molecular masses ranging from 5.2 to 185.8 kDa were detected and those of 185.8, 80, 34, 20.8, 18.8, 17.5, and 10 kDa were considered as novel proteins. Neutral lipids and phospholipids before and after gel filtration were the same, and the detected neutral lipid spots were monoacylglycerol, cholesterol, 1,2- and 1,3-disaturated diacylglycerol, 1,2- and 1,3-saturated, unsaturated diacylglycerol, whereas the detected phospholipid spots were sphingomyelin, phosphatidylcholine, phosphatidylserine, and three species of phosphatidylinositol, phosphatidylethanolamine, cerebroside, and polyglycerol phosphatide. The results suggest that premature capacitation during freeze-thaw processes could be reduced by adding desalted and lyophilized SP.

Pinacidil and levamisole prevent glutamate-induced death of hippocampal neuronal cells through reducing ROS production

Abstract Activators of both adenosine 5′-triphosphate (ATP)-sensitive K+ (KATP) channel and cystic fibrosis transmembrane conductance regulator (CFTR) Cl−  channel have significant in vivo and in vitro neuroprotection against glutamate-induced death of some neuronal cells. Here, the effect of the KATP channel activator, pinacidil, and the CFTR Cl−  channel opener, levamisole, against glutamate-induced oxidative stress were investigated in mouse hippocampal cells, HT22. The results from cell viability assay (WST-1) showed that pinacidil and levamisole weakly protected cells against glutamate-induced toxicity at 10 μM and their effect increased in a dose-dependent manner till reach maximum protection at 300 μM. Pretreatment with pinacidil or levamisole significantly suppressed the elevation of reactive oxygen species (ROS) triggered by glutamate through stabilising mitochondrial membrane potential and subsequently protected HT22 cells against glutamate-induced death. HT22 cells viability was maintained by pinacidil and levamisole in presence of glutathione inhibitor, BSO. Also, pinacidil and levamisole pretreatment did not induce recovery of glutathione levels decreased by glutamate Expectedly, this protection was abolished by the KATP and CFTR Cl−  channels blocker, glibenclamide. Thus, both pinacidil and levamisole protect HT22 cells against glutamate-induced cell death through stabilising mitochondrial membrane potential and subsequently decreasing ROS production.

Regulation of Chick Ebf1-3 Gene Expression in the Pharyngeal Arches, Cranial Sensory Ganglia and Placodes

This study was conducted to identify the regulation of the expression of the cEbf1-3 (chick early B-cell factor 1-3) genes in the pharyngeal arches (PAs), cranial sensory ganglia and placodes. cEbf1 and cEbf3 were mainly expressed in the cranial neural crest cells (NCCs) occupying the PAs, but cEbf2 was expressed in the mesenchymal core. cEbf1-3 were prominently expressed in the olfactory placodes, but cEbf1 and cEbf3 were only expressed in the otic vesicle. cEbf1 was expressed in all cranial sensory ganglia, cEbf2 (only) in the dorsolateral ganglia and cEbf3 in the trigeminal and vestibular ganglia. The removal of the source (the cranial neural tube) of the cranial NCCs before their migration to the PAs led to downregulation of cEbf1 and cEbf3 and upregulation of cEbf2 expression. Gain- and loss-of-function experiments showed that sonic hedgehog did not regulate cEbf1-3 expression in the PAs or associated ganglia. Bone morphogenetic protein 2 (Bmp2) can, however, directly and indirectly regulate cEbf1 and cEbf3 expression in the PAs and the proximal (NCC-derived) portion, but not the distal (placodal-derived) portion of the cranial sensory ganglia. Conversely, cEbf2 expression was upregulated following injection of Noggin before the migration of NCCs, but did not change after the overexpression of either Noggin or Bmp2 in the arch after NCC migration. In conclusion, Bmp2 regulates cEbf1 and cEbf3 expression in PAs and cranial sensory ganglia both directly and indirectly, via the migration of cranial NCCs. However, cEbf2 expression in the mesenchymal core of PAs is controlled by other undetermined signals.

Comparative glycoconjugates histochemistry of proventriculus of chicken, ducks and geese

A lectin histochemical study was performed on formalin-fixed paraffin-embedded tissue sections of proventriculus from three mature chickens, three mature ducks and three mature geese. The purpose of this study was to focus on the comparative glycoconjugates of chicken, ducks and geese proventriculus. The distribution of glycoconjugates in the proventriculus of chicken, ducks and geese was studied using eight (LCA, ConA, PNA, RCA 120, WGA, DBA, UEA and PHA-E4) horse raddish peroxidase labeled lectins and peroxidase activity was visualized by Dako cytomation (DAB).The neutral mucopolysaccharides were restricted on the supranuclear part of the epithelium of mucosal folds (plicae) and negative in the basal part (sulci) epithelium in all species under investigation. The examined parts of the proventriculus (plicae epithelium, sulcui epithelium, ducts of the glands and glandular tissue) were labeled to all lectins used in case of geese. The ducks proventriculus showed expression to all lectins except glandular duct epithelium with D glucose, N acetylegalactosamine and L fucose. The proventriculus of chicken was labeled to all lectins except DBA which bind to N acetyle galactosamine. Moreover, the glandular tissue and sulci epithelium were negative to D glucose and D galactose respectively. In conclusion, all investigated species showed neutral mucopolysaccharides with different sugar residues to protect the mucosa and glandular tissue from the harmful effect of the acid content.

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