Desalted and lyophilized seminal plasma increases protein tyrosine-phosphorylation of frozen-thawed bull spermatozoa incubated with a cell-permeable cyclic AMP (cAMP) analog (cBiMPS)

The present study investigates the effect of desalted seminal plasma (SP) added to semen extender on hyperactivated motility and protein tyrosine-phosphorylation (PTP) of bull spermatozoa. The SP was harvested by centrifugation and desalted using Sephadex G-25 columns in order to be added to semen extender at 0 (control), 2.5, 12.5 and 25 mg/ml. Frozen-thawed spermatozoa were incubated with a cellpermeable cyclic AMP (cAMP) analog (cBiMPS) and examined subjectively for hyperactivated motility and for PTP by Western blotting. Although, the added SP sustains sperm motility at all incubation times especially in the presence of cBiMPS but without significant difference from the control samples. Moreover, total sperm motility of 12.5 and 25 mg/ml in the presence of cBiMPS at 60, 120 and 180 min were similar (P ≥ 0.05). Surprisingly, cBiMPS-incubated spermatozoa in the presence of desalted SP were capable of exhibiting hyperactivated motility. Addition of SP increased and prolonged intracellular cAMP-induced PTP and in total 21 phosphorylated proteins with molecular weight ranging from 10 to ＞230 kDa were detected. The most prominent tyrosine-phosphorylated proteins (TPPs) were of 32, 38, 74 and 80 kDa which were more predominant in fertile bulls than subfertile bull. Furthermore, TPPs of 45 and 48 kDa were cBiMPS-dependent in fertile bulls whereas, in subfertile bull the latter was barely detectable and the former was cBiMPS-independent at only 0 min. This increase in PTP not only emphasizing the beneficial roles of desalted SP but excluding any detrimental effect of it on sperm cell functions during storage as well

Acute ammonia exposure combined with heat stress impaired the histological features of gills and liver tissues and the expression responses of immune and antioxidative related genes in Nile tilapia

Ammonia exposure can be considered more stressful for aquatic animals when it coincides with high temperature. This study was conducted to detect the effects of ammonia exposure and heat stress and their interactions on the histological features of gills and liver tissues and the expression responses of immune and antioxidative related genes in Nile tilapia. Thus, 180 fish were divided into four groups (triplicates), where the first and third groups were kept in clean water without total ammonium nitrogen (TAN) exposure. At the same time, the second and fourth groups were exposed to 5 mg TAN/L. After seven days, the water temperature was raised in the third (without ammonia toxicity) and fourth (exposed with 5 mg TAN/L) groups up to 32 °C and kept under these conditions for 24 h. While the first (without ammonia toxicity) and second (exposed with 5 mg TAN/L) groups were kept under optimum water temperature (27.28 °C) then gills and liver tissues were dissected. Marked upregulation of keap1 was seen in the gills of fish exposed to ammonia/heat stress. The expression of mRNA levels for nrf2, nqo-1, cat, and gpx genes were downregulated in all stressed groups, with the lowest was recorded in the ammonia/heat stress group. The transcription of ho-1 was upregulated in the ammonia and heat stress groups while downregulated in the ammonia/heat stress group. The transcription of the complement C3 gene was downregulated in the livers of heat stress and ammonia/heat stress groups, while the lysozyme gene was downregulated in the ammonia/heat stress group. The mRNA expression levels of nf-κB, il-1β, and tnf-α genes were higher in the ammonia group than in the heat stress group. The highest transcription level of nf-κB, il-1β, tnf-α, il-8, and hsp70 genes and the lowest C3 and lysozyme genes were observed in fish exposed to ammonia/heat stress. The co-exposure to ammonia with heat stress triggered degeneration of primary and secondary gill filaments with telangiectasia and vascular congestion of secondary epithelium while, the liver showed hepatic vascular congestion and visible necrotic changes with nuclear pyknosis. In conclusion, the combined exposure of ammonia and heat stress induced oxidative stress, immunosuppression, and inflammation in Nile tilapia

Osteoblast-activating peptide exhibits a specific distribution pattern in mouse ovary and may regulate ovarian steroids and local calcium levels

Osteoblast-activating peptide (OBAP) is a novel protein affecting osteoblast proliferation and differentiation, but its ovarian expression is yet to be reported. Osteoporosis is a common disease, caused mainly by low estrogen levels in females. We investigated whether OBAP regulates estrogen synthesis and osteoporosis. Using immunohistochemical analyses, we studied the distribution of OBAP in different parts of the mouse ovary. We also attempted to clarify the correlation of OBAP with ovarian steroids and calcium-regulating factors in the same ovarian tissues, including aromatase (CYP19), 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD), estrogen receptor (ER), progesterone receptor (PR), receptor activator of nuclear factor-kappa B (RANK), calmodulin, calbindin, and calcium-sensing receptor. The ovarian interstitial endocrine cells (IC) showed the greatest localization of OBAP, followed by the mature corpus luteum and the oocytes of mature Graafian follicles (MGF), while there were strong negative correlations of OBAP with CYP19. Strong positive correlations with 3 beta-HSD (except MGF), RANK (except IC), and calmodulin (except MGF and IC) were demonstrated. OBAP also showed partially positive correlations with ER and PR in the corpus luteum and with IC and calbindin in the MGF. We conclude that OBAP might be related to estrogen synthesis and calcium homeostasis

Garlic Alleviates the Injurious Impact of Cyclosporine-A in Male Rats through Modulation of Fibrogenic and Steroidogenic Genes

This work aimed to study the hepato-testicular protective effect of garlic in rats treated with cyclosporine A (CsA). Forty male Westar albino rats were randomly distributed in five groups (8 rats each): control, olive oil, garlic, CsA, and CsA co-treated with garlic. CsA induced an upsurge in the alanine transaminase, aspartate transaminase, and alkaline phosphatase levels and decreased albumin and total protein levels, expression of superoxide dismutase (SOD) gene, serum testosterone, triiodothyronine, and thyroxine levels compared to the control group. Additionally, there was an increase in the cholesterol, triglyceride, and low-density lipoprotein levels and a substantial reduction in the high-density lipoprotein levels compared to the control groups. Histopathological investigation of the liver showed abnormalities like hepatic cell degeneration, congestion of blood vessels, and highly active Kupffer cells in the CsA group. Histopathological examination of testes showed damaged seminiferous tubules, stoppage of the maturation of spermatogonia, and the presence of cells with irregular dense nuclei in the lumina of some tubules. For the groups treated with garlic, mitigation of the damage caused by CsA in the liver and testes, liver function tests, lipid profiles, and hormones was seen along with improved gene expression of SOD and steroidogenesis genes, and decreased gene expression of collagen I-α1 and transforming growth factor-1β. Conclusively, garlic had a positive impact on CsA-induced hepatic and sperm toxicity. It is recommended that garlic should be supplemented in transplant treatments using CsA to alleviate the cyclosporin-induced oxidative injuries and other harmful effects

Dietary effect of soybean lecithin on the growth performance, digestive enzyme activity, blood biomarkers, and antioxidative status of striped catfish, Pangasianodon hypophthalmus.

Soybean lecithin (SBL) is usually added to aquafeed as a lipid source because aquatic animals cannot synthesize phospholipids. Hence, this study aimed to investigate the role of SBL on the growth, nutrient consumption, digestive enzyme activity, blood parameters, and antioxidant capability of striped catfish. The fish were fed on five experimental diets with five grading levels of SBL (0, 2, 4, 6, and 8%) for 60 days. The final weight, weight gain, specific growth rate, feed intake, and protein efficiency ratio were markedly higher in striped catfish treated with 2-4% SBL than the control level (0% SBL). However, the lowest feed conversion ratio was in the fish-fed groups of 4-6% SBL. The carcass lipid content was significantly higher in fish fed 2-4% SBL compared to the control level (0% SBL). The lipase, amylase, and protease activities were significantly increased in the fish fed 2-6% SBL compared to 0% SBL-fed group. The gradually increased levels of SBL improved the structural appearance and increased the intestinal villi length and branching appearance. The triglycerides and total cholesterol were increased in the fish fed with 4, 6, and 8% compared to the control level, with the highest being in the fish fed with 8%. The lysozyme activity was higher in the fish fed with 2, 4, and 6% of SBL compared to the control level, with higher activity in the fish fed with 2 and 4% than 6%. Superoxide dismutase, glutathione peroxidase, and catalase activities were increased in the fish fed with 2, 4, and 6% SBL. The malondialdehyde level was lower in the fish fed with 4-6% SBL compared to the control level. The regression analysis revealed that the optimum dose of SBL is required at 3.65-4.42% for better productivity and health performances in striped catfish