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HDAC2/3 inhibitor MI192 mitigates oligodendrocyte loss and reduces microglial activation upon injury: A potential role of epigenetics

Background: During development, oligodendrocyte (OL) lineage cells are susceptible to injury, leading to life-long clinical neurodevelopmental decits, which lack effective treatments. Drugs targeting epigenetic modications that inhibit histone deacetylases (HDACs) protect from many clinical neurodegenerative disorders. Aim: This study aimed to investigate the therapeutic potential of histone deacetylase 2/3 (HDAC2/3) inhibitor MI192 on white matter (WM) pathology in a model of neonatal rat brain injury.Methods: Wistar rats (8.5-day-old, n = 32) were used to generate brain tissues. The tissues were cultured and then randomly divided into four groups and treated as following: group I (sham); the tissues were cultured under normoxia, group II (vehicle); DMSO only, group III (injury, INJ); the tissues were exposed to 20 minutes oxygen-glucose deprivation (OGD) insult, and group IV (INJ + MI192); the tissues were subjected to the OGD insult and then treated with the MI192 inhibitor. On culture day 10, the tissues were xed for biochemical and histological examinations. Results: The results showed that inhibition of HDAC2/3 activity alleviated WM pathology. Specically, MI192 treatment signicantly reduced cell death, minimized apoptosis, and mitigates the loss of the MBP+ OLs and their precursors (NG2+ OPCs). Additionally, MI192 decreased the density of reactive microglia (OX−42+). These ndings demonstrate that the inhibition of HDAC2/3 activity post-insult alleviates WM pathology through mechanism(s) including preserving OL lineage cells and suppressing microglial activation. Conclusion: The ndings of this study suggest that HDAC2/3 inhibition is a rational strategy to preserve WM or reverse its pathology upon newborn brain injury. Keywords: Brain injury, Epigenetics, MI192, Microglia, Oligodendrocyte
Mansur Ennuri Shmela, Mohamed A. Al-Griw, Emad M. Bennour(8-2021)
عرض موقع المنشور

Extraction and Purification of Short-chain Fatty Acids from Fermented Reconstituted Skim Milk Supplemented with Inulin

Short-chain fatty acids (SCFAs) acetate, propionate, butyrate and lactate were determined in 12 % reconstituted skim milk (RSM) and RSM supplemented with inulin (RSMI). The fermentation was performed with Bifidobacterium animalis subsp. lactis (BB 12) and Lactobacillus rhamnosus LGG ATCC 53013. Fermentation culture activities produced substantial amounts of SCFAs, which were detected and quantitated using a HPLC-UV technique. Using HPLC-UV, we were able to detect low concentrations of lactate and SCFAs from fermented samples; lactate, acetate, propionate and butyrate were detected at 10.10, 12.06, 14.80 and 18.06 μg/mL, respectively. The retention time of all SCFAs and lactic acid were similar to the standard quality control (±0.05), and average recovery ranged between 89.73 and 91.03 %. The experimental conditions and sample preparation were applied to preparative HPLC to isolate and purify SCFAs with concentration range between 0.09 and 2.86 mg/mL. The purity of extracted SCFAs was confirmed using atmospheric pressure chemical ionization/mass spectrometry by determining the molecular masses of target purified compounds. The scaled up validated analytical HPLC-UV method will further enhance and improve the use of this approach to produce purified large-scale SCFAs.
M. Asarat , T. Vasiljevic, M. Ravikumar, V. Apostolopoulos, O. Donkor(4-2016)
موقع المنشور

Short-Chain Fatty Acids Regulate Secretion of IL-8 from Human Intestinal Epithelial Cell Lines in vitro

Short-chain fatty acids (SCFAs) including acetate, propionate and butyrate play an important role in the physiological functions of epithelial cells and colonocytes, such as immune response regulation. Human intestinal epithelial cells (IECs) contribute in intestinal immune response via different ways, such as production of different immune factors including Interleukin (IL) IL-8, which act as chemoattractant for neutrophils, and subsequently enhance inflammation. Therefore, we aimed to evaluate the effects of SCFAs on IECs viability and production of IL-8 in vitro. SCFAs were co-cultured with either normal intestinal epithelial (T4056) or adenocarcinoma derived (HT-29) cell lines for 24–96 h in the presence of E.coli lipopolysaccharides (LPS). Cell viability, proliferation, production of IL-8 and expression of IL-8 mRNA were determined in the cell cultures. The result showed that 20 mM of SCFAs was non-cytotoxic to T4056 and enhanced their growth, whereas the growth of HT-29 was inhibited. The SCFAs down regulated LPS-stimulated IL-8 secretion with different response patterns, but no obvious effects on the release of IL-8 from non LPS- stimulated cells. In conclusion, SCFAs showed regulatory effect on release of LPS-stimulated IL-8 as well as the expression of mRNA of IL-8; these might explain the anti-inflammatory and anti-carcinogenic mechanism of SCFAs.
M. Asarat, T. Vasiljevic, V. Apostolopoulos, O. Donkor(10-2015)
موقع المنشور

Short-Chain Fatty Acids Regulate Cytokines and Th17/Treg Cells in Human Peripheral Blood Mononuclear Cells in vitro.

Short-chain fatty acids (SCFAs) have been recognized as mediators of immune responses, including pathways of cytokine production. In this study, we investigated the immune-regulatory effects of SCFAs on human peripheral blood mononuclear cells (PBMCs) from buffy coat of healthy donors. PBMCs were exposed to varying concentrations of individual SCFAs or of their mixtures of acetate, propionate and butyrate. The productions of interleukin (IL) IL-1β, IL-2, IL-6, IL-10, IL-17, IL-21, IL-23 and transforming growth factor beta 1 (TGF-β1) were assessed. T cell differentiation after exposure to SCFAs was also examined. Compared with lipopolysaccharide (LPS)-stimulated cells (controls), SCFAs slightly decreased TGF-β1 production and reduced IL-6 production; butyrate was more effective than acetate or propionate. SCFAs particularly butyrate caused the induction of CD4+CD25+ regulatory T cells (Treg) rather than Th17 cells. SCFAs may up-regulate the production of anti-inflammatory cytokines in PBMCs, resulting in the induction of CD4+CD25+ Treg cells
M. Asarat, V. Apostolopoulos, T. Vasiljevic , O. Donkor(3-2016)
موقع المنشور

Short-chain fatty acids produced by synbiotic mixtures in skim milk differentially regulate proliferation and cytokine production in peripheral blood mononuclear cells.

Short chain fatty acids (SCFAs) are major products of prebiotic fermentation and confer human health benefits such as immune-regulation. In this study, reconstituted skim milk supplemented with prebiotics (RSMP) including inulin, hi-maize or β-glucan was fermented by probiotic strains of Lactobacillus spp. and Bifidobacteria spp. After 24 h of fermentation, probiotics growth and SCFAs production were investigated and the produced SCFAs were extracted. Inulin and Lactobacillus rhamnosus GG ATCC 53013 (LGG) combination released highest concentrations of SCFAs compared to LGG and hi-maize or β-glucan. Extracted SCFAs were then used for in vitro immune modulation study in human peripheral blood mononuclear cells (PBMCs). In lipopolysaccharide (LPS)-stimulated PBMCs, SCFAs particularly butyrate down-regulated tumor necrosis factor alpha, interleukin (IL)-12, interferon gamma (IFN-γ) and transforming growth factor beta-1 (TGF-β1), and up-regulated IL-4, IL-10, while no significant effect was noted in non-LPS-stimulated PBMCs. The results indicate that SCFAs regulated cytokine milieu in LPS-stimulated PBMCs to anti-inflammatory cytokines.
M. Asarat , V. Apostolopoulos, T. Vasiljevic , O. Donkor(9-2015)
موقع المنشور

Ovarian granulosa cell tumor in a Persian cat: Case report from Tripoli, Libya

Abstract Background: Ovarian granulosa cell tumors (GCTs) are common neoplasms in domestic animals that originate from the sex-cord stromal cells of the ovary. The aim of the present paper was to report a case of ovarian GCT in a 5-year-old Persian female cat and to describe the histopathological pattern of this tumor. Case Description: The cat was brought for a routine diagnostic examination for pregnancy at Albaittar clinic in Tripoli, Libya with a history of 1 month mating before it was brought to the clinic for investigation of progressive behavioral changes. The cat external examination showed noticeable enlargement in the abdomen that was potentially suspected of pregnancy; however, the abdominal ultrasonography showed a great mass located on the right ovary. A therapeutic ovariohysterectomy was performed. The cat’s clinical signs resolved, 6 months later it was diagnosed with a mass on the right kidney suspected as metastasis and one week later the cat died. Conclusion: In this case, macroscopic and microscopic studies are discussed and histopathological examination confirmed ovarian GCT with cystic endometrial hyperplasia and suspected metastasis in one kidney. Keywords: GCT, Ovary, Histopathological finding, Persian cat, Tripoli.
Asma mohamed elbahi, Shaima A. M. Mahgiubi, Abdurraouf O. Gaja3(8-2022)
موقع المنشور

Pigmented villonodular synovitis (PVNS) in human and animals: a mini review article and conclusive view

ales and females and also in animals, it could lead to severe pain, joint deformity and decrease of joint function with a consequent lowering of the productivity. Moreover it is a diagnostic challenge, the difficulty stems from the insidious onset and nonspecific presentation of the disease, as well as its subtle radiographic findings. In addition, the disease is difficult to be differentiated from conditions such as rheumatoid arthritis, osteoarthritis and other inflammatory and neoplastic processes of the synovial lining. PVNS comes in two forms: localized and diffuse. Diffuse PVNS affects the entire synovium and typically occurs in large joints such as the knee or hip. Localized or nodular, PVNS is less common than the diffuse form and typically occurs in smaller joints such as the hands and feet. Involvement of bursa (PVNB, pigmented villonodular bursitis) is the least common. The incidence of pigmented villonodular synovitis is 1.8 cases per 1 million people per year. It is equally affect males and females. Animals are less commonly affected and the condition mostly misdiagnosed. Histopathologically, diffuse disease is characterized by a mononuclear stromal cell infiltrate in the synovial membrane.
Seham AlAzreg(1-2017)
موقع المنشور

Subacute ruminal acidosis outbreak in feedlot Barki sheep due to faulty fattening practice

Subacute ruminal acidosis (SARA) is an important disorder in producing animals characterized by prolonged periods of low ruminal pH with deleterious effects on animal’s health and productivity. SARA was studied usually experimentally due to its subclinical nonspecific presentation. We present here a field outbreak of SARA in feedlot Barki sheep and the related ruminal pathological changes. Rumens of six feedlot sheep fed on high-grain fiber- deficient diets were examined for the pH of their contents as well as any abnormalities in their morphology and tissues. The results revealed that the ruminal juice of affected animals was acidic. Grossly, the ruminal mucosa was grey, hyperemic and severely thickened with multiple areas of epithelial detachment. Histopathologically, the common findings were epithelial desquamation, hydropic degeneration and ulcer formation. In addition, submuscular venular thrombosis and multifocal lesion with eosinophilic granular proteinaceous material mixed with variable numbers of inflammatory cells were noticed. This is, according to the best of the authors’ knowledge, the first example of a non-experimental field SARA outbreak in feedlot sheep and its ruminal gross and histopathological picture.
Seham AlAzreg(1-2021)