Niu, X., C. Hu, S. Chen, J. Wen, X. Liu, Y. Yong, Z. Yu, Xingbin Ma, C. Li, M. Warda, et al., "Chitosan-gentamicin conjugate attenuates heat stress-induced intestinal barrier injury via the TLR4/STAT6/MYLK signaling pathway: In vitro and in vivo studies.", Carbohydrate polymers, vol. 321, pp. 121279, 2023. Abstract

Heat stress (HS) has a negative impact on animal health. A modified chitosan-gentamicin conjugate (CS-GT) was prepared to investigate its potential protective effects and mechanism of action on heat stress-induced intestinal mucosa injury in IPEC-J2 cells and mouse 3D intestinal organs in a mouse model. CS-GT significantly (P < 0.01) reversed the decline in transmembrane resistance and increased the FITC-dextran permeability of the IPEC-J2 monolayer fusion epithelium caused by heat stress. Heat stress decreased the expression of the tight binding proteins occludin, claudin1, and claudin2. However, pretreatment with CS-GT significantly increased (P < 0.01) the expression of these tight binding proteins. Mechanistically, CS-GT inhibited the activation of the TLR4/STAT6/MYLK signaling pathway induced by heat stress. Molecular docking showed that CS-GT can bind effectively with TLR4. In conclusion, CS-GT alleviates heat stress-induced intestinal mucosal damage both in vitro and in vivo. This effect is mediated, at least partly, by the inhibition of the TLR4/STAT6/MYLK signaling pathway and upregulation of tight junction proteins. These findings suggest that CS-GT may have therapeutic potential in the prevention and treatment of heat stress-related intestinal injury.

Wen, J., S. Chen, M. Bao, C. Hu, L. Wu, Y. Yong, X. Liu, Y. Li, Z. Yu, Xingbin Ma, et al., "Slc9a1 plays a vital role in chitosan oligosaccharide transport across the intestinal mucosa of mice.", Carbohydrate polymers, vol. 299, pp. 120179, 2023. Abstract

The mechanism underlying the intestinal transport of COS is not well understood. Here, transcriptome and proteome analyses were performed to identify potential critical molecules involved in COS transport. Enrichment analyses revealed that the differentially expressed genes in the duodenum of the COS-treated mice were mainly enriched in transmembrane and immune function. In particular, B2 m, Itgb2, and Slc9a1 were upregulated. The Slc9a1 inhibitor decreased the transport efficiency of COS both in MODE-K cells (in vitro) and in mice (in vivo). The transport of FITC-COS in Slc9a1-overexpressing MODE-K cells was significantly higher than that in empty vector-transfected cells (P < 0.01). Molecular docking analysis revealed the possibility of stable binding between COS and Slc9a1 through hydrogen bonding. This finding indicates that Slc9a1 plays a crucial role in COS transport in mice. This provides valuable insights for improving the absorption efficiency of COS as a drug adjuvant.

Kasas, E. A. H., I. M. Farag, H. R. Darwish, Y. A. SOLIMAN, E. E. M. Nagar, Marwa A Ibrahim, S. Kamel, and M. Warda, "Molecular characterization of alpha subunit 1 of sodium pump (ATP1A1) gene in Camelus dromedarius: its differential tissue expression potentially interprets the role in osmoregulation.", Molecular biology reports, vol. 49, issue 5, pp. 3849-3861, 2022. Abstract

BACKGROUND: Dromedary or one-humped camel (Camelus dromedarius) is distinctively acclimatized to survive the arid conditions of the desert environment. It has an excellent ability to compete dehydration with substantial tolerance for rapid dehydration. Therefore, it offers an excellent model for studying osmoregulation. Molecular characterization of Na/K ATPase as a central regulator of electrolyte normohemostasis affords a better understanding of this mechanism in camel. Here is the first to resolve the full-length of alpha-1 subunit of sodium pump (ATP1A1) gene with its differential expression in dromedary tissues.

RESULTS: The nucleotide sequence for the recovered full cDNA of ATP1A1was submitted to the GenBank (NCBI GenBank accession #MW628635) and bioinformatically analyzed. The cDNA sequence was of 3760 bp length with an open reading frame (ORF) of 3066 bp encoding a putative 1021 amino acids polypeptide with a molecular mass of 112696 Da. Blast search analysis revealed the shared high similarity of dromedary ATP1A1gene with other known ATP1A1genes in different species. The comparative analysis of its protein sequence confirmed the high identity with other mammalian ATP1A1 proteins. Further transcriptomic investigation for different organs was performed by real-time PCR to compare its level of expression among different organs. The results confirm a direct function between the ATP1A1 gene expression and the order of vital performance of these organs. The expression of ATP1A1 mRNA in the adrenal gland and brain was significantly higher than that in the other organs. The noticed down expression in camel kidney concomitant with overexpression in the adrenal cortex might interpret how dromedary expels access sodium without water loss with relative high ability to restrain mineralocorticoid-induced sodium retention on drinking salty water.

CONCLUSION: The results reflect the importance of sodium pump in these organs. Na/K ATPase in the adrenal gland and brain than other organs.

Chaudhry, Z. L., M. Gamal, I. Ferhati, M. Warda, and B. Y. Ahmed, "ER Stress in COVID-19 and Parkinson's Disease: In Vitro and In Silico Evidences.", Brain sciences, vol. 12, issue 4, 2022. Abstract

The outbreak of COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) signifies a serious worldwide concern to public health. Both transcriptome and proteome of SARS-CoV-2-infected cells synergize the progression of infection in host, which may exacerbate symptoms and/or progression of other chronic diseases such as Parkinson's disease (PD). Oxidative stress is a well-known cause of endoplasmic reticulum (ER) stress observed in both SARS-CoV-2 and PD. In the current study, we aimed to explore the influence of PKR-like ER kinase (PERK) stress pathway under SARS-CoV-2-mediated infection and in human cell model of PD. Furthermore, we investigated whether they are interconnected and if the ER stress inhibitors could inhibit cell death and provide cellular protection. To achieve this aim, we have incorporated in silico analysis obtained from gene set enrichment analysis (GSEA), a literature review and laboratory data. The neurotoxin, 6-hydroxy dopamine (6OHDA), was used to mimic the biochemical and neuropathological characteristics of PD by inducing oxidative stress in dopamine-containing neurons differentiated from ReNVM cell line (dDCNs). Furthermore, we explored if ER stress influences activation of caspases-2, -4 and -8 in SARS-CoV-2 and in stressed dDCNs. Our laboratory data using Western blot, immunocytochemistry and 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) analyses indicated that 6OHDA-induced toxicity triggered activation of caspases-2, -4 and -8 in dDCNs. Under SARS-CoV-2 infection of different cell types, GSEA revealed cell-specific sensitivities to oxidative and ER stresses. Cardiomyocytes and type II alveolar epithelial-like cells were more vulnerable to oxidative stress than neural cells. On the other side, only cardiomyocytes activated the unfolded protein response, however, the PERK pathway was operative in both cardiomyocytes and neural cells. In addition, caspase-4 activation by a SARS-CoV-2 was observed via in silico analyses. These results demonstrate that the ER stress pathway under oxidative stress in SARS-CoV-2 and PD are interconnected using diverse pathways. Furthermore, our results using the ER stress inhibitor and caspase specific inhibitors provided cellular protection suggesting that the use of specific inhibitors can provide effective therapeutic approaches for the treatment of COVID-19 and PD.

Liu, H., Z. Zhang, J. Li, W. Liu, M. Warda, B. Cui, and A. M. Abd El-Aty, "Oligosaccharides derived from ameliorate glycolipid metabolism and modulate the gut microbiota community and the faecal metabolites in a type 2 diabetes mouse model: metabolomic bioinformatic analysis.", Food & function, vol. 13, issue 9, pp. 5416-5429, 2022. Abstract

Herein, we assessed the effects of oligosaccharides (LBO) on the intestinal microenvironment of a type 2 diabetes (T2D) mouse model through gut microbiome and metabolomics analysis. We set high (300 mg kg), medium (200 mg kg), and low (100 mg kg) doses of LBO for intervention once a day for 4 weeks. The results showed that the intervention effect of the medium-dose group was the most significant. It reduced the symptoms of hyperglycemia, inflammation, insulin resistance, and lipid accumulation in the T2D mouse model. It restored the structure of damaged tissues and cells, such as the pancreas, liver, and kidneys. LBO increased the relative abundance of beneficial bacteria, such as , , , and , and maintained intestinal barrier integrity. The faecal metabolic map showed that the contents of glycogen amino acids, such as proline, serine, and leucine, increased. The contents of cholic, capric, and dodecanoic acid decreased. In summary, we may suggest that LBO can be used as a prebiotic for treating T2D.

Ghanem, N., N. H. Nasr, N. A. A. Elnaga, F. Abou-Hashim, S. Kamel, M. Warda, and others, "Molecular and physiochemical evaluation of buck semen cryopreserved with antioxidants", Reprod Domest Anim . , vol. 58, issue 6, pp. 813-822., 2023.
Hyun-ju, K., Kim Na-Ri, Y. Jae-Boum, P. Won-Sun, W. Mohamad, K. Eui-Yong, and H. Jin, "ROLE OF NADH/NADPH OXIDASE-DERIVED H2O2 IN ISOPROTERENOL-INDUCED CARDIAC HYPERTROPHY", Korean Journal of Physiology & Pharmacology , vol. 10, issue No. 0 , pp. 130 ~ 0, 2006.
Abdelsalam, M., R. Abdel-Gaber, M. A. Mahmoud, O. A. Mahdy, N. I. M. Khafaga, and M. Warda, "Morphological, molecular and pathological appraisal of Callitetrarhynchus gracilis plerocerci (Lacistorhynchidae) infecting Atlantic little tunny (Euthynnus alletteratus) in Southeastern Mediterranean.", Journal of advanced research, vol. 7, issue 2, pp. 317-26, 2016. Abstract

The Atlantic little tunny, Euthynnus alletteratus, is widely distributed in temperate and tropical waters of the Atlantic Ocean, Black and Mediterranean Seas. In this study, wild-caught little tunny from Egypt, were found to be naturally infected with trypanorhyncha metacestodes, and the overall prevalence rate of infection was 38.7%. The blastocysts were either loosely attached to the mesentery of infected fish, or firmly attached and deeply embedded within the hepatic parenchyma. These encysted plerocerci are identified as Callitetrarhynchus gracilis (Trypanorhyncha, Lacistorhynchidae) based on its morphological and molecular characterization. The morphological characteristics of C. gracilis including scolex shape; the bothridia groove; the presence of frontal glands; the length of post-larval (appendix); metabasal armature; the existence of 'Chainette' and satellite hooks of different size were studied and described by Light and Scanning electron microscope. The phylogenetic analysis of lsrDNA gene of plerocerci confirmed the identification of the species to be deeply embedded in genus Callitetrarhynchus. The histopathological examination revealed severe pathological changes in the affected organs, including necrosis, inflammatory reactions, fibrosis and migratory tracts of the parasitic larvae together with marked visceral organs adhesions. To the best of our knowledge, this is the first report describing the detection of C. gracilis in little tunny collected from the Abu Qir landing site in Alexandria, Egypt.

Ahmed-Farid, O. A., S. A. Haredy, R. M. Niazy, R. J. Linhardt, and M. Warda, "Dose-dependent neuroprotective effect of oriental phyto-derived glycyrrhizin on experimental neuroterminal norepinephrine depletion in a rat brain model.", Chemico-biological interactions, 2019. Abstract

The dose-dependent neuroprotective role of licorice-derived glycyrrhizin during subacute neuroterminal norepinephrine (NE) depletion was studied in rat brain. Experimental design included thirty 5-week-old male rats randomly divided into five groups. Compared to the saline-injected control group, the group receiving daily intraperitoneal injection of fusaric acid (FA; 5 mg/kg/b.w.) for 30 days showed pharmacological depletion of NE. The neuroprotective effects of three successively increasing oral doses of glycyrrhizin were examined in FA-treated rats. Neurochemical parameters and histo-/immunohistopathological changes in the hippocampus were examined. FA generated global hippocampal stress with altered neurobiochemical parameters, accompanied by immune-confirmed inflammatory tissue damage, and noticeable behavioral changes. Although glycyrrhizin after FA-induced intoxication did not correct the recorded drop in the NE level, it decreased the dopamine levels to control levels. Similarly, glycyrrhizin at a high dose restored the serotonin level to its normal value and blocked the FA-induced increase in the level of its metabolite, 5-hydroxyindoleacetic acid. The FA-induced rise in γ-aminobutyric acid (GABA) and histamine was alleviated after administration of a high dose of glycyrrhizin. This was accompanied by improvements in the bioenergetic status and neuronal regenerative capacity through recovery of ATP and brain-derived neurotrophic factor levels to the pre-intoxicated values. High doses of glycyrrhizin also ameliorated the FA-generated behavioral changes and oxidative damage, manifested by the reduction in the expression of cortical pro-apoptotic caspase 3 in the same group. This study suggests that glycyrrhizin can potentially mend most of the previously evoked neuronal damage induced by FA intoxication in the brain using a rat model.