RESUMEN
Decreased stemness and increased cellular senescence impair the ability of mesenchymal stem cells (MSCs) to renew themselves, change into different cell types, and contribute to regenerative medicine. There is an urgent need to discover new compounds that can boost MSCs' stemness and delay senescence. Therefore, this study aimed to investigate the impact of walnut kernel oil (WKO) and defatted (WKD) extracts on bone marrow (BM)-MSC stemness and senescence. Premature senescence and inflammation were induced in BM-MSCs using H2O2 and LPS, respectively. Phytochemical constituents of WKO and WKD extracts were detected by HPLC. The stemness (proliferation and migration), senescence-related markers (p53, p21, SIRT1, and AMPK), oxidative stress/antioxidant markers, inflammatory cytokines, and cell cycle of BM-MSCs were measured by MTT assay, qPCR, ELISA, and flow cytometry. WKO and WKD extracts improved rat BM-MSC stemness, as evidenced by (1) increased cell viability, (2) decreased apoptosis (low levels of Bax and caspase3 and high levels of Bcl2), (3) upregulated MMP9 and downregulated TIMP1 expression, and (4) cell cycle arrest in the G0/G1 phase and declined cell number in the S and G2/M phases. Additionally, WKO and WKD extracts reduced rat BM-MSC senescence, as indicated by (1) decreased p53 and p21 expression, (2) upregulated expression and levels of SIRT1 and AMPK, (3) reduced levels of ROS and improved antioxidant activity (higher activity of CAT, SOD, and GPx and upregulated expression of NrF2 and HO-1), and (4) declined levels of TNFα, IL1ß, and NF-κB. When compared to the WKO extract, the WKD extract had a greater impact on the induction of stemness and reduction of senescence of BM-MSCs due to its stronger antioxidant activity, which could be attributed to its higher levels of flavonoids and phenolic compounds, as detected by HPLC analysis. WKO and WKD extracts enhance rat BM-MSC stemness and protect them from senescence, suggesting their potential use as enhancers to increase MSCs' therapeutic efficacy.
Asunto(s)
Proteínas Quinasas Activadas por AMP , Juglans , Animales , Ratas , Antioxidantes/farmacología , Peróxido de Hidrógeno , Sirtuina 1/genética , Proteína p53 Supresora de TumorRESUMEN
Cryptosporidium parvum is a ubiquitous zonootic parasite causing enteritis in man and animals. Cryptosporidium infection was confirmed microscopically in neonatal calves (less than 6 weeks of age) at Kafr El Sheikh Province, Egypt. Multilocus analysis using a wide array of genetic markers was carried out to assess genetic diversity of C. parvum isolates. PCR amplification and partial sequence analysis of 70 kDa heat shock protein, dihydrofolate reductase, alpha-tubulin, elongation factor 1 alpha as well as thrombospondin-related anonymous protein of Cryptosporidium-1, and thrombospondin-related anonymous protein of Cryptosporidium-2 gene markers were achieved. Data indicated that the analyzed isolates belong to C. parvum genotype II with obvious sequence heterogeneity compared with counterparts deposited in Genebank.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Criptosporidiosis/veterinaria , Cryptosporidium parvum/clasificación , Cryptosporidium parvum/genética , ADN Protozoario/genética , Polimorfismo Genético , Proteínas Protozoarias/genética , Animales , Bovinos , Análisis por Conglomerados , Criptosporidiosis/parasitología , Cryptosporidium parvum/aislamiento & purificación , ADN Protozoario/química , Egipto , Genotipo , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
The stomach of three species of non-human primates was investigated by lectin histochemistry to clarify the staining affinity and distribution patterns of their sugar residues. All gastric regions, with little differences between the deep and superficial parts of the same region, were rich in. in N-acetylglucosamine and/or neuraminic acid. Although, the superficial regions of the gastric mucosa were scanty in N-acetylgalactosamine, a- D-glucose and a -D-mannose, the deep parts of the gastric mucosa were rich in these sugars. In conclusion, there is a difference among the mucosubstances of surface and foveolar mucous cells, mucous neck cells, and gastric gland cells. This indicates heterogeneous composition of gastric mucus, or mucus molecules with variations in the degree of glycosylation of their oligosaccharide chains in the different cells which suggest that lectin binding affinity in the gastric mucosa correlated mostly to the degree of cellular differentiation.
El estómago de tres especies de primates no humanos fue investigado por histoquímica de lectinas para determinar la afinidad de tinción y los patrones de distribución de sus residuos de azúcar. Todas las regiones gástricas, con pequeñas diferencias entre las partes profundas y superficiales de la misma región, eran ricas en N-acetilglucosamina y/o ácido neuramínico. Si bien, las regiones superficiales de la mucosa gástrica eran escasas en N-acetilgalactosamina, a-D-glucosa y a-D-manosa, las partes profundas de la mucosa gástrica eran ricas en estos azúcares. En conclusión, existe una diferencia entre las mucosustancias de la superficie y células mucosas foveolares, células mucosas del cuello y células de las glándulas gástricas. Esto indica una composición heterogénea de la mucosa gástrica, o moléculas de moco con variaciones en el grado de glicosilación de sus cadenas de oligosacáridos en las diferentes células, sugieriendo que la afinidad de union de lectinas en la mucosa gástrica se relacionada principalmente con el grado de diferenciación celular.