RESUMO
KNOWLEDGE TRANSFER STATEMENT: This article aims to acquaint clinicians treating pediatric patients with COVID-19 hazards and delineate the steps required for minimizing cross-infection in case of providing emergency treatment to children in dental offices.
Assuntos
Betacoronavirus , Infecções por Coronavirus , Pandemias , Pneumonia Viral , COVID-19 , Criança , Humanos , Odontopediatria , SARS-CoV-2RESUMO
BACKGROUND: Due to very sluggish turnover at the molecular and cellular level, the healing of chondral damages has been considered difficult. In the current study, the effects of the Kartogenin, a small heterocyclic molecule on chondrogenic differentiation of stem cells was compared to TGF-ß3. METHODS: Human Adipose-Derived Stem Cells were extracted during an elective surgery. Cell viability was estimated by MTT assay, differentiated cells evaluated by histological and immunohistochemical techniques. Expression of cartilage specific genes (SOX9, Aggrecan, type II and X collagens) assessed by real-time PCR. RESULTS: The real-time PCR assay has revealed the expression of gene marker of chondrogenesis, SOX9, Aggrecan and type II collagen, both in Kartogenin and TGFß3 groups compared to the control group, significantly (p < 0.05). A low expression level of collagen type X as a hypertrophic marker was seen in cartilage produced by using Kartogenin. Meanwhile, the level of type X collagen protein in Kartogenin group was significantly decreased (p > 0.05) compared to TGF-ß3 group. CONCLUSION: Kartogenin was suitable for successful chondrogenic differentiation of human adipose- derived stem cells and a suppressor of the consequent hypertrophy (Tab. 1, Fig. 5, Ref. 31).
Assuntos
Anilidas/farmacologia , Condrogênese/efeitos dos fármacos , Ácidos Ftálicos/farmacologia , Células-Tronco/efeitos dos fármacos , Fator de Crescimento Transformador beta3/farmacologia , Tecido Adiposo/citologia , Agrecanas/efeitos dos fármacos , Agrecanas/genética , Cartilagem , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Colágeno Tipo II/efeitos dos fármacos , Colágeno Tipo II/genética , Colágeno Tipo X/efeitos dos fármacos , Colágeno Tipo X/genética , Fibrina , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Fatores de Transcrição SOX9/efeitos dos fármacos , Fatores de Transcrição SOX9/genética , Alicerces TeciduaisRESUMO
PURPOSE: To determine the most optimal stage for antioxidant supplementation of culture medium to improve developmental competence, cryotolerance and DNA-fragmentation of bovine embryos. METHODS: Presumptive zygotes were first cultured in presence or absence of beta-mercaptoethanol (beta-ME), for 8 days. Subsequently, half of the expanded blastocysts developed in both groups were vitrified, warmed within 30 min and post-warming embryos along with their corresponding non-vitrified embryos were cultured for two further days in presence or absence of (100 microM) betaME. RESULTS: For vitrified and non-vitrified embryos, the best effect was found when betaME was added from day 1 of in vitro culture in continuation with post-warming culture period. Day 1-8 supplementation significantly increased the rates of cleavage, day 7 and day 8 blastocyst production. For non-vitrified embryos, betaME addition during day 1-8 and/or 9-10 of embryo culture improved both hatching rate and quality of hatched embryos. For vitrified embryos, however, the percentage of DNA-fragmentation (18.5%) was significantly higher (p < or = 0.05) than that of embryos developed in absence of betaME but supplemented with betaME during post-warming period (13.5%). CONCLUSIONS: Exogenous antioxidant increases the chance of embryos, even those of fair-quality, to develop to blastocyst. However, antioxidant inclusion during in vitro embryo development is not sufficient to maintain the redox state of these embryos during the critical period of post-warming embryo culture, and therefore, there should be a surplus source of exogenous antioxidant during post-warming embryo culture.
Assuntos
Antioxidantes/farmacologia , Blastocisto/efeitos dos fármacos , Criopreservação/métodos , Mercaptoetanol/farmacologia , Animais , Bovinos , Meios de Cultura , Técnicas de Cultura Embrionária , Transferência Embrionária , Feminino , CongelamentoRESUMO
During spermiogenesis, histones are replaced by protamines (P1 and P2), resulting in sperm chromatin condensation followed by a halt to gene expression in haploid spermatids and spermatozoa. As a consequence, protamine deficiency and aberrant P1/P2 ratio have a profound effect on both fertilization and embryo development. However, reports on the effect of the P1/P2 ratio on fertilization and embryo development after intracytoplasmic sperm injection (ICSI) are contradictory between human and animal studies. The question that still remains to be elucidated is which type of protamine deficiency is most common among protamine deficient samples. The present study has a direct bearing on this issue investigating the correlation of the P1/P2 ratio with protamine deficiency, fertilization, embryo quality and embryo development in ICSI patients. This study was carried out on 71 patients. Chromomycin A3 (CMA3) staining was used to determine protamine deficiency. Since this procedure does not indicate the type of protamine deficiency, the P1/P2 ratio was evaluated by nuclear protein extraction, acetic acid urea polyacrylamide gel electrophoresis and analysis of protein bands with software. Polyclonal anti-P1 and anti-P2 antibodies were used to confirm P1 and P2 presence. Results show a negative significant correlation of fertilization rate with protamine deficiency and P1/P2 ratio. No significant correlation was observed between protamine deficiency and P1/P2 ratio. Therefore, it can be concluded that altered P1/P2 ratio effects fertilization rate and embryo quality which subsequently may affect implantation and pregnancy outcome.
