RESUMEN
BACKGROUND: Vitrification is the most popular technique for the cryopreservation of oocytes and embryos, replacing slow freezing methods. MATERIALS AND METHODS: This study evaluated the efficient manufacturing methods of handmade open pulled straw (OPS) with a digital heating gun that could be proposed for vitrification. RESULTS: Production efficiency of OPS using 0.5 mL straw was detected at 0, 66.1, 90.5 and 85.7% for 1~2 s and 9.5, 33.3, 47.6 and 23.8% for 2~3 s of heating time at 250, 350, 400 and 450°C respectively. The production rate of OPS using 0.25 mL straw was perceived at 33.3, 76.2, 83.3, 95.2 and 57.6 % for 1.5~2.5 s time with the optimized heat setting at 330, 340, 350, 360 and 370°C respectively. The desired inner diameter (200~300 µm) of OPS could be varied according to the gamete size, embryo developmental stages or cell lines of different species. Based on our data, the production efficiency of OPS using 0.25-mL straw were increased beyond using a 0.5-mL straw. CONCLUSION: Handmade OPSs could be efficiently produced with a digital heating gun to generate a vitrification device for freezing gametes, embryos and cell lines.
RESUMEN
For successful clinical tumor immunotherapy outcomes, strong immune responses against tumor antigens must be generated. Cell-based vaccines compromise one strategy with which to induce appropriate strong immune responses. Previously, we established a natural killer T-cell (NKT) ligand-loaded, adenoviral vector-transduced B-cell-based anticancer cellular vaccine. To enhance tumor antigen delivery to B cells, we established a modified adenoviral vector (Ad-k35) that encoded a truncated form of the breast cancer antigen Her2/neu (Ad-k35HM) in which fiber structure was substituted with adenovirus serotype 35. We observed increased tumor antigen expression with Ad-k35HM in both human and murine B cells. In addition, an Ad-k35HM-transduced B-cell vaccine elicited strong antigen-specific cellular and humoral immune responses that were further enhanced with the additional loading of soluble NKT ligand KBC009. An Ad-k35HM-transduced, KBC009-loaded B-cell vaccine efficiently suppressed the in vivo growth of established tumors in a mouse model. Moreover, the vaccine elicited human leukocyte antigen (HLA)-A2 epitope-specific cytotoxic T-cell responses in B6.Cg (CB)-Tg (HLA-A/H2-D) 2Enge/Jat mice. These findings indicated that the Ad-k35 could be appropriate for the preclinical and clinical development of B-cell-based anticancer immunotherapies.
Asunto(s)
Linfocitos B/inmunología , Vacunas contra el Cáncer , Dependovirus/genética , Neoplasias Mamarias Experimentales/terapia , Receptor ErbB-2/genética , Animales , Linfocitos B/virología , Vacunas contra el Cáncer/inmunología , Células Cultivadas , Dependovirus/metabolismo , Femenino , Vectores Genéticos , Antígeno HLA-A2/inmunología , Humanos , Inmunoterapia , Neoplasias Mamarias Experimentales/inmunología , Ratones , Ratones Endogámicos BALB C , Células T Asesinas Naturales/inmunología , Receptor ErbB-2/metabolismo , Linfocitos T Citotóxicos/inmunología , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
BACKGROUND: The aim of this study was to evaluate the efficacy of palonosetron, the latest 5-HT3 receptor antagonist, for the prevention of postoperative nausea and vomiting (PONV) during the first 72 h after operation. METHODS: In this randomized, double-blinded, placebo-controlled study, 204 healthy inpatients who were undergoing elective surgery with general anaesthesia were enrolled. Patients were divided into two groups: the palonosetron group (palonosetron 0.075 mg i.v.; n=102) and the placebo group (normal saline i.v.; n=102). The treatments were given after the induction of anaesthesia. The incidence of nausea, vomiting, severity of nausea, and the use of rescue anti-emetics during the first 72 h after surgery were evaluated. RESULTS: The incidence of PONV was lower in the palonosetron group compared with the placebo group during the 0-24 h (33% vs 47%) and 0-72 h period (33% vs 52%) (P<0.05), but not during the 24-72 h postoperative period (6% vs 11%). The incidence of nausea was also significantly lower in the palonosetron group than in the placebo group during the 0-24 and 0-72 h period (P<0.05), but not during the 24-72 h postoperative period. However, there were no significant differences in the incidence of vomiting, and the use of rescue anti-emetics between the groups. CONCLUSIONS: Palonosetron 0.075 mg i.v. effectively reduced the incidence of PONV during the first 72 h after operation, with most of the reduction occurring in the first 24 h.
Asunto(s)
Antieméticos/uso terapéutico , Isoquinolinas/uso terapéutico , Náusea y Vómito Posoperatorios/prevención & control , Quinuclidinas/uso terapéutico , Adulto , Anciano , Antieméticos/efectos adversos , Método Doble Ciego , Femenino , Estudios de Seguimiento , Humanos , Isoquinolinas/efectos adversos , Masculino , Persona de Mediana Edad , Palonosetrón , Náusea y Vómito Posoperatorios/diagnóstico , Náusea y Vómito Posoperatorios/epidemiología , Quinuclidinas/efectos adversos , Medición de Riesgo , Tamaño de la Muestra , Procedimientos Quirúrgicos Operativos , Resultado del Tratamiento , Adulto JovenRESUMEN
OBJECTIVE: Survival of periodontal ligament cells is crucial issue after tooth replantation. To understand this matter, we introduced MTT assay, which can be used as a tool for measuring the viability of periodontal ligament cells from extracted rat molars. MATERIALS AND METHODS: The maxillary molars of 4-week-old Sprague-Dawley white female rats were used. Ten teeth of each immediate, 1 h Viaspan and 1 h dry were processed for MTT evaluation. Another 10 teeth from each group were treated in the same manner as above, but were replanted into their original socket. After 2 weeks, the animals were killed and the prevalence of resorption pits was evaluated. RESULTS: In vivo MTT assay corresponded with the histological results of the resorption pits (P Asunto(s)
Diente Molar/citología
, Ligamento Periodontal/citología
, Resorción Radicular/patología
, Animales
, Supervivencia Celular
, Femenino
, Formazáns
, Indicadores y Reactivos
, Diente Molar/patología
, Ligamento Periodontal/patología
, Ratas
, Ratas Sprague-Dawley
, Resorción Radicular/etiología
, Reimplante Dental/efectos adversos
RESUMEN
ETV1, ERM and E1A-F are members of the multigene ETS domain containing a class of transcription factors, first identified in the genome of the avian retrovirus E26. Based upon extensive homology between these genes within their ETS domain (96% identity each other), these three genes comprise a distinct sub-family of ETS genes as a human PEA3 sub-family. By analyzing somatic cell hybrid segregating human chromosomes, the genes encoding ETV1, ERM and E1A-F were localized to human chromosome 7, 3 and 17, respectively. Fluorescence in situ hybridization confirmed these assignments and allowed mapping of the genes to 7p22 (ETV1), 3q29 (ERM) and 17q12 (E1A-F). These results suggest the ancestral PEA3 gene may have duplicated first, then dispersed to other chromosomal locations.
Asunto(s)
Proteínas E1A de Adenovirus/genética , Cromosomas Humanos Par 17 , Cromosomas Humanos Par 3 , Cromosomas Humanos Par 7 , Proteínas de Unión al ADN/genética , Proteínas Proto-Oncogénicas/genética , Factores de Transcripción/genética , Animales , Mapeo Cromosómico , Humanos , Células Híbridas , Hibridación Fluorescente in Situ , Familia de Multigenes , Filogenia , Proteínas Proto-Oncogénicas c-ets , RoedoresRESUMEN
Chimeric transcription factors, created by gene fusions as the result of chromosomal translocations, have been implicated in the pathogenesis of several pathologically disparate solid tumors. The PAX3/FKHR fusion gene, formed by a t(2;13)(q35;q14) in alveolar rhabdomyosarcoma, encodes a hybrid protein that contains both PAX3 DNA binding domains, the paired box and homeodomain, linked to the bisected DNA binding domain of FKHR, a member of the forkhead family of transcription factors. Here we report that PAX3 and PAX3/FKHR display similar, but not identical transactivation activities when tested with model Pax recognition sequences. No functional role could be ascribed solely to the residual FKHR binding domain present in the fusion protein, but FKHR was found to contribute a strong carboxyl terminal activation domain replacing the one located in the unrearranged PAX3 gene. We show that the native PAX3/FKHR protein present in tumor cells with this translocation has transcriptional characteristics similar to the in vitro expressed protein. The ability of the PAX3/FKHR hybrid protein to bind DNA in a sequence specific manner and to transactivate the expression of artificial reporter genes suggests that its aberrant expression could subvert the transcriptional programs that normally control the growth, differentiation, and survival of primitive myogenic precursors in vivo.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Proteínas Recombinantes de Fusión/fisiología , Rabdomiosarcoma Alveolar/genética , Factores de Transcripción/genética , Factores de Transcripción/fisiología , Activación Transcripcional , Secuencia de Bases , Cromosomas Humanos Par 13 , Cromosomas Humanos Par 2 , Proteína Forkhead Box O1 , Factores de Transcripción Forkhead , Regulación Neoplásica de la Expresión Génica , Humanos , Técnicas In Vitro , Datos de Secuencia Molecular , Oligodesoxirribonucleótidos/química , Factor de Transcripción PAX3 , Factores de Transcripción Paired Box , ARN Mensajero/genética , Transcripción Genética , Translocación Genética , Células Tumorales CultivadasRESUMEN
Most Ewing's sarcomas or related primitive neuroectodermal tumors have the (11;22)(q24;q12) or less frequently the (21;22)(q22;q12) translocation. These rearrangements fuse the EWS gene on chromosome 22q12 to either the FLI1 or ERG genes, both members of the ETS family of transcription factors. Simple variant chromosomal translocations have been occasionally described in these tumors. We have identified a third Ewing's sarcoma translocation, the t(7;22)(p22;q12), that fuses EWS to the human homologue of the murine ETS gene ER81. This gene, designated ETV1 (for ETS Translocation Variant), is located on chromosome band 7p22. Identical EWS nucleotide sequences found in the majority of EWS-FLI1 and EWS-ERG chimeric transcripts are fused to a portion of ETV1 encoding an ETS domain with sequence specific DNA-binding activity. These findings confirm that the fusion of EWS to different ETS family members can result in a similar tumor phenotype.
Asunto(s)
Cromosomas Humanos Par 22 , Cromosomas Humanos Par 7 , Proteínas Nucleares/genética , Proteínas de Unión al ARN/genética , Ribonucleoproteínas/genética , Sarcoma de Ewing/genética , Factores de Transcripción/genética , Translocación Genética , Adulto , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Preescolar , Clonación Molecular , ADN Complementario , Proteínas de Unión al ADN/metabolismo , Femenino , Feto/metabolismo , Ribonucleoproteínas Nucleares Heterogéneas , Humanos , Ratones , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteína EWS de Unión a ARN , Proteínas Recombinantes de Fusión/genética , Homología de Secuencia de AminoácidoRESUMEN
Incidence of and contributory factors in postintubation laryngeal edema were determined in 7875 children under 17 years of age. Data were assembled in the manner of a prospective study. With an overall incidence of 1 percent, children between ages 1 and 4 were most susceptible. Excessive size of the endotracheal tube was suspect in half of the cases. Other factors that increase trauma to the larynx while an endotracheal tube is in place showed significant correlation to the total incidence of postintubation laryngeal edema. No tracheostomies were required.