RESUMO
OBJECTIVES: The aim of this study was to assess the predictive value of the Vandenberg and Kuse Mental Rotation Test (MRT) on performance of novice medical students for manipulation of a nasal endoscope on a cadaveric model. MATERIAL AND METHOD: We randomly selected 39 medical students who had never handled a nasal endoscope and subjected them to the MRT. General information including experience in manual, technical, or surgical activities and testing of anatomical knowledge were collected to exclude possible confounding factors. They were then asked to perform series of cadaveric model exercises using a nasal endoscope. Their cadaver performance was evaluated by 2 blinded observers, using a standardized scale. RESULTS: We found that medical students with higher mental rotation skills had significantly increased endoscopic sinus performance (P = .0002 using multivariate regression adjusted for specialty choice, previous surgical exposure, and anatomy knowledge). Higher anatomy knowledge was also associated with better endoscopic sinus performance (P = .0141). Other parameters had no impact on endoscopic sinus performance measured by the endoscopic scale (P > .05). CONCLUSION: The score obtained on the MRT was correlated with the practical performance of manipulating the nasal endoscope in cadaver. It could therefore be a useful spatial ability tool for directing targeted training in rhinology.
Assuntos
Seios Paranasais , Estudantes de Medicina , Cadáver , Endoscopia/educação , Humanos , Cavidade Nasal , Seios Paranasais/cirurgiaRESUMO
Prion diseases are caused by the transconformation of the host cellular prion protein PrP(c) into an infectious neurotoxic isoform called PrP(Sc). While vaccine-induced PrP-specific CD4(+) T cells and antibodies partially protect scrapie-infected mice from disease, the potential autoreactivity of CD8(+) cytotoxic T lymphocytes (CTLs) received little attention. Beneficial or pathogenic influence of PrP(c)-specific CTL was evaluated by stimulating a CD8(+) T-cell-only response against PrP in scrapie-infected C57BL/6 mice. To circumvent immune tolerance to PrP, five PrP-derived nonamer peptides identified using prediction algorithms were anchored-optimized to improve binding affinity for H-2D(b) and immunogenicity (NP-peptides). All of the NP-peptides elicited a significant number of IFNγ secreting CD8(+) T cells that better recognized the NP-peptides than the natives; three of them induced T cells that were lytic in vivo for NP-peptide-loaded target cells. Peptides 168 and 192 were naturally processed and presented by the 1C11 neuronal cell line. Minigenes encoding immunogenic NP-peptides inserted into adenovirus (rAds) vectors enhanced the specific CD8(+) T-cell responses. Immunization with rAd encoding 168NP before scrapie inoculation significantly prolonged the survival of infected mice. This effect was attributable to a significant lengthening of the symptomatic phase and was associated with enhanced CD3(+) T cell recruitment to the CNS. However, immunization with Ad168NP in scrapie-incubating mice induced IFNγ-secreting CD8(+) T cells that were not cytolytic in vivo and did not influence disease progression nor infiltrated the brain. In conclusion, the data suggest that vaccine-induced PrP-specific CD8(+) T cells interact with prions into the CNS during the clinical phase of the disease.
Assuntos
Sistema Nervoso Central/patologia , Doenças Priônicas/imunologia , Complexo Receptor-CD3 de Antígeno de Linfócitos T/imunologia , Scrapie/patologia , Linfócitos T/imunologia , Sequência de Aminoácidos , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Sobrevivência Celular , Sistema Nervoso Central/imunologia , Radioisótopos de Cromo , Imunofluorescência , Imunização , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Dados de Sequência Molecular , Peptídeos/imunologia , Plasmídeos/genética , Proteínas PrPC/genética , Proteínas PrPC/imunologia , Scrapie/imunologiaRESUMO
Although the alimentary tract has been suggested as the most likely portal of entry in natural scrapie, a growing amount of data indicates that the respiratory system and more specifically the pharyngeal tonsils serve as a natural portal of entry for scrapie. This study describes for the first time the broad cell populations in the lymphoid compartment of pharyngeal tonsils and more specifically inside the lymphoid follicles where the scrapie agent accumulates during the period of latency. Follicular dendritic cells (FDCs), stromal cells located in the light zone of the germinal centre of lymphoid follicles, seem to be the principal causal factor in the accumulation of the infectious agent in transmissible spongiform encephalopathy (TSE) diseases. Knowing that efficient lymphoreticular prion propagation requires PrPc expression, we analysed the expression of PrPc with the mouse monoclonal antibody Pri 909 both in situ and on FDC-cluster-enriched cell suspensions. In situ, a positive staining was observed in the germinal centre of pharyngeal lymph follicles. The germinal centre labelling was due to the presence of a follicular dendritic network as revealed after immunogold staining of isolated FDC clusters. Our results suggest that the pharyngeal lymphoreticular system and more specifically PrPc expressing follicular dendritic cells could serve as a prion "reservoir" during the latency phase, thus playing a key role during the scrapie lymphoinvasion.
Assuntos
Tonsila Faríngea/patologia , Células Dendríticas Foliculares/patologia , Scrapie/patologia , Tonsila Faríngea/imunologia , Animais , Anticorpos/imunologia , Células Dendríticas Foliculares/imunologia , Imuno-Histoquímica/veterinária , Proteínas PrPC/imunologia , Scrapie/imunologiaRESUMO
BACKGROUND: The thiopurine prodrug azathioprine is used extensively in cancer therapy. Exposure to this drug results in the selection of DNA mismatch repair-deficient cell clones in vitro. It has also been suggested that thiopurine drugs might constitute a risk factor for the emergence of human neoplasms displaying microsatellite instability (MSI) because of deficient DNA mismatch repair. METHODS: Azathioprine was administered via drinking water (6-20 mg/kg body weight per day) to mice that were null (Msh2â»(/)â»; n = 27), heterozygous (Msh2(+/)â»; n = 22), or wild type (Msh2(WT); n = 18) for the DNA mismatch repair gene Msh2. Control mice (45 Msh2â»(/)â», 38 Msh2(+/)â», and 12 Msh2(WT)) received drinking water lacking azathioprine. The effect of azathioprine on tumorigenesis and survival of the mice was evaluated by Kaplan-Meier curves using log-rank and Gehan-Breslow-Wilcoxon tests. Mouse tumor samples were characterized by histology and immunophenotyping, and their MSI status was determined by polymerase chain reaction analysis of three noncoding microsatellite markers and by immunohistochemistry. Msh2 status of tumor samples was assessed by loss of heterozygosity analyses and sequencing after reverse transcription-polymerase chain reaction of the entire Msh2 coding sequence. All statistical tests were two-sided. RESULTS: Most untreated Msh2(WT) and Msh2(+/)â» mice remained asymptomatic and alive at 250 days of age, whereas azathioprine-treated Msh2(WT) and Msh2(+/)â» mice developed lymphomas and died prematurely (median survival of 71 and 165 days of age, respectively). Azathioprine-treated Msh2(+/)â» mice developed diffuse lymphomas lacking Msh2 expression and displaying MSI due to somatic inactivation of the functional Msh2 allele by loss of heterozygosity or mutation. By contrast, azathioprine-treated Msh2(WT) mice displayed no obvious tumor phenotype, but histological examination showed microscopic splenic foci of neoplastic lymphoid cells that retained Msh2 expression and did not display MSI. Both untreated and azathioprine-treated Msh2â»(/)â» mice had a reduced lifespan compared with untreated Msh2(WT) mice (median survival of 127 and 107 days of age, respectively) and developed lymphomas with MSI. CONCLUSION: Azathioprine-induced carcinogenesis in mice depends on the number of functional copies of the Msh2 gene.