Dissection of inner ear neuroepithelium in zebrafish / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To establish a model by using zebrafish for the inner ear development and disease research, to explore the possibility of complete dissection of inner ear neuroepithelium in zebrafish by surface preparation of inner ear maculae.</p><p><b>METHODS</b>The inner ear samples of zebra fish were fixed followed by harvesting. After decalcified and dehydrated, the coronal cryosections were made at 3 µm thickness at -23°C using cryostat, the hematoxylin and eosin staining and immunofluorescence and histochemistry staining were performed, the saccules, utricles and lagenaes were successfully obtained by fine dissection. The sensory maculae were mounted on glass and then were stained.</p><p><b>RESULTS</b>After hematoxylin and eosin staining and immunofluorescence and histochemistry staining of cryosections, the sensory maculae of inner ear could be determined by otoliths. On the basis of distinguish of otoliths, the sensory maculae of inner ear could be finely dissected, the overall integrity of the sensory maculae could be preserved completely. After immunofluorescence and histochemistry staining, intact epithelia with strong hair cell bundle staining could be seen.</p><p><b>CONCLUSIONS</b>The neuroepithelium hair cell examination of zebra fish can be entirely attained by surface preparation of inner ear maculae.</p>

Virus detection and viral location in brain tissue of sulking mice infected recombinant rabies virus by frozen section / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To observe the existence and location of the recombinant rabies virus in the hippocampus of the sulking mice infected recombinant rabies virus.</p><p><b>METHOD</b>A group of one-day-old sulking mice and 4-week-old mice were challenged with the CTN-GFP strain by intracerebral inoculation, frozen longitudinal transect sections of hippocampus were prepared from the suckling mice in order to observe the expression of the GFP protein and the location of the recombinant rabies virus.</p><p><b>RESULTS</b>DAPI was performed to stain the cell nuclei in blue while GFP expression from CTN-GFP infected brain cells was observed under a confocal microscope.</p><p><b>CONCLUSION</b>The location of the rabies virus can be clearly observed by preparing frozen section of certain sites from the brain, and this method also provide a new tool to trace the route of spread of the rabies virus within the animal host.</p>

Co-stimulatory molecules on peripheral blood mononuclear cells and tissue infiltrating cells of skin wart and in vitro poke weed mitogen stimulation.

BACKGROUND: Skin wart is a lesion caused by human papilloma viruses (HPVs) that can infect both male and female. OBJECTIVE: Quantify the number of CD28+, CD86+, CD152+ and gammadelta+ in peripheral blood mononuclear cells (PBMCs) of subjects with skin wart. Identify CD86+ and gammagamma+ cells in skin wart cryosections. MATERIAL AND METHOD: Sixteen subjects with skin warts on face, hand, finger, knee, foot or plantar, both male and female, aged between 19-59 years-old, were recruited from Ramathibodi Hospital, Mahidol University, Bangkok. RESULTS: CD86 and CD152, on peripheral blood mononuclear cells (PBMCs) of subjects with skin wart are significantly lower compared to controls. Tissue cryosection staining for CD86+ and gammadelta+ cells showed no difference among subjects with skin wart and control. Proliferative response to poke weed mitogen of subjects with skin wart is significantly lower than control subjects. CONCLUSION: There was no difference in the number of subjects positive for CD28 and CD86 cell between normal and skin wart subject, but an increase in skin wart subjects with gammadelta+ cells.

Determinación de cloruros en leche pasteurizada consumida en el estado Mérida-Venezuela y su incidencia en el punto crioscópico / Determination of chlorides in whole pasteurized milk consumed in the state Merida-Venezuela and their incidence in the point crioscópico

Venezuela cuenta con grandes extensiones de suelos y climas propicios para el desarrollo de la ganadería, estas condiciones han permitido un incremento de la producción lechera en los últimos años. La Comisión Venezolana de Normas Industriales (COVENIN), en su norma 798: 1994 (1) contempla los requisitos físico-químicos y microbiológicos para la leche pasteurizada, siendo los valores para cloruros de 0.07 a 0.11 g por ciento y para el punto crioscópico de -0.555 a -0.540 º C. Resultados preliminares realizados por Medina (1998) (2) sobre estos parámetros en muestras de leche cruda consumidas en el Estado Mérida mostraron adulteración por aguado y valores de cloruros por encima de la norma. El objetivo de esta investigación fue la determinación del punto crioscópico y del por ciento de cloruros en leche pasteurizada y su relación. Se analizaron tres marcas comerciales de leche pasteurizada, codificadas como A, B y C; recolectadas semanalmente en los depósitos de las empresas lácteas de la ciudad de Mérida, por un período de 3 meses y llevadas al laboratorio bajo refrigeración para sus respectivos análisis. Para la determinación del punto crioscópico se utilizó el método oficial de la AOAC, mediante el crioscopio Advanced, la determinación de cloruros se realizó mediante el método conductimétrico, utilizando el Corning 920. Covenin 369:1982. Los resultados obtenidos fueron tratados estadísticamente empleando la media, la desviación Standard, el rango de variación como indicadores de centramiento y dispersión. El análisis de significancia estadística se realizó mediante un análisis de varianza para los dos parámetros, así como la t student. De los resultados obtenidos pudimos concluir, que hay valores de crioscopia que están fuera de la norma y los cloruros mostraron una tendencia hacia el límite superior, inclusive algunos valores por encima de 0.11g por ciento. La correlación simple entre la crioscopía y los cloruros demostró una fuerte asociación entre las do...

Cryo-microtome sections of coproculture larvae of Strongyloides stercoralis and Strongyloides ratti as antigen sources for the immunodiagnosis of human strongyloidiasis

Comparou-se o emprego de cortes em congelacao de larvas de S. stercoralis e de S. ratti como fonte de antigenos na reacao de imunofluorescencia indireta (RIF) para o diagnostico da estrongiloidiase humana. Os antigenos foram obtidos de coproculturas de fezes humanas e de ratos, respectivamente. Soros de 123 individuos foram analisados sendo 54 de pacientes com estrongiloidiase e 69 controles. Empregou-se conjugado anti IgG humano marcado com fluorescencia nos titulos ideais de 10 para S. stercoralise 100 para S. ratti. A sensibilidade da RIF foi de 94,4 por cento e 92,5 por cento e a especificidade de 94,2 por cento e 97,1 por cento respectivamente para os antigenos de S. stercoralis e S. ratti...

Distribution of calcium-binding proteins in the chick visual system

The calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV) have been extensively studied over the last decade since they appear to be important as buffers of intracellular calcium. In the present study we investigated the distribution of these proteins in the chick visual system be means of conventional immunocytochemistry. The results indicated that CB, CR, and PV are widely distributed in retinorecipient areas of the chick brain. In some regions, all three calcium-binding proteins were present at different intensities and often in different neurons such as in the dorsolateral thalamic complex. In other areas, such as the nucleus geniculatus lateralis ventralis, only CB and CR were detected, whereas PV was absent. These results show that these three calcium-binding proteins are differentially distributed in the visual system of the chick, with varying degreees of colocalization.

Biodynamics of the Total Human Cadaveric

Spinal trauma produced from motor vehicle accidents, diving accidents, or falls occur at high rates of loading. This study was undertaken to reproduce clinically relevant cervical spine injuries under controlled conditions. Six isolated head-T2 human cadaveric preparations were tested using an electrohydraulic piston actuator at loading rates from 295 to 813 cm/sec. The Hybrid III head-neck was tested similarly at rates from 401 to 683 cm/sec. The input forces for specimen tests were of higher magnitude and shorter duration than the distally measured forces. In contrast, the Hybrid III head-neck revealed similar magnitude and duration force traces from input to output. The specimen preparations were analyzed kinematically at 1200 frame/sec with 20 to 30 retroreflective targets fixed to each level of the cervical spine. With this technique it is possible to temporally follow cervical damage as a function of applied forces. Wedge, burst, and anterior compression fractures were documented using CT and cryosectioning techniques. This pathology can be evaluated quantitatively using the target analysis. For example, a burst fracture occurred suddenly and 2.5 msec, whereas wedge fractures were progressive and took 4 to 5 msec to occur.