HPV In situ -hybridisaation käyttöönotto

Ihmisen papilloomavirus (Human papillomavirus HPV) aiheuttaa yleisen sukupuoliteitse leviävän tartuntataudin. Virusinfektiolla on todettu olevan yhteys kohdunkaulan syöpään ja on siksi tärkeä tutkimuskohde. Kätilöopiston sairaalan patologian laboratoriossa tutkitaan HPV:ta Hybrid Capture 2 -menetelmällä, joka antaa tuloksen kvantitatiivisena. Tulos on joko positiivinen tai negatiivinen korkean riskin HPV-genotyypeille. Tutkimuskohteena oli ihmisen papilloomavirusta määrittävä in situ -hybridisaatiomenetelmä, joka on tarkoitus ottaa käyttöön Kätilöopiston sairaalan patologian laboratorioon. Näytemateriaali kerättiin HUSLAB:n Qpati-tietokannasta. 40 potilastapausta valittiin, joista kymmenen potilastapausta kuuluu kondyloma planum-, kymmenen dysplasia levis-, kymmenen dysplasia moderata- ja viimeiset kymmenen dysplasia gravis -luokitukseen. Näytteet oli valettu parafiiniin, joista leikkattiin uudet näytelasit. Näistä tehtiin HPV in situ -hybridisaatio-, p16INK4a-, Ki67- ja hematoksyliini-eosiinivärjäykset. HPV ISH:n tuloksia verrattiin negatiivisten ja positiivisten kontrollien antamiin tuloksiin. Lisäksi tutkittiin proteiinien p16INK4a ja Ki67 vastaavuutta HPV ISH:n antamiin tuloksiin. Ki67 ja p16INK4a ovat merkkiaineita, joita havaitaan HPV:n aiheuttamissa dysplastisissa muutoksissa. HE-värjäyksen avulla tarkistettiin diagnoosiluokat. HPV in situ -hybridisaationäytteissä esiintyi HPV DNA-viruskopioita 82,5 %:ssa (33/40). Näytteistä negatiivisen tuloksen antoi 12,5 % (5/40) ja 5 % (2/40) niistä ei voitu tulkita. Immunohistokemialliset värjäykset antoivat keskenään yhteneväisiä tuloksia. Tulokset olivat yhteneviä myös HPV in situ -hybridisaation antamiin tuloksiin. Tulokset olivat luotettavia, koska negatiivinen kontrolli antoi negatiivisen ja positiivinen kontrolli positiivisen tuloksen. Hematoksyliini-eosiinivärjäyksellä tarkistetuista diagnoosiluokista 20 prosenttia (8/40) oli muuttunut. Tulokset olivat luotettavia, joten voidaan todeta, että in situ -hybridisaatiomenetelmä voidaan ottaa käyttöön Kätilöopiston sairaalan patologian laboratoriolle.

Heterogeneous 40Ar* distributions in naturally deformed muscovite: in situ UV-laser ablation evidence for micro structurally controlled intragrain diffusion

Micas are commonly used in Ar-40/Ar-39 thermochronological studies of variably deformed rocks yet the physical basis by which deformation may affect radiogenic argon retention in mica is poorly constrained. This study examines the relationship between deformation and deformation-induced microstructures on radiogenic argon retention in muscovite, A combination of furnace step-heating and high-spatial resolution in situ UV-laser ablation Ar-40/Ar-39 analyses are reported for deformed muscovites sampled from a granitic pegmatite vein within the Siviez-Mischabel Nappe, western Swiss Alps (Penninic domain, Brianconnais unit). The pegmatite forms part of the Variscan (similar to 350 Ma) Alpine basement and exhibits a prominent Alpine S-C fabric including numerous mica `fish' that developed under greenschist facies metamorphic conditions, during the dominant Tertiary Alpine tectonic phase of nappe emplacement. Furnace step-heating of milligram quantities of separated muscovite grains yields an Ar-40/Ar-39 age spectrum with two distinct staircase segments but without any statistical plateau, consistent with a previous study from the same area. A single (3 X 5 mm) muscovite porphyroclast (fish) was investigated by in situ UV-laser ablation. A histogram plot of 170 individual Ar-40/Ar-39 UV-laser ablation ages exhibit a range from 115 to 387 Ma with modes at approximately 340 and 260 Ma. A variogram statistical treatment of the (40)Ad/Ar-39 results reveals ages correlated with two directions; a highly correlated direction at 310 degrees and a lesser correlation at 0 degrees relative to the sense of shearing. Using the highly correlated direction a statistically generated (Kriging method) age contour map of the Ar-40/Ar-39 data reveals a series of elongated contours subparallel to the C-surfaces which where formed during Tertiary nappe emplacement. Similar data distributions and slightly younger apparent ages are recognized in a smaller mica fish. The observed intragrain age variations are interpreted to reflect the partial loss of radiogenic argon during Alpine (similar to 35 Ma) greenschist facies metamorphism. One-dirnensional diffusion modelling results are consistent with the idea that the zones of youngest apparent age represent incipient shear band development within the mica porphyroclasts, thus providing a network of fast diffusion pathways. During Alpine greenschist facies metamorphism the incipient shear bands enhanced the intragrain loss of radiogenic argon. The structurally controlled intragrain age variations observed in this investigation imply that deformation has a direct control on the effective length scale for argon diffusion, which is consistent with the heterogeneous nature of deformation. (C) 2001 Elsevier Science B.V. All rights reserved.

Expression of T cell receptor genes in the thymus: localization of transcripts in situ and comparison of mature and immature subsets.

We have analyzed the expression of T cell receptor (TcR) genes in the thymus using in situ RNA hybridizations with probes to the constant regions of the TcR alpha, beta, gamma and delta chains. Localization of transcripts revealed low TcR alpha mRNA levels in the thymus cortex and very low levels in the subcapsular region. In contrast, TcR beta message was very abundant in the cortex. TcR gamma or delta mRNA+ thymocytes showed a scattered, predominantly cortical localization. In contrast to gamma, TcR delta transcripts were abundant in the subcapsular region. Control experiments with sorted TcR alpha/beta or gamma/delta cells revealed a detection efficiency of 75%-85% for the respective TcR mRNA and data on TcR gene expression in mature, CD3+ thymocytes were consistent with previous reports. The analysis of immature, CD3- thymocyte subsets, however, revealed a virtual absence of TcR alpha transcripts and an unexpectedly high proportion of cells (14%-29%) expressing the gene for the TcR delta chain. The data are discussed in view of current models of lineage relationships in the thymus.

Validation of methane measurement using headspace-GC-MS and quantification by a stable isotope-labeled internal standard generated in situ.

A previous study has shown the possibility to identify methane (CH4 ) using headspace-GC-MS and quantify it with a stable isotope as internal standard. The main drawback of the GC-MS methods discussed in literature for CH4 measurement is the absence of a specific internal standard necessary to perform quantification. However, it becomes essential to develop a safer method to limit the manipulation of gaseous CH4 and to precisely control the injected amount of gas for spiking and calibration by comparison with external calibration. To avoid the manipulation of a stable isotope-labeled gas, we have chosen to generate a labeled gas as an internal standard in a vial on the basis of the formation of CH4 by the reaction of Grignard reagent methylmagnesium chloride with deuterated water. This method allows precise measurement of CH4 concentrations in gaseous sample as well as in a solid or a liquid sample after a thermodesorption step in a headspace vial. A full accuracy profile validation of this method is then presented.

In situ introducer sheath dilatation for complex aortic access.

Aortic access problems due to diseased or small peripheral vessels are a major issue in endovascular aneurysm repair (EVAR). In the emergency setting, like aortic rupture after blunt trauma, or in patients with a hostile abdomen, a more proximal access to the aorta is not a pleasant perspective. We developed in situ introducer sheath dilatation as a bail-out technique for patients with difficult aortic access under various circumstances including EVAR, intra-aortic balloon pump insertion and cannulation for perfusion. The method described allows to increase the access vessel diameter by 50% (from 6 to 9 mm) or the luminal circumference from 18 to 27 F. We have used this technique in five patients without complication, very much in contrast to the traditionally practiced 'forced device insertion'.

Resultados preliminares de las mediciones de fuerza de blanco in situ de las principales especies pelágicas. Crucero BIC Humboldt 9803-05 de Tumbes a Tacna

Se han utilizado los resultados de 144 lances ejecutados en el transcurso del Crucero 9803-05. Se compararon los histogramas de TS y los de tallas en búsqueda de correlación entre ambas a fin de determinar el factor b20 de la Ecuación de Fuerza de Blanco (20 log (LT,cm) + b20). Se han determinado nuevas ecuaciones de TS para anchoveta (38 y 120 kHz), sardina (120 kHz), jurel (38 kHz) y caballa (38 y 120 kHz). Se deberá continuar con estas mediciones puesto que un cambio en las propuestas reflectivas de los peces puede afectar la correcta obtención de los estimados de biomasa.

Mediciones in situ de TS de diversas especies a finales del invierno de 1998. Crucero BIC Humboldt 9808-09

Se efectuaron mediciones de TS (Fuerza de Blanco, Target Strenght) durante los 108 lances de comprobación de ecotrazos realizados por el BIC Humboldt en el marco del Crucero 9808-09 de Evaluación Hidroacústica de Recursos Pelágicos. Se consideran para el análisis únicamente aquellos lances donde más del 90% de la captura perteneció a una sola especie. Los factores b20 de la ecuación de TS (TS=20 log L - b20) que han sido determinados, las cuales deberán ser considerados como provisionales, son las siguientes: Pez cinta (Trichiurus lepturus) = 70,95 (120 kHz); Vinciguerria o pez linterna (Vinciguerria lucetia pacifici) = 83,29 (120 kHz); Samasa (Anchoa nasus) = 86,57 (120 kHz); Caballa (Scomber japonicus) = 83,09 (120 kHz); Pez cinta (Trichiurus lepturus) = 71,41 (38 kHz); y Vinciguerria o pez linterna (Vinciguerria lucetia pacifici)= 82,04 (38kHz).

Mediciones in situ de TS para el krill (Euphasia superba) a 120 Khz. Verano austral 1998. Perú antar IX

Se realizaron mediciones de la Fuerza de Blanco (TS) del krill (Euphausia superba) durante los 29 lances efectuados en el Crucero de Evaluación Hidroacústica utilizando la ecosonda SIMRAD EK 500 a bordo del BIC Humboldt entre los días 12 y 24 de enero de 1998 a lo largo del Estrecho de Bransfield y alrededores de la Isla Elefante. Se derivaron los valores de b20 a partir de le ecuación de TS de FOOTE (1990), de las longitudes promedios de los individuos capturados durante cada uno de los lances y de las tablas de TS generadas por la ecosonda, determinándose una ecuación de TS para el krill en un rango de longitud comprendido entre 2,1 y 5,3 cm de la siguiente forma: TS = 20 log L - 89,26. Se discute el posible sesgo de la ecuación debido, entre otros aspectos a que no se consideraron los estadíos sexuales del krill.

Mediciones in situ de TS a finales de la primavera 1998 crucero BIC José Olaya Balandra 9811-12

Se efectuaron mediciones de TS (Fuerza de Blanco: Target Strength) durante los 100 lances de comprobación de ecotrazos realizados por el BIC José Olaya Balandra durante el Crucero 9811-12 de Evaluación Hidroacústica de Recursos Pelágicos. Se consideraron para el análisis únicamente aquellos lances donde más del 90% de la captura perteneció a una determinada especie y que, además, se haya observado en ellos una adecuada correlación entre los histogramas de tallas y de TS. Los factores b20 de la ecuación de TS (TS = 20 log L - b20) que han sido determinados, las cuales serán consideradas como provisionales, corresponden a anchoveta (2) y a bregmaceros (1). Estas son las siguientes: anchoveta, b20 = 81, 8 (para ejemplares entre 6 y 8,5 cm); anchoveta, b20 = 78,5 (para ejemplares entre 11,5 y 14,5 cm); bregmaceros, b20 = 80,25 (para ejemplares entre 5,5 y 8 cm).

Mediciones in situ de ts a finales del verano 1999. Crucero BIC Jose Olaya Balandra 9902-03

Se efectuaron mediciones de TS (Fuerza de Blanco: Target Strength) durante los 136 lances de comprobación de ecotrazos realizados por el BIC José Olaya Balandra durante el Crucero 9902-03 de Evaluación Hidroacústica de Recursos Pelágicos. Se consideraron para el análisis únicamente aquellos lances donde más del 90% de la captura perteneció a una especie y que, además, se haya observado en ellos una adecuada correlación entre los histogramas de tallas y de TS. Los factores b20 de la ecuación de TS (TS=20 log L - b20) que han sido determinados, las cuales serán consideradas como provisionales, corresponden a anchoveta (2) y a pota (1). Estas son las siguientes: anchoveta a 120 kHz, b20=82,2 (para ejemplares entre 5,5 y 9,5 cm); anchoveta 38 kHz, b20=81,2 (para ejemplares entre 5,5 y 10,5 cm); pota a 38 kHz, b20=87,2 (para ejemplares entre 12 y 24 cm de longitud de manto).

Fluorescence in situ hybridization of potato somatohaploids and their somatic hybrid donors using two Solanum brevidens specific sequences

Selostus: Perunan somaattisten hybridien ja niiden somatohaploidien fluoresenssi in situ -hybridisaatio Solanum brevidens -lajin spesifisten sekvenssien avulla

Particle agglomeration study in rf silane plasmas: In situ study by polarization-sensitive laser light scattering

To determine self‐consistently the time evolution of particle size and their number density in situ multi‐angle polarization‐sensitive laser light scattering was used. Cross‐polarization intensities (incident and scattered light intensities with opposite polarization) measured at 135° and ex situ transmission electronic microscopy analysis demonstrate the existence of nonspherical agglomerates during the early phase of agglomeration. Later in the particle time development both techniques reveal spherical particles again. The presence of strong cross‐polarization intensities is accompanied by low‐frequency instabilities detected on the scattered light intensities and plasma emission. It is found that the particle radius and particle number density during the agglomeration phase can be well described by the Brownian free molecule coagulation model. Application of this neutral particle coagulation model is justified by calculation of the particle charge whereby it is shown that particles of a few tens of nanometer can be considered as neutral under our experimental conditions. The measured particle dispersion can be well described by a Brownian free molecule coagulation model including a log‐normal particle size distribution.