Concordancia entre citologia cervico vaginal anormal, colposcopia y biopsia de cervix de usuarias de la Clinica Colombia

El cáncer de cuello uterino, es una de las principales causas de morbimortalidad por cáncer a nivel mundial, el cual se perfila como un problema de salud pública; que gracias a la introducción de la toma de citología cervico vaginal como prueba de tamizaje, para detección temprana de cáncer de cuello uterino, ha venido en descenso. La sociedad Americana de Cáncer en estado Unidos, estima 11.270 casos nuevos para el 2009 y a nivel mundial, casi 500.000 casos nuevos (1,2). Actualmente, según las últimas estadísticas del Instituto Nacional de Cancerología 2009, comprende un 19,1% de casos nuevos. En el siguiente estudio se realizó un análisis descriptivo de concordancia, entre la citología cervico vaginal anormal, con el reporte de colposcopia y el estudio histológico (biopsia de cérvix); así como las principales alteraciones citológicas, según la clasificación de Bethesda que se presentan en las pacientes que son remitidas a la consulta de Ginecología Oncológica de la clínica Colombia. Los resultados obtenidos del estudio, nos mostró una concordancia kappa pobre a débil entre las variables: citología cervico vaginal anormal, colposcopia y biopsia de cérvix, en la Clínica Universitaria Colombia. Se recomienda realizar nuevos estudios de concordancia, previa modificación del informe de colposcopia de la CUC, para así poder unificar todos los conceptos, con el sistema de Bethesda, y el histológico y obtener un mejor resultado de concordancia.

Carga viral de seis tipos de Virus del Papiloma Humano de alto riesgo y su asociacion con lesiones cervicales

Introducción: La infección por un tipo de Virus del Papiloma Humano de alto riesgo (VPH-AR), es el factor principal en el desarrollo de Cáncer de Cérvix (CC). La carga viral puede modular esta asociación, por lo que resulta importante su cuantificación y el establecimiento de su relación con lesiones precursoras de CC. Metodología: 60 mujeres con lesiones escamosas intraepiteliales (LEI) y 120 mujeres sin LEI, confirmadas por colposcopia, fueron incluidas en el estudio. Se determinó la carga viral de 6 tipos de VPH-AR, mediante PCR en tiempo real. Se estimaron OR crudos y ajustados para evaluar la asociación entre la carga viral de cada tipo y las lesiones cervicales. Resultados: 93.22% de mujeres con LEI y 91.23% de mujeres negativas, fueron positivas para al menos un tipo de VPH. VPH-18 y VPH-16 fueron los tipos más prevalentes, junto con VPH-31 en mujeres sin LEI. No se encontraron diferencias estadísticamente significativas de las cargas virales entre éstos dos grupos, aunque se observó un mayor carga viral en lesiones para algunos tipos virales. Una mayor frecuencia de lesiones se asoció a infecciones con carga baja de VPH-16 (ORa: 3.53; IC95%: 1.16 – 10.74), en comparación a mujeres con carga alta de VPH-16, (ORa: 2.63; IC95%: 1.09 – 6.36). En infecciones por VPH-31, la presencia de carga viral alta, se asoció con una menor frecuencia de lesiones (ORa: 0.34; IC95%: 0.15 – 0.78). Conclusiones: La prevalencia tipo-específica de VPH se corresponde con las reportadas a nivel mundial. La asociación entre la carga viral del VPH y la frecuencia de LEI es tipo específica y podría depender de la duración de la infección, altas cargas relacionadas con infecciones transitorias, y bajas cargas con persistentes. Este trabajo contribuye al entendimiento del efecto de la carga viral en la historia natural del CC; sin embargo, estudios prospectivos son necesarios para confirmar estos resultados.

Caracterizacion de nódulos tiroideos: concordancia entre estudio citopatologico y ecografia en la detección de nódulos malignos

: Los métodos imagenológicos para evaluar los nódulos tiroideos han sido motivo de estudio en las últimas décadas, especialmente la ecografía sobresale sobre las otras modalidades diagnósticas por su accesibilidad, portabilidad, y seguridad. A pesar de ello, las características ecográficas de cada nódulo han sido objeto de controversia en cuanto a su potencial detección de malignidad o benignidad. Se presenta un estudio de concordancia entre el estudio citopatológico y la ecografía para la caracterización nódulos tiroideos de naturaleza maligna y benigna, y su análisis de pruebas diagnósticas. Metodología: Se realizó un estudio descriptivo de concordancia con estudio de pruebas diagnósticas anidado. Se escogieron todos los pacientes con nódulos tiroideos a quienes se les realizó ecografía y estudio citopatológico de la lesión y se estudiaron los hallazgos ecográficos para evaluar su potencial diagnóstico para malignidad. Se incluyeron un total de 100 pacientes con nódulos tiroideos potencialmente malignos. La concordancia entre la ecografía en modo B y el estudio citopatológico fue moderada (índice kappa 0.55). La característica con mayor potencial para detectar malignidad fue la presencia de Microcalcificaciones (sensibilidad 75%, especificidad 92%).

Covariates of high-risk human papillomavirus (HPV) infections are distinct for incident CIN1, CIN2 and CIN3 as disclosed by competing-risks regression models

Background: In addition to the oncogenic human papillomavirus (HPV), several cofactors are needed in cervical carcinogenesis, but whether the HPV covariates associated with incident i) CIN1 are different from those of incident ii) CIN2 and iii) CIN3 needs further assessment. Objectives: To gain further insights into the true biological differences between CIN1, CIN2 and CIN3, we assessed HPV covariates associated with incident CIN1, CIN2, and CIN3. Study Design and Methods: HPV covariates associated with progression to CIN1, CIN2 and CIN3 were analysed in the combined cohort of the NIS (n = 3,187) and LAMS study (n = 12,114), using competing-risks regression models (in panel data) for baseline HR-HPV-positive women (n = 1,105), who represent a sub-cohort of all 1,865 women prospectively followed-up in these two studies. Results: Altogether, 90 (4.8%), 39 (2.1%) and 14 (1.4%) cases progressed to CIN1, CIN2, and CIN3, respectively. Among these baseline HR-HPV-positive women, the risk profiles of incident GIN I, CIN2 and CIN3 were unique in that completely different HPV covariates were associated with progression to CIN1, CIN2 and CIN3, irrespective which categories (non-progression, CIN1, CIN2, CIN3 or all) were used as competing-risks events in univariate and multivariate models. Conclusions: These data confirm our previous analysis based on multinomial regression models implicating that distinct covariates of HR-HPV are associated with progression to CIN1, CIN2 and CIN3. This emphasises true biological differences between the three grades of GIN, which revisits the concept of combining CIN2 with CIN3 or with CIN1 in histological classification or used as a common end-point, e.g., in HPV vaccine trials.

Correlación entre estudio citológico y estudio histopatológico en el diagnóstico de Neoplasia Tiroidea. SOLCA - Cuenca. 2009- 2013

La neoplasia tiroidea impulsa la búsqueda de métodos diagnósticos para obtener un dictamen precoz y tratamiento oportuno que permitan mayor supervivencia y mejor calidad de vida. Objetivo: determinar la correlación entre estudio citológico e histopatológico en el diagnóstico de Neoplasia Tiroidea en pacientes atendidos en SOLCA – Cuenca, periodo 2009-2013. Metodología: estudio observacional, retrospectivo, analítico y de correlación diagnóstica, elaborado con historias clínicas de pacientes en quienes se realizó punciones (PAAF) para la citología, según el Sistema Bethesda, y con histopatología, para diagnosticar neoplasia tiroidea. Resultados: investigación desarrollada con 415 pacientes con neoplasia tiroidea. Caracterizada por 89.2% de mujeres; edad promedio de 51.8 ± 15.2 años, de 41-55 años fue la mayor categoría (36,9%); 47.2% procedieron de Cuenca y el 37.8% de las provincias vecinas. Estado civil casado/a fue más frecuente, 269 (64,8%), y de profesión “amas de casa” fueron las más afectadas 231 (55,7%). El 96.4% de diagnósticos citológicos Bethesda categoría 6, fueron confirmados por histología. Hubo correlación (r = 0.49) significativa y concordancia moderada (kappa = 0.337) entre citología e histología. Sensibilidad=63% (IC95%: 58 – 69), Especificidad=94% (IC95%: 89 – 98), RVP=10.9 (IC95%: 5 – 22) y RVN=0.39 (IC95%: 0.3 – 0.4). Conclusiones: la citología por PAAF es una herramienta para el estudio, diagnóstico de pacientes con afecciones tiroideas. Una punción realizada por expertos es una técnica rápida, económica, bien tolerada, y produce resultados confiables. La categorización Bethesda representa un sistema confiable, válido para reportar citología de tiroides

Examination of Endogenous Rotund Expression and Function in Developing Drosophila Olfactory System Using CRISPR-Cas9-Mediated Protein Tagging.

The zinc-finger protein Rotund (Rn) plays a critical role in controlling the development of the fly olfactory system. However, little is known about its molecular function in vivo. Here, we added protein tags to the rn locus using CRISPR-Cas9 technology in Drosophila to investigate its subcellular localization and the genes that it regulates . We previously used a reporter construct to show that rn is expressed in a subset of olfactory receptor neuron (ORN) precursors and it is required for the diversification of ORN fates. Here, we show that tagged endogenous Rn protein is functional based on the analysis of ORN phenotypes. Using this method, we also mapped the expression pattern of the endogenous isoform-specific tags in vivo with increased precision. Comparison of the Rn expression pattern from this study with previously published results using GAL4 reporters showed that Rn is mainly present in early steps in antennal disc patterning, but not in pupal stages when ORNs are born. Finally, using chromatin immunoprecipitation, we showed a direct binding of Rotund to a previously identified regulatory element upstream of the bric-a-brac gene locus in the developing antennal disc.

The zymogen-enteropeptidase system: A practical approach to study the regulation of enzyme activity by proteolytic cleavage

The present research describes an efficient procedure to obtain high levels of trypsinogen and chymotrypsinogen by using a simple, rapid, and easily reproducible method. The extraction process and the time-course of activation of zymogens can be carried out in a single laboratory period, without sophisticated equipment. The main objective was to prepare a laboratory class that would stimulate student interest in enzyme regulation, exploring the fact that the catalytic activity of some enzymes is regulated by different mechanisms. The regulation of proteolytic enzymes requires the synthesis of an inactive zymogen and its being irreversibly switched on by specific proteolytic cleavage.

Medial prefrontal cortex endocannabinoid system modulates baroreflex activity through CB1 receptors

Ferreira-Junior NC, Fedoce AG, Alves FHF, Correa FMA, Resstel LBM. Medial prefrontal cortex endocannabinoid system modulates baroreflex activity through CB1 receptors. Am J Physiol Regul Integr Comp Physiol 302: R876-R885, 2012. First published December 28, 2011; doi: 10.1152/ajpregu.00330.2011.-Neural reflex mechanisms, such as the baroreflex, are involved in the regulation of cardiovascular system activity. Previous results from our group (Resstel LB, Correa FM. Medial prefrontal cortex NMDA receptors and nitric oxide modulate the parasympathetic component of the baroreflex. Eur J Neurosci 23: 481-488, 2006) have shown that glutamatergic synapses in the ventral portion of the medial prefrontal cortex (vMPFC) modulate baroreflex activity. Moreover, glutamatergic neurotransmission in the vMPFC can be modulated by the endocannabinoids system (eCBs), particularly the endocannabinoid anandamide, through presynaptic CB1 receptor activation. Therefore, in the present study, we investigated eCBs receptors that are present in the vMPFC, and more specifically whether CB1 receptors modulate baroreflex activity. We found that bilateral microinjection of the CB1 receptor antagonist AM251 (100 or 300 pmol/200 nl) into the vMPFC increased baroreflex activity in unanesthetized rats. Moreover, bilateral microinjection of either the anandamide transporter inhibitor AM404 (100 pmol/200 nl) or the inhibitor of the enzyme fatty acid amide hydrolase that degrades anandamide, URB597 (100 pmol/200 nl), into the MPFC decreased baroreflex activity. Finally, pretreatment of the vMPFC with an ineffective dose of AM251 (10 pmol/200 nl) was able to block baroreflex effects of both AM404 and URB597. Taken together, our results support the view that the eCBs in the vMPFC is involved in the modulation of baroreflex activity through the activation of CB1 receptors, which modulate local glutamate release.

A metalloproteinase karilysin present in the majority of Tannerella forsythia isolates inhibits all pathways of the complement system

Tannerella forsythia is a poorly studied pathogen despite being one of the main causes of periodontitis, which is an inflammatory disease of the supporting structures of the teeth. We found that despite being recognized by all complement pathways, T. forsythia is resistant to killing by human complement, which is present at up to 70% of serum concentration in gingival crevicular fluid. Incubation of human serum with karilysin, a metalloproteinase of T. forsythia, resulted in a decrease in bactericidal activity of the serum. T. forsythia strains expressing karilysin at higher levels were more resistant than low-expressing strains. Furthermore, the low-expressing strain was significantly more opsonized with activated complement factor 3 and membrane attack complex from serum compared with the other strains. The high-expressing strain was more resistant to killing in human blood. The protective effect of karilysin against serum bactericidal activity was attributable to its ability to inhibit complement at several stages. The classical and lectin complement pathways were inhibited because of the efficient degradation of mannose-binding lectin, ficolin-2, ficolin-3, and C4 by karilysin, whereas inhibition of the terminal pathway was caused by degradation of C5. Interestingly, karilysin was able to release biologically active C5a peptide in human plasma and induce migration of neutrophils. Importantly, we detected the karilysin gene in >90% of gingival crevicular fluid samples containing T. forsythia obtained from patients with periodontitis. Taken together, the newly characterized karilysin appears to be an important virulence factor of T. forsythia and might have several important implications for immune evasion.

Fiberoptic system for recording dendritic calcium signals in layer 5 neocortical pyramidal cells in freely moving rats

Calcium influx into the dendritic tufts of layer 5 neocortical pyramidal neurons modifies a number of important cellular mechanisms. It can trigger local synaptic plasticity and switch the firing properties from regular to burst firing. Due to methodological limitations, our knowledge about Ca2+ spikes in the dendritic tuft stems mostly from in vitro experiments. However, it has been speculated that regenerative Ca2+ events in the distal dendrites correlate with distinct behavioral states. Therefore it would be most desirable to be able to record these Ca2+ events in vivo, preferably in the behaving animal. Here, we present a novel approach for recording Ca2+ signals in the dendrites of populations of layer 5 pyramidal neurons in vivo, which ensures that all recorded fluorescence changes are due to intracellular Ca2+ signals in the apical dendrites. The method has two main features: 1) bolus loading of layer 5 with a membrane-permeant Ca2+ dye resulting in specific loading of pyramidal cell dendrites in the upper layers and 2) a fiberoptic cable attached to a gradient index lens and a prism reflecting light horizontally at 90 degrees to the angle of the apical dendrites. We demonstrate that the in vivo signal-to-noise ratio recorded with this relatively inexpensive and easy-to-implement fiberoptic-based device is comparable to conventional camera-based imaging systems used in vitro. In addition, the device is flexible and lightweight and can be used for recording Ca2+ signals in the distal dendritic tuft of freely behaving animals.

Sympathetic nervous system activity in rat thyroid: potential role in goitrogenesis

The role of sympathetic innervation in regulation of thyroid function is incompletely understood. We, therefore, carried out studies in rats utilizing techniques of norepinephrine turnover to assess thyroid sympathetic activity in vivo. Thyroidal sympathetic activity was increased 95% by exposure to cold (4 degrees C), 42% by chronic ingestion of an iodine-deficient diet, and 32% in rats fed a goitrogenic diet (low-iodine diet supplemented with propylthiouracil). In addition, fasting for 2 days reduced sympathetic nervous system activity in thyroid by 38%. Thyroid growth and 125I uptake were also compared in intact and decentralized hemithyroids obtained from animals subjected to unilateral superior cervical ganglion decentralization. Unilateral superior cervical ganglion decentralization led to a reduction in thyroid weight, in 125I uptake by thyroid tissue, and in TSH-induced stimulation of 125I uptake in decentralized hemithyroids. These results suggest that sympathetic activity in thyroid contributes to gland enlargement and may modulate tissue responsiveness to TSH.