Detección de un mosaico de trisomía 21 en líquido amniótico / Detection of a mosaicism of trisomy 21 in amniotic liquid

Resumen Se analizó un resultado con alteración cromosómica tomado de una base de datos conformada por un total de 4755 muestras de líquido amniótico extraídos mediante amniocentesis con indicación de su médico tratante, riesgo sérico y edad materna avanzada. En este reporte se presenta la detección de un mosaico de trisomía 21 en líquido amniótico, mediante la técnica de Banda G donde se analizaron 20 metafases. Los resultados obtenidos documentan una composición cromosómica 47, XY+21 y 46, XY con una relación 9:11 respecto a las metafases analizadas, confirmándose así el diagnóstico del Síndrome de Down secundario a mosaico.

Abstract A result with chromosomal alteration was analyzed from a database consisting of a total of 4755 samples of amniotic fluid extracted by amniocentesis with indication of the attending physician, serum risk and advanced maternal age. This report presents the detection of a mosaicism of trisomy 21 in amniotic fluid, using G- Banding where 20 metaphases were analyzed. The results obtained document a chromosomal composition 47, XY + 21 and 46, XY with a 9:11 ratio with respect to the metaphases analyzed, confirming the diagnosis of Down syndrome secondary to mosaicism.

El adipocito en el laboratorio: historia, perspectivas y reporte de experiencia / The adipocyte in the laboratory: history, perspectives and report of experience

Introducción: el tejido adiposo humano está compuesto por diferentes tipos celulares, y ha sido objeto de múltiples estudios en los últimos años debido a su acción en diversas funciones. Métodos: se realizó una búsqueda bilbiográfica en PubMed, Scielo, Science Direct and Google Scholars y se reporta la experiencia de los autores. Resultados: los primeros modelos de estudio fueron con tejido de roedores, y permitieron la comprensión del metabolismo de carbohidratos y ácidos grasos y facilitaron el estudio de la biología del adipocito, junto a estudios histológicos y el aislamiento de adipocitos maduros. Posteriormente se realizaron estudios con células precursoras (preadipocitos) que propiciaron el aislamiento de la línea celular 3T3-1L murina. En Latinoamérica, se han realizado diversos estudios con líneas celulares y con células madre mesenquimales precursoras de adipocitos para estudiar el efecto de hormonas y otras sustancias y para genotipificación. En Colombia se han realizado estudios con adipocitos 3T3-L1 para determinar los efectos de medicamentos y sustancias en estas células. En el laboratorio de Fisiología Celular de la Universidad Tecnológica de Pereira el proceso de obtención de muestras ha evidenciado dificultades por tratarse de tejido humano, pero el protocolo de aislamiento y cultivo pudo ser estandarizado a lo largo de seis años de experimentación; se aislaron preadipocitos y adipocitos maduros que permitieron estudiar los efectos de hormonas, realizar caracterización electrofisiológica y estudiar la fisiología del calcio. Conclusiones: este es un campo de investigación muy relevante debido a la implicación de este tipo celular en funciones metabólicas sistémicas y su relación con patologías de alta prevalencia como la obesidad y el síndrome metabólico.

Introduction: The human adipose tissue is composed of different cell types. It has been subject of several studies in the past years due to its role on diverse functions. Methods: A search in the PubMed, Scielo, Science direct and Google Scholars databases was performed and the authors experience was reported. Results: The first model used was rodent fat, which allowed a better comprehension on carbohydrate and fatty acid metabolism and enhanced research in adipocyte cell biology in addition with histological studies and mature adipocyte isolation. Afterwards, learning about precursor cell (pre-adipocytes) promoted the isolation of murine 3T3-L1 cell line. In Latin America research has been conducted using cell lines and adipocyte precursor mesenchymal stem cells to describe effects of hormones and perform DNA sequencing. In Colombia, studies in 3T3-L1 cell line aimed to stablish the effects of different compounds on these cells. In the Cell Physiology Laboratory of the Universidad Tecnológica de Pereira, sample collection process has shown difficulties because the source was human tissue; nevertheless isolation and cell culture protocols were standardized throughout the last six years of experimentation. Pre-adipocytes and mature adipocytes were isolated to study the effects of hormones, perform electrophysiological characterization and study calcium physiology. Conclusions: This is a relevant research field since these cells have important systemic metabolic functions and they have a clear relationship with high-prevalence pathologies such as obesity and the metabolic syndrome.

Cytotoxicity of dental alginates / Citotoxicidad de alginatos dentales

Alginate, or irreversible hydrocolloid, is one of the most accepted impression materials used in dentistry. However, some substances existing in these materials can be toxic. The aim of this study was to assess the cytotoxicity of alginates for dental applications. Fourteen different alginates were assessed: Jeltrate, Jeltrate Plus, Jeltrate Chromatic, Alga Gel, Printer Gel, Ava Gel, New Print, Kromopan 100, Tropicalgin, Cavex Orthotrace, Hydrogum, Orthoprint, Cavex Color Change, and Qualitygel. Three control groups were also used in this study: positive control group (C+) consisting of cell detergent Tween 80, negative control group (C-) consisting of PBS, and cell control group (CC) consisting of non-exposed cells. After manipulating the materials according to the manufacturer’s instructions, samples were made by using silicone rings. Next, the samples were immersed into Eagle’s minimum essential medium (MEM) for 2 minutes followed by removal of supernatants and contact with L929 fibroblasts. After contact with the medium, the cells were incubated for further 24 hours in which 100µl of 0.01 por ciento neutral red stain were added. Cells were incubated again for 3 hours so that the stain could be absorbed. After this period, the cells were fixed and viable cell counting was performed by using a spectrophotometer (BioTek, Winooski, Vermont, USA) at wavelength of 492 nm. The results demonstrated statistical differences between CC and C- groups in relation to other ones (p<0.05). No statistical differences were observed between Jeltrate Plus and Hydrogum groups, between Jeltrate and Jeltrate Chromatic, Printer Gel, Tropicalgin, and Qualitygel groups, and between Jeltrate Chromatic and Alga Gel, Ava Gel, New Print, Kromopan 100, Cavex Orthotrace, Hydrogum, Orhtoprint, and Cavex Color Change groups. One can conclude, based on the results of this study, that all alginate materials were found to be cytotoxic.

El alginato o hidrocoloide irreversible, es uno de los materiales de impresión más aceptados y utilizados en odontología. Sin embargo, algunas substancias existentes en estos materiales pueden ser tóxicas. El objetivo de este estudio fue evaluar la citotoxicidad de los alginatos para aplicaciones dentales. Fueron evaluados 14 alginatos diferentes: Jeltrate, Jeltrate Plus, Jeltrate Chromatic, Alga Gel, Printer Gel, Ava Gel, New Print, Kromopan 100, Tropicalgin, Cavex Orthotrace, Hydrogum, Orthoprint, Cavex Color Change y Qualitygel. También se utilizaron tres grupos de control también se utilizaron en este estudio: grupo control positivo (C+) que consiste en células de detergente Tween 80, el grupo de control negativo (C-) que consiste en PBS, y el grupo de células de control (CC) que consiste de las células no expuestas. Después de la manipulación de los materiales de acuerdo a las instrucciones del fabricante, las muestras fueron hechas mediante el uso de anillos de silicona. A continuación, las muestras se sumergieron en medio mínimo esencial de Eagle (MEM) durante 2 minutos, seguido de la eliminación de los sobrenadantes y el contacto con los fibroblastos L929. En caso de contacto con el medio, las células fueron incubadas durante 24 horas más en 100ml de tinción roja neutra al 0,01 por ciento. Las células se incubaron nuevamente durante 3 horas para que la tinción pueda ser absorbida. Después de este período, las células fueron fijadas y el recuento de células viables se realizó mediante un espectrofotómetro (BioTek, Winooski, Vermont, EE.UU.) a la longitud de onda de 492 nm. Los resultados demostraron diferencias estadísticamente significativas entre los grupos de CC y C- en relación con los demás (p<0,05). No se observaron diferencias estadísticas entre los grupos Jeltrate Plus y Hydrogum, entre Jeltrate y los grupos Jeltrate Chromatic, Printer Gel, Tropicalgin y Qualitygel, y entre Jeltrate Chromatic y los grupos Alga Gel, Ava Gel, New Print, ...

Citotoxicidade da Punica granatum L. (romã) sobre cultura de fibroblastos e de células de linhagem cancerígena / Cytotoxicity of Punica granatum L. (pomegranate) a fibroblast cell line and a carcinoma cell line

Embora diversos estudos já tenham sido realizados sobre a Punica granatum L., seus possíveis efeitos citotóxicos não são bem conhecidos. No presente trabalho foi avaliada a toxicidade da Punica granatum L. (PG) por meio de cultura celular com duas linhagens: fibroblastos humanos de mucosa oral (FLM) e células de carcinoma epidermóide oral humano (KB). As células foram submetidas ao teste de viabilidade celular por 24 horas em placas de 96 poços nas concentrações da PG (1%, 0,5%, 0,25%, 0,125%, 0,062% e 0,031%) e aos testes de citotoxicidade celular em placas de 24 poços, em cinco períodos diferentes (4 horas, 1, 3, 5 e 7 dias) e com quatro diferentes concentrações (1%, 0,5%, 0,25%, 0,062%). Todos os testes foram realizados em triplicata e em três momentos diferentes. Foram adicionados dois grupos, um controle negativo (Tryton 10%) e um controle positivo constituído de meio de cultura acrescido de soro bovino fetal 10% e sem o extrato de PG. A quantificação celular foi realizada pelo método do Resazurin, com avaliação por espectrofotometria óptica (espectrofotômetro UVM340 - AsysHitech GmbH) e leitura de absorbância dos comprimentos de onda de 570 nm a 600 nm. Os dados obtidos foram submetidos à ANOVA (5%). Os resultados de viabilidade mostraram que nas concentrações de 0.031%; 0,062%; 0,125%; 0,25% e 0,05% o extrato de PG inibiu a viabilidade celular (≥ 40%) principalmente das células KB. O teste de citotoxicidade mostrou que apenas as culturas tratadas com PG a 0,062% exibiram citotoxicidade para as linhagens celulares. A PG a 1% foi tóxica para as células FLM e KB. O extrato de PG inibiu a citotoxicidade celular nas demais concentrações testadas. Estes resultados podem ser relacionados a propriedade anti carcinogênica da Punica granatum L.

Several studies have been conducted on Punica granatum L., but their cytotoxic effects are not well known. The purpose of this study was to evaluate the cytotoxicity of Punica granatum L. extract (PG) in two cell lines: human oral mucosa fibroblasts (FLM) and cells of human oral squamous cell carcinoma (KB). The cells were analized to viability on 24 h using different concentrations of PG (1%, 0.5%, 0.25%, 0.125%, 0.062% and 0.031%) and to cell proliferation in five different periods (4 h, 1, 3, 5 and 7 days) using four different concentrations of PG (1%, 0.5%, 0.25%, 0.062%). All tests were performed in triplicate and in three different times. There were a negative control (Tryton 10%) and a positive control (culture medium plus 10% SBF, without PG extract). Cell quantification was performed by the Resazurin method, with evaluation by optical spectrophotometry (spectrophotometer UVM340 - Asys Hitech GmbH) and measure of absorbance at a wave length of 570 nm to 600 nm. Data were submitted to ANOVA (5%). The results showed inhibition of cell proliferation in 0,031%, 0.062%, 0.125%, 0.25% and 0.05% concentrations of PG extract (≥ 40%) specialty to KB cells in comparison to FLM cells. PG 1% was toxic to KB and FLM cells. The proliferation test showed cell proliferation only when using PG 0,062%, for both cell lines and inhibited proliferation whith all other PG concentrations. These effects might be related to the anticarcinogenic properties of PG.