Utilidad del recuento de unidades formadoras de colonia de Cryptococcus sp. en líquido cefalorraquídeo de pacientes con criptococosis meníngea asociada al sida / Cryptococcus' colony forming unit count usefulness

Antecedentes: El recuento de unidades formadoras de colonia (UFC) de Cryptococcus en el líquido cefalorraquídeo (LCR) sería un marcador fiable para el pronóstico del paciente y una herramienta simple y económica. Objetivo: Evaluar la utilidad del recuento de UFC de Cryptococcus spp. y compararlo con las variaciones de antígeno capsular de Cryptococcus (AgCr) en LCR.Materiales y métodos: Se realizó la revisión de historias clínicas de pacientes con meningoencefalitis por Cryptococcus asociada con el sida en nuestro centro, entre febrero de 2016 y julio de 2020. Se evaluaron los valores de UFC y AgCr en LCR durante la evolución de la micosis. Resultados y discusión: Se analizaron datos de 94 episodios clínicos de 85 pacientes, con un total de 297 observaciones de muestras de LCR. Se evidenció el valor del recuento de UFC por ser un marcador de viabilidad y de carga fúngica. El recuento de UFC bajo no necesariamente coexistió con un nivel bajo de AgCr. Con respecto a la evolución en el tiempo, la mayoría de los pacientes fueron diagnosticados con una alta carga fúngica y su descenso ocurrió más rápido que el del AgCr, por lo que reflejaría la mejora del paciente, permitiendo tomar conductas al respecto.Palabras clave: Criptococosis, carga fúngica, ufc/mL.

Background. The Cryptococcus' colony-forming unit (CFU) count in cerebrospinal fluid (CSF) would be a reliable marker for patient prognosis and a simple and inexpensive tool. Objectives: To evaluate the usefulness of the CFU count of Cryptococcus spp. And to compare it with the variations of Cryptococcus' capsular antigen (CrAg) in CSF.Materials and methods. Clinical records of patients with aids-related meningoencephalitis caused by Cryptococcusassisted in our center between February 2016 and July 2020 were reviewed. CFU count and CrAg values in CSF were evaluated during the evolution of the mycosis.Results and Discussion. Data from 94 clinical episodes of 85 patients with a total of 297 observations of CSF samples were analyzed.The importance of using the CFU count was evidenced as it is a viability and fungal load marker.Low CFU count did not necessarily coexist with low CrAg.Regarding the evolution over time, most of the patients were diagnosed with a high fungal load and its decrease occurred faster than that the one of AgCr. This would reflect the improvement of the patient, allowing behaviors to be taken in this regard

Avaliação microbiológica de leite pasteurizado “tipo C” comercializado na cidade de Itabuna, Bahia, Brasil / Microbiological evaluation of pasteurised milk “type C” traded in Itabuna, Bahia, Brazil

A qualidade da matéria prima é um dos maiores entraves ao desenvolvimento e consolidação da indústria de laticínios no Brasil pois , grande parte do leite produzido pelo país é oriunda de produtores rurais com baixa capacitação técnica e limitação financeira para os investimentos necessários. Diante do risco, é fundamental que as normativas estabelecidas sejam seguidas para garantir segurança ao consumidor final. Para tanto objetivou-se avaliar a qualidade microbiológica de quatro marcas distintas de leite pasteurizado tipo C, comercializado em Itabuna – Bahia. Do material coletado foram feitas analises para a contagem de unidades formadoras de colônia por mililitro, coloração de Gram, e teste de coagulase e catalase. Os resultados mostraram presença de Staphylococcus spp. e enterobactérias nas marcas de leite pasteurizado selecionadas.

Long noncoding RNA UFC1 promotes metastasis and invasion of hepatocellular carcinoma cells via GSK-3β/β-catenin axis / 南方医科大学学报

OBJECTIVE@#To explore the role of Long noncoding RNA UFC1 (lincRNA-UFC1) in modulating the metastasis and invasion of hepatocellular carcinoma (HCC) cells and the underlying mechanism.@*METHODS@#Human HCC cell line Huh7 was infected with the lentiviral vector carrying lincRNA-UFC1 to obtain a cell line with lincRNA-UFC1 overexpression. A short hairpin RNA (shRNA) targeting lincRNA-UFC1 was delivered in human HCC BEL-7402 cells via a lentiviral vector to obtain a cell line with lincRNA-UFC1 knockdown. Expression levels of lincRNA-UFC1 in the two HCC cell lines were detected using real-time PCR, and the changes in the cell invasion and migration in response to lincRNA-UFC1 overexpression or knockdown were analyzed using Transwell and wound-healing assays. The expressions of GSK-3β/β-catenin-related proteins in the cells were detected with Western blotting. XAV-939, a GSK-3β/β-catenin inhibitor, was used for assessing the impact of lincRNAUFC1 overexpression on the invasion and migration of the HCC cells through Transwell and wound-healing assays.@*RESULTS@#Overexpression of lincRNA-UFC1 significantly promoted the invasion and migration of Huh7 cells as compared with the control cells ( < 0.001), while lincRNA-UFC1 knockdown obviously suppressed the invasion and migration of BEL-7402 cells ( < 0.001). The results of Western blotting showed that the expressions of proteins associated with the cell invasion and migration, namely β-catenin and P-GSK-3β, were significantly upregulated in response to lincRNA-UFC1 overexpression, and were obviously lowered after lincRNA-UFC1 knockdown. Treatment of the cells with XAV-939 significantly reversed the effect of lincRNA-UFC1 overexpression on the cell invasion and migration ( < 0.001).@*CONCLUSIONS@#lincRNA-UFC1 overexpresison promotes cell invasion and migration through the GSK-3β/β-catenin axis in HCC cells .

Impacto do estado nutricional e do tempo de jejum nas complicações gastrointestinais e no tempo de hospitalização em pacientes cirúrgicos / Impact of the nutritional status and fasting time in gastrointestinal complications and hospitalization time in surgical patients

Introdução: O estado nutricional e o tempo de jejum pré e pós-cirúrgico impactam na morbimortalidade de pacientes submetidos a cirurgias eletivas. No Brasil, o tempo de jejum prolongado e a elevada prevalência de desnutrição são comuns nos hospitais terciários. Objetivo: Realizar levantamento do tempo de jejum médio praticado em hospital terciário e avaliar o impacto do jejum pré e pós-cirúrgico e do estado nutricional nas complicações gastrointestinais imediatas e no tempo de hospitalização em pacientes cirúrgicos. Material e métodos: Estudo prospectivo, realizado no Hospital Universitário Walter Cantídio, em Fortaleza. Foram avaliados 173 indivíduos submetidos a cirurgias eletivas de médio e grande porte entre agosto de 2016 a janeiro de 2017. Foram coletadas medidas antropométricas, hemograma e informações como tempo de jejum pré e pós-cirúrgico, permanência hospitalar e complicações gastrointestinais. As análises de correlação e associação foram realizadas no Software SPSS®, com nível de significância fixado em p< 0,05. Resultados: A maioria dos pacientes (71,1%) foi submetida à cirurgia de porte II (grande porte). Houve correlação negativa entre tempo de permanência hospitalar e índice de massa corporal [r= -0,223; p= 0,003], circunferência braquial [r= -0,335; p< 0,001], dobra cutânea tricipital [r= -0,320; p< 0,001], área muscular do braço corrigida [r= -0,253; p= 0,001], contagem total de linfócitos [r= -0,223; p= 0,008], hemoglobina [r= -0,243; p= 0,004] e relação linfócito/monócito [r= -0,308; p< 0,001]. Adicionalmente, houve correlação positiva entre tempo de permanência hospitalar com o tempo de jejum pós-cirúrgico [r= 0,456; p< 0,001]. Ausência de complicações gastrointestinais foi associada a valores de triagem (Nutritional Risk Screenin-2002) < 3 (p= 0,034) e entre menor tempo de jejum pós-cirúrgico (p= 0,033). Conclusão: Pacientes desnutridos, com maior risco nutricional e os que se submeteram a um tempo de jejum pós-cirúrgico elevado apresentaram maior tempo de permanência hospitalar e maior incidência de complicações gastrointestinais.

Introduction: The nutritional status and pre and postoperative fasting time impact the morbidity and mortality of patients undergo to elective surgeries. In Brazil, prolonged fasting time and high prevalence of malnutrition are common in tertiary hospitals. Objective: To evaluate the mean preoperative and postoperative fasting time and the impact of fasting, and nutritional status on gastrointestinal complications and on length of hospital stay in surgical patients. Material and methods: A prospective study, conducted at Walter Cantídio University Hospital, in Fortaleza. One hundred and seventy three surgery patients were evaluated between August 2016 and January 2017. The subjects were divided in two groups (medium and large size). Anthropometric measurements, blood count and information about pre and postoperative fasting time, hospital stay and gastrointestinal complications data were collected. Correlation and association test were performed by Software SPSS®. P<0.05 was considered significant. Results: The majority of the patients (71,1%) were included in size II (large) surgery. There was a negative correlation between length of stay in hospital and body mass index [r= -0,223; p= 0,003], brachial circumference [r= -0,335; p<0,001], triceps skinfold thickness [r= -0,320; p<0,001], arm muscle area [r= -0,253; p= 0,001], total lymphocyte count [r= -0,223; p= 0,008], hemoglobin [r= -0,243; p= 0,004] and lymphocyte/monocyte ratio [r= -0,308; p<0,001]. In addition, there was a positive correlation between length of stay in hospital and postoperative fasting time [r= 0,456; p<0,001]. Absence of gastrointestinal complications was associated with Nutrition Risk Screening-2002 <3 (p= 0,034) and shorter post-operative fasting time (p= 0,033). Conclusion: Malnourished patients, higher nutritional risk and higher postoperative fasting time were associated to longer hospital stay and a higher incidence of gastrointestinal complications.

Cuantificación de bacterias cultivables mediante el método de Goteo en Placa por Sellado (o estampado) Masivo / Quantification of cultivable bacteria by the Massive Stamping Drop Plate method

Cuando se desea cuantificar el número de bacterias presentes en múltiples muestras, los procedimientos de rutina suelen consumir mucho tiempo. En ese periodo las muestras podrían sufrir modificaciones en su población. En el presente trabajo se evaluó una metodología alternativa para cuantificar bacterias cultivables de forma masiva, rápida y económica en la que se implica el sellado, estampado o impresión de diluciones seriadas de muestras de diversa procedencia. El tiempo requerido para preparar 22 muestras para su sellado en placa es de 15 minutos. El método se basó en realizar diluciones seriadas (base 10) de las muestras líquidas originales contenidas en una placa multipozos con la ayuda de una pipeta multicanal. Después, con un replicador se tomó un volumen (aproximadamente 1,65 µl) de muestra de cada pozo, que se inoculó por sellado en un medio de crecimiento gelificado de interés. Las placas se incubaron el tiempo necesario, se contó el número de colonias presentes en la dilución contable y se calculó el número de Unidades Formadoras de Colonia por mililitro (UFC/ml) para cada muestra. La metodología se denominó "Goteo por Sellado en Placa Masivo" (GSPM) y ha sido aplicada para cuantificar exitosamente bacterias provenientes de diferentes muestras de laboratorio, por ejemplo, de cultivos líquidos, muestras clínicas (como exudados y secreciones) y bacterias presentes en la rizósfera de plantas de maíz. Sin embargo, la metodología GSPM podría aplicarse para contabilizar masivamente a bacterias de cualquier otra procedencia.

In an attempt to quantify the number of bacteria present in a high number of samples, routine procedures are usually very time-consuming. During this period of time, bacterial population could be modified. In this work, an alternative for a massive, quick and economic method was evaluated in order to count viable bacteria, consisting in the sealing or stamping of serial dilutions performed in samples from different origins. The time required to prepare 22 samples for plate stamping is only 15 minutes. The quantification was based in performing serial dilutions (10-fold) of the original liquid samples contained in a multiwell plate using a multichannel micropipette. Afterwards, using a replicator, the same volume of each sample (approximately 1,65 µl) was recovered from each well, and then it was inoculated and sealed in a solid growth media of interest. Plates were incubated as needed, colonies were counted in the quantifiable dilution and Colony Forming Units per milliliter (CFU/ml) was calculated for each sample. We called this method "Massive Stamping Drop Plate" (MSDP) and it has been successfully applied to count bacteria from different lab samples, including liquid cultures, clinical samples (exudates and secretions) and bacteria recovered from the rhizosphere of corn plants. However, MSDP could also be applied to massively count bacteria from any other source.

Medición de cambios cuantitativos de la microbiota subgingival posterior a la remoción de placa bacteriana supragingival / Measurement of quantitative changes of the microbiota subgingival after to removal of bacterial plaque supragingival

Uno de los campos de interés en el estudio de la microbiología periodontal para muchos investigadores ha sido identificar si es posible que los microorganismos responsables del origen y progresión de la enfermedad periodontal que habitan sobre el margen gingival (supragingival) y bajo este (subgingival) tengan una relación directa que permita mantener interacciones influyentes en el crecimiento y desarrollo de las diferentes especies bacterianas que habitan en los tejidos periodontales. Por lo tanto, al remover los microorganismos que se ubican supragingivalmente sería posible encontrar cambios en el medio subgingival al no existir un intercambio entre los ambientes aerobios (supragingival) y anaerobios (subgingival) una vez desorganizada la placa bacteriana supragingival. Para demostrar esta relación se seleccionaron 7 individuos con diagnóstico de periodontitis crónica moderada y severa a los cuales se les realizó un destartraje supragingival de boca completa para lograr desorganizar la placa bacteriana supragingival. A su vez se tomaron muestras microbiológicas de los sacos periodontales más profundos de cada cuadrante de estos individuos, siendo la primera muestra tomada previo al destartraje supragingival considerada como muestra basal (día 0), luego se tomaron a las 24 horas, a los 7 y 21 días de removida la placa bacteriana supragingival. De los resultados del presente estudio pudimos concluir que al desorganizar el biofilm supragingival se observa una disminución en la cantidad total de microorganismos subgingivales, así como también disminuye de manera considerable la proporción de Porphyoromona gingivalis presente en el medio subgingival. Lo cual permitiría establecer la existencia de una relación directa y dependiente entre los microorganismos que habitan el medio supragingival y subgingival.

One of the fields of interest in the study of the microbiology periodontal for many investigators has been to identify if it is possible that the microorganisms responsible for the origin and progression of the disease periodontal that live on the margin gingival (supragingival) and under this (subgingival) they have a direct relation that allows to support influential interactions in the growth and development of the different bacterial species that they live in the tissue periodontal.Therefore having removed the microorganisms that are located supragingivalmente would be possible to find changes in the way subgingival when an exchange not to exist between the aerobic environments (supragingival) and anaerobic (subgingival) once disorganized the bacterial plate supragingival. To demonstrate this relation 7 individuals selected with diagnosis of periodontitis chronicle moderate and severe to which they there was realized a destartraje supragingival of complete mouth to achieve supragingival to disorganize the bacterial plate. In turn microbiological samples of the sacks took periodontales deeper of every quadrant of these individuals, being the first taken sample before the destartraje supragingival considered as sample basal (the 0th), then they took at to 1, 7 and 21 days of removed the bacterial plate supragingival anaerobios (subgingival) once disorganized the bacterial plate supragingival Of the results of the present study we could conclude that on having disorganized the biofilm supragingival a decrease is observed in the total quantity of microorganisms subgingivales, as well as also it diminishes in a considerable way the proportion of present Porphyoromona gingivalis in the way subgingival. Which would lead to thinking that there exists a direct and dependent relation between the microorganisms that live the way supragingival and subgingival.