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Aim: To determine if the artificial staining with black tea (BT) influences the enamel microhardness before in-office bleaching and if BT staining is necessary to evaluate the efficacy of bleaching with 35% hydrogen peroxide Methods: Enamel/dentin blocks were randomized into groups according to the staining protocol (n=5/group): (CO) control ­ maintained in artificial saliva solution (AS); (BT4) immersed in black tea solution for 4 h; (BT24) immersed in black tea solution for 24 h. After the staining protocols, all specimens were kept in AS for one week, followed by bleaching (three sessions of HP application for 40 min). Knoop surface microhardness (kgF/mm2) was determined at baseline (T0), after staining (T1), after 7 days of storage in AS (T2), and after bleaching (T3). The color (∆E00) and coordinate changes (∆L, ∆a, ∆b) were measured using a digital spectrophotometer at T0 and T3. Data were submitted to one-way (∆E00, ∆L, ∆a, ∆b) or two-way ANOVA repeated measures (kgF/mm2) and Tukey's test (a=5%). Results: The staining protocols (BT4 and BT24) promoted significantly lower microhardness (T1 and T2, p<0.05) than CO, whereas CO was the only group to maintain microhardness values over time. Bleaching promoted perceptible ∆E00 without a significant difference among the groups regardless of the staining protocol (p=0.122). CO and BT4 showed no differences in terms of ∆L and ∆a (p>0.05), but BT4 displayed a higher ∆b than CO. Conclusion:The artificial staining with BT negatively affected the enamel surface microhardness and was not essential to evaluate the efficacy of 35% hydrogen peroxide bleaching

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INTRODUCTION: A weak venous wall is one of the major reasons contributing to vein graft failure after coronary artery bypass grafting (CABG). We investigated whether adventitial collagen cross-linking by glutaraldehyde reinforces venous wall, preserving the endothelium of veins during high-pressure distention. METHODS: Human saphenous veins (SVs) were collected from 40 patients undergoing CABG, and adventitia cross-linking was performed with 0.3% glutaraldehyde for five minutes. The cross-linked SVs were accessed by biodegradation assay, immunofluorescent staining, and tensile test. Native SVs and cross-linked SVs from another 20 patients received the 200 mmHg pressure distention for two minutes. Pressure-induced injury of SVs were accessed by immunohistochemistry and electron microscopy. RESULTS: Time to digestion was 97±13 minutes for native SVs and 720±0 minutes for cross-linked SVs (P<0.05). After adventitial cross-linking, the collagen I fibres of the vein remarkably presented with compact and nonporous arrangement. In the high-stretch region (stretch ratio 1.4-1.8), the Young's elastic modulus of stress-stretch ratio curve in cross-linked SVs was larger than that in native SVs (13.88 vs. 5.83, P<0.05). The cross-linked SVs had a lower extent of endothelial denudation without fibre fracture during high-pressure distension than native SVs. Comparing with the non-cross-linked SVs, the percentage of endothelial nitric oxide synthase staining length on the endothelium of cross-linked SVs was significantly preserved after high-pressure distension (85.2% vs. 64.7%, P<0.05). CONCLUSION: Adventitial collagen cross-linking by glutaraldehyde reinforced venous wall by increasing stiffness and decreasing extensibility of SVs and mitigated the endothelial damage under high-pressure distension.

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ABSTRACT Introduction: A weak venous wall is one of the major reasons contributing to vein graft failure after coronary artery bypass grafting (CABG). We investigated whether adventitial collagen cross-linking by glutaraldehyde reinforces venous wall, preserving the endothelium of veins during high-pressure distention. Methods: Human saphenous veins (SVs) were collected from 40 patients undergoing CABG, and adventitia cross-linking was performed with 0.3% glutaraldehyde for five minutes. The cross-linked SVs were accessed by biodegradation assay, immunofluorescent staining, and tensile test. Native SVs and cross-linked SVs from another 20 patients received the 200 mmHg pressure distention for two minutes. Pressure-induced injury of SVs were accessed by immunohistochemistry and electron microscopy. Results: Time to digestion was 97±13 minutes for native SVs and 720±0 minutes for cross-linked SVs (P<0.05). After adventitial cross-linking, the collagen I fibres of the vein remarkably presented with compact and nonporous arrangement. In the high-stretch region (stretch ratio 1.4-1.8), the Young's elastic modulus of stress-stretch ratio curve in cross-linked SVs was larger than that in native SVs (13.88 vs. 5.83, P<0.05). The cross-linked SVs had a lower extent of endothelial denudation without fibre fracture during high-pressure distension than native SVs. Comparing with the non-cross-linked SVs, the percentage of endothelial nitric oxide synthase staining length on the endothelium of cross-linked SVs was significantly preserved after high-pressure distension (85.2% vs. 64.7%, P<0.05). Conclusion: Adventitial collagen cross-linking by glutaraldehyde reinforced venous wall by increasing stiffness and decreasing extensibility of SVs and mitigated the endothelial damage under high-pressure distension.

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Abstract Introduction: This study's objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats. Methods: Sprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1β, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry. Results: The expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1β, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group. Conclusion: Downregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.

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INTRODUCTION: This study's objective is to investigate the effect of downregulation of micro ribonucleic acid (miR)-124a on myocardial injury after ischemia reperfusion (I/R) in rats. METHODS: Sprague Dawley (SD) rats (n=20) were divided into four groups - sham, I/R, I/R+miR-124a antagomir (I/R+ant-miR-124a), and I/R+ant-normal control (NC). The pathomorphological and infarct size variance of injured myocardial tissues with IR were conducted with hematoxylin (HE) and triphenyltetrazolium chloride (TTC) staining. The expression levels of miR-124a, BAX, nuclear factor kappa B (NF-KB), Notch1, and Hes1 were examined by quantitative real-time polymerase chain reaction or Western blot in myocardium. The inflammatory cytokines interleukin (IL)-6, IL-1ß, and tumor necrosis factor alpha (TNF-α) were detected by the enzyme-linked immunosorbent assay, as well as the activity of lactate dehydrogenase (LDH) and creatine kinase (CK) in serum by colorimetry. RESULTS: The expression of miR-124a was increased in the I/R group. Compared with I/R and I/R+ant-NC groups, after downregulating miR-124a, the expression of IL-6, IL-1ß, TNF-α, BAX, NF-KB, LDH, and CK were decreased, but the expression of Notch1 and Hes1 were increased. In HE staining, myocardial tissue edema, red blood cell exudation, and myocardial fiber arrangement disorder were accompanied by inflammatory cell infiltration and local necrosis in the I/R group. However, the pathological injury of myocardial tissue was alleviated after downregulating miR-124a. Additionally, TTC results showed that the myocardial infarction area was decreased in the I/R+ant-miR-124a group. CONCLUSION: Downregulation of miR-124a expression through Notch pathway can significantly reduce myocardial damage after 24 hours of I/R in SD rats. Therefore, miR-124a may become a potential therapeutic target for I/R injury.

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The pigmentation of the resin cement at the tooth/ceramic interface compromises the esthetic and longevity of ultra-thin ceramic veneers. Aim: The aim of this study was to evaluate marginal staining of ultra-thin ceramic veneers cemented to intact enamel (non-prepared) and prepared enamel. Methods:Thirty-two (32) permanent central incisors were selected and randomly divided into two groups: intact enamel (IE) and prepared enamel (PE). The ceramic veneers of PE group were bonded to the prepared enamel and the ceramic veneers of IE group were cemented directly onto the intact enamel, with no preparation. Both preparation and cementation were standardized and performed by a single operator. Each group was subdivided into two subgroups (n = 8) with different immersion media - coffee and water. After an immersion period of 10 days, stereomicroscope images were made at 20X magnification of the mesial, distal, cervical and incisal surface of each specimen. Three blinded, trained and calibrated examiners evaluated the images of the resin cement interface of each surface. The data were subjected to KruskalWallis and MannWhitney statistical analysis. Immersion media and enamel preparation influenced the marginal staining of the tooth/ceramic interface. Results: When immersed in coffee, prepared interfaces presented greater marginal staining than unprepared interfaces. When immersed in water, there was no statistically significant difference between the groups. Conclusion: The cementation of ultra-thin ceramic veneers onto intact enamel is associated with less marginal staining and, consequently, improved esthetics

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Abstract Objective: To examine the effects of classical technique, electrocautery, and ultrasonic dissection on endothelial integrity, function, and preparation time for harvesting the radial artery (RA) during coronary artery bypass grafting (CABG). Methods: Forty-five patients who underwent isolated CABG and whose RA was suitable for use were studied and divided into three groups: Group 1, classical method (using sharp dissection); Group 2, electrocautery; and Group 3, ultrasonic cautery. Levels of prostacyclin and nitric oxide derivatives were examined biochemically; vascular cell adhesion molecule 1 (VCAM-1) and endothelial nitric oxide synthetase (eNOS) values were assessed using immunohistochemical staining. RA preparation time, RA length/harvesting time ratio, and drainage amounts at the site of RA removal were compared. Results: Differences in RA preparation time (Group 1: 25±6 min, Group 2: 18±3 min, Group 3: 16±3 min, P<0.001) and length/harvesting time ratio (Group 1: 0.76±0.19 cm/min, Group 2: 0.98±0.16 cm/min, Group 3: 1.13±0.09 cm/min, P<0.001) were statistically significant among the groups. Levels of prostacyclin and nitric oxide derivatives were not statistically significant different, VCAM-1 and eNOS expressions were observed to be similar among the groups, and endothelial damage was detected in only one patient per group. Conclusion: Use of ultrasonic cautery during RA preparation considerably reduces the preparation time and postoperative drainage amount. However, the superiority of one method over the others could not be demonstrated when the presence of endothelial damage with both biochemical and histopathological evaluations was considered.

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OBJECTIVE: To examine the effects of classical technique, electrocautery, and ultrasonic dissection on endothelial integrity, function, and preparation time for harvesting the radial artery (RA) during coronary artery bypass grafting (CABG). METHODS: Forty-five patients who underwent isolated CABG and whose RA was suitable for use were studied and divided into three groups: Group 1, classical method (using sharp dissection); Group 2, electrocautery; and Group 3, ultrasonic cautery. Levels of prostacyclin and nitric oxide derivatives were examined biochemically; vascular cell adhesion molecule 1 (VCAM-1) and endothelial nitric oxide synthetase (eNOS) values were assessed using immunohistochemical staining. RA preparation time, RA length/harvesting time ratio, and drainage amounts at the site of RA removal were compared. RESULTS: Differences in RA preparation time (Group 1: 25±6 min, Group 2: 18±3 min, Group 3: 16±3 min, P<0.001) and length/harvesting time ratio (Group 1: 0.76±0.19 cm/min, Group 2: 0.98±0.16 cm/min, Group 3: 1.13±0.09 cm/min, P<0.001) were statistically significant among the groups. Levels of prostacyclin and nitric oxide derivatives were not statistically significant different, VCAM-1 and eNOS expressions were observed to be similar among the groups, and endothelial damage was detected in only one patient per group. CONCLUSION: Use of ultrasonic cautery during RA preparation considerably reduces the preparation time and postoperative drainage amount. However, the superiority of one method over the others could not be demonstrated when the presence of endothelial damage with both biochemical and histopathological evaluations was considered.

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ABSTRACT BACKGROUND: Esophageal cancer is the eighth most common cancer. The prognosis is bleak in patients with advanced stages. Patients with early disease have a better prognosis than those with advanced stage. There are several techniques for the screening of premalignant and superficial lesions including chromoendoscopy. OBJECTIVE: This article aimed to determine the effectiveness of chromoendoscopy with toluidine blue combined with Lugol's solution for diagnosis of esophageal premalignant and superficial neoplastic lesions in high risk patients. METHODS: Routine white light upper endoscopy was performed. Toluidine blue was sprayed from the gastroesophageal junction to 20 cm of the dental arch. Then the uptake dye areas were characterized. Later Lugol's solution was sprayed. Areas with less-intense staining were characterized. Biopsy of the toluidine blue capturing areas and areas with less-intense staining of Lugol's solution were taken. In the cases where lesions were not evidenced after application of dyes, biopsies four quadrants of the esophageal mucosa were taken. The samples were evaluated by a digestive pathologist. RESULTS: Barrett's esophagus was the most common premalignant lesion and the early neoplastic lesion was adenocarcinoma with a sensitivity of 100%, specificity 85.7%, positive predictive value 30%, negative predictive value 100%, positive likelihood ratio 7 negative likelihood ratio 0. CONCLUSION: Chromoendoscopy with toluidine blue combined with Lugol's solution is a useful tool in the screening of esophageal premalignant lesions and superficial neoplasms.

RESUMO CONTEXTO: O câncer de esôfago é o oitavo câncer mais comum. O prognóstico é sombrio em pacientes com estágios avançados. Pacientes com doença precoce têm um melhor prognóstico do que aqueles com estágio avançado. Existem várias técnicas para a triagem de lesões pré-malignas e superficiais, incluindo cromoendoscopia. OBJETIVO: Este artigo objetivou determinar a efetividade da cromoendoscopia com azul de toluidina combinada com a solução de Lugol para o diagnóstico de lesões neoplásicas pré-malignas e superficiais esofágicas em pacientes de alto risco. MÉTODOS - A endoscopia de luz branca de rotina foi realizada de forma rotineira. O azul do toluidina foi pulverizado desde a junção gastroesofágica até 20 cm da arcada dentária. As áreas impregnadas pela tintura da tomada foram então caracterizadas. Mais adiante a solução de Lugol foi pulverizada. Áreas com coloração menos intensa foram caracterizadas. Foram realizadas biópsias das áreas de captura de azul de toluidina e áreas com coloração menos intensa da solução de Lugol. Nos casos onde as lesões não foram evidenciadas após a aplicação das tinturas, foram feitas biópsias em quatro quadrantes da mucosa esofágica. As amostras foram avaliadas por um patologista especializado. RESULTADOS: O esôfago de Barrett foi a lesão pré-maligna mais frequente e a lesão neoplásica precoce foi adenocarcinoma com sensibilidade de 100%, especificidade de 85,7%, valor preditivo positivo de 30%, valor preditivo negativo 100%, razão de verossimilhança positiva 7 e razão de verossimilhança negativa 0. CONCLUSÃO: A cromoendoscopia com azul de toluidina combinada com a solução de Lugol é uma ferramenta útil na triagem de lesões pré-malignas esofágicas e neoplasias superficiais.

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INTRODUCTION: The workflow in clinical flow cytometry laboratories must constantly be reviewed to develop technical procedures that improve quality and productivity and reduce costs. Using the Beckman Coulter dry coating technology, we customized a ten-color tube with dried antibody reagents, designated the Duraclone screening tube (DST), for screening hematological malignancies. Here, we compared the applicability, clinical and numerical equivalence, and cost and time required for the technical procedures between the liquid reagents and the DST. METHODS: The DST contains CD4 + Kappa-FITC, CD8 + Lambda-PE, CD3 + CD14-ECD, CD33-PE-Cy5.5, CD20 + CD56-PE-Cy7, CD34-APC, CD19-APC-AlexaFluor700, CD10-APC-AlexaFluor750, CD5-Pacific Blue, and CD45-Krome Orange. We evaluated 20 bone marrow samples, 13 peripheral blood samples, 6 lymph node biopsy samples, 5 fine-needle aspirate samples, 5 cerebrospinal fluid samples, and 1 pleural fluid sample. RESULTS: The DST was useful for more than 60% of our samples. It was able to enumerate the majority of the populations in all types of samples with a statistically acceptable correlation with the liquid reagents. The use of the DST translated into significant time and cost savings of 15.8% and 12.3%, respectively, compared with the use of the liquid reagent. The cost was reduced by $14.36 per sample. CONCLUSIONS: The DST is an efficient solution for screening hematological malignancies with improved quality, productivity, standardization, and sustainability. These improvements could benefit patients by providing faster diagnoses using a higher quality and lower cost reagent.

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Resumen: el laboratorio de microbiología es un sistema complejo que implica muchos pasos para cada actividad, requiere de diversas personas y cuyos resultados analíticos generados deben ser lo más exactos posible. De no ser así, las consecuencias se pueden volver muy significativas, entre ellas, la realización de procedimientos innecesarios al paciente, los retrasos en la obtención de un resultado correcto y la ejecución de pruebas diagnósticas adicionales que se podían evitar. Estas consecuencias incrementan los gastos, tanto en términos económicos como en tiempo y esfuerzos del personal. La mayoría de las técnicas analíticas implican diversos procedimientos que están sujetos a cierto grado de imprecisión y posibilidad de error. Por tal razón, es importante la realización de un control de calidad interno que asegure que los productos finales o valores analíticos, que son producidos por un laboratorio, sean lo suficientemente fiables y adecuados para la finalidad que persiguen. De esta manera, un laboratorio microbiológico debe evaluar y documentar el desempeño de todos los aspectos de sus procedimientos, que incluye la calidad de la muestra, la eficiencia de los reactivos y antisueros, las coloraciones, los medios de cultivo, el funcionamiento de instrumentos o equipos, las cepas de referencia y la verificación o validación de los resultados obtenidos. El objetivo de este manuscrito es presentar una revisión bibliográfica sobre algunos elementos del laboratorio de microbiología susceptibles de control de calidad interno, como parte integral del trabajo diario del personal que labora en este. (AU)

Abstract: The microbiology laboratory is a complex system, which involves many steps for each activity and different people, and where analytical results must be as accurate as possible. If the results are inaccurate, the consequences can be significant, including the performance of unnecessary procedures, delays in obtaining a proper result and implementation additional diagnostic tests that could be avoided. These consequences increase spending both economics terms as time and efforts of the staff. Most analytical techniques involve several procedures that are subject to some degree of imprecision and possibility of error. Therefore, it's important to apply an internal quality control that ensure that finished products or analytical values which are produced by a laboratory are sufficiently reliable and appropriate to the objective pursued. In this way, a microbiological laboratory must assess and document the performance of all aspects of its procedures that includes the quality of the sample, the efficiency of reagents and antisera, the staining, the culture media, the operation of instruments or equipment, the strains of reference, and the verification or validation of the results obtained. The aim of this manuscript is to present a literature review about some elements of microbiology laboratory that are susceptible of internal quality control, as an integral part of the daily work of the staff working in this one. (AU)

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Introducción: la detección del género Corynebacterium en flujo vaginal representa un reto para el estudio de su posible papel patogénico en el tracto genital femenino. La coloración de Gram es la técnica recomendada para la detección de corinebacterias en muestras clínicas; no obstante, algunas características biológicas de estas bacterias dificultan la identificación por medio de esta. Objetivo: evaluar el desempeño de la coloración de Albert para la identificación de Corynebacterium spp. en muestras de flujo vaginal. Materiales y métodos: se evaluó la tinción de Albert como prueba diagnóstica para la identificación de Corynebacterium spp. en 451 flujos vaginales en comparación de la coloración de Gram. Se calculó la sensibilidad, la especificidad, los valores predictivos, las razones de verosimilitud y el índice J de Youden con el software Epidat 3.1. Resultados: la coloración de Albert presentó sensibilidad y especificidad de 61,5% y 74,6%, respectivamente, valor predictivo positivo de 78,3%, valor predictivo negativo de 56,5%, razón de verosimilitud positiva de 2,4 y negativa de 0,5 y un Índice de Youden de 0,4. Conclusiones: la tinción de Albert no mostró un mejor rendimiento que la coloración de Gram en la identificación del género Corynebacterium en muestras de flujo vaginal. Por lo tanto, se recomienda el uso en conjunto de las coloraciones de Gram y Albert para la identificación de corinebacterias en flujo vaginal. (AU)

Introduction: The detection of Corynebacterium genus in vaginal discharge represents a challenge for the study of its possible pathogenic role in the female genital tract. Gram's stain is recommended for corynebacteria detection in clinical samples, however, some biological characteristics of these bacteria difficult their identification by this technique. Objetive: to evaluate Albert's stain performans to identify Corynebacterium spp. in vaginal discharge. Materials and methods: Albert's stain was evaluated as a diagnostic test for the identification of Corynebacterium spp. in 451 vaginal swabs in comparison to Gram's stain. Sensitivity, specificity, predictive values, likelihood ratio, as well as Youden's J index were calculated using Epidat software 3.1 version. Results: Albert's stain had a sensitivity and specificity of 61.5% and 74.6%, respectively, positive predictive value of 78.3 %, negative predictive value of 56.5 %, positive likelihood ratio of 2.4 and negative likelihood ratio of 0.5, as well as Youden index of 0.4. Conclusions: Albert's stain did not show better performance than Gram's stain for identification of corynebacteria in vaginal swabs. Thereforeit is recommended the concomitant use of the Gram and Albert stains for the identification of corynebacteria in vaginal discharge. (AU)

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The present study aimed to evaluate the Fine Needle Aspiration Biopsy in different staining techniques in nodular lesions of the oral cavity and head and neck region, as their sensitivity, specificity and accuracy, staining with Panoptic, Papanicolaou and Hematoxylin-Eosin (H&E) stains. 46 patients who sought the Clinic of the Discipline of Clinical Stomatology at FOUSP were selected consecutively, with nodular lesions in the oral cavity and head and neck region. The material obtained by FNAB was sent on 6 different slides, stained by the method of Panoptic, Papanicolaou and H&E, to the same pathologist only with the clinical diagnosis. After the final report of FNAB, the biopsy report was issued, serving as gold standard. After the calculations, the results of sensitivity, specificity and accuracy for Panoptic staining were 28.6%, 76% and 15.4%, respectively. The result of sensitivity, specificity and accuracy for Papanicolaou staining were 71.4%, 76.7% and 23.3%, respectively. The result of sensitivity, specificity and accuracy for H&E staining were 82.1%, 23.3%, 28.6%, respectively. We can conclude, according to the methodology of this study that, H&E and Papanicolaou stains showed the same sensitivity of diagnosing malignant neoplasms. H&E stain showed a better specificity for diagnosing benign neoplasms, compared with Papanicolaou and Panoptic stains. H&E stain showed better accuracy, to give definitive diagnosis, followed by Papanicolaou and Panoptic stains.

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A mola hidatiforme (MH) é a forma mais comum de doença trofoblástica gestacional e representa uma condição benigna que em alguns casos pode sofrer malignização. Todas as pacientes diagnosticadas com doenças molares são acompanhadas por pelo menos seis meses para detecção precoce da neoplasia trofoblástica gestacional. No momento, existem poucas ferramentas para avaliação prognóstica da mola hidatiforme. Foi descrita a expressão diferencial de diversos fatores em tecido molar em comparação ao trofoblasto não neoplásico. Essas moléculas podem estar relacionadas com o comportamento agressivo da MH e consequentemente poderiam servir para melhor entendimento do processo de malignização e como preditoras da evolução da doença trofoblástica gestacional.

The hydatidiform mole (HM) is the most common form of gestational trophoblastic disease and a benign condition that in some cases may undergo malignant transformation. All patients diagnosed with molar diseases are monitored for at least six months for early detection of gestational trophoblastic neoplasia. Currently, there are few prognostic tools for the prediction of hydatidiform mole evolution. Differential expression on molar tissue of different molecular factors have been described when compared to non-neoplastic trophoblast. These markers may be associated with aggressive behavior of HM and therefore could serve as predictors of the development of gestational trophoblastic disease and to better understand molar malignant transformation. This review article will summarize and evaluate prognostic molecular markers of HM.

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Different applications of trypan blue (TB) for intraocular surgery have been reported, with very high levels of safety and efficacy. We describe the use of TB as an alternative vital dye for staining the ocular surface to assess the integrity of superficial cell layers of the cornea and the surface environment. This facilitates the diagnosis of various ocular surface disorders, including screening for dry eye disease (DED) among refractive and cataract patients. TB staining properties are different from fluorescein and both are stable in a solution, so that a double staining technique is introduced.

Diferentes aplicações do azul de tripano (AT) foram descritas para cirurgia intraocular, com elevados patamares de eficácia e segurança. Neste relato, é descrito a aplicação do AT como corante vital para superfíce ocular, de modo a estudar a integridade das células da superfície corneana e conjuntival na superfície ocular. Tal abordagem permite um diagnóstico mais sensível de desordens da superfície ocular, destacando-se disfunção lacrimal ou síndrome do olho seco. O AT tem propriedades distintas da fluoresceína, com a qual se mantém estável em solução permitindo a técnica de coloração dupla que é introduzida.

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Background: Vaginal infection is the commonest cause of genital symptoms and has obstetric and gynecological implications. Aim: To compare the Nugent and Spiegel methods for the diagnosis of bacterial vaginosis (BV) and to analyze discordant specimens using Ison and Hay (Ison/Hay) criteria. Material and Methods: After discardingcases with Candidiasis, deficientspecimens or those lacking bacteria, a total of348 Gram-stained smears vaginal specimens receivedfor the diagnosis of BV, were analyzed. Results: Vaginal microbiota was classified as normal in 203 and 237 samples (58 and 68 percent of samples), accordingto Nugent and Spiegel criteria, respectively One hundred andfive (30 percent) and 111 samples (32 percent), were classified as VB accordingto Nugent and Spiegel criteria, respectively. Both criteria were concordant in 308 samples (88.5 percent). The 40 (11.5 percent) discordant specimens were classified as intermedíate microflora by the Nugent system and as normal or BV by Spiegel. Among these, the Ison/Hay procedure identified four categories of microbiota. Ten (25 percent) specimens were classified as grade II microbiota, confirming their categorization by Nugent as intermedíate microbiota, six (15 percent) were classified in the III category, confirming the diagnosis ofBV by Spiegel, 13 (32.5 percent) corresponded to the category III, that does not exist in the Nugent and Spiegel categorization systems. Finally, 11 specimens could not be assigned to one category due to microscopic limitations to distinguish bacterial morphotypes. Conclusions: The systems proposed by Spiegel, Nugent and Ison I Hay are comparable for the diagnosis of BV. However, we recommend the use of Ison/ Hay procedure to evalúate vaginal microbiota, due to its wider range of categories, allowing a better discrimination ofthe vaginal microbiota.

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Objective: to evaluate the effects of pre- and/or postnatal exposure to particulate air pollution on fertilization, embryo development, and cell lineage segregation in preimplantation blastocysts during late-life reproductive period, using the in vitro fertilization (IVF) mouse model. Materials and methods: five-month-old mice underwent superovulation and were pre- and/or postnatally exposed to filtered air (FA-FA), filtered-ambient air (FA-AA), or ambient air (AA-AA) in exposure chambers, 24 hours per day, during six months. Results: ovarian response was not affected by the exposure protocol. A multivariate effect for pre- and/or postnatal exposure to ambient air on blastocyst differential staining, but not on IVF and embryo development, was found. Cell counts in inner cell mass (ICM) and ICM/trophectoderm ratios in blastocysts produced in FA-FA protocol were significantly higher than in blastocysts produced in FA-AA and AA-AA protocols. Conclusions: our study suggests that the exposure to particulate air pollution of a large urban center does not affect ovarian function but may negatively affect the female reproductive health in the late-life period by disrupting the lineage specification at the blastocyst stage.

Objetivo: avaliar os efeitos da exposição pré e/ou pós-natal ao ar ambiente no final da vida reprodutiva sobre a fertilização, desenvolvimento embrionário e segregação das linhagens celulares em blastocistos pré-implantacionais, utilizando o modelo de fertilização in vitro (FIV) de camundongo. Material e métodos: fêmeas de camundongo com idade de cinco meses tiveram a ovulação estimulada e, no período pré e/ou pós-natal, foram expostas ao arfiltrado (AF-AF), ar filtrado-ar ambiente (AF-AA) ou ar ambiente-ar ambiente (AA-AA) em câmaras de exposição, 24 horas por dia durante seis meses. Resultados: a resposta ovariana não foi afetada pelo protocolo de exposição. Um efeito multivariável para a exposição pré e/ou pós-natal ao material particulado fino ambiente sobre coloração diferencial dos blastocistos, mas não sobre a FIV e o desenvolvimento embrionário, foi observado. A contagem celular na massa celular interna (MCI) e a razão MCI/trofoectoderma dos blastocistos produzidos no protocolo AF-AF foram significativamente maioresdo que aquelas em blastocistos produzidos nos protocolos FA-AA e AA-AA. Conclusões: nosso estudo sugere que a exposição à poluição ambiental particulada de um grande centro urbano não altera a função ovariana, mas pode afetar negativamente a saúde reprodutiva feminina no período final do menacme, em razão da alteração da especificação das linhagens celulares do embrião no estágio de blastocisto.

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