Method development and validation of ursodiol and its major metabolites in human plasma by HPLC-tandem mass spectrometry.

BACKGROUND: Ursodeoxycholic acid (UDCA) and its metabolites tauroursodeoxycholic acid (TUDCA) and glycoursodeoxycholic acid (GUDCA) have been the subject of several pharmacological studies. The objective of this study was to develop an innovative method of quantification by HPL-tandem mass spectrometry (LC-MS/MS), with a lower cost and suitable, for application in bioequivalence studies. METHODS: The procedure involved liquid-liquid extraction for quantification of UDCA/GUDCA and precipitation extraction for TUDCA, using deuterated substances as internal standards (ISs) and Phenomenex Luna 250×4.6 mm 5µ C18 100A column. The mobile phase used was acetonitrile/ammonium acetate 30 mM (420: 580 v/v pH 7) for UDCA, acetonitrile/ammonium acetate 10 mM/ammonium hydroxide (400:600: 0.5 v/v/v pH 9) for GUDCA, and acetonitrile/ammonium acetate 10 mM (570: 430 v/v pH 7) for TUDCA. Ions were monitored by the electrospray ion source (ESI) mass spectrometer, operating in a negative ionization mode. Compound determination was performed by LC-MS/MS system using a calibration curve of 15-10,000 ng/mL for UDCA/GUDCA and 5-500 ng/mL for TUDCA. The method was developed and validated according to the Brazilian National Health Surveillance Agency (ANVISA) of Brazil norms harmonized with the main international guidelines as a prerequisite for conducting in vivo study in human volunteers. RESULTS: The method did not present matrix effect and residual effect, showing to be selective for studied molecules, with adequate accuracy and precision. In addition, the method was considered sensitive presenting a coefficient of variation less than 20% for the lower limit of quantification of each compound. CONCLUSION: This method can be applied in bioequivalence studies to determine ursodiol and its metabolites reproducibly, simply, and effectively with the use of readily accessible analytical materials and instrumentation.

Efeito da fotobiomodulação no músculo masseter de criança com paralisia cerebral: relato de caso / Photobiomodulation effect on the masseter muscle in children with cerebral palsy: a case report

A espasticidade acarreta hipertonia nos músculos mastigatórios dos indivíduos com paralisia cerebral (PC), interferindo na amplitude de abertura bucal, dificultando a realização da higiene oral predispondo estes indivíduos a condições consideradas de risco para o desenvolvimento de doenças bucais. Objetivo: Avaliar o efeito da fotobiomodulação com laser de diodo, de baixa intensidade na espessura do músculo masseter em uma criança com PC do tipo espástico. Método: O relato do cuidador era que a criança apresentava grande dificuldade na realização da higiene bucal e com movimentos de fuga da cabeça quando a escova dental tocava a região de molares superiores. Com relação ao desconforto da criança, a mãe referiu como extremo. A 1ª avaliação ultrassonográfica foi realizada na avaliação inicial, e a 2ª avaliação após 6 sessões de aplicação de fobioestimulação. Foi empregado o Laser infravermelho, de Diodo, de baixa intensidade, As-Ga-Al, (? = 808 ± 3 nm, 120 mW; Twin Flex Evolution Laser MMOptics São Paulo, Brazil), usando 5,0 J/cm2 energia dose/local, com 20 segundos de exposição/local. A área do músculo masseter irradiado bilateralmente foi o ponto médio no sentido da sua extensão e largura. Foram realizadas seis sessões, com intervalo de 7 dias entre elas. Resultados: Ao final da sexta sessão da fotobioestimulação, a responsável relatou que a criança dormia melhor, apresentava redução no número de movimentos involuntários realizados pela mandíbula e a realização da higiene bucal era possível sem expressão dolorosa da criança. Durante a palpação observou-se menor hipertonia em masseter bilateral, ganho em espessura, e aumento na distância inter-incisal de 7 mm. Conclusão: A fotobioestimulação com laser de diodo parece ser efetiva na redução da espasticidade no músculo masseter de crianças com PC do tipo espástico

Spasticity causes stiffness in the masticatory muscles of individuals with cerebral palsy (CP), affecting the amplitude of mouth opening, making oral hygiene conditions difficult and predisposing these individuals to the risk of developing oral diseases. Objective: To evaluate the effect of a low-intensity photobiomodulation diode laser on the thick part of the masseter muscle in a child with spastic type CP. Method: The caregiver reported that the child had great difficulty in performing the oral hygiene, making avoidance movements of the head when the toothbrush touched the mucosa of the upper molar region in the mouth. The mother described the child's discomfort as extreme. The first ultrasound evaluation was performed at the first appointment, and the second evaluation after 6 sessions of photobiostimulation. The low-intensity Infrared Laser Diode, Ga-As-Al, was employed (? = 808 ± 3 nm, 120 mW; Twin Flex Evolution Laser MMOptics São Paulo, Brazil), using 5.0 J/cm2 dose energy/location, with 20 seconds exposure/site. The area of the masseter muscle was bilaterally irradiated towards the midpoint of its length and width. Six sessions were performed, with an interval of 7 days between them. Results: At the end of the sixth session of photobiostimulation, the mother reported that the child slept better, had reduction in involuntary movements performed by the jaw, and oral hygiene was possible now with no painful expression of the child. During palpation there was less stiffness in the bilateral masseter and an increased masseter thickness and increase in the amplitude of the mouth opening of 7 mm. Conclusion: The diode laser photobiostimulation appears to be effective in reducing spasticity in the masseter muscle of children with spastic type CP

The new noninvasive occlusion spectroscopy hemoglobin measurement method: a reliable and easy anemia screening test for blood donors.

BACKGROUND: The tests used for anemia screening in blood donors are based on fingerstick samples, leading to discomfort and complaints. The aim of this study was to analyze the feasibility of occlusion spectroscopy method in blood banks and to compare the method with fingerstick hemoglobinometer and hemoglobin (Hb) determination on an automatic blood analyzer. STUDY DESIGN AND METHODS: The study enrolled 205 consecutive volunteer blood donors. Samples were collected by fingerstick and venous punction to determine Hb level by a Hemocue Hb201+ (Hb-F) and automatic blood analyzer (Hb-V) and compare to the noninvasive Hb determination by occlusion spectroscopy using NBM200 system (Hb-NI). The percentage errors of Hb-F and Hb-NI of all donors as well as stratified by sex, weight, and age levels were compared to Hb-V as reference values using Wilcoxon signed rank test. RESULTS: The results obtained with Hb-F showed significant errors (p<0.001) in the general group as well as when stratified by sex, weight, and age groups, above values obtained with Hb-V. Hb-NI showed significant errors only in females (p=0.026) and weight level of 61 to 70kg (p=0.034), below Hb-V values. CONCLUSIONS: Hb-NI seems to be a good method in terms of precision and feasibility for anemia screening of blood donors as well as being much more comfortable for donors.

Translation, cultural adaptation, and validation of the screening for occult renal disease (SCORED) questionnaire to Brazilian Portuguese.

UNLABELLED: Screening Chronic Kidney Disease (CKD) allows early interventions, which may alter the natural course of the disease, including cardiovascular morbidity and mortality. Screening for Occult Renal Disease (SCORED) is questionnaire with nine questions with different weights, and predicts a 20% chance for CKD if a individual score > 4 points. AIM: Translate to Portuguese, perform the adaptation to the Brazilian culture and validate the original version of SCORED questionnaire. METHODS: Steps of the process: Translation from English into Brazilian Portuguese; back-translation into English; application to a population sample; and Proof-reading and completion. The translations and reviews were made by professionals experts in Portuguese and English. The questionnaire was applied to 306 participants and CKD was diagnosed as suggested by the NKF KDOQI™. RESULTS: The participants mean age was 49 ± 13 years, 61% were women, 69% were white, and 68% had education below high school, 38.5% had hypertension, and 12.3% diabetics. The final Brazilian Portuguese version of the SCORED questionnaire was well understood. CKD was diagnosed in 20 (6.5%) of the participants. The Brazilian version of the SCORED questionnaire showed sensitivity of 80%, specificity of 65%, positive predictive value of 14%, negative predictive value of 97%, and accuracy of 66%. CONCLUSION: The steps used for the translation, transcultural adaptation, and validation allowed a Brazilian Portuguese version of the SCORED questionnaire which was well understood, acceptable and costless, characteristics that make it a useful tool in the identification of people that chance of having CKD.

Tradução, adaptação cultural e validação do questionário Rastreamento da Doença Renal Oculta (Screening For Occult Renal Disease - SCORED) para o português brasileiro / Translation, cultural adaptation, and validation of the Screening For Occult Renal Disease (SCORED) questionnaire to Brazilian Portuguese

INTRODUÇÃO: Identificar a Doença Renal Crônica (DRC) em seus estágios iniciais permite intervenções com potencial de alterar a evolução natural da doença e de diminuir a mortalidade precoce. O Screening For Occult Renal Disease (SCORED) é um questionário de nove questões com pesos diferentes e prevê uma chance de 20% para DRC em caso de pontuação > 4 pontos. OBJETIVO: Traduzir, adaptar transculturalmente e validar o questionário SCORED para o português brasileiro. MÉTODOS: Etapas do processo: 1. Tradução do inglês para o português brasileiro; 2. Retrotradução para o inglês; 3. Avaliação das versões por comitê de especialistas, gerando uma versão consensual; 4. Validação da versão final para a cultura brasileira. O questionário foi aplicado em 306 indivíduos avaliados para DRC segundo os critérios do NKF KDOQI™. RESULTADOS: A idade média dos participantes foi de 49 ± 13 anos, 61% eram mulheres, 69% eram brancos, 68% apresentavam escolaridade até o ensino médio, 38,5% tinham hipertensão arterial e 12,3% Diabetes Mellitus. A versão final do questionário SCORED em português brasileiro não apresentou dificuldades de compreensão. A DRC foi diagnosticada em 20 (6,4%) participantes. A versão brasileira do questionário SCORED apresentou sensibilidade de 80%, especificidade de 65%, valor preditivo positivo de 14%, valor preditivo negativo de 97% e acurácia de 66%. CONCLUSÃO: As etapas cumpridas no processo de adaptação transcultural permitiram desenvolver a versão brasileira do questionário SCORED, ferramenta que, por ser de fácil compreensão, boa aceitação e de baixíssimo custo, poderá constituir importante instrumento de rastreio de pessoas com chance de apresentar DRC.

ABSTRACT: Screening Chronic Kidney Disease (CKD) allows early interventions, which may alter the natural course of the disease, including cardiovascular morbidity and mortality. Screening for Occult Renal Disease (SCORED) is questionnaire with nine questions with different weights, and predicts a 20% chance for CKD if a individual score > 4 points. AIM: Translate to Portuguese, perform the adaptation to the Brazilian culture and validate the original version of SCORED questionnaire. METHODS: Steps of the process: Translation from English into Brazilian Portuguese; back-translation into English; application to a population sample; and Proof-reading and completion. The translations and reviews were made by professionals experts in Portuguese and English. The questionnaire was applied to 306 participants and CKD was diagnosed as suggested by the NKF KDOQI™. RESULTS: The participants mean age was 49 ± 13 years, 61% were women, 69% were white, and 68% had education below high school, 38.5% had hypertension, and 12.3% diabetics. The final Brazilian Portuguese version of the SCORED questionnaire was well understood. CKD was diagnosed in 20 (6.5%) of the participants. The Brazilian version of the SCORED questionnaire showed sensitivity of 80%, specificity of 65%, positive predictive value of 14%, negative predictive value of 97%, and accuracy of 66%. CONCLUSION: The steps used for the translation, transcultural adaptation, and validation allowed a Brazilian Portuguese version of the SCORED questionnaire which was well understood, acceptable and costless, characteristics that make it a useful tool in the identification of people that chance of having CKD.

Mobilization of reserve proteins and activities of cysteine peptidases during germinative and post-germinative events of cowpea seeds.

Cysteine peptidases are the best characterized peptidases among those involved with storage protein mobilization during seed germination. In the present work we show two major groups of cysteine peptidase activities, one of higher (55 to 97 kDa) and other with lower (15 to 20 kDa) molecular masses which are temporally activated after 24 and 48 HAI, respectively, in germinating cowpeas. The former group is found both in protein bodies and in cytoplasmic fraction, while the latter is mostly present outside protein bodies. A third cysteine peptidase activity of ~37 kDa was specifically active at quiescent cotyledons and at 12 and 60 hours after imbibition (HAI). Main peptidase activities of albumin fractions were synchronizedly detected with radicle emergence at 36 HAI. Major vicilin mobilization was more pronounced from 60 HAI onwards and steadily increased until 144 HAI, when low levels of the smallest vicilin subunit were present. Cysteine peptidases were susceptible to iodoacetamide, E-64, iodoacetic acid, pCMB and ß-mercaptoethanol, except for the ~37 kDa peptidase, which was not affected by any of the inhibitors. By a two-dimensional native/SDS-PAGE combination it was observed an apparent linear arrangement of protein breakdown products as well as of peptidase activity spots. The finding may indicate a complex set of sequential proteolytic events where peptidases induce or activate new peptidases, which may act upon different aggregates or zymogens, and these hydrolysis products appear in a line of constant decreasing Rf x Mr ratio.

Effect of two axial wall height preparations on the retentive properties of full crowns using three different cements.

This study evaluated how different tooth preparation heights and luting cements affected the retention of metallic full crowns. Tooth preparations were made on 60 extracted maxillary premolars with occluso-cervical heights of 4.0 or 5.0 mm. The nickel chromium (NiCr) metallic crowns in each group were subdivided randomly into three groups (n = 10); the samples in each group were cemented with either a self-adhesive resin cement, a conventional resin cement, or a zinc phosphate cement. Cementation was performed by applying 5.0 kg pressure in 50% relative humidity. Specimens were stored in 100% humidity at 37 degrees C for seven days, at which time they were subjected to a vertical tensile test with a universal testing machine at 0.5 mm/min. Data (Kgf) were analyzed by two-way ANOVA and Tukey's multiple comparisons tests (alpha = 0.01). There was a significant difference in mean force in the height preparations (P < 0.001) and in tensile strength (P < 0.001) among the resin cements. Regardless of the cement used, 5.0 mm high preparations showed significantly higher mean forces than the 4.0 mm high preparations. The self-adhesive resin cement showed significantly higher mean tensile strength than the conventional resin cement; the zinc phosphate cement displayed the lowest retention values. The self-adhesive resin cement displayed retention rates twice that of the zinc phosphate cement.

A wounding-induced PPO from cowpea (Vigna unguiculata) seedlings.

Polyphenol oxidases (PPO) are induced in cowpea plants by wounding. The highest activity levels were detected 48h after this stimulus in both wounded and neighbor-to-wounded unifoliates of cowpea seedlings; the increase of activity was in the order of 13 to 15-fold, respectively, in comparison to control unifoliates. Multiple molecular forms of active PPO (Mrs 58, 73 and congruent with220kDa) were detected by partially denaturing SDS-PAGE. Wounding-induced cowpea PPO were extracted and purified through (NH4)2SO4 precipitation and ion-exchange chromatography. The effects of substrate specificity, pH, thermal stability and sensitivity to various inhibitors - resorcinol, EDTA, sodium azide and tropolone - of partially purified soluble PPO were investigated. Purified wounding-induced cowpea PPO (wicPPO) showed the highest activities towards 4-methylcatechol (Km=9.86mM, Vmax=24.66 EU [DeltaAmin(-1)]) and catechol (Km=3.44mM, Vmax=6.64 EU [DeltaAmin(-1)]); no activity was observed towards l-tyrosine, under the assay conditions used. The optimum pH for wound-induced cowpea PPO was 6.0 with 4-methylcatechol as substrate. The enzyme was optimally activated by 10 mM SDS and was highly stable even after 5 min at 80 degrees C. The most effective inhibitor was tropolone, whereas addition of 10mM of resorcinol, EDTA and sodium azide were able to reduce PPO activities by 40%, 15% and 100%, respectively.

O mal-estar na cultura da pulsão de morte: a psicanálise, a ética e a sexualidade / The discontent in the death-drive civilization: psychoanalysis, ethics and sexuality

O artigo debate os desafios que o discurso da ciência e do laço social dominante apresenta neste novo século. Os temas centrais de discussão são a ética e a sexualidade(AU)

The article debates the challenges that the scientific discourse and the dominant social link bring forth in this new century. The central discussion themes are ethics and sexuality(A)

Tubo gástrico isoperistáltico no tratamento paliativo do carcinoma do esôfago / Tube gastric isoperistaltic in the palliative treatment of the carcinoma of the esophagus

São apresentados os resultados do desvio do trânsito esofágico com tubo gástrico isoperistáltico no tratamento paliativo de 15 doentes portadores de carcinoma espinocelular do esôfago com disfagia grave e tumor inextirpável. Os doentes eram do sexo masculino e a média etária foi 55 anos. Dez doentes referiam afagia e cinco disfagia para alimentos líquidos. Em todos os casos os tumores ultrapassavam 10cm de extensão, cinco doentes apresentavam paralisia das cordas vocais e dois invasão traqueo-brônquica. A cirurgia realizada foi o desvio esofágico por tubo gástrico isoperistáltico confeccionado a partir do fundo gástrico próximo à grande curvatura e transposto por via retro-esternal. Nove doentes (60 por cento) evoluíram com algum tipo de complicação, sendo a mais comum a fístula cervical (9 casos). A letalidade pós-operatória foi de 33 por cento (5 doentes). Os dez doentes que sobreviveram apresentaram alívio significativo da disfagia no seguimento ambulatorial que foi de quatro meses. Concluímos que esse método apresenta alta morbidade e letalidade com pouco tempo de sobrevivência nos pacientes com tumor irressecável, estando então reservado para um número restrito de casos

Ferimentos penetrantes do esôfago torácico / Penetrating injury of the thoracic esophagus

Foram analisados nove doentes portadores de ferimentos penetranes do esôfago torácico. Todos os doentes eram do sexo masculino, com média etária de 31 anos (25 a 44 ) e vítimas de ferimentos por arma de fogo. Na avaliaçäo do estado hemodinâmico na sala de admissäo, cinco doentes (56 por cento) encontravam-se estáveis, dois (22 por cento) em choque e dois (22 por cento) agônicos. Sete doentes (78 por cento) apresentavam lesöes associadas e apenas dois tinham ferimento esofágico exclusivo. A endoscopia, realizada em cinco doentes, mostrou-se efetiva no diagnóstico pré-operatório das perfuraçöes do esôfago. O tratamento foi sutura primária em sete doentes e esofagectomia subtotal em dois. A mortalidade global foi 44 por cento, tendo ocorrido pelas lesöes associadas nos doentes agônicos ou por complicaçöes infecciosas nos doentes com mais de 12 horas de perfuraçäo. Os índices de trauma (RTS, PTTI, ISS, TRISS) foram efetivos como avaliaçäo prognóstica apenas nos doentes com instabilidade hemodinâmica à admissäo

Lesäo diafragmática isolada por ferimento penetrante tratada por videolaparoscopia / Diaphragmatic penetrating injury repaired by videolaparoscopy

Penetrating trauma to the thoraco-abdominal region is a difficult clinical problem, in particular or the detection of diaphragmatic injuries. While patients with hemodynamic instability or peritonitis undergo laparotomy, clinically stable patients with equivocal peritoneal signs pose a challenge in management. This repor summarizes our preliminary experience with therapeutic laparoscopy in the assesment of four patients presenting penetrating wounds in lower chest abdomen, with isolated diaphragmatic injuries. The patients were stable on admission, with normal thoracic X-rays. The diaphragmatic injuries were repaired by laparoscopic endosuture and all the patients presented a satisfactory recovery