New human case reports of cutaneous leishmaniasis by Leishmania (Viannia) naiffi in the Amazon region, Brazil / Novos relatos de casos de leishmaniose cutânea por Leishmania (Viannia) naiffi na região amazônica, Brasil

Few cases of human cutaneous leishmaniasis (CL) caused by Leishmania naiffi were described in the medical literature. The aim of this study was to report and analyze new cases of L. naiffi in the period between the years 1992 to 2011. The strains were characterized by isoenzyme analysis. All patients assisted had small lesions; ranging from 1.0x1.0 mm and 13.5x11.5 mm. The lesions observed were widely distributed: 55.5% on the lower limb, 5.5% in the abdominal area, 16.6% on upper limb and 22.2% in upper limb and back. Seventy-two percent of patients had ulcerated lesions. Clinical course of the disease varied from 1 to 10 months. According to gender, most infected individuals were men (83.3%). The patients came from Amazonas (10), Pará (01) and Rondônia (01), north States of Brazil. Five patients were infected in experimental stations of the National Institute of Amazonian Research (INPA). Although the results of this study were similar to other reports in the literature, some of the patients had more of one skin lesion. It is also reported the first case of CL caused by L. naiffi in the State of Rondônia and identified an area of disease transmission in the experimental station of INPA.

Poucos casos de leishmaniose cutânea humana (LC) causada por Leishmania naiffi foram descritos na literatura médica. Assim, o objetivo deste estudo foi relatar e analisar novos casos de infecção por L. naiffi no período compreendido entre os anos de 1992 a 2011. As cepas foram caracterizadas por análise de isoenzimas. Todos os pacientes atendidos apresentavam lesões pequenas; variando entre 1,0x1,0 mm e 13,5x11,5 mm. As lesões observadas foram amplamente distribuídas: 55,5% no membro inferior, 5,5% na área abdominal, 16,6% no membro superior e 22,2% no membro superior e para trás. Setenta e dois por cento dos pacientes tiveram lesões ulceradas. O curso clínico da doença variou de 1 a 10 meses. De acordo com o sexo, a maioria dos indivíduos infectados eram homens (83,3%). Os pacientes vieram de Amazonas (10), Pará (01) e Rondônia (01), estados do norte do Brasil. Cinco pacientes foram infectados em estações experimentais do Instituto Nacional de Pesquisas da Amazônia (INPA). Embora os resultados encontrados neste estudo fossem semelhantes às outras descrições na literatura, alguns dos pacientes apresentavam mais de uma lesão cutânea. Também é relatado o primeiro caso de LC causada por L. naiffi no Estado de Rondônia e identificada uma área de transmissão da doença na estação experimental do INPA.

Increased salivary aldehyde dehydrogenase 1 in non-reticular oral lichen planus

Abstract: Background: Oral lichen planus is a potentially malignant disorder. One of the malignant transformation markers is cancer stem cells. One of the proposed marker for the detection of cancer stem cells's in head and neck cancer is aldehyde dehydrogenase. Recently it is shown that aldehyde dehydrogenase 1 expression in tissue samples is associated with oral lichen planus malignant transformation. Objective: This study evaluates salivary aldehyde dehydrogenase 1 in oral lichen planus. Method: Thirty patients and 30 age and sex-matched healthy volunteers were recruited. Oral lichen planus was diagnosed based on the modified World Health Organization criteria. Subjects in the case group were divided into reticular and non-reticular forms. Unstimulated salivary samples were collected at 10-12 AM. Saliva concentrations of aldehyde dehydrogenase 1 were measured by ELISA. Results: The differences between aldehyde dehydrogenase levels in the oral lichen planus group compared with the control group were not significant but aldehyde dehydrogenase in non-reticular oral lichen planus was significantly higher than that of the reticular form. Limitations of the study: This is a cross-sectional study, thus longitudinal studies in oral lichen planus may present similar or different results. Conclusions: The mechanism of malignant transformation in oral lichen planus is not defined. Previous analyses revealed that the aldehyde dehydrogenase 1 expression is significantly correlated with increased risk of transformation. This finding is consistent with our results because in the erosive and ulcerative forms of oral lichen planus, which have an increased risk of transformation, salivary aldehyde dehydrogenase 1 was overexpressed. A higher salivary aldehyde dehydrogenase level in non-reticular oral lichen planus can be a defensive mechanism against higher oxidative stress in these groups. Aldehyde dehydrogenase may be one of the malignant transformation markers in oral lichen planus. Further studies are needed for introducing aldehyde dehydrogenase as a prognostic indicator in certain lesions.

The effects of binge-pattern alcohol consumption on orthodontic tooth movement

OBJECTIVE: This study aimed to assess tissue changes during orthodontic movement after binge-pattern ethanol 20% exposure. METHODS: Male Wistar rats (n = 54) were divided into two groups. The control group (CG) received 0.9% saline solution, while the experimental group (EG) received 20% ethanol in 0.9% saline solution (3 g/kg/day). On the 30th day, a force of 25 cN was applied with a nickel-titanium closed coil spring to move the maxillary right first molar mesially. The groups were further divided into three subgroups (2, 14 and 28 days). Tartrate-resistant acid phosphatase and picrosirius were used to assess bone resorption and neoformation, respectively. Data were compared by two-way ANOVA, Tukey's HSD, Games-Howell and chi-square test. Significance level was set at 5%. RESULTS: There was a decrease in the number of osteoclasts in EG at day 28. The percentage of collagen showed no interaction between group and time. CONCLUSION: Binge-pattern 20% ethanol promoted less bone resorption at the end of tooth movement, thereby suggesting delay in tooth movement. .

OBJETIVO: objetivou-se avaliar as alterações teciduais decorrentes da administração de etanol a 20% no padrão binge, durante o movimento ortodôntico. MÉTODOS: foram utilizados ratos Wistar machos (n = 54), divididos em dois grupos, sendo Grupo Controle (GC), com administração de soro fisiológico a 0,9%; e Grupo e Experimental (GE), com administração de etanol a 20% em soro fisiológico a 0,9%, no volume de 3g/kg/dia. Após o 30º dia de administração, foi aplicada força de 25cN com mola fechada de níquel-titânio para mover o primeiro molar superior direito para mesial. Os grupos foram subdivididos nos subgrupos 2, 14 e 28 dias, correspondendo ao número de dias de movimentação dentária. Utilizou-se as colorações de fosfatase ácida-tartarato resistente e picrosírius para avaliar reabsorção óssea e neoformação óssea, respectivamente. Os dados foram comparados por ANOVA a dois critérios, Tukey HSD e Games-Howell, ao nível de significância de 5%. RESULTADOS: verificou-se diminuição no número de osteoclastos no GE II no 28º dia. A percentagem de colágeno não demonstrou alteração na interação grupo x tempo. CONCLUSÕES: o etanol no padrão binge a 20% promoveu menor reabsorção óssea no final da movimentação dentária, sugerindo atraso na movimentação dentária. .

Liver cancer stem cells are selectively enriched by low-dose cisplatin

Accumulating evidence has indicated the importance of cancer stem cells in carcinogenesis. The goal of the present study was to determine the effect of low-dose cisplatin on enriched liver cancer stem cells (LCSCs). Human hepatoblastoma HepG2 cells were treated with concentrations of cisplatin ranging from 1 to 5 μg/mL. Cell survival and proliferation were evaluated using a tetrazolium dye (MTT) assay. LCSCs were identified using specific markers, namely aldehyde dehydrogenase-1 (ALDH1) and CD133. The percentage of ALDH1+ or CD133+ cells was examined by flow cytometric analysis. The expression of ALDH1 and/or CD133 in HepG2 cells was determined by immunocytochemical analysis. Low-dose cisplatin treatment significantly decreased cell survival in HepG2 cells after 24 or 72 h. However, the percentage of LCSCs in the surviving cells was greatly increased. The percentage of ALDH1+ or CD133+ cells was increased in a time- and dose-dependent manner after treatment with 1-4 μg/mL cisplatin, whereas 5 μg/mL cisplatin exposure slightly reduced the number of positive cells. These findings indicate that low-dose cisplatin treatment may efficiently enrich the LCSC population in HepG2 cells.

On the origin and diversification of Venezuelan freshwater fishes: the genus Gephyrocharax (Ostariophysi: Characidae) a case study

We conducted a phylogeographic analysis of the genus Gephyrocharax in Venezuela to evaluate geomorphologic evidence for the formation of the country's main watersheds and to establish a biogeographical hypothesis of possible diversification mechanisms of the Neotropical freshwater fish fauna. We assayed eight enzyme systems and general proteins to estimate genetic variability (H, P), intraspecific structuring in several Gephyrocharax valencia and G. venezuelae populations (FIS, FIT, and FST), and a phylogenetic approach for the three species of Gephyrocharax in Venezuela, using Corynopoma riisei as the external group. Fourteen presumptive loci indicate that populations of the three species of Gephyrocharax analyzed show a clear genetic inter-specific differentiation, determined by four loci with fixed alleles (GPI-B*, IDH*, ME-1*, and ME-2*). The resulting cladogram shows two major clades: a monophyletic group consisting of Gephyrocharax n. sp. and G. venezuelae (restricted to the northwest of the country) and a group formed exclusively by G. valencia (distributed along the largest geographic range). Speciation of the Venezuelan lineages of the genus Gephyrocharax could be explained by the origin and course movements of the present Orinoco River together with geomorphologic processes that have occurred in northern Venezuela since the Miocene.

Foi feita uma análise filogeográfica do gênero Gephyrocharax na Venezuela a fim de avaliar as evidências geomorfológicas que levaram à formação dos principais sistemas hidrográficos do país, além de estabelecer uma hipótese biogeográfica com os possíveis mecanismos de diversificação da fauna de peixes de água doce Neotropical. Foram analisados oito sistemas enzimáticos e proteínas gerais para conhecer a variabilidade genética (H, P), estruturação intraespecífica em populações de Gephyrocharax valencia e G. venezuelae (FIS, FIT e FST), e uma abordagem filogenética com base na análise isozimática para as três espécies de Gephyrocharax na Venezuela, com Corynopoma riisei como grupo externo. Quatorze loci presumíveis indicam que as populações das três espécies de Gephyrocharax analisados revelam uma diferenciação inter-específica genética, determinada por quatro loci com alelos fixos (GPI-B*, IDH*, ME-1* e ME-2*). O cladograma resultante apresenta dois clados principais: um grupo monofilético composto por Gephyrocharax n. sp. e G. venezuelae (restrita ao noroeste do país) e um grupo formado exclusivamente por G. valencia (distribuídos ao longo da maior área geográfica). A especiação das linhagens de Gephyrocharax na Venezuela poderia ser explicada pela origem e movimentos do curso atual da bacia do rio Orinoco, associado a processos geomorfológicos que ocorrem no norte da Venezuela desde o Mioceno.

Influencia de la dieta sobre las actividades de fosforiladoras de glucosa en hígado e intestino de rata / Influence of diet on glucose phosphorylating activity of rat liver and small intestine

La fosforilación de la glucosa en los mamíferos, es catalizada por una familia de isoenzimas (hexoquinasas I-IV; HQ) de diferente Km para el azúcar. En los hepatocitos y células b-pancreáticas se encuentra la glucoquinasa (GQ; HQ IV) de Km elevado (12-20 mM). Hemos observado que GQ está presente en el intestino delgado y podría contribuir a la producción de lactato durante la absorción del azúcar. En este trabajo se determinó el efecto de la dieta (ratarina R; 60% de glucosa G; sacarosa S; almidón A; caseína C), suministrada ad libitum, sobre las actividades de HQ y GQ en homogenatos de hígado y mucosa intestinal de rata. El suministro de glucosa (5%) en el agua de beber (SG) también fue evaluado en las dietas con R y G. Las actividades de HQ (Glucosa 1 mM) y la capacidad fosforilativa total (CFT: Glucosa = 100 mM) se determinaron enzimáticamente. GQ se estimó por diferencia. En el grupo control (R) y en S, A y C, la GQ hepática fue un 85% de la CFT, mientras que en G, GSG y RSG un 66%. La HQ intestinal alcanzó en los grupos R, GSG, A y C un 87% y en RSG un 30% de la CFT. La GQ en G, S, aumentó, pero una menor magnitud. La presencia de GQ en el intestino delgado y su expresión diferencial de acuerdo a la dieta, abren la posibilidad de que dicho órgano contribuya al metabolismo inicial de la glucosa procedente de la dieta y provea al hígado de un precursor (lactato) muy eficaz para sus procesos anabólicos.

Glucose phosphorylation in mammals, is catalyzed by a family of isoenzymes (hexokinases I- IV; HQ) of different Km for the sugar. In hepatocytes and pancreatic b- cells are glucokinase (GQ ; HQ IV) of high Km (12-20 mM). We observed that GQ is present in the small intestine and may contribute to the production of lactate during the absorption of sugar. In this work, the effect of diet (ratarina R, G 60% glucose, sucrose S; starch A; casein C) provided ad libitum , on the activities of HQ and GQ in liver homogenates of rat intestinal mucosa . The supply of glucose (5%) in the drinking water ( SG ) was also evaluated in the diets with R and G. HQ activities (Glucose 1 mM) and phosphorylating full capacity ( CFT : Glucose = 100 mM ) were determined enzymatically . GQ was estimated by difference. In the control group (R) and S, A and C, the GQ liver was about 85% of CFT, whereas G, GSG and RSG 66%. The intestinal HQ reached in the R groups, GSG, A and C by 87% and 30% RSG the CFT. The GQ in G, S , increased , but a lower magnitude. the presence of glucokinase in the small intestine and its differential expression according to diet, open the possibility that this structure contributes to initial metabolism of glucose and provide to the liver a precursor (lactate) very effective for their anabolic processes.

Valor pronóstico de las isoenzimas de LDH en pacientes con linforma no hodgkin / Prognostic value of LDH isoenzymes in patients with non-Hodgkin linforma

La enzima lactato deshidrogenasa (LDH) es un factor pronóstico en Linfoma No Hodgkin (LNH). El objetivo del trabajo consistió en evaluar prospectivamente el valor pronóstico de las isoenzimas de LDH en pacientes con LNH. Se estudiaron 67 pacientes de primera consulta con diagnóstico de LNH, sin tratamiento previo, VIH negativo y sin otras enfermedades, tiempo promedio de seguimiento 30 meses (rango 3-48 meses). Las muestras de suero se recolectaron previas al tratamiento. La LDH total (LDHT) e isoenzimas de LDH se determinaron respectivamente por método cinético y electroforesis de proteínas en gel de agarosa. Se procesaron muestras de 122 controles sanos para establecer los valores de referencia de las isoenzimas de LDH. 49(73%) LNH agresivos y 18(27%) LNH indolentes y según el Índice Pronóstico Internacional (IPI), 60 (90%) bajo riesgo y 7(10%) alto riesgo. Las isoenzimas LDH1, LDH2, LDH3, LDH4 y LDH5 presentaron niveles absolutos significativamente elevados en 25 (37%), 29 (43%), 32 (48%), 20 (39%) y 11 (16%) de los casos respectivamente (p<0,0001). La actividad porcentual de LDH4 en los pacientes con LNH agresivos fue significativamente superior respecto al grupo de LNH indolentes (p=0,01). En el análisis univariado, valores absolutos elevados de LDH1 se asociaron significativamente con una sobrevida global disminuida (p=0,0064) en el grupo total de pacientes. LDH1 conservó su valor pronóstico aún en el grupo de pacientes con valores normales de LDHT (p=0,04). En pacientes con LNH agresivos, valores elevados de LDHT e IPI alto riesgo se asociaron significativamente con una menor sobrevida global (p<0,05). En el análisis multivariado la LDHT e IPI resultaron factores pronósticos independientes de la sobrevida. Alteraciones específicas del patrón de isoenzimas de LDH sugieren la relación de LDH4 con la biología del tumor y su actividad proliferativa en LNH agresivos y el valor pronóstico de LDH1 como factor adverso de la sobrevida en el análisis univariado.

Lactate dehydrogenase (LDH) is a prognostic factor in non-Hodgkin lymphoma (NHL). Our objective was to evaluate prospectively the prognostic value of LDH isoenzymes in patients with NHL. We studied 67 newly diagnosed NHL patients, previously untreated, HIV-negative and free from other disease, median follow-up of 30 month (range 3-48 month). Before starting treatment serum samples were collected for the determination of total LDH (LDHT) and LDH isoenzymes that were respectively assayed by kinetic method and protein electrophoresis in agarose gel. In order to set reference values of LDH isoenzymes samples from122 healthy controls were processed. Results: 49(73%) of the patients were aggressive NHL and 18(27%) indolent NHL and according to the International Prognostic Index (IPI), 60(90 %) low risk and 7(10%) high risk. High absolute values of LDH1, LDH2, LDH3, LDH4 and LDH5 isoenzymes were significantly elevated in 25 (37%), 29 (43%), 32 (48%), 20 (39%) and 11 (16%) of cases respectively (p<0,0001). The percentage value of LDH4 activity in aggressive NHL patients was significantly higher compared to indolent NHL group (p=0,01). In univariate analysis increased LDH1 absolute values were significantly associated with decreased overall survival in the total group of patients (p = 0.0064). LDH1 remained a prognostic factor for survival even when considering the group of patients with normal serum LDHT values (p = 0.04). In patients with aggressive NHL increased values of LDHT and high risk IPI were significantly associated with decreased overall survival (p<0.05). In a multivariate analysis LDHT and IPI score were independent prognostic factor for survival.

Comparison of microsatellites and isozymes in genetic diversity studies of Oryza glumaepatula (Poaceae) populations

The study of the genetic structure of wild plant populations is essential for their management and conservation. Several DNA markers have been used in such studies, as well as isozyme markers. In order to provide a better comprehension of the results obtained and a comparison between markers which will help choose tools for future studies in natural populations of Oryza glumaepatula, a predominantly autogamous species, this study used both isozymes and microsatellites to assess the genetic diversity and genetic structure of 13 populations, pointing to similarities and divergences of each marker, and evaluating the relative importance of the results for studies of population genetics and conservation. A bulk sample for each population was obtained, by sampling two to three seeds of each plant, up to a set of 50 seeds. Amplified products of eight SSR loci were electrophoresed on non-denaturing polyacrylamide gels, and the fragments were visualized using silver staining procedure. Isozyme analyses were conducted in polyacrylamide gels, under a discontinuous system, using six enzymatic loci. SSR loci showed higher mean levels of genetic diversity (A=2.83, p=0.71, A P=3.17, Ho=0.081, He=0.351) than isozyme loci (A=1.20, p=0.20, A P=1.38, Ho=0.006, He=0.056). Interpopulation genetic differentiation detected by SSR loci (R ST=0.631, equivalent to F ST=0.533) was lower than that obtained with isozymes (F ST=0.772). However, both markers showed high deviation from Hardy-Weinberg expectations (F IS=0.744 and 0.899, respectively for SSR and isozymes). The mean apparent outcrossing rate for SSR ( =0.14) was higher than that obtained using isozymes ( =0.043), although both markers detected lower levels of outcrossing in Amazonia compared to the Pantanal. The migrant number estimation was also higher for SSR (Nm=0.219) than isozymes (Nm=0.074), although a small number for both markers was expected due to the mode of reproduction of this species, defined ...

El estudio de la estructura genética de poblaciones de plantas silvestres es esencial para su manejo y conservación. Varios marcadores de ADN e isoenzimas se han utilizado en este tipo de análisis. Con el fin de proporcionar una mejor comprensión de los resultados obtenidos y saber que marcador codominante elegir para futuros estudios en poblaciones naturales de Oryza glumaepatula, este trabajo busco evaluar y comparar dos marcadores de ADN, isoenzimas y microsatélites, en la diversidad y estructura genética de 13 poblaciones, destacando las similitudes y divergencias de cada marcador, así como la importancia relativa de los resultados en genética de poblaciones y conservación. Para los SSR, ocho loci SSR fueron evaluados, y los fragmentos se visualizaron utilizando el procedimiento de coloración con plata. Los análisis de isoenzimas se realizaron en geles de poliacrilamida, en los seis loci enzimáticos. Los loci SSR mostraron mayores niveles de diversidad genética que los loci isoenzimáticos, en promedio. La diferenciación genética entre los loci SSR (R ST=0.631, equivalente a F ST=0.533) fue inferior a la obtenida con las isoenzimas (F ST=0.772). Ambos marcadores mostraron alta desviación del equilibrio de Hardy-Weinberg (F IS=0.744 y 0.899, respectivamente, para SSR e isoenzimas). La tasa media aparente de cruzamiento para SSR ( =0.14) fue mayor que la obtenida con isoenzimas ( =0.043), aunque ambos marcadores detectaron niveles más bajos en la tasa de fecundación cruzada para la Amazonia, en comparación con la región del Pantanal. La estimación de número de migrantes también fue mayor para los SSR (Nm=0.219) que en isoenzimas (Nm=0.074). No se obtuvo ninguna correlación entre las distancias genéticas y geográficas para los SSR, y para las isoenzimas se obtuvo una correlación positiva entre las distancias genéticas y geográficas. Llegamos a la conclusión de que estos marcadores son divergentes en la detección de los parámetros de la diversidad genética en O. glumaepatula y que los microsatélites son más eficientes para detectar la información a nivel intra-poblacional, mientras que las isoenzimas son más potentes para detectar la diversidad entre poblaciones.

Perfil de las isoenzimas de LDH en linfoma no Hodgkin: correlación con marcadores tumorales / Profiles of LDH isoenzymes in non-Hodking`s lymphoma: correlation with tumor markers

Lactate dehydrogenase (LDH) consists of five isoenzymes and is an important prognostic factor in non-Hodgkin’s lymphoma (NHL) patients. Our objective was to study the pattern of LDH isoenzymes in patients with NHL and its correlation with clinical pathological and biological tumor markers. We evaluated 67 newly diagnosed NHL patients clinically and histologically confirmed, previously untreated, HIV-negative and free from other diseases, during the period 1999-2004, the average follow-up time of 30 months (range 3-48), median age of 55 years (range 18-79), the International Prognostic Index (IPI ) 60 (90%) of low risk and 7 (10%) of high risk. Serum and whole blood samples were collected for the determination of LDH, LDH isoenzymes, enzymes (AST, ALT, phosphatase alkaline), Beta 2 Microglobulin, CA125, and IL-6, sRáIL-2, C Reactive protein, serum albumin and ESR. Serum samples were processed from healthy controls in order to set reference values of LDH isoenzymes. Serum levels of LDH and absolute values of LDH isoenzymes were significantly higher in patients compared to controls (p<0.001). Frequencies of high absolute values of LDH1, LDH2, LDH3, LDH4 and LDH5 isoenzymes were significantly elevated in 25 (37%), 29 (43%), 32 (48%), 20 (39%) and 11 (16%) of cases, respectively. LDH1 activity associated and correlated with adverse clinical pathological, biological factors and high risk IPI, suggets that it is an indicator of cell turnover and disease activity. LDH2 changes reflected its association and correlation with clinical pathological and biological factors which are indicators of disease progression, tumor proliferative activity, adverse IPI and the patient’s response to the illness. LDH3 was elevated with greater frecuency and its activity was associated with clinical pathological and biological prognostic factors reflecting the patient’s response against the tumor, as well as inflammatory activity and disease extension...

La enzima lactato deshidrogenasa (LDH) está conformada por cinco isoenzimas y es un importante factor pronóstico en pacientes con Linfoma No Hodgkin (LNH). Nuestro objetivo fue estudiar el patrón de las isoenzimas de LDH en pacientes con LNH y su correlación con marcadores tumorales clínico-patológicos y biológicos. Se evaluaron 67 pacientes de primera consulta con diagnóstico de LNH confirmados clínica e histopatológicamente, sin tratamientos previo, VHI negativo y sin presentar otras enfermedades, durante el período 1999-2004, siendo el tiempo promedio de seguimiento de 30 meses (rango 3-48 meses), edad promedio 55 años (rango 18-79), Índice Pronóstico Internacional (IPI) 60 (90 por ciento) bajo riesgo y 7 (10 por ciento) alto riesgo. Se recolectaron muestras de sangre, para la obtención de suero y sangre total para la determinación de LDH, Isoenzimas de LDH, enzimas (AST, ALT, fosfatasa alcalina), Beta 2 Microglobulina, CA125,IL-6,sRIL-2, Proteína C Reactiva, Albúmina sérica y VSG. Se procesaron muestras de suero de controles sanos para establecer los valores de referencia de las isoenzimas de LDH. Los nieveles séricos de la LDH y los valores absolutos de las isoenzimas de LDH fueron significativamente superiores en los pacientes respecto al grupo control (p<0,001). Las frecuencias de las isoenzimas LDH1, LDH2, LDH3, LDH4 y LDH5 resultaron con niveles absolutos significativamente elevados en 25 (37 por ciento), 29 (43 por ciento), 32 (48 por ciento), 20 (39 por ciento) y 11 (16 por ciento) de los casos respectivamente. La asociación y correlación de la actividad de LDH1 con los factores clínico-patológicos adversos, IPI alto riesgo y marcadores biológicos alterados , sugieren su expresión como un indicador de recambio celular y actividad de la enfermedad. Los cambios en el patrón de la LDH2 reflejaron su asociación y correlación con factores clínico-patológico y biológicos indicadores de la pregresión de la enfermedad...

Typing Candida albicans oral isolates from healthy Brazilian schoolchildren using multilocus enzyme electrophoresis reveals two highly polymorphic taxa

The genetic diversity of C. albicans oral isolates from 75 healthy schoolchildren from eight schools located in different geographic areas of Piracicaba city, São Paulo state, Brazil, was established using isoenzymes marker (Multilocus Enzyme Electrophoresis - MLEE) and cluster analysis. Patterns of monoclonal and polyclonal oral colonization by C. albicans within and between groups of schoolchildren were identified. However, significant divergence between the observed and the expected genotypic frequencies (Hardy-Weinberg equilibrium test) was not detected in the geographically adjacent groups, suggesting the hypothesis that populations of healthy schoolchildren do not correspond to the selection factor (differential survival) of strains. Two highly polymorphic and distantly genetically related taxa (A and B) were identified within the total population of yeasts, each contained subgroups (A1, A2, A3, A4, B1 and B2) and clusters of moderately related strains (from I to X), suggesting the existence of strains restricted or not to certain groups of geographically limited, healthy students. However, the coexistence of identical strains in healthy schoolchildren from the same school (geographically related) reinforces the hypothesis of oral transmission, where the sources of propagation could be explored. Furthermore, this could also be used in current and retrospective analyses of C. albicans isolated from immunocompetent and immunocompromised people, in order to detect commensal or potentially pathogenic yeast groups, predominantly in candidiasis, and in the development of strategies to prevent transmission or human propagation.

Overlapping Presence of Macroamylasemia and Hyperamylasemia in Acute Pancreatitis / 대한진단검사의학회지

No abstract available.

Biomarkers for identifying the early phases of osteoarthritis secondary to medial patellar luxation in dogs

The levels of tartrate resistant acid phosphatase (TRAP), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) in synovial fluid (SF) and serum in cases of canine osteoarthritis (OA) were measured. OA was induced by a surgically-created medial patellar luxation in the left stifle of 24 dogs. SF and blood samples were collected at 1.5- and 3-month intervals, respectively. Every 3 months, one dog was euthanatized to collect tissue samples from both stifles. TRAP levels in SF and serum were measured using a spectrophotometer, and TRAP-positive cells in joint tissues were identified by enzyme histochemistry. MMP-2 and TIMP-2 in SF and serum were detected by Western blotting and ELISA, respectively. TRAP in SF from the stifles and serum was significantly increased (p < 0.05) after 3 months. TIMP-2 in SF and serum was significantly decreased (p < 0.05), whereas MMP-2 in SF was significantly increased (p < 0.05) during the progression of OA. Histochemistry revealed an increased number of TRAP-positive cells in tissues from OA-affected joints. Assays measuring TRAP, MMP-2, and TIMP-2 in SF and serum, and methods that detect increased numbers of TRAP-positive cells in the joint tissues can play an important role in identifying the early phases of degenerative changes in canine joint components.

Caracterização isoenzimática de isolados humanos de Leishmania sp (Kinetoplastida: Trypanosomatidae) dos municípios de Rio Preto da Eva e Manaus, Estado do Amazonas / Isoenzymatic characterization of human isolates of Leishmania sp (Kinetoplastida: Trypanosomatidae) from the municipalities of Rio Preto da Eva and Manaus, State of Amazonas

Foram caracterizados/identificados por eletroforese de isoenzimas 23 isolados de Leishmania sp de pacientes dos municípios de Rio Preto da Eva e Manaus, analisando-se o grau de similaridade entre os organismos. Os resultados indicaram ocorrência de Leishmania guyanensis e Leishmania naiffi nestes dois ambientes e a heterogeneidade das amostras de Leishmania naiffi.

Twenty-three isolates of Leishmania sp from patients in the municipalities of Rio Preto da Eva and Manaus were characterized and identified by means of isoenzyme electrophoresis and the degree of similarity between the organisms was analyzed. The results indicated that Leishmania guyanensis and Leishmania naiffi were present in these two environments and that the Leishmania naiffi samples were heterogenous.

Population dynamics of Triatoma vitticeps (Stål, 1859) in Itanhomi, Minas Gerais, Brazil

Taxonomic markers (head structure morphometry, isoenzymes and randon amplified polymorphism of DNA - RAPD) were used to understand the population dynamics of Triatoma vitticeps, predominant triatomine species in Itanhomi district, using samples obtained from domestic, peridomiciliary and sylvatic habitats. Morphometric analysis revealed sexual dimorphism within the three samples although specimens could not be separated according to the habitat in which they were captured. Forty-two bands were analyzed from RAPD profiles generated using four primers. A dendrogram constructed from Dice's similarity coefficient values showed that migration of the insects between the habitats has occurred, without structuring of populations. Moreover, the dendrogram obtained from the genetic distance values showed an important gene flow between the sylvatic and domestic habitats. No polymorphism was found in the electrophoretic mobility of proteins for the ten enzymes studied. Our results revealed movement of triatomines between the three habitats, suggesting that the presence of T. vitticeps in houses should not be ignored. As invasion of houses by sylvatic insects is frequent and the natural infection indices of this species are among the highest known, epidemiological vigilance studies may reveal possible changes in T. vitticeps behaviour which could present future risks to public health.

Genetic variability in three Amazon parrot species

Parrots of the genus Amazona are among the most threatened species of the Order Pscittaciformes. This work describes allozyme polymorphisms in three Amazon parrot species - the Blue-fronted Amazon (Amazona aestiva), the Orange-winged Amazon (Amazona amazonica), and the Festive Amazon (Amazona festiva) -, and provides useful data for the evaluation of their genetic variability. We electrophoretically analyzed blood samples from 68 wild-caught individuals, maintained in captivity in three Brazilian zoos. Eight of the ten studied enzyme loci exhibited polymorphism. Glucosephosphate isomerase (Gpi) proved to be a diagnostic locus for the identification of these Amazon species. The expected average heterozygosity of the Blue-fronted Amazon (0.060) differed significantly from the expected heterozygosities of the Orange-winged Amazon and the Festive Amazon (0.040 and 0.039, respectively). This result was discussed as a consequence of hybridization between two geographic A. aestiva subspecies, and alternatively as a particular trait of this species. Genetic variability of the Blue-fronted Amazon compared to birds in general is not low on a species-wide level, despite the fact that this parrot is one of the most illegally traded species. Allozyme analysis proved to be an useful tool in monitoring the genetic variation within the genus Amazona and can be applied in the management program of other threatened species of this genus.

Papagaios do gênero Amazona estão entre as espécies mais ameaçadas da Ordem Psittaciformes. O presente trabalho descreve polimorfismos enzimáticos em três espécies de papagaios do gênero Amazona: o papagaio verdadeiro (Amazona aestiva), o papagaio do mangue (Amazona amazonica) e o papa-cacau (Amazona festiva). Estes dados foram utilizados para avaliação da variabilidade genética dessas espécies. Foram analisadas, através de eletroforese, amostras de sangue de 68 indivíduos capturados na natureza e mantidos em cativeiro em três zoológicos brasileiros. Oito dentre dez locos enzimáticos analisados exibiram polimorfismo. O loco da Glicose Fosfato Isomerase (Gpi) demonstrou ser um loco diagnóstico para a identificação dessas espécies de papagaios. A heterozigosidade média esperada para A. aestiva (0,060) diferiu significativamente das heterozigosidades esperadas para A. amazonica e A. festiva (0,042 e 0,039, respectivamente). Este resultado foi discutido como uma conseqüência de hibridização entre duas subespécies geográficas de A. aestiva e, alternativamente, como uma característica particular da espécie. Comparada a aves em geral, a variabilidade genética de A. aestiva não é baixa, apesar deste papagaio ser uma das espécies mais comercializadas ilegalmente. A análise alozímica demonstrou ser uma ferramenta útil para o monitoramento da variabilidade genética do gênero Amazona, podendo ser aplicada em programas de manejo destas e de outras espécies ameaçadas pertencentes ao mesmo gênero.

Single and concomitant experimental infectionsby Endotrypanum spp. and Leishmania (Viannia) guyanensis (Kinetoplastida: Trypanosomatidae) in the Neotropical sand fly Lutzomyia longipalpis (Diptera: Psychodidae)

Lutzomyia longipalpis females received single and mixed infections with Endotrypanum and Leishmania. Two biological parameters were analyzed: the percentage of infected females and the distribution of flagellates in the gut of the females. The principal comparisons were performed between (1) two strains of Endotrypanum, (2) cloned versus primary sample of one strain of Endotrypanum, (3) Endotrypanum versus Leishmania guyanensis, and (4) the pattern of flagellates behaviour by optical microscopy in females with single or mixed infection versus the identification of parasites isolated from digestive tracts by isoenzyme electrophoresis. Flagellates of Endotrypanum showed distinct patterns of infection suggesting that there is variation between and within strains. The distribution of Endotrypanum and L. guyanensis differed significantly in relation to the colonization of the stomodeal valve. In co-infection with L. guyanensis, a large number of flagellates were seen to be plentifully infecting the stomodeal valve in significantly more specimens than in females infected by Endotrypanum only. However, the electrophoretic profiles of isoenzymes of parasites recovered from all co-infected specimens corresponded to Endotrypanum. This suggests that the mere correlation sand fly infection-biochemical analysis of isolates may induce parasitological incorrect consideration.

Molecular characterization of human Trypanosoma cruzi isolates from endemic areas in Panama

The present work provides information on Trypanosoma cruzi genotype circulating in endemic areas of Chagas disease in Panama. A total of 26 crude stocks of T. cruzi, isolated from the blood of persons with different clinical profiles of Chagas disease were collected and crio-conserved until used. Most of the stocks had been characterized by means of isoenzyme electrophoresis on cellulose acetate membranes. The clinical profiles of infected persons included 9 (34.6 percent) asymptomatic and 17 acute (65.4 percent) including 5 (19.2 percent) fatal cases, 2 under 5 years old and 3 adults. A multiplex-PCR assay based on the amplification of the non-transcribed spacer of the mini-exon gene was performed. All stocks of T. cruzi included in the study were found to correspond to Tc I group. This result supports the predominance of T. cruzi-I in the transmission cycles affecting the human population in the Republic of Panama.

Variabilidade genética em algumas criações comerciais brasileiras de escargots (Helix aspersa, Müller, 1774) / Genetic variation at eight isoenzyme loci in subpopulations of the edible snail (Helix aspersa, Müller, 1774)

Descreveram-se os marcadores isoenzimáticos e estimou-se a variabilidade genética de 20 subpopulações brasileiras de escargots (Helix aspersa). O estudo dos oito locos foi feito por eletroforese em gel de amido, em amostras com 30 indivíduos cada, obtidas em criatórios dos estados de Santa Catarina, São Paulo e Rio de Janeiro (uma, duas e 17 amostras, respectivamente). Observou-se polimorfismo nos locos das enzimas LAP, 6-PGD, PEP 2, PEP 1 e MDH, com três alelos nos três primeiros locos e dois nos demais. Os locos da ME, da SOD e da PGI apresentaram-se monomórficos. As freqüências gênicas de sete amostras ajustaram-se ao modelo de Hardy-Weinberg (P<0,05), e as de outras seis amostras ajustaram-se ao modelo de Wright (P<0,05), indicando que elas estão submetidas a diferentes regimes reprodutivos. Os desvios da panmixia para toda a população (F IT ) e dentro das subpopulações (F IS) não foram significativos (P³0,05). O desvio entre as subpopulações (F ST=0,0485) foi significativo (P<0,05) e apontou pequena diferenciação entre elas. As estimativas de diversidade total (Ht), entre subpopulações (Dst) e dentro das subpopulações (Hs), indicaram que a diversidade genética é reduzida e sua maior parte encontra-se dentro das subpopulações, sugerindo uma base genética estreita para essa população. As distâncias genéticas também foram pequenas, não permitindo a construção de um dendrograma.

Reinfections with strains of Trypanosoma cruzi, of different biodemes as a factor of aggravation of myocarditis and myositis in mice

Reinfeccões pelo Trypanosoma cruzi em pacientes de áreas endêmicas têm sido mencionadas como fator agravante das manifestacões cardíacas na doenca de Chagas. No presente estudo, a influência da tríplice infeccão com cepas de diferentes biodemas, sobre as lesões do miocárdio e de músculo esquelético foi investigada experimentalmente. Cinqüenta e oito camundongos cronicamente infectados com a cepa Colombiana do Trypanosoma cruzi (Biodema Tipo III) foram sucessivamente reinoculadas como a seguir: 1º grupo - reinfectados com a cepa 21 SF (Tipo II) seguido pela cepa Y (Tipo I); 2º grupo - reinfeccão com a cepa Y seguida pela cepa 21SF. A análise isoenzimática dos parasitas das hemoculturas obtidas dos animais com tríplice infeccão, revelou os padrões dos diferentes zimodemas no mesmo animal. Cada cepa do Trypanosoma cruzi foi re-isolada após quatro passagens em camundongos no 7º, no 14º, ou no 30º dia após a inoculacão com o sangue de camundongos com tríplice infeccão. Resultados da histopatologia demonstraram uma significante exacerbacão das lesões inflamatórias de miocárdio e músculo esquelético, confirmadas pela avaliacão morfométrica. Não foi detectada acentuacão do parasitismo. A possibilidade de aumento da resposta celular nos animais com tríplice infeccão é sugerida.

Ocorrência de leishmaniose tegumentar americana no Estado do Mato Grosso do Sul associada à infeccão por Leishmania (Leishmania) amazonensis / Occurrence of American tegumentary leishmaniasis in the Mato Grosso do Sul State associated to the infection for Leishmania (Leishmania) amazonensis

São relatados nove casos de leishmaniose tegumentar americana ocorridos no ano de 2001 em uma unidade de treinamento militar localizada no município de Bela Vista, Estado de Mato Grosso do Sul. Parasitas obtidos de lesões de seis pacientes foram isolados em cultura e posteriormente identificados através da análise de isoenzimas como sendo Leishmania (Leishmania) amazonensis. Esta é a primeira evidência da presenca desta espécie de parasita em Mato Grosso do Sul.