The morphometric acclimation to depth explains the long-term resilience of the seagrass Cymodocea nodosa in a shallow tidal lagoon.

Cadiz Bay is a shallow mesotidal lagoon with extensive populations of the seagrass Cymodocea nodosa at intertidal and shallow subtidal elevations. This work aims to understand the mechanisms behind the resilience of this species to gradual sea level rise by studying its acclimation capacity to depth along the shallow littoral, and therefore, to gradual variations in the light environment. To address this objective, these populations have been monitored seasonally over a 10 year period, representing the longest seasonal database available in the literature for this species. The monitoring included populations at 0.4, -0.08 and -0.5 m LAT. The results show that C. nodosa has a strong seasonality for demographic and shoot dynamic properties - with longer shoots and larger growth in summer (high temperature) than in winter (low temperature), but also some losses. Moreover, shoots have different leaf morphometry depending on depth, with small and dense shoots in the intertidal areas (0.4 m) and sparse large shoots in the subtidal ones (-0.08 and 0.5 m). These differences in morphometry and shoot dynamic properties, combined with the differences in shoot density, explain the lack of differences in meadow production balance (i.e. meadow growth - meadow losses) between the intertidal (0.4 m) and the deepest population (-0.5 m), supporting the long term resilience of Cymodocea nodosa in Cadiz Bay. This study contributes to the understanding of the mechanisms behind seagrass stability and resilience, which is particularly important towards predicting the effects of climate change on these key coastal ecosystems, and also highlights the value of continuous long-term monitoring efforts to evaluate seagrass trajectories.

Long-term effects of elevated CO2 on the population dynamics of the seagrass Cymodocea nodosa: Evidence from volcanic seeps.

Population reconstruction techniques was used to assess for the first time the population dynamics of a seagrass, Cymodocea nodosa, exposed to long-term elevated CO2 near three volcanic seeps and compared them with reference sites away from the seeps. Under high CO2, the density of shoots and of individuals (apical shoots), and the vertical and horizontal elongation and production rates, were higher than at the reference sites. Nitrogen limitation effects on rhizome elongation and production rates and on biomass were more evident than CO2 as these were highest at the location where the limitation of nitrogen was highest. At the seep where the availability of CO2 was highest and nitrogen lowest, density of shoots and individuals were highest, probably due to CO2 effects on shoot differentiation and induced reproductive output, respectively. At the three seeps, there was higher short- and long-term shoot recruitment than at the reference sites, and growth rates was around zero, indicating that elevated CO2 increases the turnover of C. nodosa shoots.

Chemometric methods in antimalarial drug design from 1,2,4,5-tetraoxanes analogues.

A set of 23 steroidal 1,2,4,5-tetraoxane analogues were studied using quantum-chemical method (B3LYP/6-31 G*) and multivariate analyses (PCA, HCA, KNN and SIMCA) in order to calculate the properties and correlate them with antimalarial activity (log RA) against Plasmodium falciparum clone D-6 from Sierra Leone. PCA results indicated 99.94% of the total variance and it was possible to divide the compounds into two classes: less and more active. Descriptors responsible for separating were: highest occupied molecular orbital energy (HOMO), bond length (O1-O2), Mulliken electronegativity (χ) and Bond information content (BIC0). We use HCA, KNN and SIMCA to explain relationships between molecular properties and biological activity of a training set and to predict antimalarial activity (log RA) of 13 compounds (#24-36) with unknown biological activity. We apply molecular docking simulations to identify intermolecular interactions with a selected biological target. The results obtained in multivariate analysis aided in the understanding of the activity of the new compound's design (#24-36). Thus, through chemometric analyses and docking molecular study, we propose theoretical synthetic routes for the most promising compounds 28, 30, 32 and 36 that can proceed to synthesis steps and in vitro and in vivo assays.

Effectiveness of Reciproc Blue in removing canal filling material and regaining apical patency.

AIM: To compare the efficiency of M-Wire Reciproc and Reciproc Blue instruments in the removal of root filling material and in their ability to regain apical patency through micro-computed tomographic (micro-CT) analysis. METHODOLOGY: Ten anatomically matched pairs of teeth, with a single oval-shaped straight canal, were selected and scanned in a micro-CT device. The root canals were prepared with M-Wire Reciproc R25 instruments and filled with gutta-percha and AH Plus sealer. The root canals were then randomly allocated into two groups (n = 10), according to the instrument used: M-Wire Reciproc and Reciproc Blue. The canals were retreated up to instrument sizes 25 and then 40 in both groups. The surface area and volume of residual filling material were assessed using micro-CT imaging after the retreatment procedures. The time required to remove the root fillings was recorded. Data were analysed statistically using t-test with a significance level of 5%. RESULTS: No significant difference (P > 0.05) was observed between the instruments in terms of the volume of residual root filling material. Apical enlargement from size 25 to 40 significantly improved the removal of filling materials (P < 0.05). It was possible to regain apical patency in all specimens from both M-Wire Reciproc and Reciproc Blue groups. No difference was observed in the time required to perform the retreatment between the instruments. CONCLUSIONS: Both M-Wire Reciproc and Reciproc Blue instruments were effective in removing filling materials from oval-shaped straight canals. Apical patency was reestablished in all specimens.

Torsional fatigue resistance of R-Pilot and WaveOne Gold Glider NiTi glide path reciprocating systems.

AIM: To evaluate the torsional properties of the NiTi glide path reciprocating instruments R-Pilot (VDW, Munich, Germany) and WaveOne Gold Glider (Dentsply Sirona, Ballaigues, Switzerland). METHODOLOGY: Ten R-Pilot (size 12.5, .04 taper) and 10 WaveOne Gold Glider (size 15, .02v taper) instruments were used. The torque and angle of rotation at failure of new instruments (n = 10) were measured according to ISO 3630-1. Three millimetres of each instrument tip was clamped to a small load cell by a lever arm linked to the torsion axis. The fracture surface of all fragments was examined with a scanning electron microscope. Data were analysed using the Student t-test at a significance level of P < 0.05. RESULTS: The maximum torsional strength of the R-Pilot instruments was greater than the WaveOne Gold Glider (P < 0.05). In contrast, WaveOne Gold Glider instruments had significantly higher angular rotation to fracture than R-Pilot instruments (P < 0.05). The scanning electron microscopy of the fracture surfaces revealed similar and typical features of torsional failure (ductile type) for both brands. The torsional failure displayed two zones: the first with microvoids in the centre of the instruments (core) and the second one represented by plastic deformation around the microvoids; this configuration occurred because of the shear stresses originated during the twisting. CONCLUSIONS: R-Pilot instruments had a lower angle of rotation to fracture but a higher torque to failure than WaveOne Gold Glider instruments.

Parâmetros hematológicos e bioquímicos de cabras lactantes alimentadas com dietas contendo glicerina bruta oriunda da produção de biodiesel proveniente de óleo de fritura / Hematological and biochemical parameters of lactating goats fed diets containing crude glycerin from biodiesel production from waste frying oil

Objetivou-se analisar os efeitos da inclusão da glicerina bruta (GB), oriunda da produção de biodiesel proveniente de óleos residuais de fritura, sobre o consumo, e os parâmetros hematológicos e bioquímicos em cabras em lactação. Oito cabras com 42,06±3,5kg de peso corporal foram utilizadas, recebendo dietas contendo 0, 7, 14 e 21% de GB com base na MS total da dieta. O experimento foi conduzido em quadrado latino duplo 4×4, com duração de 80 dias, sendo quatro períodos de 20 dias, dos quais 15 para a adaptação e cinco para a coleta de dados. As amostras de sangue foram coletadas pela punção da veia jugular. Os consumos de MS, proteína bruta e matéria mineral decresceram linearmente, enquanto o de extrato etéreo aumentou quadraticamente. As concentrações do hematócrito, hemoglobina, hemácias e fósforo reduziram linearmente. Tendência quadrática foi observada para as concentrações séricas de cálcio. Não foram verificados efeitos para as concentrações de magnésio, glicose, colesterol total, triglicerídeos, proteínas plasmáticas totais, albumina, globulinas, ureia, creatinina, aspartato aminotransferase e fosfatase alcalina. Conclui-se que a GB pode ser utilizada em até 7% da MS total da dieta, sem promover problemas metabólicos.(AU)

The aim of this study was to analyze the effects of the inclusion of crude glycerin (CG) from biodiesel production from waste frying oils on nutrients intake, hematological and biochemical parameters in lactating goats. Eight goats with 42.06±3.5kg of body weight were used, receiving diets containing 0, 7, 14 and 21% of CG based on total dietary DM. The experiment was carried out in a 4×4 double Latin square, with a duration of 80 days, four periods of 20 days, of which 15 days were for adaptation and five days for data collection. Blood samples were collected by puncture of the jugular vein. Dry matter, crude protein, and mineral matter intakes decreased linearly, while intake of ether extract increased quadratically. The concentrations of hematocrit, hemoglobin, red blood cells, and phosphorus reduced linearly. A quadratic tendency for serum calcium concentrations was observed. No effects were observed for concentrations of magnesium, glucose, total cholesterol, triglycerides, total plasmatic protein, albumin, globulins, urea, creatinine, aspartate aminotransferase, and alkaline phosphatase. It can be concluded that CG can be used in up to 7% of total dietary DM without promoting metabolic problems.(AU)

Hypercholesterolemia and Ecto-enzymes of Purinergic System: Effects of Paullinia cupana.

Hypercholesterolemia is a metabolic disorder characterized by high levels of low-density lipoprotein and blood cholesterol, causing inflammatory lesion. Purinergic signaling modulates the inflammatory and immune responses through adenine nucleotides and nucleoside. Guaraná has hypocholesterolemic and antiinflammatory properties. Considering that there are few studies demonstrating the effects of guaraná powder on the metabolism of adenine nucleotides, we investigated its effects on the activity of ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase) and ecto-adenosine deaminase activity in lymphocytes of rats with diet-induced hypercholesterolemia. The rats were divided into hypercholesterolemic and normal diet groups. Each group was subdivided by treatment: saline, guaraná powder 12.5, 25, or 50 mg/kg/day and caffeine concentration equivalent to highest dose of guaraná, fed orally for 30 days. An increase in adenosine triphosphate hydrolysis was observed in the lymphocytes of rats with hypercholesterolemia and treated with 25 or 50 mg/kg/day when compared with the other groups. The hypercholesterolemic group treated with the highest concentration of guaraná powder showed decreased ecto-adenosine deaminase activity compared with the normal diet groups. Guaraná was able to reduce the total cholesterol and low-density lipoprotein cholesterol to basal levels in hypercholesterolemic rats. High concentrations of guaraná associated with a hypercholesterolemic diet are likely to have contributed to the reduction of the inflammatory process.

Maternal mood and concordant maternal and infant salivary cortisol during heel lance while in kangaroo care.

BACKGROUND: Maternal kangaroo care (MKC) is a naturalistic intervention that alleviates neonatal pain, and mothers are assumed to play a stress regulatory role in MKC. Yet, no MKC infant pain study has examined relationship between maternal and infant stress reactivity concurrently, or whether post-partum depression and/or anxiety (PPDA) alters maternal and neonatal stress response and the regulatory effects of MKC. OBJECTIVES: To examine the concordance of salivary cortisol reactivity between 42 mothers and their stable preterm infants during routine infant heel lance (HL) while in MKC and to compare salivary cortisol between groups of mothers with and without PPDA and their infants. METHODS: Maternal and infant salivary cortisol samples were collected pre-HL and 20 min post-HL with two additional maternal samples at night and in the morning. Mothers and infants were allocated to with PPDA versus without PPDA study groups on the basis of maternal post-natal mental health assessment scores. RESULTS: Higher mothers' cortisol pre-HL was weakly associated with higher infants' salivary cortisol in response to the HL procedure. Maternal depression and/or anxiety were not associated with infants' cortisol. During HL, both groups of mothers and infants showed no change in salivary cortisol. CONCLUSIONS: Concordance between mother and infant salivary cortisol supports the maternal stress regulatory role in MKC. MKC may have stress regulatory benefits for mothers and their preterm infants during HL independent of PPDA. Future MKC studies that target mothers with altered mood will help to build on these findings.

Proposed method for dimensionality reduction based on framework in gene expression domain.

The excessive use of attributes may affect the search for patterns and extraction of useful knowledge, because they harm the learning performance of algorithms in both speed and success rate. The use of dimensionality reduction methods is therefore an important alternative; however, these methods do not deal with the reduction of attributes in a specific area. This article presents a method based on framework concepts of domain for reducing attributes in a domain. The input method is a set of databases related to a domain, and the main process is the identification of common and variable attributes, plus the reduction of attributes in the original database. The proposed method was applied in the gene expression domain, using databases. The method can be used to analyze the most relevant attributes in a specific domain, granting greater confidence for models created for the application of a data mining task, thus, a previously known method in data mining. Attribute selection was also applied in the three databases for the comparison of the results. Analyses of the results using the criterion of cross-validation revealed that the employment of the methods resulted in the improvement of success rates compared to the databases containing the full range of attributes.

Preparo e caracterização de tinturas das folhas de chá verde [Camellia sinensis (L.) O. Kuntze] Theaceae / Preparation and characterization of Theaceae green tea [Camellia sinensis (L.) O. Kuntze] leaf tinctures

Tinturas preparadas com as folhas de Camellia sinensis (chá verde), foram caracterizadas visando a obtenção de extratos enriquecidos em polifenóis. As tinturas foram obtidas por meio de maceração estática da droga pulverizada, com diferentes misturas de 60, 70, 80 e 94,5% de etanol em água. Em seguida foram filtradas e, após 8 dias de extração, realizaram-se as analises: organoléptica, pH, densidade, determinação do resíduo seco, perfil cromatográfico por camada delgada, e teor de polifenóis. Os resultados demonstraram que o solvente (mistura hidroalcoólica a 60 ou 70%) extraiu a maior quantidade de constituintes químicos do chá verde; também foi o mais seletivo ao extrair, especificamente os polifenóis. Estes dados sugerem que a utilização de álcool etílico a 60 ou 70% conduz à obtenção de tinturas mais ricas em polifenóis, a partir das folhas do chá verde (C. sinensis).

In this work, we prepared and characterized Camellia sinensis leaf tinctures aiming to obtain a polyphenol-enriched extract. The tinctures were prepared through the static maceration process of the powdered drug, at different mixtures 60, 70, 80 and 94.5% of ethanol in water. The tinctures were filtered, and after 8 days of extraction, submitted to organoleptic analysis, determination of pH, density analysis, dry residue analysis, thin layer chromatographic profile and polyphenol percentage analysis. The results showed that the 60 or 70% alcohol-water mixture has the best extraction of the constituents of green tea and it was more selective to extract, specifically, the polyphenols of the plant. These data suggest the use of 60 or 70% ethanol to carry out polyphenol-enriched tinctures from green tea leaves (C. sinensis).

Does native bromeliads represent important breeding sites for Aedes aegypti (L.) (Diptera: Culicidae) in urbanized areas?

This study evaluates the importance of native bromeliads growing on rocky outcrops interspersed with urbanized areas as breeding sites for the Aedes aegypti (L.) in Vitória, state of Espírito Santo, Brazil. Oviposition traps were installed in backyards of houses in two separate zones. In the first zone houses were up to 50 m away from the rocky outcrops, while in the second zone they were at least at 200 m from the rocky outcrops. Aedes aegypti was significantly more abundant in the latter zone. The finding was that rocky outcrops with native bromeliads, even with the greater availability of potential breeding sites, do not play an important role as breeding sites for A. aegypti. This conclusion supports the hypothesis that the macrobiota of native bromeliads plays an important role in the natural control of A. aegypti. Besides, the interspecific competition between species of mosquitoes and the attractiveness of bromeliads could also be important factors.

A influência postural do salto alto em mulheres adultas: análise por biofotogrametria computadorizada / Postural influence of high heels among adult women: analysis by computerized photogrammetry

INTRODUÇÃO: Em nossa sociedade, temos observado uma oferta cada vez maior de modelos, cores, estilos, altura e diversos tipos de salto. OBJETIVO: Avaliar se o uso de calçados de salto alto influencia nas alterações posturais com base em um conjunto de variáveis mensuradas por meio da fotogrametria computadorizada. MÉTODOS: Vinte indivíduos que utilizam salto alto com freqüência (grupo 1) e 20 indivíduos que utilizam salto alto esporadicamente (grupo 2) foram fotografados no plano frontal anterior e sagital em três momentos: a) sem utilização de calçado, b) utilizando salto agulha e c) utilizando salto plataforma, sendo estas fotografias aleatorizadas e analisadas por um experimentador cego por meio da fotogrametria. A análise estatística foi realizada a partir da análise de variância em esquema fatorial 2x3, ou seja, comparando-se a freqüência do uso de salto com o tipo de calçado, com 5 por cento de significância. RESULTADOS: Apenas o ângulo protrusão da cabeça apresentou diferença quando comparados grupo 1 e 2 (p<0,01). O efeito do tipo de calçado ocorreu na variável alinhamento do joelho direito, sendo que houve diferença apenas entre o sapato agulha e os pés descalços (p=0,03); também para a variável ângulo tibiotársico, o efeito esteve presente em todos os tipos de calçado. Os demais ângulos avaliados não apresentaram diferenças entre a freqüência no uso de salto e os outros tipos de sapato. CONCLUSÕES: A freqüência do uso de salto e o tipo de salto praticamente não modificam a postura estática avaliada pela fotogrametria.

INTRODUCTION: In our society, it is observed an increasing number of models, colors, styles, heights and types of high heels. OBJECTIVE: To evaluate whether the use of high heel shoes results in postural changes, based on a set of variables measured through computerized photogrammetry. METHODS: Twenty individuals who often used high heels (group 1) and 20 individuals who only used high heels sporadically (group 2) were photographed in the frontal and sagittal planes at three conditions: a) without using footwear; b) using stiletto heels; and c) using high platform heels. These photographs were randomized and analyzed by a blinded examiner, by means of photogrammetry. Statistical analysis was performed, using a 2x3 factorial analysis of variance to compare the frequency of high heel use with the type of shoe, at the 5 percent significance level. RESULTS: Only the head protrusion angle showed a difference between groups 1 and 2 (p<0.01). The effect of the type of shoe observed in the alignment of the right knee, which only showed a difference between stiletto heels and barefoot (p=0.03). For the tibiotarsal angle variable, the effect was also observed for all types of footwear. The other angles evaluated did not present any differences regarding the frequency of high heel use and the types of shoe. CONCLUSIONS: The frequency and type of high heel practically did not change the static posture evaluated by photogrammetry.

Neuropsychological rehabilitation of memory deficits and activities of daily living in patients with Alzheimer's disease: a pilot study.

Patients with Alzheimer's disease (AD) gradually lose their cognitive competence, particularly memory, and the ability to perform daily life tasks. Neuropsychological rehabilitation is used to improve cognitive functions by facilitating memory performance through the use of external aids and internal strategies. The effect of neuropsychological rehabilitation through memory training - motor movements, verbal association, and categorization - and activities of daily living (ADL) training was tested in a sample of 5 elderly out-patients (mean age: 77.4 +/- 2.88 years), with mild AD (Mini-Mental State Examination score: 22.20 +/- 2.17) and their caregivers. All patients had been taking rivastigmine (6-12 mg/day) for at least 3 months before being assigned to the rehabilitation sessions, and they continued to take the medication during the whole program. Just before and after the 14-week neuropsychological rehabilitation program all patients were assessed by interviewers that did not participate in the cognitive training, using the Mini-Mental State Examination, Montgomery-Alsberg Depression Rating Scale, Hamilton Anxiety Scale, Interview to Determine Deterioration in Functioning in Dementia, Functional Test, Memory Questionnaire of Daily Living for patient and caregiver, Quality of Life Questionnaire for patient and caregiver, and a neuropsychological battery. The results showed a statistically significant improvement in ADL measured by Functional Test (P = 0.04), and only a small improvement in memory and psychiatric symptoms. Our results support the view that weekly stimulation of memory and training of ADL is believed to be of great value in AD treatment, not only delaying the progress of the disease, but also improving some cognitive functions and ADL, even though AD is a progressively degenerative disease.

Neuropsychological rehabilitation of memory deficits and activities of daily living in patients with Alzheimer's disease: a pilot study

Patients with Alzheimer's disease (AD) gradually lose their cognitive competence, particularly memory, and the ability to perform daily life tasks. Neuropsychological rehabilitation is used to improve cognitive functions by facilitating memory performance through the use of external aids and internal strategies. The effect of neuropsychological rehabilitation through memory training - motor movements, verbal association, and categorization - and activities of daily living (ADL) training was tested in a sample of 5 elderly out-patients (mean age: 77.4 ± 2.88 years), with mild AD (Mini-Mental State Examination score: 22.20 ± 2.17) and their caregivers. All patients had been taking rivastigmine (6-12 mg/day) for at least 3 months before being assigned to the rehabilitation sessions, and they continued to take the medication during the whole program. Just before and after the 14-week neuropsychological rehabilitation program all patients were assessed by interviewers that did not participate in the cognitive training, using the Mini-Mental State Examination, Montgomery-Alsberg Depression Rating Scale, Hamilton Anxiety Scale, Interview to Determine Deterioration in Functioning in Dementia, Functional Test, Memory Questionnaire of Daily Living for patient and caregiver, Quality of Life Questionnaire for patient and caregiver, and a neuropsychological battery. The results showed a statistically significant improvement in ADL measured by Functional Test (P = 0.04), and only a small improvement in memory and psychiatric symptoms. Our results support the view that weekly stimulation of memory and training of ADL is believed to be of great value in AD treatment, not only delaying the progress of the disease, but also improving some cognitive functions and ADL, even though AD is a progressively degenerative disease.

Epidemiological and clinical features of Leishmania (Viannia) braziliensis American cutaneous and mucocutaneous leishmaniasis in the State of Espírito Santo, Brazil

Between 1985 and 2000, epidemiological surveys of the American tegumentary leishmaniasis (ATL) were carried out in several rural and urban communities in Espírito Santo, Brazil. A total of 100 stocks of Leishmania (comprising isolates from both human and canine hosts with ATL) were identified by two methods of molecular characterization, using specific monoclonal antibodies and multilocus enzyme electrophoresis. Parasite isolates from 19 municipalities were found to belong to the same zymodeme and serodeme type as of the Leishmania (Viannia) braziliensis reference strain. In contrast, our genotyping studies have shown intra-specific variation among these parasites (comparisons of the variability of the internal transcribed spacers between the small and large subunits of the rRNA genes of the 22 stocks studiedrevealed at least 11 genotypes). Two main clusters of L. (V.) braziliensis genotypes were observed, representing parasites collected from different endemic regions in the state, where transmission reflects distinct eco-epidemiological features. Infection with this pathogen was associated with the characteristic disease forms, but neither the clinical outcome nor the response to treatment could be related to the genetic polymorphism of the isolates, as defined by using the proposed methodology.

Epidemiological and clinical features of Leishmania (Viannia) braziliensis American cutaneous and mucocutaneous leishmaniasis in the State of Espírito Santo, Brazil.

Between 1985 and 2000, epidemiological surveys of the American tegumentary leishmaniasis (ATL) were carried out in several rural and urban communities in Espírito Santo, Brazil. A total of 100 stocks of Leishmania (comprising isolates from both human and canine hosts with ATL) were identified by two methods of molecular characterization, using specific monoclonal antibodies and multilocus enzyme electrophoresis. Parasite isolates from 19 municipalities were found to belong to the same zymodeme and serodeme type as of the Leishmania (Viannia) braziliensis reference strain. In contrast, our genotyping studies have shown intra-specific variation among these parasites (comparisons of the variability of the internal transcribed spacers between the small and large subunits of the rRNA genes of the 22 stocks studied revealed at least 11 genotypes). Two main clusters of L. (V.) braziliensis genotypes were observed, representing parasites collected from different endemic regions in the state, where transmission reflects distinct eco-epidemiological features. Infection with this pathogen was associated with the characteristic disease forms, but neither the clinical outcome nor the response to treatment could be related to the genetic polymorphism of the isolates, as defined by using the proposed methodology.

A new PCR assay useful for screening of FRAXE/FMR2 mental impairment among males.

FRAXE (FMR2) is a fragile site associated with mental impairment located in Xq28, 600 kb distal to FRAXA (FMR1), the fragile X syndrome fragile site. The FRAXE mutation is an expansion of a CCG repeat that results in methylation of a nearby CpG island. FRAXE alleles could be divided into four categories: normal (6-30 CCG repeats), intermediate (31-60 CCG repeats), premutation (61-200 CCG repeats), and full mutation (over 200 repeats). We have developed a non-isotopic polymerase chain reaction (PCR)-based assay for the identification of FRAXE full mutation alleles among mentally impaired men. In this novel PCR test for the FRAXE locus, we used three primers to permit an amplification of a 223 bp monomorphic internal control fragment in addition to the amplification of a 419 bp (CCG)(16) FRAXE locus band. A linear series of 93 male patients referred for FRAXE testing but found to be negative for the (CCG)(n) expansion in the FMR2 gene by Southern blotting analysis were retested by our PCR technique. In addition, we analyzed two positive controls consisting of a FRAXE fully mutated male and one male with a Xq terminal deletion. The developed PCR test showed accuracy of 100% in the normal individuals retested by PCR analysis, as well as in the two positive control samples utilized, in which the strategy of multiplex amplification worked as expected. Although not suitable for medical diagnosis of females and mosaics, it constitutes an important strategy for PCR typing and for FRAXE population screening.

Chromosomal investigations in patients with mental retardation and/or congenital malformations

Neste estudo investigamos a organizaçäo cromossômica de pacientes com retardo mental e/ou malformaçöes congênitas, visando a avaliaçäo de causas genéticas associadas a estes distúrbios. Os padröes de bandas GTG e CBG foram estudados a partir da cultura de linfócitos de sangue periférico, estimulados por fitohemaglutinina M. Dentre os 98 indivíduos portadores de retardo mental e/ou malformaçöes congênitas analisados, diagnosticamos as seguintes síndromes: 12 casos de Down, dois de Edwards, um de Patau, cinco de Turner, dois de Klinefelter, um de "cri-du-chat", e um caso de translocaçäo balanceada entre os cromossomos 13 e 14, um caso de cromossomo derivado e um outro de cromossomo marcador. Encontramos anomalias cromossômicas em 26 por cento dos pacientes, das quais 82 por cento eram alteraçöes numéricas e o restante (18 por cento) representou rearranjos estruturais. Este percentual significativo enfatiza o uso da cariotipagem de rotina em pacientes com retardo mental e/ou malformaçöes congênitas.

Differential expression of the plasma membrane Mg2+ ATPase and Ca2+ ATPase activity during adhesion and interiorization of Leishmania amazonensis in fibroblasts in vitro.

With ultrastructural cytochemistry we localized the activity of the plasma membrane enzyme markers Mg2+ ATPase and Ca2+ ATPase during the interaction between Leishmania amazonensis and in vitro primary culture fibroblasts. The expression of the enzymes was followed during the parasite adhesion and its interiorization. After the interiorization step, a striking difference was seen between the two enzymes studied when the parasite was found within the parasitophorous vacuole in the fibroblast cytoplasm. The activity of the Ca2+ ATPase found at the Leishmania amazonensis plasma membrane during the attachment step of the infection remained also present inside the phagosome, whereas the Mg2+ ATPase activity disappeared. So far, all the reports in the literature referred the presence of Ca2+ ATPase in Leishmania parasite only in the crude ghost plasma membrane. The Ca2+ ATPase present at the parasite plasma membrane may be involved in the regulation of calcium levels inside the phagosome. Further characterization of this Ca2+ ATPase at the plasma membrane of the parasite, when still inside the phagosome, should permit a better understanding of its functional role in maintaining the parasite surface membrane structure necessary for its existence.