Biodiversity or ultra-processed food: an analysis of school meals offered in a riverside school in the Brazilian Amazon.

BACKGROUND: Rural schools in Amazonas, Brazil, often offer ultra-processed foods in school meals for students, which can lead to health problems and loss of regional food culture. We show an analysis of the menu offered in a riverside school in the Brazilian Amazon and the acceptability of students regarding the consumption of the food they are served with. METHODS: Data were collected in situ, in a riverside school in southern Amazonas, through the analysis of the school menu and the application of an investigative questionnaire to 37 students in the 9th grade of Junior High School. FINDINGS: The research revealed that the foods most consumed by students in school meals are canned beef, canned meatballs, canned sardines, sausage, biscuits, juice, rice porridge, corn porridge, pasta, meat soup, and rice with beans. In the questionnaire that was applied to students, there is a wide variation in the acceptability of the foods offered. However, 57% of students reported not liking the lunch offered at the educational institution. INTERPRETATION: To tackle this problem, it is essential that, local food culture and biodiversity food can be more valued, elements that are often excluded from school menus. This work showed that is also essential to fully adhere to the National School Meal Program (PNAE) in Brazil, which recommends that at least 30% of food intended for school meals must come from family farming, highlighting that quality food is crucial for cognitive development of students. Therefore, the meals offered in the chosen riverside school not only do not meet the PNAE guidelines but are also not well accepted by students. This study shows a significant need to consider the direct relationship between planetary health, school meals food security, and food sovereignty, given the various negative effects of foods that are rich in fat, sodium, preservatives, and other substances. Furthermore, it is imperative to integrate food into the students' context, valuing regional products from the Amazon region. FUNDING: FAPEAM (Amazonas State Research Foundation).

The significance of the two cytosolic glutamine synthetase enzymes, GLN1;3 and GLN1;5, in the context of seed development and germination in Arabidopsis thaliana.

Glutamine synthetase (GS), an initial enzyme in nitrogen (N) plant metabolism, exists as a group of isoenzymes found in both cytosolic (GS1) and plastids (GS2) and has gathered significant attention for enhancing N use efficiency and crop yield. This work focuses on the A. thaliana GLN1;3 and GLN1;5 genes, the two predicted most expressed genes in seeds, among the five isogenes encoding GS1 in this species. The expression patterns were studied using transgenic marker line plants and qPCR during seed development and germination. The observed patterns highlight distinct functions for the two genes and confirm GLN1;5 as the most highly expressed GS1 gene in seeds. The GLN1;5, expression, oriented towards hypocotyl and cotyledons, suggests a role in protein turnover during germination, while the radicle-oriented expression of GLN1;3 supports a function in early external N uptake. While the single mutants exhibited a normal phenotype, except for a decrease in seed parameters, the double gln1;3/gln1;5 mutant displayed a germination delay, substantial impairment in growth, nitrogen metabolism, and number and quality of the seeds, as well as a diminishing in flowering. Although seed and pollen-specific, GLN1;5 expression is upregulated in the meristems of the gln1;3 mutants, filling the lack of GLN1;3 and ensuring the normal functioning of the gln1;3 mutants. These findings validate earlier in silico data on the expression patterns of GLN1;3 and GL1;5 genes in seeds, explore their different functions, and underscore their essential role in plant growth, seed production, germination, and early stages of plant development.

JAGGER localization and function are dependent on GPI anchor addition.

KEY MESSAGE: GPI anchor addition is important for JAGGER localization and in vivo function. Loss of correct GPI anchor addition in JAGGER, negatively affects its localization and function. In flowering plants, successful double fertilization requires the correct delivery of two sperm cells to the female gametophyte inside the ovule. The delivery of a single pair of sperm cells is achieved by the entrance of a single pollen tube into one female gametophyte. To prevent polyspermy, Arabidopsis ovules avoid the attraction of multiple pollen tubes to one ovule-polytubey block. In Arabidopsis jagger mutants, a significant number of ovules attract more than one pollen tube to an ovule due to an impairment in synergid degeneration. JAGGER encodes a putative arabinogalactan protein which is predicted to be anchored to the plasma membrane by a glycosylphosphatidylinositol (GPI) anchor. Here, we show that JAGGER fused to citrine yellow fluorescent protein (JAGGER-cYFP) is functional and localizes mostly to the periphery of ovule integuments and transmitting tract cells. We further investigated the importance of GPI-anchor addition domains for JAGGER localization and function. Different JAGGER proteins with deletions in predicted ω-site regions and GPI attachment signal domain, expected to compromise the addition of the GPI anchor, led to disruption of JAGGER localization in the cell periphery. All JAGGER proteins with disrupted localization were also not able to rescue the polytubey phenotype, pointing to the importance of GPI-anchor addition to in vivo function of the JAGGER protein.

[Evaluation of the Filaria Detect™ IgG4 kit employing recombinant Wb123 antigen for diagnosis of lymphatic filariasis in BrazilEvaluación del estuche Detect™ para pruebas de anticuerpos antifiláricos IgG4, fabricado con el antígeno recombinante Wb123, para el diagnóstico de la filariasis linfática en Brasil]. / Avaliação do kit Filaria Detect™ IgG4 produzido com o antígeno recombinante Wb123 para diagnóstico da filariose linfática no Brasil.

The Global Programme to Eliminate Lymphatic Filariasis, launched by the World Health Organization in the year 2000, proposes the use of circulating filarial antigen tests as a diagnostic tool to assess and monitor initiatives to control filarial infection. However, despite a high sensitivity, these tests are not efficient to detect infection at early stages, before worms have reached the adult stage. Considering this limitation, anti-filarial antibody testing has been suggested as an alternative, given that the antibodies produced against the larvae are detectable before the presence of circulating filarial antigen. The objective of the present study was to determine the diagnostic cut-off and the accuracy of the Filaria Detect™ IgG4 kit employing recombinant Wb123 antigen for diagnosis of lymphatic filariasis in Brazil. For that, we performed a diagnostic evaluation study in which 256 serum samples were analyzed: 79 (30.9%) obtained from microfilaremic individuals and 177 (60.1%) from amicrofilaremic individuals who tested negative with the Bm14 CELISA and Og4C3 ELISA immunologic tests. The ideal cutoff as well as the Filaria Detect™ IgG4 kit accuracy were determined based on ROC curve analyses, with an optical density of 0.239 identified as the cutoff with the best performance, with 81.0% sensitivity and 96.6% specificity. The results show that the Filaria Detect™ IgG4 kit is a promising tool for investigation and monitoring of areas undergoing mass drug administration for lymphatic filariasis.

En el programa mundial de eliminación de la filariasis linfática, puesto en marcha por la Organización Mundial de la Salud en el año 2000, se propone el uso de pruebas de detección del antígeno filárico circulante como instrumento de diagnóstico para la evaluación y el seguimiento de las medidas de control de la parasitosis. Sin embargo, esas pruebas, a pesar de tener un alto grado de sensibilidad, no permiten detectar con eficiencia la infección en su fase inicial, cuando todavía no existen helmintos adultos. En vista de esa limitación, se ha señalado como una opción el estudio de anticuerpos antifiláricos, puesto que los anticuerpos producidos contra las larvas infectantes del parásito se detectan antes de la existencia de antígeno filárico circulante. El objetivo de este estudio fue definir el punto de corte y evaluar la exactitud del estuche Detect™ para pruebas de anticuerpos antifiláricos IgG4, fabricado con el antígeno recombinante Wb123, para el diagnóstico de la filariasis linfática en Brasil. Para ello, se realizó un estudio de evaluación de la prueba diagnóstica, en el cual se utilizaron 256 muestras de suero, a saber, 79 (30,9%) obtenidas de personas microfilarémicas y 177 (60,1%) de personas amicrofilarémicas, que arrojaron resultados seronegativos en las pruebas inmunológicas CELISA Bm14 y ELISA Og4C3. La definición del punto de corte ideal y de la exactitud del estuche Detect™ se obtuvo con la construcción de curvas de la característica operativa del receptor (ROC); una densidad óptica de 0,239 marcó el mejor nivel de desempeño de la prueba, con una sensibilidad de 81,0% y una especificidad de 96,6%. Los resultados obtenidos demostraron que el estuche Detect™ es un instrumento prometedor para la investigación y el seguimiento de las regiones donde se realiza un tratamiento masivo de la filariasis linfática.

Physiological, Biochemical and Molecular Assessment of UV-A and UV-B Supplementation in Solanum lycopersicum.

Daily UV-supplementation during the plant fruiting stage of tomato (Solanum lycopersicum L.) growing indoors may produce fruits with higher nutraceutical value and better acceptance by consumers. However, it is important to ensure that the plant's performance during this stage is not compromised by the UV supplement. We studied the impact of UV-A (1 and 4 h) and UV-B (2 and 5 min) on the photosynthesis of greenhouse-grown tomato plants during the fruiting/ripening stage. After 30 d of daily irradiation, UV-B and UV-A differently interfered with the photosynthesis. UV-B induced few leaf-necrotic spots, and effects are more evidenced in the stimulation of photosynthetic/protective pigments, meaning a structural effect at the Light-Harvesting Complex. UV-A stimulated flowering/fruiting, paralleled with no visible leaf damages, and the impact on photosynthesis was mostly related to functional changes, in a dose-dependent manner. Both UV-A doses decreased the maximum quantum efficiency of photosystem II (Fv/Fm), the effective efficiency of photosystem II (ΦPSII), and gas exchange processes, including net carbon assimilation (PN). Transcripts related to Photosystem II (PSII) and RuBisCO were highly stimulated by UV supplementation (mostly UV-A), but the maintenance of the RuBisCO protein levels indicates that some protein is also degraded. Our data suggest that plants supplemented with UV-A activate adaptative mechanisms (including increased transcription of PSII peptides and RuBisCO), and any negative impacts on photosynthesis do not compromise the final carbohydrate balances and plant yield, thus becoming a profitable tool to improve precision agriculture.

Development of an ecotoxicological test procedure for soil microalgae.

Since the 80s, ISO and OECD organizations have been developing guidelines for assessing the toxicity of new and existing chemical substances to soil biota. Up to now, any of these guidelines had soil algae as test organisms. Nevertheless, microalgae are relevant components of soil microbial communities and soil biological crusts (BSC) with a great contribution to different soil functions and ecosystem services. In an attempt to bridge the gap, the present work aimed to develop, describe and validate a standard operating procedure for an ecotoxicological test with soil microalgae. Three phases were performed, each one with specific objectives. First, soil microalgae and cyanobacteria were isolated from BSC and then genetically and morphologically characterized. The green microalga Micractinium inermum was selected because it is a species with a wide geographic distribution. Secondly, M. inermum growth curves were obtained in liquid (BG11 and Woods-Hole MBL) and solid media (OECD artificial soil) to determine test duration. The growth curves were also used to analyze the reproducibility of the test's endpoint and to propose a validation criterion. Ultimately, a range of concentrations of two reference substances (glyphosate and copper) were tested, both in soil and liquid media, to assess procedure's reproducibility. The tests made in liquid medium followed the standard guideline for ecotoxicological tests with freshwater microalgae and cyanobacteria (OECD 201:2011). The results obtained prove that when the artificial soil is used, as a test substrate, the sensitivity of M. inermum increases. The tests performed with both reference substances demonstrate that the procedure described for testing in soil was reproducible. Additionally, it will be relevant to test with other reference substances and adjust the procedure for natural soils. It will be also interesting to validate the test procedure with soil cyanobacteria.

Avaliação do kit Filaria Deteção; IgG4 produzido com o antígeno recombinante Wb123 para diagnóstico da filariose linfática no Brasil / Evaluation of the Filaria Detection; IgG4 kit employing recombinant Wb123 antigen for diagnosis of lymphatic filariasis in Brazil / Evaluación de pruebas de anticuerpos antifiláricos IgG4, fabricado con el antígeno recombinante Wb123, para el diagnóstico de la filariasis linfática en Brasil

RESUMO O Plano Global de Eliminação da Filariose Linfática, lançado pela Organização Mundial da Saúde em 2000, propõe o uso de testes de detecção de antígeno circulante filarial como ferramenta diagnóstica para avaliação e monitoramento das ações de controle da parasitose. Entretanto, esses testes, apesar de apresentarem alta sensibilidade, não conseguem detectar com eficiência a infecção em seu estágio inicial, quando ainda não existe a presença de helmintos adultos. Considerando essa limitação, a pesquisa de anticorpos antifilariais tem sido apontada como uma alternativa, uma vez que os anticorpos produzidos contra as larvas infectantes do parasito são detectados antes da presença de antígeno circulante filarial. O objetivo deste estudo foi definir o ponto de corte e avaliar a acurácia do kit Filaria Detect™ IgG4 produzido com o antígeno recombinante Wb123 para diagnóstico da filariose linfática no Brasil. Para isso, foi realizado um estudo de avaliação de teste diagnóstico, no qual foram utilizadas 256 amostras de soro: 79 (30,9%) obtidas de indivíduos microfilarêmicos e 177 (60,1%), de indivíduos amicrofilarêmicos e que testaram negativo para os testes imunológicos Bm14 CELISA e Og4C3 ELISA. A definição do ponto de corte ideal, bem como da acurácia do kit Filaria Detect™ IgG4, foi obtida através da construção de curvas ROC, sendo a densidade óptica de 0,239 aquela na qual o teste obteve melhor desempenho, com sensibilidade de 81,0% e especificidade de 96,6%. Os resultados obtidos demonstraram que o kit Filaria Detect™ IgG4 é uma ferramenta promissora para investigação e monitoramento de áreas submetidas ao tratamento em massa para filariose linfática.

ABSTRACT The Global Programme to Eliminate Lymphatic Filariasis, launched by the World Health Organization in the year 2000, proposes the use of circulating filarial antigen tests as a diagnostic tool to assess and monitor initiatives to control filarial infection. However, despite a high sensitivity, these tests are not efficient to detect infection at early stages, before worms have reached the adult stage. Considering this limitation, anti-filarial antibody testing has been suggested as an alternative, given that the antibodies produced against the larvae are detectable before the presence of circulating filarial antigen. The objective of the present study was to determine the diagnostic cut-off and the accuracy of the Filaria Detect™ IgG4 kit employing recombinant Wb123 antigen for diagnosis of lymphatic filariasis in Brazil. For that, we performed a diagnostic evaluation study in which 256 serum samples were analyzed: 79 (30.9%) obtained from microfilaremic individuals and 177 (60.1%) from amicrofilaremic individuals who tested negative with the Bm14 CELISA and Og4C3 ELISA immunologic tests. The ideal cutoff as well as the Filaria Detect™ IgG4 kit accuracy were determined based on ROC curve analyses, with an optical density of 0.239 identified as the cutoff with the best performance, with 81.0% sensitivity and 96.6% specificity. The results show that the Filaria Detect™ IgG4 kit is a promising tool for investigation and monitoring of areas undergoing mass drug administration for lymphatic filariasis.

RESUMEN En el programa mundial de eliminación de la filariasis linfática, puesto en marcha por la Organización Mundial de la Salud en el año 2000, se propone el uso de pruebas de detección del antígeno filárico circulante como instrumento de diagnóstico para la evaluación y el seguimiento de las medidas de control de la parasitosis. Sin embargo, esas pruebas, a pesar de tener un alto grado de sensibilidad, no permiten detectar con eficiencia la infección en su fase inicial, cuando todavía no existen helmintos adultos. En vista de esa limitación, se ha señalado como una opción el estudio de anticuerpos antifiláricos, puesto que los anticuerpos producidos contra las larvas infectantes del parásito se detectan antes de la existencia de antígeno filárico circulante. El objetivo de este estudio fue definir el punto de corte y evaluar la exactitud del estuche Detect™ para pruebas de anticuerpos antifiláricos IgG4, fabricado con el antígeno recombinante Wb123, para el diagnóstico de la filariasis linfática en Brasil. Para ello, se realizó un estudio de evaluación de la prueba diagnóstica, en el cual se utilizaron 256 muestras de suero, a saber, 79 (30,9%) obtenidas de personas microfilarémicas y 177 (60,1%) de personas amicrofilarémicas, que arrojaron resultados seronegativos en las pruebas inmunológicas CELISA Bm14 y ELISA Og4C3. La definición del punto de corte ideal y de la exactitud del estuche Detect™ se obtuvo con la construcción de curvas de la característica operativa del receptor (ROC); una densidad óptica de 0,239 marcó el mejor nivel de desempeño de la prueba, con una sensibilidad de 81,0% y una especificidad de 96,6%. Los resultados obtenidos demostraron que el estuche Detect™ es un instrumento prometedor para la investigación y el seguimiento de las regiones donde se realiza un tratamiento masivo de la filariasis linfática.

Arabidopsis thaliana mutants devoid of chloroplast glutamine synthetase (GS2) have non-lethal phenotype under photorespiratory conditions.

Chloroplast located Glutamine Synthetase (GS2) is believed to play a major role in the reassimilation of ammonium generated by photorespiration, being GS2 knockout mutants unable to grow under photorespiratory conditions (low-CO2 atmosphere) in the species characterized so far (Barley, Lotus). To investigate the importance of GS2 in A. thaliana nitrogen metabolism mutant plants devoid of this GS isoenzyme were characterized. It was shown that GS2 mutants although smaller, slightly chlorotic and with the nitrogen metabolism impaired, were able to grow and complete their life cycle under ordinary air conditions. Surprisingly, GS2 mutants were more tolerant to salt stress than wild-type plants. The lack of GS2 seems to be compensated by higher expression of some GS cytosolic isogenes, namely GLN1;2 and GLN1;3 and by glutamate dehydrogenase, whose activity and expression is enhanced in the GS2 mutant plants and might account for the increased tolerance to salt stress. Under conditions that minimize photorespiration (CO2-enriched atmosphere) plant growth and ammonium assimilation impairment is less evident in the GS2 mutant plants and is accompanied by an adjustment of levels of expression of the cytosolic isogenes, with an increase in the expression of GLN1;3 and a decrease in the expression of the GLN1;1 and GLN1;2. Altogether the results confirm a major role of GS2 in the assimilation of ammonium released during photorespiration, but suggest a redundancy of activity with cytosolic GSs and GDH and further support the involvement of the chloroplastic isoenzyme in primary nitrogen assimilation and plant growth and development in A. thaliana.

Quantification of GHB and GHB-GLUC in an 1,4-butanediol intoxication: A case report.

Gamma-hydroxybutyric acid (GHB) is an endogenous compound with known action at the neural level. Its psychoactive effects led to an illicit use context including recreational purposes, muscle building effects in bodybuilders and drug-facilitated crimes, specifically in sexual assaults. Besides the misuse of the main compound, there are precursors like Gammabutyrolactone (GBL) and 1,4-butanediol (1,4-BD), usually non controlled substances, becoming a much easier way to obtain the target-compound. The authors present the first reported intoxication case in Portugal with 1,4-Butanediol, including the quantification of GHB and GHB-GLUC in serum, by GC-MS/MS TQD. A suspicious liquid and a serum sample were sent by an hospital ER and analysed by GC-MS-single quadrupole and GC-MS/MS TQD, respectively. A methodology including protein precipitation and GC-MS/MS TQD analysis was used to detect and quantify GHB and GHB-GLUC in serum. Toxicological analysis revealed the presence of 1,4-Butanediol in the liquid and GHB [171 mg/L] and GHB-GLUC [13,7 mg/L] in serum. The victim reverted the coma with no neurological sequelae. This was the first detected case, in Portugal, with 1,4-Butanediol, suggesting that it is important to be aware that consumers have different options to obtain illicit compounds, such as GHB. On the other hand, GHB-GLUC was identified and quantified for the first time in a real case, due to intoxication. This case highlights the importance of analysing all samples for active compounds, precursors and metabolites that can lead to the main intoxication origin.

Effectiveness of annual single doses of diethylcarbamazine citrate among bancroftian filariasis infected individuals in an endemic area under mass drug administration in Brazil.

The Global Program to Eliminate Lymphatic Filariasis has achieved extraordinary success in reducing transmission and preventing morbidity through mass drug administration (MDA) to the population at-risk. Brazil is the only currently using diethylcarbamazine citrate (DEC) alone for MDA, so an assessment of its effectiveness is needed. We report the trends of filarial markers in a cohort of 175 individuals infected with Wuchereria bancrofti in areas that underwent MDA in the city of Olinda, Northeastern Brazil. The prospective study was conducted between 2007 and 2012 (corresponding to five annual MDA rounds). The quantification of microfilaraemia (QMFF) was assessed by filtration. Circulating filarial antigen (CFA) was detected through immunochromatographic point-of-care test (POCT-ICT) and Og4C3-ELISA whereas antifilarial antibody titres (IgG4) were assessed through Bm14 assay. The CFA and IgG4 titres were measured by Optical Density (OD). The main characteristics at baseline, MDA coverage and the trend of filarial infection markers during follow up were described. The trend of filarial markers in relation to time (years of MDA), sex and age were analysed through Generalized Estimating Equations (GEE) models. The models demonstrated a significant decrease in all markers during MDA. The probability of remaining positive by QMFF and POCT-ICT diminished 70% and 46%, respectively, after each MDA round. There was a significant annual drop in CFA (-0.290 OD) and IgG4 antibodies titres (-0.303 OD). This study provides evidence that MDA with DEC alone can be effective in the elimination of LF in Brazil.

A fast and reliable method for quantitation of THC and its 2 main metabolites in whole blood by GC-MS/MS (TQD).

An analytical method was developed for the simultaneous determination of Δ9- tetrahydrocannabinol (THC), 11-hydroxy-Δ9-tetrahydrocannabinol (11-OH-THC) and l1-nor-9-carboxy-Δ9-tetrahydrocannabinol (THC-COOH) in whole blood samples, for identification and quantitation purposes. Samples were prepared using solid-phase extraction, followed by gas chromatography/tandem mass spectrometry (GC-MS/MS) analysis in multiple reaction monitoring mode, with a total run time of 7.6min. MS/MS detection was achieved with two ion transitions per substance. The method was fully validated, including selectivity and capacity of identification, limit of detection and limit of quantitation, recovery, carryover, linearity (1-100ng/mL), intra-assay precision, inter-assay accuracy. The obtained results allowed its use in routine forensic analysis, with the application to real samples, both clinical and post-mortem.

Evaluation of the recombinant antigens Wb14 and WbT for the capture antibody diagnosis of lymphatic filariasis.

BACKGROUND: Lymphatic filariasis (LF) is a parasitic disease caused mainly by the Wuchereria bancrofti worm and that affects up to 120 million people worldwide. LF is the second cause of chronic global deformity, responsible for 15 million people with lymphedema (elephantiasis) and 25 million men with scrotal hydrocele. Its diagnosis is still associated with numerous difficulties, such as the sample collection periods (microfilaria nocturnal periodicity) and limited diagnostic kits. OBJECTIVES: The aim of this work was to evaluate two recombinant antigens (Wb14 and WbT) as part of an enzyme-linked immunosorbent assay (ELISA) based antibody capture tests for LF. METHODS: The recombinant antigens rWb14 and rWbT were expressed in Escherichia coli BL21 and an antibody capture ELISA was performed. For this, sera were used from microfilaremic individuals with W. bancrofti (MF), chronic pathology (CP), individuals infected with Strongyloides (SP) and healthy controls from endemic (EN) and non-endemic (NE) areas. FINDINGS: Both tests showed similar results, with 90% sensitivity and 96.6% specificity. In comparison with the BM14 ELISA commercial test, the Wb14 and WbT antigens performed with identical sensitivity but greater specificity. Reduced positivity with the CP suggested a potential to monitor cure. This was not confirmed, however, when sera from individuals up to seven years after treatment were assayed. MAIN CONCLUSIONS: The Wb14 and WbT ELISAs were considered efficient and promising diagnostic tests. Due to the importance of antibody capture analysis to evaluate the Global Program to Eliminate Lymphatic Filariasis (GPELF), the tests proposed here appear as great alternatives to the available commercial system.

Mechanisms of social synchrony between circadian activity rhythms in cohabiting marmosets.

In marmosets, social synchrony between circadian profiles of activity is stronger in animals that cohabit in a family. The activity of three breeding pairs was recorded by actiwatches to investigate the mechanisms involved in the synchrony between the circadian activity profiles during cohabitation in marmoset reproductive pairs. The dyads were submitted to LD 12:12 (21 days) and LL: 1) cohabitation (24 days), 2) removal of the cage mate (20 days), 3) reintroduction of the mate into the cage of the 1st situation (30 days) and 4) removal of the cage mate (7 days). Next, they were rejoined and maintained in LD 12:12 (11 days). In conditions involving cohabitation of pair, the general and maximum correlation indexes between circadian profiles were higher in cage mates compared to animals of the same or different sex with which they maintain only acoustic and olfactive contact. This strong synchrony between rhythms was accompanied by a stable phase relationship at the activity onset and offset, with identical circadian periods between mates. When the pairs were separated, there was a break in stability in the phase relationships between activity profiles with different circadian periods and a greater phase angle difference between rhythms of cage mates. During separation, two females and one male progressively anticipated the activity onset and offset in a phase similar to that in previous conditions, expressing entrainment to the mate. During the first reintroduction, two pairs exhibited signs of masking in rhythm. Although modulation in the rhythm of some animals has been observed through acoustic cues from animals outside the colony, we suggest that cohabitation favors strong synchrony between the circadian activity profiles of marmoset reproductive pairs involving synchronization by entrainment and masking. Further studies in the absence of external social cues are necessary to clarify the role of these mechanisms on social synchronization in marmosets.

A fast method for GHB-GLUC quantitation in whole blood by GC-MS/MS (TQD) for forensic purposes.

γ-Hydroxybutyric acid (GHB) is an endogenous compound with a historical use, both in licit and illicit terms. Importantly, the post-mortem behavior of GHB has been studied due to the possibility of using this compound as a biomarker for estimating the post-mortem interval (PMI). However, the post-mortem behavior of the recently discovered glucuronated GHB metabolite (GHB-GLUC) has not been studied. Nevertheless, GHB-GLUC may also have potential both to assist in PMI determination and also to increase the window of detection of GHB consumption. In this work, for the first time, a reliable method using GC-MS/MS for the quantification of GHB-GLUC in whole blood samples was developed and validated, with a simple, fast and cheap sample pretreatment. The method proved to be specific, precise, linear in a work range between 200 and 5000ng/mL, with LOD and LOQ of 52.65ng/mL and 200ng/mL, respectively, and an extraction recovery of 51%. Furthermore, the method was applied to a set of real post-mortem blood samples non-related with GHB intoxication and the obtained results were also discussed.

Evaluation of the recombinant antigens Wb14 and WbT for the capture antibody diagnosis of lymphatic filariasis

BACKGROUND Lymphatic filariasis (LF) is a parasitic disease caused mainly by the Wuchereria bancrofti worm and that affects up to 120 million people worldwide. LF is the second cause of chronic global deformity, responsible for 15 million people with lymphedema (elephantiasis) and 25 million men with scrotal hydrocele. Its diagnosis is still associated with numerous difficulties, such as the sample collection periods (microfilaria nocturnal periodicity) and limited diagnostic kits. OBJECTIVES The aim of this work was to evaluate two recombinant antigens (Wb14 and WbT) as part of an enzyme-linked immunosorbent assay (ELISA) based antibody capture tests for LF. METHODS The recombinant antigens rWb14 and rWbT were expressed in Escherichia coli BL21 and an antibody capture ELISA was performed. For this, sera were used from microfilaremic individuals with W. bancrofti (MF), chronic pathology (CP), individuals infected with Strongyloides (SP) and healthy controls from endemic (EN) and non-endemic (NE) areas. FINDINGS Both tests showed similar results, with 90% sensitivity and 96.6% specificity. In comparison with the BM14 ELISA commercial test, the Wb14 and WbT antigens performed with identical sensitivity but greater specificity. Reduced positivity with the CP suggested a potential to monitor cure. This was not confirmed, however, when sera from individuals up to seven years after treatment were assayed. MAIN CONCLUSIONS The Wb14 and WbT ELISAs were considered efficient and promising diagnostic tests. Due to the importance of antibody capture analysis to evaluate the Global Program to Eliminate Lymphatic Filariasis (GPELF), the tests proposed here appear as great alternatives to the available commercial system.

[Clinical and epidemiological characteristics of strongyloidiasis in patients with comorbidities]. / Características clínico-epidemiológicas de la estrongiloidiasis en pacientes portadores de co-morbilidades.

The strongyloidiasis is a parasitic disease that poses as a serious public health problem, mainly in tropical and subtropical countries. Over the years, some conditions, such as advances in corticosteroid treatment and immunosuppressive diseases, have improved not only the increase in cases of strongyloidiasis, but also the emergence of severe forms of the disease and / or deaths. For these reasons, the objective of this study is to make a critical analysis of the occurrence of strongyloidiasis in patients with comorbidities, describing clinical and epidemiological characteristics associated with these diseases that can highlight the importance of monitoring this parasitosis in most susceptible groups.

Características clínico-epidemiológicas de la estrongiloidiasis en pacientes portadores de co-morbilidades / Clinical and epidemiological characteristics of strongyloidiasis in patients with comorbidities

The strongyloidiasis is a parasitic disease that poses as a serious public health problem, mainly in tropical and subtropical countries. Over the years, some conditions, such as advances in corticosteroid treatment and immunosuppressive diseases, have improved not only the increase in cases of strongyloidiasis, but also the emergence of severe forms of the disease and / or deaths. For these reasons, the objective of this study is to make a critical analysis of the occurrence of strongyloidiasis in patients with comorbidities, describing clinical and epidemiological characteristics associated with these diseases that can highlight the importance of monitoring this parasitosis in most susceptible groups.

La estrongiloidiasis es una parasitosis que representa un grave problema de salud pública, principalmente en países ubicados en regiones tropicales y subtropicales. A lo largo de los años, algunas condiciones, como por ejemplo, avances en el tratamiento con corticosteroides y enfermedades que evolucionan con inmunosupresión, han favorecido no solamente al aumento de casos de estrongiloidiasis, sino también al surgimiento de formas graves de la enfermedad y/u decesos. Por lo expuesto, el objetivo del presente estudio fue realizar un análisis crítico de la ocurrencia de la estrongiloidiasis en portadores de co-morbilidades, describiendo las características clínico-epidemiológicas de esa asociación que puedan resaltar la importancia de vigilar esta parasitosis en grupos considerados más susceptibles.

Postmortem in vitro ethanol production-It could be more common than we think!

The blood alcohol concentration (BAC) is the most frequent determination in a Forensic Toxicology Laboratory. Despite its apparent simplicity, the results interpretation can be complex and always have relevant social and legal implications, particularly in postmortem analysis. In the present report we describe the case of a 55-year-old male with an apparent natural death by myocardial infarction, whose initial BAC was 0.18g/L but, in repeated determinations prompted by discrepancies observed in the first two, it rapidly increased to 0.85g/L three days later, leading to the suspicion of in vitro ethanol production. A microbiological examination of the sample revealed the presence of the bacteria Escherichia coli and Enterococcus faecalis, and yeast Candida parapsilosis, known for their involvement in ethanol production. Although this is a case report and it is not meant to be generalizable, we discuss an existing large body of scientific literature showing the difficulties, limitations and some relevant medico-legal questions regarding BAC determinations in postmortem samples and their interpretation, particularly in the context of plausible in vitro ethanol production. The key conclusion is that evaluating a postmortem BAC is a complex and multifactorial process that always deserves a thorough analysis and a careful interpretation.

Pentobarbital in the context of possible suicides: Analysis of a Case.

Pentobarbital is a barbiturate, acting as a central nervous system depressant (CNS), being used for its anticonvulsant, sedative, hypnotic and anaesthetic properties. Barbiturates were replaced by benzodiazepines, leading to a decrease in poisoning cases with these compounds. However, pentobarbital is still used in many countries as an anaesthetic in veterinary medicine. Due to its properties, this compound is sought after by people who wish to commit suicide, acquiring it on the black market. The authors present an unusual fatal pentobarbital intoxication case, in a 37 years-old male salesperson, with no known connection with the veterinary field, being more difficult to obtain this compound. Toxicological results in cardiac blood revealed the presence of pentobarbital (111mg/L), ethanol (0.94g/L), diazepam (33ng/mL), nordiazepam (50ng/mL), oxazepam (3.3ng/mL), temazepam (5.3ng/mL), and metoclopramide. No illicit drugs were detected. Pentobarbital analysis in urine and gastric content was also positive, as well as its presence in the glass powder and in the bottle residue sent to the laboratory. In the present case, it was possible to conclude that the death was a suicide due to pentobarbital intoxication in association with other depressants of the CNS (benzodiazepines and ethanol). It is important to search pentobarbital in routine toxicological analyses, since it is one of the drugs most frequently mentioned by entities defending "painless death", advising the simultaneous use of metoclopramide for emesis avoidance.