Low sensitivity of nested PCR using Plasmodium DNA extracted from stained thick blood smears: an epidemiological retrospective study among subjects with low parasitaemia in an endemic area of the Brazilian Amazon region

BACKGROUND: The success of PCR technique depends on many factors, such as high quality DNA pellets obtained from blood samples, good reagents and adequate conditions of amplification. Taking these limitations into account, a retrospective epidemiological study for malaria diagnosis was conducted in a mesoendemic area in the Brazilian Amazon. METHODS: A nested PCR protocol with DNA extracted from two blood storage devices obtained from Giemsa-stained thick blood smears and filter-papers was used for malaria diagnosis. The extracted DNA was used as a template to amplify approximately 100 bp species-specific sequences of the small subunit of the ribosomal RNA (18S SSU rRNA) of Plasmodium sp. The prevalence of single and mixed infections was examined in a cross-sectional survey carried out among 369 miners living in the district of Apiacás, Mato Grosso State. The parasitemia levels detected by microscopic examination were compared to the PCR results. RESULTS: DNA samples isolated from blood on filter-paper allowed the detection and identification of Plasmodium in 165 (44.7%) of the 369 individuals evaluated, while only 62 (16.8%) had positive results using DNA obtained from thick smears, a similar rate observed by microscopic examination. The sensitivities of PCR using DNA from blood smears and filter-papers were 65% and 73.0%, respectively. Low parasite infections (below 20 parasites/µL blood) were not detected when thick blood smears were used as a DNA source. CONCLUSIONS: Although the blood preserved as thick blood smears provides an alternative and useful tool for malaria molecular diagnosis, its relatively poor performance at low level parasitemias impairs the correct determination of malaria prevalence in epidemiological studies. However, the results obtained in the present study confirm that the use of filter-paper to collect blood is useful for field studies

The binding protein BiP attenuates stress-induced cell death in soybean via modulation of the N-Rich protein-mediated signaling pathway

The molecular chaperone binding protein (BiP) participates in the constitutive function of the endoplasmic reticulum (ER) and protects the cell against stresses. In this study, we investigated the underlying mechanism by which BiP protects plant cells from stress-induced cell death. We found that enhanced expression of BiP in soybean (Glycine max) attenuated ER stress- and osmotic stress-mediated cell death. Ectopic expression of BiP in transgenic lines attenuated the leaf necrotic lesions that are caused by the ER stress inducer tunicamycin and also maintained shoot turgidity upon polyethylene glycol-induced dehydration. BiP-mediated attenuation of stress-induced cell death was confirmed by the decreased percentage of dead cell, the reduced induction of the senescence-associated marker gene GmCystP, and reduced DNA fragmentation in BiP-overexpressing lines. These phenotypes were accompanied by a delay in the induction of the cell death marker genes N-RICH PROTEIN-A (NRP-A), NRP-B, and GmNAC6, which are involved in transducing a cell death signal generated by ER stress and osmotic stress through the NRP-mediated signaling pathway. The prosurvival effect of BiP was associated with modulation of the ER stress- and osmotic stress-induced NRP-mediated cell death signaling, as determined in transgenic tobacco (Nicotiana tabacum) lines with enhanced (sense) and suppressed (antisense) BiP levels. Enhanced expression of BiP prevented NRP- and NAC6-mediated chlorosis and the appearance of senescence-associated markers, whereas silencing of endogenous BiP accelerated the onset of leaf senescence mediated by NRPs and GmNAC6. Collectively, these results implicate BiP as a negative regulator of the stress-induced NRP-mediated cell death response

Use of active films in the minimally processed potato conservation

Batatas da variedade “Monalisa”, foram fatiadas e minimamente processadas e, após, cobertas por filmes ativos, empregados para minimizar o escurecimento enzimático e o crescimento de microrganismos. Utilizaram-se filmes celulósicos incorporados com aditivos (2% de ácido cítrico (AC), 0,5% de monocloridrato de L-cisteína (CIS), 7% de ácido sórbico (AS) e mistura dos compostos mencionados (MIS)) ou sem a incorporação destes (Filme FP), que foram intercalados às batatas fatiadas. Como controle, foram utilizadas batatas mantidas sem filme intercalado (SF). Todas as amostras foram acondicionadas em bandejas de poliestireno expandido, envoltas em filme de PVC e armazenadas por 9 dias a 8ºC ± 2ºC. A cada três dias, as amostras de batatas foram avaliadas quanto à cor, pH, atividade de água (Aa) e qualidade microbiológica (contagem de Coliformes totais e a 45ºC, Fungos filamentosos e leveduras, Psicrotróficos, Bacillus cereus e Staphylococcus coagulase positiva). Observou-se que para os mesmos tempos de estocagem, os tratamentos não diferiram significativamente (p>0,05) para a maioria dos parâmetros avaliados, com exceção da diferença de cor, onde diferiram significativamente aos 10% de probabilidade. Conclui-se que os filmes ativos anti escurecimento têm potencial de serem usados em vegetais que apresentam deterioração por escurecimento enzimático.    Potatoes of the variety “Monalisa” were minimally processed like slices and submitted to different treatments, employed to decrease the enzymatic browning and growth of microorganisms. These treatments consisted on pure cellulose films (FP) or incorporated with active compounds (2% of citric acid (AC), 0,5% of L-cysteine Hydrochloride Monohydrate (CIS), 7% of sorbic acid (AS) and film incorporated with all of them (MIS)) which were intercalated to sliced potatoes. For control, potatoes were conditioned without film. All of the samples were conditioned in expanded polystyrene trays, wrapped up in PVC film and stored 8ºC ± 2ºC by nine days. Every three days, they were evaluated for color, pH, water activity (Aw) and microbiological quality (growth of total Coliformes and to 45ºC, Molds and yeasts, Psychrotrophics, Bacillus cereus and Staphylococcus positive coagulase). The treatments didn’t differ significantly (p>0,05) for most of the evaluated parameters, except for the color difference, that presented significant difference to the 10% of probability. In conclusion, the active films can minimize the enzymatic browning of vegetables

Quality of life in children and teenagers with atopic dermatitis Qualidade de vida das crianças e adolescentes com dermatite atópica

BACKGROUND: Atopic Dermatitis is a disease which has increased during the past years despite our improved understanding of it. OBJECTIVE: To assess the impact of Atopic Dermatitis in the quality of life of children and teenagers and their family. METHOD: A descriptive cross-sectional method with prospective data collection of 50 children and teenagers diagnosed with Atopic Dermatitis ranging in age from 5-16 years. Fifty parents and/or guardians answered the quality of life questionnaires The Children's Dermatology Life Quality Index and Family Dermatitis Impact Questionnaire. The socio-demographic and clinical variables were evaluated by a clinical record chart designed specifically for the research and socioeconomic standardized questionnaire by the Brazilian Association of Research Enterprises, which evaluates assets acquired and the educational level of the head of the household. RESULTS: Thirty-five out of the 50 patients were female (70%), and 28 (56%) of them were from social class C. The Questionnaire Children's Dermatology Life Quality Index showed that 19 (38%) patients ranged from 7 to 12 points (moderate impact of atopic dermatitis) and 17 patients (34%) ranged from 13 to 30 points (high impact of atopic dermatitis). The Family Dermatitis Impact Questionnaire revealed that 15 (30%) families had scores between 7 and 12 points and 22 families (44%) scored between 13 and 30 points. CONCLUSION: The results show that there is a very high impact on the QoL for atopic dermatitis patients and their families. This makes us suggest the importance of including the quality of life study in clinical evaluations.<br>FUNDAMENTOS: A dermatite atópica é uma doença cuja prevalência vem aumentando nos últimos anos apesar do conhecimento crescente sobre a mesma. OBJETIVO: Avaliar a qualidade de vida das crianças e adolescentes com dermatite atópica e de suas famílias. MÉTODO: Estudo transversal descritivo com coleta prospectiva de dados de 50 crianças e adolescentes de 5 a 16 anos, com diagnóstico de DA e 50 pais ou responsáveis dos mesmos através da utilização de dois questionários de qualidade de vida, o Qualidade de Vida na Dermatologia Infantil e o Impacto da Dermatite Atópica na Família. As variáveis sócio-demográficas e clínicas foram avaliadas por uma ficha clínica elaborada especificamente para a pesquisa, os aspectos socioeconômicos, pelo questionário padronizado da Associação Brasileira de Empresas de Pesquisa, que avalia bens adquiridos e o grau de instrução do chefe da família. RESULTADOS: Havia 35/50 (70%) pacientes do sexo feminino; 28(56%) da classe social C. Através do questionário Qualidade de Vida na Dermatologia Infantil observou-se que: 19 (38%) pacientes ficaram na faixa de 7 a 12 pontos (impacto moderado da dermatite atópica) e 17 (34%), na faixa de 13 a 30 pontos (impacto elevado da dermatite atópica). Pelo questionário Impacto da Dermatite Atópica na Família observou-se: 15 (30%) famílias apresentaram escores entre 7 e 12 pontos e 22 (44%) entre 13 e 30 pontos. CONCLUSÃO: Os resultados mostram que a repercussão da doença na vida dos pacientes com dermatite atópica e de suas famílias é alta, o que nos faz sugerir que seria importante inserir o estudo da qualidade de vida na avaliação clínica dos mesmos

ALKALOIDS FROM LEAVES OF GUATTERIA POGONOPUS (ANNONACEAE) AND THEIR CYTOTOXICITIES

The phytochemical investigation of the alkaloid-rich fraction obtained from the leaves of Guatteria pogonopus Mart. (Annonaceae) allowed the isolation and identification for the first time in this species of: (+)-nornuciferine (1), a mixture of 1 and (+)-anonaine (2), (+)-isocorydine (3), (+)-nuciferine (4), (+)-roemerine (5), (-)-tetrahydropseudocolumbamine (6), a mixture of 6, liriodenine (9) and lysicamine (10), a mixture of 1,2,9-trimethoxy-10-hydroxyaporphine (7) and bulbocapnine (8), 9, 10, and (+)-N-methyllindicarpine (11). Compounds 6, 7, 8, and 11 have not been previously reported in the family Annonaceae. Furthermore, the formerly synthetic 1,2,9-trimethoxyaporfin-10-ol (7) is described for the first time as a natural aporphine alkaloid herein. The chemical structures were established by 1D and 2D NMR as well as in comparison with data previously reported in the literature. The cytotoxic activity of the alkaloids was evaluated against tumor (B16-F10, HepG2, HL-60, and K562) and non-tumor (PBMC) cell lines. Alkaloid 1 presented significant activity against HepG2 cell lines with IC50 of 9.60 µmol L-1 while the mixture of 6, 9 and 10 displayed strong cytotoxic activity against HL-60 and K562 cell lines with IC50 values of 3.41 an 8.50 µmol L-1, respectively