Abstract Twenty six sweet cassava clones were evaluated in two agricultural years, in randomized block design, with 3 replications and plots with 25 plants. The harvests were at 12 months after planting, in both years. The traits evaluated were root yield (RY), dry matter content (DMC), total carotenoids content (TC), β-carotene content (BC), trans-β-carotene content (TrBC) and total cyanide content (TCy). Genotype was significant (P<0.05) in all traits. Broad-sense heritability estimates at the plot level ( h m g 2 ) ranged from 77% (RY) to 93% (TC and TrBC), while the accuracy ranged from 0.88 (RY) to 0.96 (BC and TrBC). The genetic correlations among TC, BC and TrBC were high (0.96 to 0.99) and significant, and the genetic correlations involving these traits and TCy were negative. These results demonstrate the existence of great genetic variability in characteristics important to cassava biofortification and, consequently, great perspectives in the breeding for this purpose.

ABSTRACT Objective: Low levels of neutrophils can be an intrinsic condition, with no clinical consequences or immunity impairment. This condition is the benign constitutional neutropenia (BCN), defined as an absolute neutrophils count (ANC) ≤2000 cells/mm. Diagnosis of BCN is of exclusion where patients are submitted to blood tests and possibly to invasive diagnostic search until secondary causes of neutropenia are ruled out. The natural history of the disease suggests benign evolution and Brazilian study showed an overall frequency of 2.59%. The main mechanisms include reduced neutrophil production, increased marginalization, extravasation to the tissues and immune destruction. Genetic studies showed strong association between the single nucleotide variant rs2814778 located on chromosome 1q23.2 in the promoter region of the atypical chemokine receptor 1 (Duffy blood group system) gene (ACKR1, also termed DARC) and BCN. The aim of this study is to evaluate FY phenotypes and genotypes including the analysis of the rs2814778 SNP in Brazilian patients with BCN in order to determine an effective diagnostic tool, allowing reassurance of the patient and cost reduction in their care. Methods: Case control study, with 94 individuals (18 patients and 76 controls). Phenotyping was performed by gel test and genotyping was performed by PCR-RFLP. Results: White blood cell (WBC) and absolute neutrophils (AN) counts showed lower levels in patients compared to controls. In the patient group 83.3% were genotyped as FY*B/FY*B. The SNP rs2814778 (-67T > C) was identified in 77.8% of the patients genotyped as FY*B-67C/FY*B-67C. In the control group, 72.7% were homozygous for the wild type and 23.3% were heterozygous. Conclusion: This study reinforces that FY phenotyping and genotyping can be used to detect most people with BCN, avoiding excessive diagnostic investigation. Besides, this procedure may reduce health costs and be reproductible in clinical practice.

Abstract We evaluated the efficacy of xylitol varnishes on the remineralization of newly erupted permanent and deciduous teeth in vitro and in situ. Human enamel specimens were randomly allocated to 8 groups (n = 15/group). Artificial caries lesions were produced and enamel alterations were quantified by surface/subsurface hardness and transverse microradiography. The blocks were then treated with the following varnishes: DuraphatTM; 20 wt% xylitol (146 μm) varnish; 20 wt% milled xylitol (80 μm) varnish, and placebo varnish, and removed after 6 h of immersion in artificial saliva. In vitro the blocks were subjected to pH-cycles for 8 days. In situ fifteen subjects wore palatal appliances containing four pre-demineralized and treated enamel specimens, for 5 days. Data were statistically analyzed by ANOVA/Tukey and Kruskal-Wallis/Tukey’s test (p < 0.05). The %SHR in both studies was significantly increased by xylitol and Duraphat™ varnishes when compared to placebo. Considering subsurface remineralization, only the xylitol varnishes were able to significantly reduce the enamel lesion. Xylitol varnishes can be promising alternatives to promote enamel remineralization of newly erupted permanent and deciduous teeth.

RESUMO: A perfusão regional intravenosa (PRI) em membros de equinos é um método comprovadamente eficiente para o tratamento de infecções sinoviais, porém ainda existem lacunas quanto à dose, volume e intervalos de administração a serem utilizados. Neste estudo, objetivou-se avaliar a concentração intra-articular da gentamicina administrada por PRI em equinos, ao longo de 24 horas pós-administração, bem como avaliar a influência do volume total perfundido. Vinte equinos foram distribuídos em dois grupos experimentais, Grupo Gentamicina 60 (G60) e Grupo Gentamicina 250 (G250), representados por 10 equinos em cada grupo que receberam, por PRI, 6,6 mg/kg de gentamicina diluída em Ringer lactato para um total de 60 ml (G60) ou 250 ml (G250). As colheitas de líquido sinovial foram realizadas, na articulação metacarpo-falangeana, antes do início do experimento e após 4, 8, 12, 16, 20, e 24 horas. Difusão em ágar foi o método para doseamento das concentrações de gentamicina. Quatro horas após a PRI, a concentração intra-articular da gentamicina era de 47,58±49,21 µg/ml no G60 e 10,92±6,11 µg/ml no G250. Durante as 24 horas de colheita, a concentração intra-articular de gentamicina permaneceu acima da concentração inibitória mínima (CIM) (2 μg/ml) em ambos os grupos experimentais. No G60, as concentrações de gentamicina no líquido sinovial atingiram a concentração máxima e mantiveram-se 8 a 10 vezes mais elevadas que a CIM (Cmáx: 16 a 20 μg/ml) durante 12 horas após a PRI. Conclui-se que a PRI em membros, com 6,6 mg/kg de gentamicina diluída para o volume de 60 ml, promove concentrações intra-articulares mais elevadas que o volume de 250 ml, permanecendo acima da CIM durante 24 horas e em concentração máxima durante 12 horas.

ABSTRACT: Intravenous regional limb perfusion (IRLP) is an efficient method to treat horses with synovial infections. However, information on the dose, volume, and intervals of administration is lacking. The aim of this study was to evaluate the intra-articular concentration of gentamicin administered by IRLP in horses over 24 h post-administration and evaluate the influence of the total perfused volume. Twenty horses were assigned to two treatment groups. Gentamicin 60 group (G60, n=10) and Gentamicin 250 group (G250, n=10) received IRLP of 6.6 mg/kg of gentamicin diluted in Ringer’s lactate for a total volume of 60 ml (G60) and 250 ml (G250), respectively. Synovial fluid harvests were performed in the metacarpophalangeal joint before (0) and 4, 8, 12, 16, 20, and 24 hours after IRLP. Agar diffusion was the assay for measuring gentamicin concentrations. Four hours after IRLP, the intra-articular concentration of gentamicin was 47.58±49.21 μg/ml in G60 and 10.92±6.11 μg/ml in G250. During the 24 h of harvest, the intra-articular concentration of gentamicin remained above the minimum inhibitory concentration for horses (MIC; 2 μg/ml) in both experimental groups. In G60, the concentration of gentamicin in synovial fluid reached the maximal concentration and remained 8 to 10-fold higher than the MIC (Cmax: 16 to 20 μg/ml) during 12 h after IRLP. These findings support that IRLP with 6.6 mg/kg of gentamicin diluted to a volume of 60 ml promotes intra-articular concentrations higher than 250 ml, remaining above the MIC for 24 h and at the maximum concentration for 12 h.

In this study, a rapid and simple analytical method was proposed, based on protein precipitation as sample preparation for simultaneous determination of tetracycline, oxytetracycline, penicillin G and ceftiofur in raw milk by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The method was applied to raw milk samples from dairy cows medicated with tetracycline for subclinical mastitis. Samples were collected from the adjacent teat to the teat treated with tetracycline of eight different cows at 24, 48, 72 and 96 h after treatment. The limits of quantification of the proposed method ranged between 1 and 5 ng g-1 and limits of detection ranged between 0.1 and 0.5 ng g-1. The recoveries ranged from 61 to 111% and the linear range was 1 to 2064 ng g-1 for tetracycline and oxytetracycline, and 5 to 2064 ng g-1 for penicillin G and ceftiofur. Approximately 75 and 63% of the treated animals revealed more tetracycline than legally recommended at 72 and 96 h since last treatment, respectively.

RESUMO Objetivo: Determinar o perfil de liberação, no humor aquoso, de moxifloxacino encapsulado em lipossomas como um sistema de liberação controlada para aplicação intracameral. Métodos: Lipossomas contendo moxifloxacino foram obtidos através do método de hidratação do filme lipídico e caracterizados por tamanho da partícula e eficiência de encapsulação. Utilizaram-se coelhos fêmeas foram para o estudo do perfil de liberação in vivo. Lipossomas contendo moxifloxacino foram injetados na câmara anterior do olho direito de cada animal. Os coelhos foram divididos em cinco grupos, e uma amostra de humor aquoso foi coletada 2, 4, 8, 24 e 48 h após a administração de lipossomas contendo moxifloxacino. As concentrações de moxifloxacino no humor aquoso foram analisadas usando cromatografia líquida de alta eficiência. Resultados: O tamanho médio dos lipossomas contendo moxifloxacino foi de 60,5 ± 0,72 nm com uma distribuição de tamanho de partícula de 0,307. A eficiência de encapsulação de moxifloxacino nos lipossomas foi de 92,24 ± 0,24. Os resultados de um estudo de liberação in vivo de lipossomas contendo moxifloxacino, mostraram que a concentração máxima de moxifloxacino foi atingida dentro das primeiras 2 h após sua administração (5,27 ± 1,09 mg/mL) e foi seguida de um decréscimo na concentração intracameral (0,35 ± 0,05 mg/mL) até a marca de 24 h. Conclusão: Os experimentos in vivo resultaram em lipossomas contendo moxifloxacino que eram homogêneos em tamanho e exibiam alta eficiência de encapsulação do fármaco. Os resultados indicam que lipossomas contendo moxifloxacino oferecem um perfil de liberação de humor aquoso satisfatório após a aplicação intracameral.

ABSTRACT Purpose: To determine the release profile of moxifloxacin encapsulated in liposomes in the aqueous humor as a controlled release system for intracameral application. Methods: Liposomes containing moxifloxacin were obtained using the lipid film hydration method and were characterized by particle size and encapsulation efficiency. Female rabbits were used for the in vivo profile release study. Liposomes containing moxifloxacin was injected into the anterior chamber of the right eye of each animal. The rabbits were divided into five groups, and a sample of aqueous humor was collected 2, 4, 8, 24, and 48 h after administration of liposomes containing moxifloxacin administration. Moxifloxacin concentrations in the aqueous humor were analyzed using high-performance liquid chromatography. Results: The average size of the liposomes containing moxifloxacin was 60.5 ± 0.72 nm with a particle size distribution of 0.307. The encapsulation efficiency of moxifloxacin in liposomes was 92.24 ± 0.24%. The results of an in vivo release study of liposomes containing moxifloxacin, showed that the maximum moxifloxacin concentration was achieved within the first 2 h after administration (5.27 ± 1.09 mg/mL) and was followed by a decrease in intracameral concentration (0.35 ± 0.05 mg/mL) until the 24 h mark. Conclusions: The in vivo experiments resulted in liposomes containing moxifloxacin that were homogenous in size and exhibited high drug encapsulation efficiency. The results indicate that liposomes containing moxifloxacin offers a satisfactory aqueous humor release profile after intracameral application.

Resumo Objetivo: Verificar a associação entre a depressão pós-parto e a ocorrência do aleitamento materno exclusivo. Método: Estudo de corte transversal feito nos estados da Região Nordeste, durante a campanha de vacinação de 2010. A amostra consistiu de 2.583 binômios mães-crianças entre 15 dias e três meses. Usou-se a Escala de Depressão Pós-Parto de Edimburgo para rastrear a depressão pós-parto. O desfecho consistiu da ausência de aleitamento materno exclusivo nas 24 horas que antecederam a entrevista. A depressão pós-parto foi variável explanatória de interesse e as covariáveis foram: condições socioeconômicas e demográficas, assistência pré-natal, ao parto e pós-natal e fatores da criança. Fez-se análise de regressão logística multivariada com o objetivo de controlar possíveis fatores de confusão. Resultados: A amamentação exclusiva foi observada em 50,8% das crianças e 11,8% das mulheres apresentaram sintomatologia indicativa de depressão pós-parto. Na análise de regressão logística multivariada foi verificada uma maior chance de ausência do aleitamento materno exclusivo entre as mães com sintomas de depressão pós-parto (OR = 1,67; p < 0,001). Conclusões: A depressão pós-parto contribuiu para redução da prática do aleitamento materno exclusivo. Assim, esse transtorno deveria ser incluído nas orientações de apoio desde o pré-natal e nos primeiros meses pós-parto, especialmente em mulheres de baixo nível socioeconômico.

Abstract Objective: To investigate the association between postpartum depression and the occurrence of exclusive breastfeeding. Method: This is a cross-sectional study conducted in the states of the Northeast region, during the vaccination campaign in 2010. The sample consisted of 2583 mother-child pairs, with children aged from 15 days to 3 months. The Edinburgh Postnatal Depression Scale was used to screen for postpartum depression. The outcome was lack of exclusive breastfeeding, defined as the occurrence of this practice in the 24 h preceding the interview. Postpartum depression was the explanatory variable of interest and the covariates were: socioeconomic and demographic conditions; maternal health care; prenatal, delivery, and postnatal care; and the child's biological factors. Multivariate logistic regression analysis was conducted to control for possible confounding factors. Results: Exclusive breastfeeding was observed in 50.8% of the infants and 11.8% of women had symptoms of postpartum depression. In the multivariate logistic regression analysis, a higher chance of exclusive breastfeeding absence was found among mothers with symptoms of postpartum depression (OR = 1.67; p < 0.001), among younger subjects (OR = 1.89; p < 0.001), those who reported receiving benefits from the Bolsa Família Program (OR = 1.25; p = 0.016), and those started antenatal care later during pregnancy (OR = 2.14; p = 0.032). Conclusions: Postpartum depression contributed to reducing the practice of exclusive breastfeeding. Therefore, this disorder should be included in the prenatal and early postpartum support guidelines for breastfeeding, especially in low socioeconomic status women.

ABSTRACT INTRODUCTION: Schistosomiasis, a parasitic disease caused by trematode flatworms of the genus Schistosoma, affects more than 200 million people worldwide, and its control is dependent on a single drug, praziquantel. Here, we report the in vitro effect of rotundifolone, a monoterpene isolated from Mentha x villosa (Lamiaceae), on Schistosoma mansoni adult worms. METHODS: The in vitro effect of rotundifolone on adult Schistosoma mansoni was evaluated by analysis of behavior and mortality and through a scanning electron microscopic analysis of ultrastructural changes in the tegument of the worms. RESULTS: At concentrations of 3.54 and 7.09μg/mL-1 rotundifolone, no worm mortality was observed at any of the sampling intervals. A minor reduction in movement of the tail, suckers, and gynecophoral canal membrane was observed after 96 h of exposure to 7.09μg/mL-1 rotundifolone. At 70.96μg/mL-1, a lack of movement was observed from 72h onwards and all worms were deemed dead; similar effects were observed at 48h with 177.4μg/mL-1, and at 24h with 354.8μg/mL-1 and 700.96μg/mL-1. Rotundifolone also caused death of all parasites and separation of coupled pairs into individual males and females after 24h at 354.8μg/mL-1. CONCLUSIONS: The main changes in the tegument induced by the different ROT treatments were: after 24h incubation, bubble lesions spread over the entire body and loss of tubercles occurred in some regions of the ventral region.

Introduction: Schistosomiasis is a chronic disease caused by trematode flatworms of the genus Schistosoma and its control is dependent on a single drug, praziquantel (PZQ), but concerns over PZQ resistance have renewed interest in evaluating the in vitro susceptibility of recent isolates of Schistosoma mansoni to PZQ in comparison with well-established strains in the laboratory. Material and methods: The in vitro activity of PZQ (6.5-0.003 µg/mL) was evaluated in terms of mortality, reduced motor activity and ultrastructural alterations against S. mansoni. Results: After 3 h of incubation, PZQ, at 6.5 µg/mL, caused 100% mortality of all adult worms in the three types of recent isolates, while PZQ was inactive at concentrations of 0.08-0.003 µg/mL after 3 h of incubation. The results show that the SLM and Sotave isolates basically presented the same pattern of susceptibility, differing only in the concentration of 6.5 µg/mL, where deaths occurred from the range of 1.5 h in Sotave and just in the 3 h range of SLM. Additionally, this article presents ultrastructural evidence of rapid severe PZQ-induced surface membrane damage in S. mansoni after treatment with the drug, such as disintegration, sloughing, and erosion of the surface. Conclusion: According to these results, PZQ is very effective to induce tegument destruction of recent isolates of S. mansoni.

Os objetivos deste trabalho foram: avaliar a qualidade fisiológica de sementes de cedro (Cedrela fissilis), procedentes de localidades do sul do Brasil, através de testes de vigor e avaliar diferentes tratamentos nas sementes para controle de patógenos e para promoção da germinação da espécie. Para tanto, foram utilizadas seis amostras com procedências dos Estados do Rio Grande do Sul, Santa Catarina e Paraná, as quais foram submetidas à determinação de teor de água, teste de germinação e primeira contagem, envelhecimento acelerado (testando períodos de exposição às condições de envelhecimento), emergência em viveiro e tratamento de sementes. Os tratamentos utilizados para controle de patógenos foram: testemunha (T0); físico com calor seco a 70°C±3°C por 48 horas (T1); extrato aquoso de alho (Allium sativum) (T2); biológico à base de Trichoderma spp. - Agrotich Plus® (T3); e químico com fungicida protetor Captan (T4). A germinação variou de 56 a 87%; o período de 48 h sob temperatura de 41 °C foi o mais eficiente para estratificar as amostras em níveis de vigor; a emergência variou de 51 a 88% e as variáveis de desempenho de plântulas analisadas conseguiram estratificar as amostras em níveis de vigor. Quanto ao tratamento de sementes, o calor seco e o tratamento à base de extrato de alho se mostram eficientes no controle de microrganismos em semente de cedro, sem prejuízos ao vigor destas.

The objectives of this study were to evaluate the physiological quality of Cedrela fissilis seeds, from different origins, using different vigor tests and evaluating different seeds treatments for control of pathogens and to promote the germination of the species. Therefore, six samples from different places in states of Rio Grande do Sul, Santa Catarina and Paraná were used, which were subjected to determination of water content, germination and first count of germination, accelerated aging (testing different periods), emergence, with assessments seedlings and seed treatment. The treatments were the following: control (T0); physical treatment with dry heat at 70°C±3°C for 48 hours (T1); aqueous extract of Allium sativum (T2); biological with product based on Trichoderma spp. - Agrotich Plus® (T3); and chemic with fungicide Captan (T4). Germination ranged 56-87%, the period of 48 hours at a temperature of 41 ° C has been more efficient to stratify the samples at levels of vigor; emergence ranged 51-88% and the performance of seedlings variables were able to stratify the samples analyzed at levels of vigor. Regarding seed treatments, dry heat and treatment based on extracts of A. sativum are effective in controlling micro-organisms in C. fissilis seeds, without damage to the vigor of them.

PROPOSTA: Placas vulneráveis são caracterizadas por matriz mixomatosa, centro lipídico necrótico, espécies reativas de oxigênio e alto níveis de microorganismos. Micróbios aeróbicos como Chlamydophila pneumoniae e Mycoplasma pneumoniae usualmente não sobrevivem em meio de estresse oxidativo. Arquéias são microorganismos anaeróbicos com poderosas enzimas anti-oxidantes que permitem detoxificação de radicais livres e a presença delas poderia favorecer a sobrevivência de micróbios aeróbicos. Pesquisamos por elementos de arquéia em placas vulneráveis e sua possível associação com degeneração mixomatosa da matriz e aumento do número de clamídias e micoplasmas. MÉTODOS: Vinte e nove amostras de 13 produtos de aterotomia de lesões grandes e excêntricas de óstio ou bifurcação de coronárias foram estudadas pela microscopia óptica e eletrônica. Agentes compatíveis com arquéia, clamídia e micoplasma foram semiquantificados pela microscopia eletrônica e correlacionados com quantidade de tecido fibromuscular, matriz mixomatosa e células xantomatosas. Seis casos foram também submetidos à reação em cadeia da polimerase com oligonucleotídeos de arquéia. RESULTADOS: Os 13 casos foram positivos para estruturas sugestivas de arquéia, micoplasma ou clamídia, em pelo menos uma amostra. Houve correlação positiva entre intensidade de matriz mixomatosa versus arquéia (r=0.44, p=0.02); arquéia versus micoplasma (r=0.41, p=0.03) e clamídia versus células xantomatosas r=0,42; 0.03). PCR foi positiva para DNA de arqueia em 4 dos 6 fragmentos. DISCUSSÃO: DNA e formas compatíveis com arquéia estão presentes em placas vulneráveis e podem ter papel fundamental na proliferação de micoplasma e clamídia. Este parece ser o primeiro relato de arquéia aparentemente patogênica em lesões de órgãos internos humanos.

PURPOSE: Vulnerable plaques are characterized by a myxoid matrix, necrotic lipidic core, reactive oxygen species, and high levels of microorganisms. Aerobic microbes such as Chlamydophila pneumoniae and Mycoplasma pneumoniae usually do not survive in oxidative stress media. Archaea are anaerobic microbes with powerful anti-oxidative enzymes that allow detoxification of free radicals whose presence might favor the survival of aerobic microorganisms. We searched for archaeal organisms in vulnerable plaques, and possible associations with myxoid matrix, chlamydia, and mycoplasma bodies. METHODS: Twenty-nine tissue samples from 13 coronary artherectomies from large excentric ostial or bifurcational lesions were studied using optical and electron microscopy. Infectious agents compatible with archaea, chlamydia, and mycoplasma were semiquantified using electron micrographs and correlated with the amounts of fibromuscular tissue, myxoid matrix, and foam cells, as determined from semi-thin sections. Six of the cases were also submitted to polymerase chain reaction with archaeal primers. RESULTS: All 13 specimens showed archaeal-compatible structures and chlamydial and mycoplasmal bodies in at least 1 sample. There was a positive correlation between extent of the of myxoid matrix and archaeal bodies (r = 0.44, P = 0.02); between archaeal and mycoplasmal bodies (r = 0.41, P = 0.03), and between chlamydial bodies and foam cells (r = 0.42; P = 0.03). The PCR test was positive for archaeal DNA in 4 of the 6 fragments. DISCUSSION: DNA and forms suggestive of archaea are present in vulnerable plaques and may have a fundamental role in the proliferation of mycoplasma and chlamydia. This seems to be the first description of apparently pathogenic archaea in human internal organ lesions.