Probiotics, prebiotics, and synbiotics in childhood diarrhea

Abstract Acute diarrhea is the second leading cause of morbidity and mortality attributed to infections in children under five years of age worldwide, with 1.7 million annual estimated cases and more than 500,000 deaths. Although hydroelectrolytic replacement is the gold standard in treating diarrhea, it does not interfere with the restoration of the intestinal microbiota. Several studies have searched for an adequate alternative in restructuring intestinal homeostasis, finding that treatments based on probiotics, prebiotics, and synbiotics are effective, which made such treatments increasingly present in clinical practice by reducing illness duration with minimal side effects. However, there are still controversies regarding some unwanted reactions in patients. The diversity of strains and the peculiarities of the pathogens that cause diarrhea require further studies to develop effective protocols for prevention and treatment. Here, we provide a descriptive review of childhood diarrhea, emphasizing treatment with probiotics, prebiotics, and synbiotics.

Phagocytic activity of monocytes and neutrophils in patients with periodontitis, whether or not associated to type 2 diabetes / Atividade fagocítica de monócitos e neutrófilos em pacientes com periodontite, associada ou não ao diabetes tipo 2

ABSTRACT Phagocytic functions by neutrophils/ monocytes and biochemical parameters were assessed in peripheral blood of patients with periodontitis, whether or not associated to type 2 diabetes, or patients with type 2 diabetes, or systemically healthy people. Fifty-eight participants were divided into four groups: Control - systemically and periodontally healthy patients (C, n=16), Periodontitis (P, n=14), Type 2 Diabetes (DM, n=11) and Periodontitis associated with type 2 diabetes (DMP, n=17). Blood samples were used to analyze phagocytic activity and the production of superoxide anion using optical microscopy. Significantly lower phagocytic activity of neutrophils was observed in non-opsonized samples (p = 0.008, Kruskal- Wallis) of the periodontitis group and in opsonized samples (p = 0.029, Kruskal-Wallis) of the periodontitis associated with type 2 diabetes group when these groups were compared to the healthy individuals when a 20:1 yeast: phagocyte stimulus was used. Periodontitis patients, whether associated (p = 0.0007, sensitized; Kruskal-Wallis, 20:1) or not with diabetes (p = 0.018 and 0.0007, in the proportions 5:1 and 20:1 yeast: monocyte respectively in sensitized samples; Kruskal-Wallis) also showed lower phagocytic function of monocytes compared to the control group. There was no significant difference in the production of superoxide anion among the evaluated groups. Severe clinical attachment loss was associated with lower levels of HDL in periodontitis patients and a higher percentage of A1C in diabetes with periodontitis patients (p<0.05; Pearson and Spearman correlations, respectively). Patients with both associated diseases had higher levels of triglycerides and CRP (p<0.001, Kruskal-Wallis) compared to patients with diabetes only. The results of the present study suggest that periodontitis negatively interferes with the innate immune response and may represent a major risk of systemic complications such as cardiovascular disease in diabetic patients or even in healthy individuals.

RESUMO As funções fagocíticas de neutrófilos/monócitos e parâmetros bioquímicos foram avaliados no sangue periférico de pacientes com periodontite com ou sem diabetes do tipo 2, ou em pacientes com diabetes tipo 2, ou em pessoas saudáveis sistemicamente. 58 participantes foram divididos em quatro grupos: Controle - pacientes sistemicamente e periodontalmente saudáveis (C, n = 16), Periodontite (P, n = 14), Diabetes Tipo 2 (DM, n = 11) e Periodontite associada a diabetes tipo 2 (DMP, n = 17). Amostras de sangue foram usadas para analisar a atividade fagocítica e a produção de ânion superóxido por microscopia óptica. Observou-se menor atividade fagocítica dos neutrófilos em amostras não opsonizadas (p = 0,008, Kruskal-Wallis) do grupo periodontite e em amostras opsonizadas (p = 0,029, Kruskal-Wallis) do grupo periodontite associada ao diabetes tipo 2 quando esses grupos foram comparados aos indivíduos saudáveis sob um estímulo de levedura:monócito de 20:1. Pacientes com periodontite associada (p = 0,0007, sensibilizados; Kruskal-Wallis, 20: 1) ou não com diabetes (p = 0,018 e 0,0007, nas proporções 5: 1 e 20: 1 de levedura: monócito, respectivamente, em amostras sensibilizadas; Kruskal- Wallis) também demonstraram menor função fagocítica dos monócitos em comparação com o grupo controle. Não houve diferença significativa na produção de ânion superóxido entre os grupos avaliados. A perda de inserção clínica grave foi associada a níveis mais baixos de HDL na periodontite e maior percentual de A1C nos pacientes com periodontite associada ao diabetes (p<0,05; correlações de Person e Spearman, respectivamente). Os pacientes com ambas as doenças associadas apresentaram níveis mais altos de triglicerídeos e PCR (p<0,001, Kruskal- Wallis) em comparação aos pacientes com somente diabetes. Os resultados do presente estudo sugerem que a periodontite interfere negativamente na resposta imune inata e pode representar um risco maior para complicações sistêmicas, como a doença cardiovascular, em pacientes com diabetes ou mesmo em indivíduos saudáveis.

Efeito dos níveis de lisina/proteína ideal sobre desempenho, características de carcaça, digestibilidade e parâmetros sanguíneos de suínos machos, castrados, da raça Duroc, de 30kg a 50kg / [Effects of lysine/ideal protein level on performance, carcass characteristics, digestibility and blood parameters of Duroc barrows from 30 to 50kg]

Objetivou-se avaliar níveis de proteína e aminoácidos, mantendo-se as relações entre os aminoácidos para suínos machos, castrados, de30kg a 50kg. Foram utilizados 50 suínos machos, castrados, com peso inicial de 30,35±1,96kg, distribuídos em delineamento experimental inteiramente ao acaso, com cinco tratamentos e cinco repetições com dois animais por unidade experimental. Os tratamentos consistiram em níveis de lisina digestível, mantendo-se a relação com os demais aminoácidos digestíveis: 0,73%; 0,83%; 0,93%; 1,03% e 1,13% na dieta. Avaliou-se desempenho, avaliação de carcaça, parâmetros sanguíneos e digestibilidade das dietas. Houve efeito quadrático para ganho de peso, conversão alimentar e níveis de creatinina em função dos níveis de lisina, com níveis ótimos estimados em 0,92%, 0,93% e 0,93%, respectivamente. As características de carcaça não foram influenciadas significativamente pelos tratamentos. Constatou-se efeito linear positivo para digestibilidade aparente da proteína bruta, da proteína total e da ureia sérica. Conclui-se que os níveis de proteína e lisina digestível recomendados para dietas de suínos machos, castrados, da raça Duroc, na fase de crescimento I, são de 16,70% e 0,93%, respectivamente, pois esses níveis proporcionaram melhorias no ganho de peso, na conversão alimentar e na creatinina sérica.(AU)

The objective of this study was to evaluate digestible lysine levels, keeping the relation among amino acids for Duroc barrows from 30 to 50kg. Fifty Duroc barrows (30.35±1.96kg live weight) were allotted in a completely randomized experimental design, divided in five treatments with five replicates and two animals in each experimental unit. The treatments consisted of digestible lysine levels (0.73%; 0.83%; 0.93%; 1.03% and 1.13%), keeping the relation with other essential amino acids. Performance, carcass characteristics, blood parameters and digestibility of the diets were evaluated. There was a quadratic response on weight gain, feed conversion and creatinine serum concentration as a function of the digestible lysine levels, with the greater levels obtained at 0.92%, 0.93% and 0.93%, respectively. The carcass characteristics were not influenced by the treatments. There was a linear increase of apparent digestibility of crude protein, total serum protein and urea. Results suggest that the requirement of protein and digestible lysine was 16.70% and 0.93%, respectively, providing improvements on weight gain, feed conversion and creatinine serum concentration of Duroc barrows in the growth phase.(AU)

Defining the landscape of ATP-competitive inhibitor resistance residues in protein kinases.

Kinases are involved in disease development and modulation of their activity can be therapeutically beneficial. Drug-resistant mutant kinases are valuable tools in drug discovery efforts, but the prediction of mutants across the kinome is challenging. Here, we generate deep mutational scanning data to identify mutant mammalian kinases that drive resistance to clinically relevant inhibitors. We aggregate these data with subsaturation mutagenesis data and use it to develop, test and validate a framework to prospectively identify residues that mediate kinase activity and drug resistance across the kinome. We validate predicted resistance mutations in CDK4, CDK6, ERK2, EGFR and HER2. Capitalizing on a highly predictable residue, we generate resistance mutations in TBK1, CSNK2A1 and BRAF. Unexpectedly, we uncover a potentially generalizable activation site that mediates drug resistance and confirm its impact in BRAF, EGFR, HER2 and MEK1. We anticipate that the identification of these residues will enable the broad interrogation of the kinome and its inhibitors.

Influência de diferentes sistemas e curvas de congelamento na congelabilidade e fertilidade do sêmen equino / Influence of systems and freezing curve in equine semen freezability and fertility

Avaliou-se o efeito de curvas de congelação nos parâmetros espermáticos e na fertilidade, usando sêmen de alta e baixa congelabilidade. Experimento 1 - utilizou-se sêmen de quatro garanhões resistentes à congelação: grupo 1, palhetas refrigeradas até 5°C e congeladas com curva de -8°C/min; grupos 2 e 3, palhetas refrigeradas até 5°C (0,5°C/min.) e congeladas com curvas de -20°C/min e -10°C/min, respectivamente. Experimentos 2 e 3 - utilizaram-se cinco garanhões (Mangalarga Marchador), respectivamente, de alta e baixa congelabilidade: grupo 4, a mesma metodologia descrita no grupo 1; grupos 5 e 6, palhetas refrigeradas até 5°C (0,5°C/min) e congeladas com curva de -20°C/min, entre 5°C e -60°C, e -10°C/min, entre -60°C e -100ºC (grupo 5), e -25°C/min, de 5°C até -100°C (grupo 6). O sêmen foi avaliado após descongelamento pelo método computadorizado. No experimento 1, não houve diferença nos parâmetros avaliados. No experimento 2, os parâmetros motilidade total (MT) e motilidade progressiva foram superiores aos do grupo 6 em relação ao grupo 4. No experimento 3, a MT foi superior no grupo 6 em relação ao grupo 4. As curvas de congelação mais rápidas apresentaram melhores parâmetros de cinética espermática, após a descongelação, para o sêmen de garanhões da raça Mangalarga Marchador.(AU)

The effect of freezing curves on sperm parameters and fertility, using resistant and sensitive semen to cryopreservation, was evaluated. In experiment 1, Semen from 4 stallions resistant to freezing was used: Group 1, straws were cooled to 5°C and frozen with a curve of - 8°C/min; Groups 2 and 3, straws were cooled to 5°C (0.5°C/min) and frozen with curves of - 20°C / min and - 10°C/min, respectively. In experiments 2 and 3, 5 stallions (Mangalarga Marchador) presenting respectively resistant and sensitive sperm to cryopreservation were used: Group 4, same methodology described for Group 1 was performed; Groups 5 and 6, straws were cooled to 5°C (0.5°C/min) and frozen with a curve of - 20°C/min. between 5°C and - 60°C and -10°C/min. between - 60°C and - 100°C (Group 5) and - 25°C/min. 5°C to - 100°C (Group 6). Thawed-semen was evaluated by the computerized method CASA. In Experiment 1, there was no difference in the evaluated parameters. In Experiment 2, total motility (MT) and progressive motility (PM) were higher in Group 6 compared to Group 4. In Experiment 3, TM was higher in Group 6 than Group 4. The faster freezing curves showed better parameters of sperm kinetics after thawing, for the Mangalarga Marchador stallion semen.(AU)

A new setup for the quantitative analysis of drying by the use of gas-phase FTIR-spectroscopy.

Drying rates are important for the manufacture of thin films and in specific for the production of electrodes used in electrochemical devices such as fuel cells and electrolyzers. The known procedures to investigate time-dependent sample compositions and selective evaporation rates are insufficient to obtain mean information about the full area instead of a single point analysis. Therefore, a new setup is presented using gas-phase Fourier-transform infrared spectroscopy. This method analyzes the gas-phase composition to recalculate the layer composition in electrode fabrication at any time during drying. According to the golden rule of measurement technology, manufacturer specifications are often overestimated. Therefore, our alternative procedures were used to evaluate the precision of devices used. The calculated measurement precision is confirmed by validation. The expected deviation is quantified to be less than 2% for the common application. Further on, the relative test-retest standard deviation is determined to be 0.3%-0.4%. As a result of the error propagation, the measurement precision is limited by the background gas flow rate precision for common application. At low volume fractions, the influence of the substance flow rate deviations becomes significant. However, further studies will focus on increasing the gas flow rate precision.

Clustering patterns of oral and general health-risk behaviours in Brazilian adolescents: Findings from a national survey.

OBJECTIVES: To evaluate how oral and general health-risk behaviours cluster among Brazilian adolescents. METHODS: The study comprised a total of 109 104 adolescents (52.2% female) participating in the Brazilian National School-based Student Health Survey (PeNSE). Seventeen behaviours (including diet; oral and hand hygiene; frequency of dental visits; tobacco, alcohol and drug use; sexual behaviour; physical activity, and risk for external causes) were measured using a self-reported questionnaire. Pairwise correlations between the health-risk behaviours were performed, and clustering was assessed by the hierarchical agglomerative cluster analysis (HACA), which was used to identify stable cluster solutions of the health-risk behaviours. RESULTS: All health-risk behaviours were correlated with at least 1 behaviour (P < .01). HACA indicated 2 broad stable clusters (n = 105 604). The first cluster included current smoking, illegal drug use, no hand washing before meals, unprotected sex, no helmet use, less frequent toothbrushing, no seatbelt use, physical fighting, skipping breakfast, current drinking, high sugar intake and, at the final stage, no dental visits. The second cluster included insufficient physical activity, eating while watching TV or studying, and low fruit intake. CONCLUSIONS: The health-risk behaviours clustered into 2 specific patterns among Brazilian adolescents. One cluster gathered a combination of lack of adherence to preventive behaviours and the undertaking of risky conduct, while the second reflected an unhealthy lifestyle (sedentary habits and low fruit diet). Knowledge about the clustering patterns of oral and general health behaviours in adolescents can better direct the integration of oral and general health promotion interventions.

Effectiveness of tranexamic acid for reducing intraoperative bleeding in palatoplasties: A randomized clinical trial.

BACKGROUND: Few data are available regarding blood loss during cleft palate surgery. This study assessed the effect of using tranexamic acid in the reduction of intraoperative bleeding. METHODS: A double-blind, randomized clinical trial compared intraoperative bleeding and the incidence of oronasal fistulas after palatoplasties in a control group that was given placebo and an intervention group that was given 10 mg/kg tranexamic acid followed by a continuous infusion of 1 mg/kg/h of the same until the end of surgery. Patients who underwent primary palatoplasty with no known or suspected coagulation disorders at our institution during the study period were included in the study. RESULTS: 70 patients were allocated, 66 received the intervention. Blood loss was reduced by 11.9%, without statistical significance. The incidence of fistulas in the intervention and control groups was 12.9% and 18.75%, respectively. The reduction of 5.8% (CI 95%: 12%-23.8%) was not significant. CONCLUSIONS: The reduction of intraoperative blood loss was lower than expected and not statistically significant; a larger sample is needed to confirm the observed reduction. The drug did not seem to have negative effects on flap viability.

Taxanes-induced cutaneous eruption: another histopathologic mimicker of malignancy.

BACKGROUND: Paclitaxel and docetaxel are antineoplastic drugs that bind the microtubules, producing the arrest of mitoses, which may be seen histopathologically. These histopathologic changes may simulate an intraepidermal keratinocytic malignant neoplasm, and an accurate diagnosis may be only established by clinicopathological correlation. OBJECTIVES: We report six cases of cutaneous eruptions by taxanes in which a striking cytotoxic effect was evident histopathologically. METHODS: Cutaneous biopsies were obtained in each patient. RESULTS: Atypical starburst-like or ring-like mitoses and dyskeratosis on basal and suprabasal layers of the epidermis. Areas of squamous syringometaplasia were also seen in one case. DISCUSSION: These findings were interpreted as expression of mitotic arrest due to taxanes. Similar changes have been described in association with other chemotherapeutic drugs such as vincristine, podophyllin and its derivative etoposide; colchicine, busulfan and maytansine, but cases like ours due to taxanes are exceptional or under-reported. CONCLUSION: Dermatopathologists should be aware of these effects in order to interpret carefully cutaneous biopsy specimens of patients receiving taxanes.

Treatment options for visceral artery aneurysms: ten year experience.

AIM: Open surgical repair (OSR) and endovascular techniques (ET) are both described in the literature for treating visceral artery aneurysms (VAAs). Aim of this study is to report a two-center experience of patients treated for a VAA using either OSR or ET, analyzing perioperative outcomes. METHODS: Clinical data of 32 VAAs in 32 patients treated between January 2001 and May 2011 were retrospectively reviewed and outcomes analyzed. RESULTS: Eighteen patients were men (56.3%). Median age was 64 years (range 26-79). Sixteen aneurysms were symptomatic: half of them were ruptured causing hemoperitoneum or gastrointestinal bleeding. ET were employed in 19 cases (59%) using covered stents (7 patients), coil embolization (5), plug placement (1), thrombin injection (2) and multiple associated techniques (4). OSR consisted in aneurysmectomy with end to end anastomoses (5 patients) or interposition graft (1), aneurysm ligation (4), splenectomy (2). One patient died during open surgery for hemoperitoneum due to VAA rupture (3%). OSR and ET had similar perioperative complication rates (5.2% vs. 15.3%, P=0.76). OSR had a longer in-hospital stay than ET (8 vs. 4 days, P=0.04). The presence of pancreatitis and alcohol abuse were more frequent in patients who presented with VAAs rupture. Clinical presentation with hemoperitoneum or aneurysm rupture were associated with higher mortality, regardless of the type of treatment. CONCLUSION: Both OSR and ET offered a safe way to treat VAAs in our experience.

Impact of chronic kidney disease on quality of life, lung function, and functional capacity / Repercussão da doença renal crônica na qualidade de vida, função pulmonar e capacidade funcional

OBJECTIVES: To evaluate the impact of the chronic kidney disease (CKD) on quality of life, from the children's and their parents' perspective, respiratory muscle strength, lung function, and functional capacity in children and adolescents. METHOD: Cross-sectional study of children with CKD aged 8 to 17 years. Those incapable of taking the tests were excluded. After an interview, quality of life by Pediatric Quality of Life Inventory) (PedsQLTM), muscular strength, pulmonary function tests, and the 6-minute walking test (6MWT) were applied. Student's t-test, ANOVA (difference in means), and Pearson's coefficient of correlation were used. The level of significance was set at 5%. RESULTS: Of the 40 patients, the mean distance walked at the 6MWT was 396 meters, and the mean final score at the quality of life test as perceived by the children and parents was 50.9 and 51, respectively. From the children's perspective, the transplanted patients had a higher quality of life score when compared to those undergoing hemodialysis (p < 0.001); those who practiced physical activity had better quality of life when compared to the sedentary children (p < 0.001). From the children's and the parents' perspectives, the male gender had a higher quality of life score (p < 0.05). There was a positive correlation between the distance walked at the 6MWT and age, height, final PedsQLTM, forced vital capacity (FVC), and forced expiratory volume in the first second (FEV1), as well as a negative correlation between FEV1/FVC and the distance walked. CONCLUSION: A significant reduction in the quality of life and the functional capacity was observed in children with CKD, influenced by the type of treatment, gender, and sedentary life style. .

OBJETIVOS: Avaliar repercussões da doença renal crônica (DRC) sobre a qualidade de vida na percepção das crianças e dos pais, força muscular respiratória, função pulmonar e capacidade funcional em crianças e adolescentes. MÉTODO: Estudo transversal de crianças e adolescentes com DRC de oito a 17 anos. Excluídas as incapazes de realizar os testes. Após entrevista, aplicou-se questionário de qualidade de vida (PedsQLTM), testes de força muscular, função pulmonar e teste de caminhada de 6 minutos (TC6 min). Foi utilizado o teste t de Student e ANOVA (diferenças de médias) e o coeficiente de correlação de Pearson. Considerou-se nível de significância de 5%. RESULTADOS: Dentre os 40 pacientes, a média da distância percorrida no TC6 min foi de 396 ± 71 metros, e a média do escore final de qualidade de vida percebida pelas crianças e pelos pais de 50,9 e 51, respectivamente. Na percepção das crianças, os transplantados apresentaram maior escore de qualidade de vida, comparados aos em hemodiálise (p < 0,001), e aos com atividade física e melhor qualidade de vida, comparadas às sedentárias (p < 0,001). Na percepção das crianças e dos pais, o sexo masculino apresentou maior escore de qualidade de vida (p < 0,05). Houve correlação positiva entre a distância percorrida no TC6 min e as variáveis idade, altura, PedsQLTM final da criança, capacidade vital forçada (CVF) e volume expiratório forçado no primeiro segundo (VEF1) e negativa entre VEF1/CVF e a distância percorrida. CONCLUSÃO: Observou-se redução significativa na qualidade de vida e na capacidade funcional em crianças com DRC influenciadas pelo tipo de tratamento, sexo e sedentarismo. .

Long-term results of PTFE trilaminate graft versus venous graft and composite graft for below-the-knee revascularization.

AIM: We report our experience with the use of PTFE trilaminate graft Advanta SST (Atrium Maquet™) versus autologous great saphenous vein (AGSV) and composite PTFE trilaminate-vein graft for below-the-knee (BTK) revascularization. METHODS: Data of all consecutive patients who underwent either a BTK or a distal femoro-popliteal bypass from January 2004 to March 2013 using Advanta SST, AGSV or composite Advanta SST-vein graft were retrospectively reviewed and outcomes were analyzed. Kaplan-Meier method with Log-rank test was used to estimate survival, limb salvage and patency rates. Chi-square test was used to evaluate risk factors affecting outcomes. P value <0.05 was considered significant. RESULTS: Sixty-one patients (48 males, 78.7%; median age 73 years, IQR 66-80) underwent a BTK/distal revascularization for peripheral artery disease causing critical limb ischemia (52 patients, 85.2%) or disabling claudication (9 patients). Advanta SST graft was used in 17 patients (group A), AGSV in 30 (Group B) and composite Advanta-AGSV graft in the remaining 14 (group C). Patients' demographics and risk factors were similar among the three groups, being hypertension and smoke the most frequent comorbidities. When Advanta SST was employed, the median duration of intervention was significantly lower than using vein or composite grafts (212 minutes, IQR 177-257; 270 minutes, IQR 220-375, P=.02; 327.5 minutes, IQR 252.5-405, P=0.003 respectively). At 30-days, wound complications tended to be significantly higher in Group B (13.3%) and C (11.7%) than in group A (0%), P=0.02. At long term, survival, primary assisted and secondary patency rates did not differ significantly among the three groups. Limb salvage was similar as well among group A, B and C (P=0.29), being 81.2+9.7%, 89.4+5.8% and 67.7+13.5% respectively at 1 year and 81.2+9.7%, 83.4%+7.9% and 54.2%+16.2% respectively at 3 years. Primary patency rate at 1 year was significantly better for group B than for group A and C (71.2+8.6%, 49.6+12.7% and 47.6+14.1% respectively, P=0.02), but after 1 year the patency rate for group B and A was similar (55.5+10.6% and 49.6+12.7% respectively), being for group C significantly worse (19+11.8%). A history of previous or current smoke affected significantly primary patency rate in group A (RR 0.39, 95%CI 0.08-0.95, P=0.03). CONCLUSION: Long-term results of the use of Advanta SST graft for BTK/distal revascularization seems to be promising, with significant lower duration of the operation and wound complications. At long term, survival, limb salvage, primary assisted and secondary patency rates did not differ significantly among the three groups. The AGSV still remains the best graft in terms of primary patency at 1 year, but after one year primary patency rates using the Advanta SST compare favorably to those of AGSV, while composite bypass grafts have the worst performance. These results need to be increased further.

Catechin attenuates behavioral neurotoxicity induced by 6-OHDA in rats.

This study was designed to investigate the beneficial effect of catechin in a model of Parkinson's disease. Unilateral, intrastriatal 6-hydroxydopamine (6-OHDA)-lesioned rats were pretreated with catechin (10 and 30 mg/kg) by intraperitoneal (i.p.) injection 2h before surgery and for 14 days afterwards. After treatments, apomorphine-induced rotations, locomotor activity, working memory and early and late aversive memories were evaluated. The mesencephalon was used to determine the levels of monoamines and measurement of glutathione (GSH). Immunohistochemical staining was also used to evaluate the expression of tyrosine hydroxylase (TH) in mesencephalic and striatal tissues. Catechin administration attenuated the increase in rotational behavior and the decrease in locomotor activity observed in lesioned rats. Although catechin did not rescue the impairment of late aversive memory, it protected the animals against 6-OHDA-induced working memory deficits. Furthermore, catechin treatment restored GSH levels, and significantly increased dopamine and DOPAC content, and TH-immunoreactivity in 6-OHDA-lesioned rats. Catechin protected 6-OHDA-lesioned rats due to its antioxidant action, indicating that it could be useful as an adjunctive therapy for the treatment of Parkinson's disease.

Recurrent pulmonary embolism secondary to popliteal vein aneurysm with intraluminal wall ulcer.

The popliteal vein is the most frequent site of venous aneurysm. Surgical treatment is indicated above a 2.5 cm diameter to prevent complications, notably deep venous thrombosis and pulmonary embolism (PE). Here we report a case of recurrent episodes of severe PE, leading to cardio-circulatory shock caused by a popliteal vein aneurysm (PVA) despite oral anticoagulant therapy. When surgical correction of the aneurysm was performed, we found an ulcerative lesion in the inner aspect of the vein that was acting as a 'thrombogenic focus' inside the aneurysm. An accurate inspection of the intimal wall is always important during surgery of PVA, particularly when tangential resection is performed.

Prevalence of DNA-HPV in male sexual partners of HPV-infected women and concordance of viral types in infected couples.

Investigation of HPV infection in men remains important due to its association with genital warts and anorectal cancer, as well as to the role men play in HPV transmission to their female sexual partners. Asymptomatic men (n = 43), whose sexual partners had presented cervical HPV infection, were enrolled in this study. Among the 43 men, 23 had their female partner included and tested for HPV-DNA, totaling 23 couples. HPV-DNA was detected by PCR. Type specific PCR to detect HPV 16, 18, 31, 33, 45 and 6/11 was performed. At least one type of HPV was detected in 86.0% (37/43) of the male patients and more than one HPV type was identified in 39.5% (17/43) of the samples, including high and low risk HPV. HPV-16 proved to be the most prevalent viral type in both male and female samples. Concordance of at least one viral type was observed in 56.5% (13/23) of the couples. Among couples that have shown concordance of viral types, 84.6% (11/13) of the men had the same high risk viral type presented by the female sexual partner. These data suggest that HPV infected men is an important reservoir, contributing to a higher transmission to women and maintenance of infection, and consequently, a higher risk of developing cervical cancer. HPV vaccination in men will protect not only them but will also have implications for their sexual partners.

New multilocus sequence typing of MRSA in São Paulo, Brazil

An increased incidence of nosocomial and community-acquired infections caused by methicillin-resistant Staphylococcus aureus (MRSA) has been observed worldwide. The molecular characterization of MRSA has played an important role in demonstrating the existence of internationally disseminated clones. The use of molecular biology methods in the surveillance programs has enabled the tracking of MRSA spread within and among hospitals. These data are useful to alert nosocomial infection control programs about the potential introduction of these epidemic clones in their areas. Four MRSA blood culture isolates from patients hospitalized at two hospitals in the city of São Paulo, Brazil, were analyzed; one of them was community acquired. The isolates were characterized as SCCmec, mecA and PVL by PCR, pulsed-field gel electrophoresis (PFGE) profile and molecular sequence typing (MLST) genotyping. The isolates presented type IV SCCmec, and none proved to be positive for PVL. The isolates showed a PFGE profile similar to the pediatric clone. MLST genotyping demonstrated that the isolates belonged to clonal complex 5 (CC5), showing a new yqiL allele gene, resulting in a new sequence typing (ST) (1176). Our results showed that strains of MRSA carrying a new ST are emerging in community and nosocomial infections, including bacteremia, in São Paulo, Brazil.

Viral and atypical bacterial detection in acute respiratory infection in children under five years.

BACKGROUND: Acute respiratory infection (ARI) is a leading cause of morbidity and mortality in children worldwide. This study aimed to determine the viral and atypical bacterial causes of different severities and clinical manifestations of ARI in preschool children from low-income families in North-East Brazil. METHODS: Clinical/demographic data and nasopharyngeal aspirates (NPA) were prospectively collected from children <5 years presenting with ARI over one year to a paediatric A&E department. Disease severity was grouped according to presence of lower respiratory tract signs, need for hospital admission and need for oxygen. Clinical manifestation of ARI was based on discharge diagnosis from hospital with four conditions predominating: bronchiolitis, pneumonia, episodic viral wheeze/asthma and upper respiratory tract infection. Multiplex PCR was used to detect 17 common respiratory viral and atypical bacterial pathogens in NPA. FINDINGS: 407 children with a median age of eight months were recruited. Pathogens were detected in 85·5% samples with co-infection being particularly common (39·5%). Respiratory Syncytial Virus (RSV; 37%), Adenoviruses (AdV; 25%), Rhinoviruses (hRV; 19%), Bocavirus (hBoV; 19%), human Meta-pneumovirus (hMPV; 10%) and Mycoplasma pneumoniae (Mpp; 10%) were most prevalent. Detection and co-infection rates were similar in all severities and clinical manifestations of ARI apart from RSV, which was associated with more severe disease and specifically more severe cases of bronchiolitis, and Mpp, which was associated with more severe cases of pneumonia. Mpp was detected in 17% of children admitted to hospital with pneumonia. INTERPRETATION: This study underlines the importance of viral and atypical bacterial pathogens in ARI in pre-school children and highlights the complex epidemiology of these pathogens in this age group. Generally, viruses and atypical bacteria were detected in all severities and clinical manifestations of ARI but RSV and Mpp were associated with more severe cases of bronchiolitis and pneumonia respectively.

Retroviral vector production under serum deprivation: The role of lipids.

The use of retroviral vectors for gene therapy applications demands high titer preparations and stringent quality standards. However, the manufacturing of these vectors still represents a highly challenging task due to the low productivity of the cell lines and reduced stability of the vector infectivity, particularly under serum-free conditions. With the objective of understanding the major limitations of retroviral vector production under serum deprivation, a thorough study of viral production kinetics, vector characterization and cell growth and metabolic behavior was conducted, for 293 FLEX 18 and Te Fly Ga 18 producer cell lines using different serum concentrations. The reduction of serum supplementation in the culture medium resulted in pronounced decreases in cell productivity of infectious vector, up to ninefold in 293 FLEX 18 cells and sevenfold in Te Fly Ga 18 cells. Total particles productivity was maintained, as assessed by measuring viral RNA; therefore, the decrease in infectious vector production could be attributed to higher defective particles output. The absence of the serum lipid fraction was found to be the major cause for this decrease in cell viral productivity. The use of delipidated serum confirmed the requirement of serum lipids, particularly cholesterol, as its supplementation not only allowed the total recovery of viral titers as well as additional production increments in both cell lines when comparing with the standard 10% (v/v) FBS supplementation. This work identified lower production ratios of infectious particles/total particles as the main restraint of retroviral vector production under serum deprivation; this is of the utmost importance concerning the clinical efficacy of the viral preparations. Lipids were confirmed as the key serum component correlated with the production of infective retroviral vectors and this knowledge can be used to efficiently design medium supplementation strategies for serum-free production. Biotechnol. Bioeng. 2009; 104: 1171-1181. (c) 2009 Wiley Periodicals, Inc.

Thermosensitivity of the reverse transcription process as an inactivation mechanism of lentiviral vectors.

Lentiviral vectors are an important tool for gene transfer research and gene therapy purposes. However, the low stability of these vectors affects their production, storage, and efficacy in preclinical and clinical settings. In the present work the mechanism underlying the thermosensitivity of lentiviral vectors was evaluated. For lentiviral vectors pseudotyped with amphotropic and RDpro envelopes, the capacity to perform reverse transcription was lost rapidly at 37 degrees C, in high correlation with the loss of infectivity. The vector with RDpro envelope presented a higher level of stability than that with amphotropic envelope for both the reverse transcription process and viral infectivity. Reverse transcriptase enzyme inactivation and viral template RNA degradation were not implicated in the loss of the viral capacity to perform reverse transcription. Furthermore, early entry steps in the infection process do not determine the rate of viral inactivation, as the amount of viral RNA and p24 protein entering the cells decreased slowly for both vectors. Taken together, it can be concluded that the reverse transcription process is thermolabile and thus determines the rate of lentiviral inactivation. Strategies to stabilize the reverse transcription process should be pursued to improve the applicability of lentiviral vectors in gene therapy.