Structural studies of BmooMP alpha-I, a non-hemorrhagic metalloproteinase from Bothrops moojeni venom

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prática da atividade física e a prevalência de doenças cardio-metabólicas no estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação do desenvolvimento de biofilme e da atividade antibiofilme, pH e solubilidade de cimentos endodônticos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Determinação de puberdade em cutias (Rodentia:Dasyproctidae) criadas em cativeiro, através da quantificação das células espermatogênicas

A cutia (Dasyprocta spp) é um roedor de médio porte, que apresenta o hábito de enterrar parte de seus alimentos, principalmente sementes, é considerado um dispersor em potencial, pois contribui com o reflorestamento natural. Por ser uma espécie muito procurada para consumo na área rural da Amazônia, o manejo em cativeiro e o conhecimento dos seus aspectos reprodutivos são formas alternativas para a criação de programas de produção e preservação dessa espécie com potencial econômico. O objetivo do presente trabalho, foi determinar o período que ocorre a puberdade, caracterizar os estádios do Ciclo do Epitélio Seminífero (CES), determinar a freqüência relativa dos estádios, calcular o rendimento geral da espermatogênese e o índice das células de sertoli. Foram utilizados 7 grupos com idade variando de 4 a 17 meses, os animais foram divididos em G1 (4 e 5 meses, n=4), G2 (6 e 7 meses, n=4), G3 (8 e 9 meses, n=4), G4 (10 e 11 meses, n=3), G5 (12 e 13 meses, n=4), G6 (14 e 15 meses, n=3) e G7 (16 e 17 meses, n=2). Ao atingir a idade programada os animais foram castrados, sob anestesia, e as amostras testiculares após biometria foram fixadas em ALFAC por 24 horas, submetidas ao processamento histológico de rotina, foram realizados cortes de 5 mm de espessura e os tecidos obtidos corados com HE. As fases de desenvolvimento reprodutivo, incluindo a puberdade, foram determinadas através da quantificação das células espermatogênicas de 10 túbulos seminíferos/animal com o contorno circular, os quais se encontravam no estádio I do CES, previamente caracterizado pelo método da morfologia tubular, que apresentou oito estádios do CES, e de 20 túbulos seminíferos/animal que não apresentavam espermatogênese completa, o número real das células foi obtido através da correção dos números brutos pelo diâmetro nuclear/nucleolar médio e espessura do corte histológico. Para determinar a freqüência relativa, 100 túbulos seminíferos de cada animal que já havia atingido a puberdade foram analisados. Os pesos corporal e testicular apresentaram correlações significativas com a idade e entre si. O peso corporal e biometria testicular aumentaram significativamente (P<0.05) até a maturidade sexual, onde na puberdade o PC e PT foram (1.903 + 0.55 Kg; 1.8 + 1.4 g, respectivamente) e na fase adulta foram (2.825 + 0.11Kg; 6.1 + 0.37g). Os grupos analisados foram classificados como G1 impúbere (13.21 + 1.6 mm); G2 pré-puberdade (7.09 + 1.7 mm); G3 puberdade (93.82 + 58.7 mm); G4 Pós-puberdade 1 (164.03 + 10.03 mm); G5 e G6 Pós-puberdade 2 de fase longa (173.8 + 10.5 mm; 185.9 + 1.5 mm) e G7 Adulto (238.8 + 72.6 mm). As células de sertoli, diminuiram significativamente (P<0.05) da fase impúbere (21.7 + 3.5 mm) até a puberdade (9.7 + 4.2 mm) onde iniciaram sua estabilização até a fase adulta (9.03 + 0.01mm). As células espermatogênicas apresentaram correlações altas e significativas com o peso testicular. As médias das seções transversais do diâmetro tubular aumentaram significativamente (P<0.05) entre os grupos analisados, G1 (109.5 + 4.6 mm); G2 (119.7 + 10 mm); G3 (174.6 + 24.1 mm); G4 (240.9 + 14.1 mm); G5 (219.6 + 9.8 mm); G6 (221.1 + 7.9 mm) e G7 (258.1 + 55.3 mm). Após a análise de 1600 túbulos seminíferos, os oito estádios caracterizados pelo método da morfologia tubular apresentaram as seguintes freqüências relativas: I (16.8 + 2.3%); II (18.8 + 3.4%); III (7.6 + 1.3%); IV (11.1 + 1.2%); V (21.2 + 4.2%); VI (10.4 + 3.4%); VII (7.9 + 2.4%); VIII (6.4 + 1.4%). A freqüência conjunta dos estádios foi pré-meiótica (43.5%), meiótica (10.7%) e pós-meiótica (45.8%). Portanto, a puberdade em cutias é alcançada no período de 8 e 9 meses, onde se observa maior produção das células espermatogênicas e estabelecimento das células de sertoli, a partir dessa fase inicia-se uma tentativa de estabilização das células espermatogênicas, o qual é acompanhado por uma tendência a estabilização do crescimento testicular.

Genetic Instability Persists in Non-Neoplastic Urothelial Cells from Patients with a History of Urothelial Cell Carcinoma

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of a Denture Adhesive on the Satisfaction and Kinesiographic Parameters of Complete Denture Wearers: A Cross-Over Randomized Clinical Trial

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Organic phosphorus fractions in soil fertilized with cattle manure

Inorganic phosphorus (Pi) usually controls the P availability in tropical soils, but the contribution of organic P (Po) should not be neglected, mainly in systems with low P input or management systems that promote organic matter accumulation. The aims of this study were to evaluate the changes in the Po fractions over time in soil fertilized and not fertilized with cattle manure and to correlate Po forms with available P extracted by anion exchange resin. The experiment was carried out under field conditions, in a sandy-clay loam Haplustox. The experimental design was a 2 x 9 randomized complete block factorial design, in which the first factor was manure application (20 t ha(-1)) or absence, and the second the soil sampling times (3, 7, 14, 21, 28, 49, 70, 91, and 112 days) after manure incorporation. Labile, moderately labile and non-labile Po fractions were determined in the soil material of each sampling. Manure fertilization increased the Po levels in the moderately labile and non-labile fractions and the total organic P, but did not affect the Po fraction proportions in relation to total organic P. On average, 5.1 % of total Po was in the labile, 44.4 % in the moderately labile and 50.5 % in the non-labile fractions. Available P (resin P) was more affected by the manure soluble Pi rather than by the labile Po forms. The labile and non-labile Po fractions varied randomly with no defined trend in relation to the samplings; for this reason, the data did not fit any mathematical model.

Trasmissione post-natale del citomegalovirus attraverso il latte materno al neonato VLBW

Introduction Postnatal human cytomegalovirus (CMV) infection is usually asymptomatic in term babies, while preterm infants are more susceptible to symptomatic CMV infection. Breastfeeding plays a dominant role in the epidemiology of transmission of postnatal CMV infection, but the risk factors of symptomatic CMV infection in preterm infants are unknown. Patients and Methods Between December 2003 and August 2006, eighty Very Low Birth Weight (VLBW) preterm infants (gestational age ≤ 32 weeks and birth weight < 1500 g), admitted to the Neonatal Intensive Care Unit of St Orsola-Malpighi General Hospital, Bologna were recruited. All of them were breastfed for at least one month. During the first week of life, serological test for CMV was performed on maternal blood. Furthermore, urinary CMV culture was performed in all the infants in order to exclude a congenital CMV infection. Urine samples from each infant were collected and processed for CMV culture once a week. Once every 15 days a blood sample was taken from each infant to evaluate the complete blood count, the hepatic function and the C reactive protein. In addition, samples of fresh breast milk were processed weekly for CMV culture. A genetic analysis of virus variant was performed in the urine of the infected infants and in their mother’s milk to confirm the origin of infection. Results We evaluated 80 VLBW infants and their 68 mothers. Fifty-three mothers (78%) were positive for CMV IgG antibodies, and 15 (22%) were seronegative. In the seronegative group, CMV was never isolated in breast milk, and none of the 18 infants developed viruria; in the seropositive group, CMV was isolated in 21 out of 53 (40%) mother’s milk. CMV was detected in the urine samples of 9 out of 26 (35%) preterm infants, who were born from 21 virolactia positive mothers. Six of these infants had clinically asymptomatic CMV infection, while 3 showed a sepsis-like illness with bradycardia, tachypnea and repeated desaturations. Eight out of nine infants showed abnormal hematologic values. The detection of neutropenia was strictly related to CMV infection (8/9 infected infants vs 17/53 non infected infants, P<.005), such as the detection of an increase in conjugated bilirubin (3/9 infected infants vs 2/53 non infected infants, P<.05). The degree of neutropenia was not different between the two groups (infected/non infected). The use of hemoderivatives (plasma and/or IgM–enriched immunoglobulin) in order to treat a suspected/certain infection in newborn with GE< 28 ws was seen as protective against CMV infection (1/4 infected infants vs 18/20 non infected infants [GE<28 ws]; P<.05). Furthermore, bronchopulmonary dysplasia (defined both as oxygen-dependency at 30 days of life and 36 ws of postmenstrual age) correlated with symptomatic infection (3/3 symptomatic vs 0/6 asymptomatic: P<.05). Conclusion Our data suggest that CMV infection transmitted to preterm newborn through human milk is always asymptomatic when newborns are clinically stable. Otherwise, the infection can worsen a preexisting disease such as bronchopulmonary dysplasia. Human milk offers many nutritional and psychological advantages to preterm newborns: according to our data, there’s no reason to contraindicate it neither to pasteurize the milk of all the mothers of preterm infants who are CMV seropositive.

Mastocitosi cutanee: il mastocitoma in età pediatrica

Background: Mastocytosis is a rare disease involving mast cells (MC) and their CD34+ progenitors. According to the WHO consensus classification, cutaneous mastocytosis (CM) is considered a benign disease confined to the skin, preferentially seen in young children with a marked tendency to regress spontaneously. Aim of our study was the long-term assessment of the outcome of solitary (SM) and multiple (MM) mastocytomas in a pediatric population. Materials and methods: From January 1996 to December 2010, 241 pediatric patients with a diagnosis of CM were followed-up at the outpatient division of pediatric dermatology of the University of Bologna. We focused our retrospective evaluation on patients affected by SM or MM. We collected, through the analysis of medical records and with a telephone questionnaire for patients and their families, information on clinical aspects of the disease evolution and on the efficacy of topical steroid therapy. Results: Over the 241 considered patients we recorded: SM or MM in 176 (73%) pts., urticaria pigmentosa in 53 (22%) pts., telangiectasia macularis eruptiva perstans in 9 (4%) pts., diffuse CM in 2 (0,9%) pts. and polymorph CM in 1 (0,4%) pt. On 176 children affected by SM or MM (97 M vs. 79 F), 130 (74%) patients were followed-up with a mean of 56,3 (r. 4-142) months. A satisfactory outcome was recorded in 99 (76%) cases of whom 52 (53%) treated with topic steroids. Mean time to complete regression was 16.4 m. on treated patients vs. 34.7 m. on non treated patients (p=0,001). Conclusions: From our study emerged that resolution of the disease is independent from therapy, but the time to regression and to complete recovery of the coetaneous lesions is faster and favored by the application of topic steroid with an improvement of the quality of life for children and their families.

Long-term clinical outcomes of biodegradable polymer biolimus-eluting stents versus durable polymer sirolimus-eluting stents in patients with coronary artery disease (LEADERS): 4 year follow-up of a randomised non-inferiority trial

Background The effectiveness of durable polymer drug-eluting stents comes at the expense of delayed arterial healing and subsequent late adverse events such as stent thrombosis (ST). We report the 4 year follow-up of an assessment of biodegradable polymer-based drug-eluting stents, which aim to improve safety by avoiding the persistent inflammatory stimulus of durable polymers. Methods We did a multicentre, assessor-masked, non-inferiority trial. Between Nov 27, 2006, and May 18, 2007, patients aged 18 years or older with coronary artery disease were randomly allocated with a computer-generated sequence to receive either biodegradable polymer biolimus-eluting stents (BES) or durable polymer sirolimus-eluting stents (SES; 1:1 ratio). The primary endpoint was a composite of cardiac death, myocardial infarction, or clinically-indicated target vessel revascularisation (TVR); patients were followed-up for 4 years. Analysis was by intention to treat. This trial is registered with ClinicalTrials.gov, number NCT00389220. Findings 1707 patients with 2472 lesions were randomly allocated to receive either biodegradable polymer BES (857 patients, 1257 lesions) or durable polymer SES (850 patients, 1215 lesions). At 4 years, biodegradable polymer BES were non-inferior to durable polymer SES for the primary endpoint: 160 (18·7%) patients versus 192 (22·6%) patients (rate ratios [RR] 0·81, 95% CI 0·66–1·00, p for non-inferiority <0·0001, p for superiority=0·050). The RR of definite ST was 0·62 (0·35–1·08, p=0·09), which was largely attributable to a lower risk of very late definite ST between years 1 and 4 in the BES group than in the SES group (RR 0·20, 95% CI 0·06–0·67, p=0·004). Conversely, the RR of definite ST during the first year was 0·99 (0·51–1·95; p=0·98) and the test for interaction between RR of definite ST and time was positive (pinteraction=0·017). We recorded an interaction with time for events associated with ST but not for other events. For primary endpoint events associated with ST, the RR was 0·86 (0·41–1·80) during the first year and 0·17 (0·04–0·78) during subsequent years (pinteraction=0·049). Interpretation Biodegradable polymer BES are non-inferior to durable polymer SES and, by reducing the risk of cardiac events associated with very late ST, might improve long-term clinical outcomes for up to 4 years compared with durable polymer SES. Funding Biosensors Europe SA, Switzerland.

IL28B alleles associated with poor hepatitis C virus (HCV) clearance protect against inflammation and fibrosis in patients infected with non-1 HCV genotypes

Genetic polymorphisms near IL28B are associated with spontaneous and treatment-induced clearance of hepatitis C virus (HCV), two processes that require the appropriate activation of the host immune responses. Intrahepatic inflammation is believed to mirror such activation, but its relationship with IL28B polymorphisms has yet to be fully appreciated. We analyzed the association of IL28B polymorphisms with histological and follow-up features in 2335 chronically HCV-infected Caucasian patients. Assessable phenotypes before any antiviral treatment included necroinflammatory activity (n = 1,098), fibrosis (n = 1,527), fibrosis progression rate (n = 1,312), and hepatocellular carcinoma development (n = 1,915). Associations of alleles with the phenotypes were evaluated by univariate analysis and multivariate logistic regression, accounting for all relevant covariates. The rare G allele at IL28B marker rs8099917-previously shown to be at risk of treatment failure-was associated with lower activity (P = 0.04), lower fibrosis (P = 0.02) with a trend toward lower fibrosis progression rate (P = 0.06). When stratified according to HCV genotype, most significant associations were observed in patients infected with non-1 genotypes (P = 0.003 for activity, P = 0.001 for fibrosis, and P = 0.02 for fibrosis progression rate), where the odds ratio of having necroinflammation or rapid fibrosis progression for patients with IL28B genotypes TG or GG versus TT were 0.48 (95% confidence intervals 0.30-0.78) and 0.56 (0.35-0.92), respectively. IL28B polymorphisms were not predictive of the development of hepatocellular carcinoma.

Alteration in 5HT1A Receptor Activity from a Prenatal Exposure to Dexamethasone in a Stressed and Non-Stressed Adult Male Rat

Synthetic glucocorticoids (GC) are used as a clinical therapeutic to stimulate lung development in fetuses that present the risk of preterm delivery. Previous studies have shown that a prenatal exposure to Dexamethasone (DEX) causes a disturbance in normal GC mediation of neuritic outgrowth, cell signaling, and serotonergic systems. Our hypothesis is that a prenatal exposure to DEX during the third trimester of pregnancy alters 5HT1A receptor function. Pregnant dams were injected daily with 150μg/ml/kg of DEX from gestation day 14 through 19. Control dams were treated with and equal volume of saline. Swim stress followed by elevated plus maze testing was conducted on male rats an hour and a half prior to being sacrificed to induce postnatal acute stress. The non-stressed group was also tested and allowed to return to baseline before sacrifice. Hippocampi were analyzed using a radioligand-receptor binding assay and GTPγS35 incorporation (3H-MPPF antagonist and 8-OH-DPAT agonist, respectively). A significant increase in Kd was found in non-stressed DEX-exposed animals compared to non-stressed controls (p

LH and IGF-1 release during oestrus and early luteal phase in lactating and non-lactating horse mares

The aim of the present study was to determine effects of lactation on basal LH and IGF-1 concentrations and on the LH response to a GnRH-analogue at different stages of the oestrous cycle in mares. A total of 17 cyclic Haflinger mares were included in the study. Experiments were performed on lactating mares in first postpartum oestrus, the subsequent early luteal phase, and second postpartum oestrus. Non-lactating mares were used in oestrus and early luteal phase. Blood samples were taken for 1 h at 15 min intervals. Mares were then injected with the GnRH-analogue buserelin (GnRHa; 5 microg i.v.) and blood samples were drawn every 15 min for further 2 h. LH in all samples and basal IGF-1-concentrations were determined by RIA. In lactating mares, basal LH concentrations during the early luteal phase tended to be lower (p = 0.07) and the LH response to GnRHa, calculated as area under the curve, was significantly less pronounced compared to non-lactating mares (p < 0.01). As well in lactating mares, the basal LH concentration between first early luteal phase and second oestrus differed significantly (p < 0.05) and the net response to GnRHa was significantly lower between first oestrus as well as second oestrus and first early luteal phase (p < 0.05) but not between first and second oestrous postpartum. Within the group of non-lactating mares, the LH response to GnRHa was as well significantly lower during oestrus than during early luteal phase (p < 0.01). IGF-1 concentrations differed neither between groups nor stages of the cycle within groups. In conclusion, basal and GnRHa-stimulated LH release in lactating mares is lower than in non-lactating mares. This difference, however, occurs only in the early luteal phase. In lactating mares, concentrations of LH appear adequate to allow ovulation to occur.

rCBF in hemorrhagic, non-hemorrhagic and mixed contusions after severe head injury and its effect on perilesional cerebral blood flow

Intracerebral contusions can lead to regional ischemia caused by extensive release of excitotoxic aminoacids leading to increased cytotoxic brain edema and raised intracranial pressure. rCBF measurements might provide further information about the risk of ischemia within and around contusions. Therefore, the aim of the presented study was to compare the intra- and perilesional rCBF of hemorrhagic, non-hemorrhagic and mixed intracerebral contusions. In 44 patients, 60 stable Xenon-enhanced CT CBF-studies were performed (EtCO2 30 +/- 4 mmHg SD), initially 29 hours (39 studies) and subsequent 95 hours after injury (21 studies). All lesions were classified according to localization and lesion type using CT/MRI scans. The rCBF was calculated within and 1-cm adjacent to each lesion in CT-isodens brain. The rCBF within all contusions (n = 100) of 29 +/- 11 ml/100 g/min was significantly lower (p < 0.0001, Mann-Whitney U) compared to perilesional rCBF of 44 +/- 12 ml/100 g/min and intra/perilesional correlation was 0.4 (p < 0.0005). Hemorrhagic contusions showed an intra/perilesional rCBF of 31 +/- 11/44 +/- 13 ml/100 g/min (p < 0.005), non-hemorrhagic contusions 35 +/- 13/46 +/- 10 ml/100 g/min (p < 0.01). rCBF in mixed contusions (25 +/- 9/44 +/- 12 ml/100 g/min, p < 0.0001) was significantly lower compared to hemorrhagic and non-hemorrhagic contusions (p < 0.02). Intracontusional rCBF is significantly reduced to 29 +/- 11 ml/100 g/min but reduced below ischemic levels of 18 ml/100 g/min in only 16% of all contusions. Perilesional CBF in CT normal appearing brain closed to contusions is not critically reduced. Further differentiation of contusions demonstrates significantly lower rCBF in mixed contusions (defined by both hyper- and hypodense areas in the CT-scan) compared to hemorrhagic and non-hemorrhagic contusions. Mixed contusions may evolve from hemorrhagic contusions with secondary increased perilesional cytotoxic brain edema leading to reduced cerebral blood flow and altered brain metabolism. Therefore, the treatment of ICP might be individually modified by the measurement of intra- and pericontusional cerebral blood.

Image registration for triggered and non-triggered DTI of the human kidney: Reduced variability of diffusion parameter estimation

BACKGROUND: To investigate if non-rigid image-registration reduces motion artifacts in triggered and non-triggered diffusion tensor imaging (DTI) of native kidneys. A secondary aim was to determine, if improvements through registration allow for omitting respiratory-triggering. METHODS: Twenty volunteers underwent coronal DTI of the kidneys with nine b-values (10-700 s/mm2 ) at 3 Tesla. Image-registration was performed using a multimodal nonrigid registration algorithm. Data processing yielded the apparent diffusion coefficient (ADC), the contribution of perfusion (FP ), and the fractional anisotropy (FA). For comparison of the data stability, the root mean square error (RMSE) of the fitting and the standard deviations within the regions of interest (SDROI ) were evaluated. RESULTS: RMSEs decreased significantly after registration for triggered and also for non-triggered scans (P < 0.05). SDROI for ADC, FA, and FP were significantly lower after registration in both medulla and cortex of triggered scans (P < 0.01). Similarly the SDROI of FA and FP decreased significantly in non-triggered scans after registration (P < 0.05). RMSEs were significantly lower in triggered than in non-triggered scans, both with and without registration (P < 0.05). CONCLUSION: Respiratory motion correction by registration of individual echo-planar images leads to clearly reduced signal variations in renal DTI for both triggered and particularly non-triggered scans. Secondarily, the results suggest that respiratory-triggering still seems advantageous.J. Magn. Reson. Imaging 2014. (c) 2014 Wiley Periodicals, Inc.