Showing papers by "Federal University of São Paulo published in 2000"

Comparison of Upper Gastrointestinal Toxicity of Rofecoxib and Naproxen in Patients with Rheumatoid Arthritis

Abstract: Background Each year, clinical upper gastrointestinal events occur in 2 to 4 percent of patients who are taking nonselective nonsteroidal antiinflammatory drugs (NSAIDs). We assessed whether rofecoxib, a selective inhibitor of cyclooxygenase-2, would be associated with a lower incidence of clinically important upper gastrointestinal events than is the nonselective NSAID naproxen among patients with rheumatoid arthritis. Methods We randomly assigned 8076 patients who were at least 50 years of age (or at least 40 years of age and receiving long-term glucocorticoid therapy) and who had rheumatoid arthritis to receive either 50 mg of rofecoxib daily or 500 mg of naproxen twice daily. The primary end point was confirmed clinical upper gastrointestinal events (gastroduodenal perforation or obstruction, upper gastrointestinal bleeding, and symptomatic gastroduodenal ulcers). Results Rofecoxib and naproxen had similar efficacy against rheumatoid arthritis. During a median follow-up of 9.0 months, 2.1 confirmed ga...

The genome sequence of the plant pathogen Xylella fastidiosa

Abstract: Instituto Ludwig de Pesquisa sobre o Câncer, Rua Prof. Antonio Prudente, 109-4 andar, 01509-010, Sao Paulo-SP

Mudanças na composição e adequação nutricional da dieta familiar nas áreas metropolitanas do Brasil (1988-1996)

Abstract: OBJETIVO: Atualizar a tendencia secular (1962-1988) da composicao e da adequacao nutricional da dieta familiar praticada nas areas metropolitanas do Brasil, com base em nova pesquisa sobre orcamentos familiares realizada em meados da decada de 90. METODOS: Utilizaram-se como fontes de dados as pesquisas sobre orcamentos familiares (POF) da Fundacao IBGE, realizadas entre marco de 1987 e fevereiro de 1988 (13.611 domicilios) e entre outubro de 1995 e setembro de 1996 (16.014 domicilios), tendo ambas como universo de estudo as areas metropolitanas do Brasil. Nas duas pesquisas chegou-se a disponibilidade domiciliar diaria per capita de alimentos, dividindo-se o total de alimentos adquiridos no mes pelo numero de pessoas residentes no domicilio e pelo numero de dias do mes. O padrao alimentar foi caracterizado com base na participacao relativa de grupos selecionados de alimentos e de nutrientes na disponibilidade calorica total. A comparacao entre as duas pesquisas levou em conta o conjunto das areas metropolitanas do Pais e estratos dessas areas correspondentes as regioes menos desenvolvidas (Norte e Nordeste) e mais desenvolvidas (Centro-Oeste, Sudeste e Sul). RESULTADOS: Observou-se intensificacao do consumo relativo de carnes, de leites e de seus derivados (exceto manteiga) em todas as areas metropolitanas, enquanto o consumo de ovos passou a declinar, sobretudo no Centro-Sul do Pais. Leguminosas, raizes e tuberculos prosseguiram sua trajetoria descendente, mas cereais e derivados tenderam a se estabilizar no Centro-Sul ou mesmo a se elevar ligeiramente no Norte-Nordeste. A participacao relativa de acucar refinado e refrigerantes cresceu em todas as areas, sendo que a participacao de oleos e gorduras vegetais manteve-se constante no Norte-Nordeste e declinou intensamente no Centro-Sul. CONCLUSOES: A tendencia ascendente da participacao relativa de lipidios na dieta do Norte e do Nordeste, o aumento no consumo de acidos graxos saturados em todas as areas metropolitanas do Pais, ao lado da reducao do consumo de carboidratos completos, da estagnacao ou da reducao do consumo de leguminosas, verduras, legumes e frutas e do aumento no consumo ja excessivo de acucar sao os tracos marcantes e negativos da evolucao do padrao alimentar entre 1988 e 1996. Mudancas que podem indicar a adesao da populacao a dietas mais saudaveis -- declinio no consumo de ovos e recuo discreto da elevada proporcao de calorias lipidicas -- foram registradas apenas no Centro-Sul do Pais.

Shotgun sequencing of the human transcriptome with ORF expressed sequence tags

Abstract: Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. Systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome.

Randomized, double-blind clinical trial, controlled with placebo, of the toxicology of chronic melatonin treatment.

Abstract: The objective of the present study was to assess the toxicology of melatonin (10 mg), administered for 28 days to 40 volunteers randomly assigned to groups receiving either melatonin (N = 30) or placebo (N = 10) in a double-blind fashion. The following measurements were performed: polysomnography (PSG), laboratory examinations, including complete blood count, urinalysis, sodium, potassium and calcium levels, total protein levels, albumin, blood glucose, triglycerides, total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL), urea, creatinine, uric acid, glutamic-oxalacetic transaminase (GOT), glutamic-pyruvate transaminase (GPT), bilirubin, alkaline phosphatase, gama-glutamic transaminase (GGT), T3, T4, TSH, LH/FSH, cortisol, and melatonin serum concentrations. In addition, the Epworth Somnolence Scale (ESS) and a sleep diary (SD) were also applied to the volunteers 1 wk before each PSG. In addition, the volunteers were asked about possible side effects (SE) that appeared during the treatment. The study was carried out according to the following timetable: Visit 0, filling out the term of consent and inclusion criteria; Visit 1, PSG, laboratory examinations, ESS, SD, melatonin serum concentrations; Visit 2, SD, melatonin serum concentrations, SE; Visit 3, melatonin serum concentrations, PSG, ESS, SE; Visit 4, laboratory examinations, SE, melatonin serum concentrations, SD; and Visit 5, PSG, ESS, SE. Analysis of the PSG showed a statistically significant reduction of stage 1 of sleep in the melatonin group. No other differences between the placebo and melatonin groups were obtained. In the present study we did not observe, according to the parameters analyzed, any toxicological effect that might compromise the use of melatonin at a dose of 10 mg for the period of time utilized in this study.

Highly purified glycosylphosphatidylinositols from Trypanosoma cruzi are potent proinflammatory agents.

Abstract: Intracellular protozoan parasites are potent stimulators of cell‐mediated immunity. The induction of macrophage proinflammatory cytokines by Trypanosoma cruzi is considered to be important in controlling the infection and the outcome of Chagas9 disease. Here we show that the potent tumour necrosis factor‐α‐, interleukin‐12‐ and nitric oxide‐inducing activities of T.cruzi trypomastigote mucins were recovered quantitatively in a highly purified and characterized glycosylphosphatidylinositol (GPI) anchor fraction of this material. The bioactive trypomastigote GPI fraction was compared with a relatively inactive GPI fraction prepared from T.cruzi epimastigote mucins. The trypomastigote GPI structures were found to contain additional galactose residues and unsaturated, instead of saturated, fatty acids in the sn ‐2 position of the alkylacylglycerolipid component. The latter feature is essential for the extreme potency of the trypomastigote GPI fraction, which is at least as active as bacterial endotoxin and Mycoplasma lipopeptide and, therefore, one of the most potent microbial proinflammatory agents known.

Host Cell Invasion by TRYPANOSOMA cRUZI Is Potentiated by Activation of Bradykinin B2 Receptors

Abstract: The parasitic protozoan Trypanosoma cruzi employs multiple molecular strategies to invade a broad range of nonphagocytic cells. Here we demonstrate that the invasion of human primary umbilical vein endothelial cells (HUVECs) or Chinese hamster ovary (CHO) cells overexpressing the B2 type of bradykinin receptor (CHO-B2R) by tissue culture trypomastigotes is subtly modulated by the combined activities of kininogens, kininogenases, and kinin-degrading peptidases. The presence of captopril, an inhibitor of bradykinin degradation by kininase II, drastically potentiated parasitic invasion of HUVECs and CHO-B2R, but not of mock-transfected CHO cells, whereas the B2R antagonist HOE 140 or monoclonal antibody MBK3 to bradykinin blocked these effects. Invasion competence correlated with the parasites' ability to liberate the short-lived kinins from cell-bound kininogen and to elicit vigorous intracellular free calcium ([Ca2+]i) transients through B2R. Invasion was impaired by membrane-permeable cysteine proteinase inhibitors such as Z-(SBz)Cys-Phe-CHN2 but not by the hydrophilic inhibitor 1-trans-epoxysuccinyl-l-leucyl-amido-(4-guanidino) butane or cystatin C, suggesting that kinin release is confined to secluded spaces formed by juxtaposition of host cell and parasite plasma membranes. Analysis of trypomastigote transfectants expressing various cysteine proteinase isoforms showed that invasion competence is linked to the kinin releasing activity of cruzipain, herein proposed as a factor of virulence in Chagas' disease.

Human antibodies against a purified glucosylceramide from Cryptococcus neoformans inhibit cell budding and fungal growth.

Abstract: A major ceramide monohexoside (CMH) was purified from lipidic extracts of Cryptococcus neoformans. This molecule was analyzed by high-performance thin-layer chromatography (HPTLC), gas chromatography coupled with mass spectrometry, and fast atom bombardment-mass spectrometry. The cryptococcal CMH is a beta-glucosylceramide, with the carbohydrate residue attached to 9-methyl-4,8-sphingadienine in amidic linkage to 2-hydroxyoctadecanoic acid. Sera from patients with cryptococcosis and a few other mycoses reacted with the cryptococcal CMH. Specific antibodies were purified from patients' sera by immunoadsorption on the purified glycolipid followed by protein G affinity chromatography. The purified antibodies to CMH (mainly immunoglobulin G1) bound to different strains and serological types of C. neoformans, as shown by flow cytofluorimetry and immunofluorescence labeling. Transmission electron microscopy of yeasts labeled with immunogold-antibodies to CMH and immunostaining of isolated cell wall lipid extracts separated by HPTLC showed that the cryptococcal CMH predominantly localizes to the fungal cell wall. Confocal microscopy revealed that the beta-glucosylceramide accumulates mostly at the budding sites of dividing cells with a more disperse distribution at the cell surface of nondividing cells. The increased density of sphingolipid molecules seems to correlate with thickening of the cell wall, hence with its biosynthesis. The addition of human antibodies to CMH to cryptococcal cultures of both acapsular and encapsulated strains of C. neoformans inhibited cell budding and cell growth. This process was complement-independent and reversible upon removal of the antibodies. The present data suggest that the cryptococcal beta-glucosylceramide is a fungal antigen that plays a role on the cell wall synthesis and yeast budding and that antibodies raised against this component are inhibitory in vitro.

Mucin-like molecules form a negatively charged coat that protects Trypanosoma cruzi trypomastigotes from killing by human anti-alpha-galactosyl antibodies

Abstract: In the presence of sialic acid donors Trypanosoma cruzi acquires up to 10(7) sialic acid residues on its surface, in a reaction catalyzed by its unique trans-sialidase. Most of these sialic acid residues are incorporated into mucin-like glycoproteins. To further understand the biological role of parasite sialylation, we have measured the amount of mucin in this parasite. We found that both epimastigote and trypomastigote forms have the same number of mucin molecules per surface area, although trypomastigotes have less than 10% of the amount of glycoinositol phospholipids, the other major surface glycoconjugate of T. cruzi. Based on the estimated surface area of each mucin, we calculated that these molecules form a coat covering the entire trypomastigote cell. The presence of the surface coat is shown by transmission electron microscopy of Ruthenium Red-stained parasites. The coat was revealed by binding of antibodies isolated from Chagasic patients that react with high affinity to alpha-galactosyl epitopes present in the mucin molecule. When added to the trypomastigote, these antibodies cause an extensive structural perturbation of the parasite coat with formation of large blebs, ultimately leading to parasite lysis. Interestingly, lysis is decreased if the mucin coat is heavily sialylated. Furthermore, addition of MgCl2 reverses the protective effect of sialylation, suggesting that the sialic acid negative charges stabilize the surface coat. Inhibition of sialylation by anti-trans-sialidase antibodies, found in immunized animals, or human Chagasic sera, also increase killing by anti-alpha-galactosyl antibodies. Therefore, the large amounts of sialylated mucins, forming a surface coat on infective trypomastigote forms, have an important structural and protective role.

Validacao do questionario do Hospital Saint George na doenca respiratoria (SGRQ) em pacientes portadores de doenca pulmonar obstrutiva cronica no Brasil

Abstract: Introducao: O termo qualidade de vida tem adquirido cada vez mais importância no contexto cientifico. O presente estudo descreve a adaptacao para as lingua e cultura brasileiras de um questionario doenca-especifico desenvolvido por Paul Jones et al. em 1991(1): o Questionario do Hospital Saint George na Doenca Respiratoria (SGRQ), para a avaliacao de qualidade de vida em pacientes portadores de doenca pulmonar obstrutiva cronica (DPOC). Esse questionario contem tres componentes (sintomas, atividade e impactos) divididos em 76 itens. E auto-administrado e pode ser lido para pacientes analfabetos. Objetivo: Verificar se o SGRQ e um instrumento valido para medir qualidade de vida em pacientes portadores de DPOC no Brasil. Metodos: Para a validacao deste questionario no Brasil, realizou-se, inicialmente, uma versao da lingua inglesa para o portugues; em seguida, foi realizada a traducao retrograda (back translation), do portugues para o ingles, e uma versao final foi aplicada em 30 pacientes com diagnostico de DPOC, estaveis clinicamente e baseado em criterios de espirometria e oximetria. Os pacientes responderam ao questionario por duas vezes, num intervalo de 15 dias. O tempo de resposta foi cronometrado e as duvidas apontadas pelos pacientes, anotadas. Foi utilizado o teste estatistico de Wilcoxon para calculo de probabilidade de r e calculado o coeficiente de correlacao intraclasse para testar a fidedignidade e a confiabilidade do questionario. Resultados: Dos 30 pacientes que participaram do estudo, 10 eram do sexo feminino e 20 do masculino. A media de idade foi de 65,9 anos. A maioria dos pacientes encontrava-se no estadio 2 (56,7%) da DPOC, segundo a classificacao da American Thoracic Society. O coeficiente de correlacao intraclasse para a pontuacao total do questionario foi a = 0,79 e o resultado do teste de Wilcoxon p = 0,2110 (nao significante estatisticamente). O tempo medio de resposta dos dois dias de entrevista foi, respectivamente, 11 minutos e 50 segundos e 10 minutos e 31 segundos. Em relacao as duvidas, as questoes mais frequentemente referidas foram as das secoes 4 e 5, que contem uma frase cada na forma negativa. Conclusao: Pode-se concluir que a versao brasileira do Questionario do Hospital Saint George na Doenca Respiratoria (SGRQ) e um instrumento valido e fidedigno para medir qualidade de vida em pacientes portadores de DPOC no Brasil.

Prevalence of asthma symptoms in Latin America: The international study of asthma and allergies in childhood (ISAAC)

Abstract: The prevalence of respiratory symptoms indicative of asthma in children from Latin America has been largely ignored. As part of the International Study of Asthma and Allergies in Childhood (ISAAC), 17 centers in 9 different Latin American countries participated in the study, and data from 52,549 written questionnaires (WQ) in children aged 13-14 years and from 36,264 WQ in 6-7 year olds are described here. In children aged 13-14 years, the prevalence of asthma ever ranged from 5.5-28%, and the prevalence of wheezing in the last 12 months from 6.6-27%. In children aged 6-7 years, the prevalence of asthma ever ranged from 4.1-26.9%, and the prevalence of wheezing in the last 12 months ranged from 8.6-32.1%. The lower prevalence in centers with higher levels of atmospheric pollution suggests that chronic inhalation of polluted air in children does not contribute to asthma. Furthermore, the high figures for asthma in a region with a high level of gastrointestinal parasite infestation, and a high burden of acute respiratory infections occurring early in life, suggest that these factors, considered as protective in other regions, do not have the same effect in this region. The present study indicates that the prevalence of asthma and related symptoms in Latin America is as high and variable as described in industrialized or developed regions of the world.

Protective effect of oleuropein, an olive oil biophenol, on low density lipoprotein oxidizability in rabbits

Abstract: On the basis of the results obtained with pilot studies conducted in vitro on human low density lipoprotein (LDL) and on cell cultures (Caco-2), which had indicated the ability of certain molecules present in olive oil to inhibit prooxidative processes, an in vivo study was made of laboratory rabbits fed special diets. Three different diets were prepared: a standard diet for rabbits (diet A), a standard diet for rabbits modified by the addition of 10% (w/w) extra virgin olive oil (diet B), a modified standard diet for rabbits (diet C) differing from diet B only in the addition of 7 mg kg−1 of oleuropein. A series of biochemical parameters was therefore identified, both in the rabbit plasma and the related isolated LDL, before and after Cu-induced oxidation. The following, in particular, were selected: (i) biophenols, vitamins E and C, uric acid, and total, free, and ester cholesterol in the plasma; (ii) proteins, triglycerides, phospholipids, and total, free, and ester cholesterol in the native LDL (for the latter, the dimensions were also measured); (iii) lipid hydroperoxides, aldehydes, conjugated dienes, and relative electrophoretic mobility (REM) in the oxidized LDL (ox-LDL). In an attempt to summarize the results obtained, it can be said that this investigation has not only verified the antioxidant efficacy of extra virgin olive oil biophenols and, in particular, of oleuropein, but has also revealed a series of thus far unknown effects of the latter on the plasmatic lipid situation. In fact, the addition of oleuropein in diet C increased the ability of LDL to resist oxidation (less conjugated diene formation) and, at the same time, reduced the plasmatic levels of total, free, and ester cholesterol (−15, −12, and −17%, respectively), giving rise to a redistribution of the lipidic components of LDL (greater phospholipid and cholesterol amounts) with an indirect effect on their dimesions (bigger by about 12%).

Mitochondria in Ca2+ signaling and apoptosis.

Abstract: Cellular Ca2+ signals are crucial in the control of most physiological processes, cell injuryand programmed cell death; mitochondria play a pivotal role in the regulation of such cytosolicCa2+ ([Ca2+]c) signals Mitochondria are endowed with multiple Ca2+ transport mechanismsby which they take up and release Ca2+ across their inner membrane These transport processesfunction to regulate local and global [Ca2+]c, thereby regulating a number of Ca2+-sensitivecellular mechanisms The permeability transition pore (PTP) forms the major Ca2+ effluxpathway from mitochondria In addition, Ca2+ efflux from the mitochondrial matrix occursby the reversal of the uniporter and through the inner membrane Na+/Ca2+ exchanger Duringcellular Ca2+ overload, mitochondria take up [Ca2+]c, which, in turn, induces opening of PTP,disruption of mitochondrial membrane potential (ΔΨm) and cell death In apoptosis signaling,collapse of ΔΨ;m and cytochrome c release from mitochondria occur followed by activationof caspases, DNA fragmentation, and cell death Translocation of Bax, an apoptotic signalingprotein from the cytosol to the mitochondrial membrane, is another step during thisapoptosis-signaling pathway The role of permeability transition in the context of cell death in relationto Bcl-2 family of proteins is discussed

Angiotensin II antagonists for hypertension: are there differences in efficacy?

Abstract: We compared the antihypertensive efficacy of available drugs in the new angiotensin-II-antagonist (AIIA) class. The antihypertensive efficacy of losartan, valsartan, irbesartan, and candesartan was evaluated from randomized controlled trials (RCT) by performing a metaanalysis of 43 published RCT. These trials involved AIIA compared with placebo, other antihypertensive classes, and direct comparisons between AIIA. A weighted-average for diastolic and systolic blood pressure reduction with AIIA monotherapy, dose titration, and with addition of low-dose hydrochlorothiazide (HCTZ) were calculated. Weighted-average responder rates were also determined. The metaanalysis assessed a total of 11,281 patients. The absolute weighted-average reductions in diastolic (8.2 to 8.9 mm Hg) and systolic (10.4 to 11.8 mm Hg) blood pressure reductions (not placebo-corrected) for AIIA monotherapy were comparable for all AIIA. Responder rates for AIIA monotherapy were 48% to 55%. Dose titration resulted in slightly greater blood pressure reduction and an increase in responder rates to 53% to 63%. AIIA/hydrochlorothiazide combinations produced substantially greater reduction in systolic (16.1 to 20.6 mm Hg) and diastolic (9.9 to 13.6 mm Hg) blood pressure reductions than AIIA monotherapy and responder rates for AIIA/HCTZ combinations were 56% to 70%. This comprehensive analysis shows comparable antihypertensive efficacy within the AIIA class, a near-flat AIIA-dose response when titrating from starting to maximum recommended dose, and substantial potentiation of the antihypertensive effect with addition of HCTZ.

Overweight and obesity in Brazilian adolescents.

Abstract: OBJECTIVE: This study aimed to describe the prevalence of overweight and obesity (OW+O) among Brazilian adolescents and to identify risks for subpopulations defined according to the five country macro-regions and situation (urban–rural) of the domiciles, income, years of school attendance, age and sex. DESIGN: A nationwide home-based survey representative of the Brazilian civilian noninstitutionalized population, performed in 1989. METHODS: The sampling plans followed a stratified, multistage, probability cluster design in The National Research of Health and Nutrition sample, which collected anthropometric data of 14,455 domiciles. In all, 13,715 adolescents ranging from 10 to 19 y of age were studied. The OW+O was defined from a body mass index (BMI) equal or superior to the 85th percentile of the reference population of the NCHS. The prevalences in the different studied groups were compared using the adjusted odds ratio in logistic regression models. RESULTS: The prevalence of OW+O was of 7.7%, reaching 10.6% within the female group and 4.8% within the male group. A direct relation could be established between the socioeconomic level and OW+O. Adolescents of the most industrialized region of the country presented a risk of OW+O 1.86 (95% CI 1.51–2.30) times higher than that found in the least developed region. Male youngsters who lived in urban areas were more liable (OR=1.71, 95% CI 1.30–2.25) to overweight than their counterparts of rural areas. The occurrence of menarche increased two and a half times (OR=2.58, 95% CI 2.11–3.15) the risk of OW+O within the female group of adolescents. CONCLUSIONS: The results demonstrate a low prevalence of OW+O among Brazilian adolescents when compared with adolescents of more industrialized regions. The OW+O is twice as high within the female group, which represents a much greater difference than the one encountered in industrialized countries, probably owing to the muscular work carried out preponderantly by male adolescents of lower socioeconomic levels. Higher prevalences in subpopulations of higher socioeconomic level and of more industrialized regions show the great need for differentiated actions to control overweight and obesity in the country.

Production of IFN-? is impaired in patients with paracoccidioidomycosis during active disease and is restored after clinical remission

Abstract: Cellular immunity is usually suppressed during paracoccidioidomycosis (PCM) and is restored after treatment In this study we evaluated the induction of a type 1 (interferon γ (IFN-γ)), a type 2 (interleukin (IL)-10) and a primarily macrophage derived cytokine (tumor necrosis factor (TNF)-α) in peripheral blood mononuclear cells (PBMC) from patients with PCM Eight male patients with active PCM, nine male patients with clinical remission of the disease and 10 healthy control subjects were enrolled in the study Cytokines were induced with non-specific stimuli -- phytohaemagglutin (PHA) (induces IL-10 and IFN-γ), Lipopolysaccharide (induces TNF-α) -- and Paracoccidioides brasiliensis antigen (PbAg) (induces IL-10, IFN-g and TNF-a) Induction of IFN-g with PHA differed among the three groups (P<0·01; Kruskal-Wallis test) and with PbAg was lower in patients with active disease compared to those in clinical remission (P=0·05; Mann-Whitney) Induction of IL-10 and of TNF-α was similar in the three groups T

Differences in pain expression between male and female newborn infants.

Abstract: The study of neonatal gender differences in pain expression is important since neonatal pain behavior occurs prior to any learned reaction pattern. The objective of this study was to verify the presence of gender differences in pain expression in preterm and term newborn infants. Sixty-five consecutive neonates (37 female and 28 male infants) with gestational age between 28 and 42 weeks and with 25–120 h of life were studied. Healthy term neonates required a capillary puncture for PKU screening and clinically stable premature infants needed a capillary puncture for glucose dosage. The Neonatal Facial Coding System (NFCS) and the Neonatal Infant Pain Scale (NIPS) were evaluated at bedside prior to the puncture, when patients were at rest, during foot heating; during capillary puncture; and at 1, 3, and 5 min after heel lancing. Results were analyzed by repeated-measures ANOVA followed by the Multiple Comparison Method of Bonferroni. A significant difference among the mean NFCS scores during the six study periods was noted for the whole group of neonates ( P P =0.025). Regarding NIPS, ANOVA showed only a significant difference among the mean NIPS scores during the six study periods for the whole group of neonates ( P

Peptidase specificity characterization of C- and N-terminal catalytic sites of angiotensin I-converting enzyme.

Abstract: Quenched fluorescence peptides were used to investigate the substrate specificity requirements for recombinant wild-type angiotensin I-converting enzyme (ACE) and two full-length mutants bearing a single functional active site (N- or C-domain). We assayed two series of bradykinin-related peptides flanked by o-aminobenzoic acid (Abz) and N-(2,4-dinitrophenyl)ethylenediamine (EDDnp), namely, Abz-GFSPFXQ-EDDnp and Abz-GFSPFRX-EDDnp (X = natural amino acids), in which the fluorescence appeared when Abz/EDDnp are separated by substrate hydrolysis. Abz-GFSPFFQ-EDDnp was preferentially hydrolyzed by the C-domain while Abz-GFSPFQQ-EDDnp exhibits higher N-domain specificity. Internally quenched fluorescent analogues of N-acetyl-SDKP-OH were also synthesized and assayed. Abz-SDK(Dnp)P-OH, in which Abz and Dnp (2,4-dinitrophenyl) are the fluorescent donor-acceptor pair, was cleaved at the D-K(Dnp) bond with high specificity by the ACE N-domain (k(cat)/K(m) = 1.1 microM(-)(1) s(-)(1)) being practically resistant to hydrolysis by the C-domain. The importance of hydroxyl-containing amino acids at the P(2) position for N-domain specificity was shown by performing the kinetics of hydrolysis of Abz-TDK(Dnp)P-OH and Abz-YDK(Dnp)P-OH. The peptides Abz-YRK(Dnp)P-OH and Abz-FRK(Dnp)P-OH which were hydrolyzed by wild-type ACE with K(m) values of 5.1 and 4.0 microM and k(cat) values of 246 and 210 s(-)(1), respectively, have been shown to be excellent substrates for ACE. The differentiation of the catalytic specificity of the C- and N-domains of ACE seems to depend on very subtle variations on substrate-specific amino acids. The presence of a free C-terminal carboxyl group or an aromatic moiety at the same substrate position determines specific interactions with the ACE active site which is regulated by chloride and seems to distinguish the activities of both domains.

Cell damage and neurogenesis in the dentate granule cell layer of adult rats after pilocarpine- or kainate-induced status epilepticus.

Abstract: Dentate granule cells are generally considered to be relatively resistant to excitotoxicity and have been associated with robust synaptogenesis after neuronal damage. Synaptic reorganization of dentate granule cell axons, the mossy fibers, has been suggested to be relevant for hyperexcitability in human temporal lobe epilepsy and animal models. A recent hypothesis suggested that mossy-fiber sprouting is dependent on newly formed dentate granule cells. However, we recently demonstrated that cycloheximide (CHX) can block the mossy-fiber sprouting that would otherwise be induced by different epileptogenic agents and does not interfere with epileptogenesis in those models. Here, we investigated cell damage and neurogenesis in the dentate gyrus of pilocarpine- or kainate-treated animals with or without coadministration of CHX. Dentate granule cells were highly vulnerable to pilocarpine induced-status epilepticus (SE), but were hardly damaged by kainate-induced SE. CHX pretreatment markedly reduced the number of injured neurons after pilocarpine-induced SE. Induction of SE dramatically increased the mitotic rate of KA- and KA + CHX-treated animals. Induction of SE in animals injected with pilocarpine alone led to 2-7-fold increases in the mitotic rate of dentate granule cells as compared to 5- and 30-fold increases for pilocarpine + CHX animals. We suggest that such increased mitotic rates might be associated with a protection of a vulnerable precursor cell population that would otherwise degenerate after pilocarpine-induced SE. We further suggest that mossy-fiber sprouting and neurogenesis of granule cells are not necessarily linked to one another.

The relationship between oral habits and malocclusion in preschool children

Abstract: OBJETIVO: Avaliar como o desenvolvimento de habitos bucais deleterios e os problemas de fala afetam a oclusao dentaria em pre-escolares. METODOS: Foi constituida a amostra probabilistica por 2.139 criancas de ambos os sexos, na faixa etaria de 3 a 5 anos, matriculadas em instituicoes publicas ou privadas do Municipio de Bauru, SP, Brasil. Foi desenvolvido estudo transversal em duas etapas: exame de oclusao e questionario socioeconomico. A classificacao de Angle foi adotada para avaliacao de aspectos morfologicos da oclusao, observando-se tambem trespasse horizontal e vertical, espacamento/apinhamento, mordida aberta anterior, mordida cruzada total, mordida cruzada anterior e mordida cruzada posterior uni ou bilateral. Uma subamostra de 618 criancas apresentou resposta ao questionario sobre habitos bucais, saude infantil e informacoes sobre condicoes socioeconomicas. RESULTADOS: A prevalencia de ma oclusao foi de 51,3% para o sexo masculino e 56,9% para o sexo feminino, sem variacao quanto ao sexo. A maior prevalencia de ma oclusao foi verificada no grupo etario de tres anos, decrescendo significantemente com a idade (p<0,05). CONCLUSOES: Entre os fatores ambientais estudados, o habito de succao de chupeta foi o mais importante na associacao com ma oclusao (OR=5,46), seguido da succao digital (OR=1,54). Dificuldades na fala nao apresentaram relacao com a ma oclusao.

A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth.

Abstract: Activin A (Act.A), a member of the transforming growth factorβ family of secreted proteins, has been implicated in the regulation of growth and differentiation of various cell types. Betacellulin (BTC), a member of the epidermal growth factor family, converts exocrine AR42J cells to insulin-expressing cells when combined with Act.A. We have used primary cultures of human fetal pancreatic tissue to identify the effects of Act.A and/or BTC on islet development and growth. Exposure to Act.A resulted in a 1.5-fold increase in insulin content (P < 0.005) and a 2-fold increase in the number of cells immunopositive for insulin (P < 0.005). The formation of islet-like cell clusters, containing mainly epithelial cells, during a 5-day culture, was stimulated 1.4-fold by BTC (P < 0.05). BTC alone caused a 2.6-fold increase in DNA synthesis (P < 0.005). These data suggest that Act.A induces endocrine differentiation, whereas BTC has a mitogenic effect on human undifferentiated pancreatic epithelial cells.