Altered inflammasome activation in neonatal encephalopathy persists in childhood.

Neonatal encephalopathy (NE) is characterized by altered neurological function in term infants and inflammation plays an important pathophysiological role. Inflammatory cytokines interleukin (IL)-1ß, IL-1ra and IL-18 are activated by the nucleotide-binding and oligomerization domain (NOD)-, leucine-rich repeat domain (LRR)- and NOD-like receptor protein 3 (NLRP3) inflammasome; furthermore, we aimed to examine the role of the inflammasome multiprotein complex involved in proinflammatory responses from the newborn period to childhood in NE. Cytokine concentrations were measured by multiplex enzyme-linked immunosorbent assay (ELISA) in neonates and children with NE in the absence or presence of lipopolysaccharide (LPS) endotoxin. We then investigated expression of the NLRP3 inflammasome genes, NLRP3, IL-1ß and ASC by polymerase chain reaction (PCR). Serum samples from 40 NE patients at days 1 and 3 of the first week of life and in 37 patients at age 4-7 years were analysed. An increase in serum IL-1ra and IL-18 in neonates with NE on days 1 and 3 was observed compared to neonatal controls. IL-1ra in NE was decreased to normal levels at school age, whereas serum IL-18 in NE was even higher at school age compared to school age controls and NE in the first week of life. Percentage of LPS response was higher in newborns compared to school-age NE. NLRP3 and IL-1ß gene expression were up-regulated in the presence of LPS in NE neonates and NLRP3 gene expression remained up-regulated at school age in NE patients compared to controls. Increased inflammasome activation in the first day of life in NE persists in childhood, and may increase the window for therapeutic intervention.

Self-amplifying memory based on multiple cascading four-wave mixing via recoil-induced resonance.

We report on a new, to the best of our knowledge, type of optical memory that allows for the amplification of the optical signal carrying the stored information during its reading process. The memory mechanism is demonstrated in an ensemble of cold cesium atoms and is based on the multiple parametric four-wave mixing exploring the external atomic degrees of freedom via recoil-induced resonances. We have particularly demonstrated the storage of light carrying orbital angular momentum with a fourfold amplifying factor for the retrieved signal during the reading process. Memory lifetimes of the order of hundreds of microseconds have been measured, and possible applications for this self-amplifying memory are discussed.

Barbatic acid from Cladia aggregata (lichen): Cytotoxicity and in vitro schistosomicidal evaluation and ultrastructural analysis against adult worms of Schistosoma mansoni.

Schistosomiasis is one of the most significant neglected tropical diseases, affecting around 260 million people worldwide, and Praziquantel is currently the only available drug for the treatment of infected persons. Thus, the search for new schistosomicidal compounds is urgent. The objective of this study was to investigate of the schistosomicidal effect of barbatic acid, a lichen metabolite, on adult worms of Schistosoma mansoni. The in vitro schistosomicidal effect was evaluated through the assessment of motility and mortality, cellular viability of the worms and ultrastructural analysis through scanning electron microscopy. To evaluate the cytotoxicity of barbatic acid, a cell viability assay was performed with human peripheral blood mononuclear cells. Barbatic acid showed a schistosomicidal effect after 3 h of exposure. At the end of 24 h the concentrations of 50-200 µM presented lethality on the worms. Motility changes were observed at sublethal concentrations. The IC50 obtained by the cell viability assay for S. mansoni was 99.43 µM. Extensive damage to the worm's tegument was observed from 25 µM. No cytotoxicity was observed on human peripheral blood mononuclear cells. This report provides data showing the schistosomicidal effect of barbatic acid on S. mansoni, causing death, motility changes and ultrastructural damage to worms. In addition, barbatic acid was shown to be non-toxic to human peripheral blood mononuclear cells at concentrations that are effective against S. mansoni.

Data set of the toxic effects of divaricatic acid depside on Biomphalaria glabrata and Schistosoma mansoni cercariae.

In this study, the molluscicidal and antiparasitic activities of divaricatic acid was evaluated, targeting the mollusc Biomphalaria glabrata and cercariae of the helminth Schistosoma mansoni. Divaricatic acid showed high toxicity against both adult snails (5.5 µg/mL) and embryos (20 µg/mL after 6 h of exposure). Similar activity was observed in S. mansoni cercariae after only a short exposure time. The divaricatic acid proved to be a promising substance for the control of the snail B. glabrata, an intermediate host of schistosomiasis, as well as the cercariae of the pathogen.

Radiosensitizer effect of usnic acid on Biomphalaria glabrata embryos.

PURPOSE: Some phytochemicals have shown the potential of being radiomodifiers, especially phenolic compounds, such as lichenic secondary metabolites. To evaluate the phytochemical usnic acid as a radiomodifier, embryonic cells of molluscs have been used due to their ease of collection, high sensitivity to physical and chemical agents, well-known embryology and low cost for analysis. MATERIALS AND METHODS: This study aimed to assess the radiosensitizing action of usnic acid on Biomphalaria glabrata embryos. Samples were irradiated with 4 Gy of gamma rays from a 60Co source (dose rate 2.906 Gy/h). An acute toxicity test was performed using B. glabrata embryos in the blastula stage, in order to determine the toxicity of usnic acid and to establish the lethal Concentration for 50% (LC50). Subsequently, the radiomodifing capacity of usnic acid was estimated using assays with B. glabrata embryos. RESULTS: Irradiation increased the number of non-viable embryos compared to unirradiated controls. Additionally, it was observed that embryos exposed to a non-toxic concentration of usnic acid (0.6 µg/mL) before irradiation showed a further enhancement in non-viable embryos when compared with exposure to ionizing radiation alone. CONCLUSION: The results presented here indicate that usnic acid makes cells more sensitive to the damaging effects of radiation.

Laboratory assessment of divaricatic acid against Biomphalaria glabrata and Schistosoma mansoni cercariae.

In this study, the molluscicidal and antiparasitic activities of divaricatic acid was evaluated, targeting the mollusc Biomphalaria glabrata and cercariae of the helminth Schistosoma mansoni. In addition, the environmental toxicity of divaricatic acid was assessed by bioassay using the microcrustacean Artemia salina. Divaricatic acid showed high toxicity against both adult snails (5µg/mL) and embryos (20µg/mL after 6h of exposure). Similar activity was observed in Schistosoma mansoni cercariae after only a short exposure time (10µg/mL after 30min of exposure). The divaricatic acid did not show toxicity in the acute test using Artemia salina at concentrations equal to or below 200µg/mL. The divaricatic acid proved to be a promising substance for the elimination of the snail Biomphalaria glabrata, an intermediate host of schistosomiasis, as well as the cercariae of the pathogen, while being non-toxic to the Artemia salina at the same concentrations. This is the first experimental observation of the molluscicidal and cercaricide activity of divaricatic acid.

Alterations in circulating lymphoid cell populations in systemic small vessel vasculitis are non-specific manifestations of renal injury.

Innate lymphocyte populations, such as innate lymphoid cells (ILCs), γδ T cells, invariant natural killer T (iNK T) cells and mucosal-associated invariant T (MAIT) cells are emerging as important effectors of innate immunity and are involved in various inflammatory and autoimmune diseases. The aim of this study was to assess the frequencies and absolute numbers of innate lymphocytes as well as conventional lymphocytes and monocytes in peripheral blood from a cohort of anti-neutrophil cytoplasm autoantibody (ANCA)-associated vasculitis (AAV) patients. Thirty-eight AAV patients and 24 healthy and disease controls were included in the study. Patients with AAV were sampled both with and without immunosuppressive treatment, and in the setting of both active disease and remission. The frequencies of MAIT and ILC2 cells were significantly lower in patients with AAV and in the disease control group compared to healthy controls. These reductions in the AAV patients remained during remission. B cell count and frequencies were significantly lower in AAV in remission compared to patients with active disease and disease controls. Despite the strong T helper type 2 (Th) preponderance of eosinophilic granulomatosis with polyangiitis, we did not observe increased ILC2 frequency in this cohort of patients. The frequencies of other cell types were similar in all groups studied. Reductions in circulating ILC2 and MAIT cells reported previously in patients with AAV are not specific for AAV, but are more likely to be due to non-specific manifestations of renal impairment and chronic illness. Reduction in B cell numbers in AAV patients experiencing remission is probably therapy-related.

Gallic acid as the major antioxidant in pequi (Caryocar brasiliense Camb. ) fruit peel / Ácido gálico como principal antioxidante da casca do fruto do pequi (Caryocar brasiliense Camb. )

ABSTRACTIn this study, ethanol-water extracts of pequi fruit peel were fractionated in order to identify and quantify the major antioxidant present in it. The fractions were subjected to liquid-liquid phase extraction and silica-gel column chromatography, and antioxidant activity was monitored using the 2,2-diphenyl-1-picrylhydrazyl radical-scavenging assay. The purity of the fractions was evaluated using thin-layer chromatography and high-performance liquid chromatography (HPLC). The substance with antioxidant property was identified through the analysis in a liquid chromatography-mass spectroscopy fragmentation and was quantified using HPLC. After the Silica-gel fractionation, it was identified a fraction with high antioxidant activity and purity, which contained gallic acid as the main compound. The gallic acid was found at the amount of 26.54 ± 1.13 mg/g of the dry mass of the pequi fruit peel. Because the quantifications were performed using crude ethanol-water extract, it was suspected that gallic acid was present in a free form. Thus, pequi fruit peel may serve as an attractive alternative of feedstock for natural antioxidant production. Moreover, the results obtained in this study emphasize the value of the pequi plant, and suggests improved opportunities for families that use this fruit`s products.

RESUMOExtratos hidroetanólicos da casca do fruto do pequi foram fracionados a fim de identificar e quantificar o principal antioxidante presente. Frações do extrato foram submetidas ao particionamento líquido-líquido e fracionamento em coluna de sílica gel. As atividades antioxidantes das frações foram monitoradas usando ensaio de redução do radical 2,2-difenil-1-picrilhidrazila e a pureza das frações foi avaliada em cromatografia de camada delgada e cromatografia líquida de alta eficiência (CLAE). A substância com propriedades antioxidantes foi identificada através da análise em sistema de cromatografia líquida associada à espectrometria de massas e foi quantificada em HPLC. Após o fracionamento identificou-se uma fração com alta atividade antioxidante e pureza, contendo ácido gálico como o composto principal. Ácido gálico foi encontrado em concentrações de 26,54 ± 1,13 mg/g de massa seca. Devido às quantificações terem sido realizadas no extrato hidroetanólico bruto, acredita-se que o ácido gálico esteja presente na forma livre. Assim, a casca do fruto pequi pode servir como interessante alternativa de matéria prima para a produção desse antioxidante natural. Além disso, esse resultado enfatiza o valor da planta do pequi e sugere oportunidades para as famílias que utilizam produtos de pequi.

Maternal high-fat diet consumption modulates hepatic lipid metabolism and microRNA-122 (miR-122) and microRNA-370 (miR-370) expression in offspring.

Maternal consumption of a high-fat diet (HFD) during pregnancy and lactation is closely related to hepatic lipid accumulation, insulin resistance and increased serum cytokine levels in offspring and into their adulthood. MicroRNA (miRNA) have been implicated in cholesterol biosynthesis and fatty acid metabolism. We evaluated the modulation of hepatic fatty acid synthesis (de novo), ß-oxidation pathways, and miRNA-122 (miR-122) and miRNA-370 (miR-370) expression in recently weaned offspring (day 28) of mouse dams fed a HFD (HFD-O) or a standard chow (SC-O) during pregnancy and lactation. Compared with SC-O mice, HFD-O mice weighed more, had a larger adipose tissue mass and were more intolerant to glucose and insulin (P< 0·05). HFD-O mice also presented more levels of serum cholesterol, TAG, NEFA and hepatic IκB kinase and c-Jun N-terminal kinase phosphorylation compared with SC-O mice (P< 0·05). Protein levels of fatty acid synthase, acetyl-CoA carboxylase and 3-hydroxy-3-methylglutaryl-CoA reductase were similar in HFD-O and SC-O mice, whereas expression levels of SCD1 mRNA and protein were more abundant in HFD-O mice than in SC-O mice (P< 0·05). Interestingly, mRNA expression levels of the ß-oxidation-related genes ACADVL and CPT1 were decreased in HFD-O mice (P< 0·05). Furthermore, the expression of miR-122 was reduced but that of miR-370 was increased in HFD-O mice compared with that in SC-O mice (P< 0·05). Changes in hepatic lipid metabolism were accompanied by increased mRNA content of AGPAT1 and TAG deposition in HFD-O mice (P< 0·05). Taken together, the present results strongly suggest that maternal consumption of a HFD affects the early lipid metabolism of offspring by modulating the expression of hepatic ß-oxidation-related genes and miRNA that can contribute to metabolic disturbances in adult life.

Low-dose gamma irradiation of food protein increases its allergenicity in a chronic oral challenge.

Few chronic food protein models have described the relationship between allergenicity and the molecular structure of food protein after physical processing. The effect of γ-radiation on the structure of food protein was measured by fluorescence, circular dichroism and microcalorimetry. BALB/c mice were intraperitoneally sensitized and then given non-irradiated and irradiated Con-A by daily gavage for 28days. The tendency to form insoluble amorphous aggregates and partially unfolded species was observed after irradiation. The administration of non-irradiated and irradiated samples at low-dose significantly increased weight loss as well as plasma levels of eotaxin in animals repeatedly exposed to Con-A. Significant lymphocytic infiltrate filling completely the stroma of microvilli and tubular glands was observed in the small intestinal of the group given Con-A irradiated at a low dose. This phenotype was not observed in animals treated with Con-A irradiated at a high dose.

HTR1B and HTR2C in autism spectrum disorders in Brazilian families.

Autism spectrum disorders (ASD) is a group of behaviorally defined neurodevelopmental disabilities characterized by multiple genetic etiologies and a complex presentation. Several studies suggest the involvement of the serotonin system in the development of ASD, but only few have investigated serotonin receptors. We have performed a case-control and a family-based study with 9 polymorphisms mapped to two serotonin receptor genes (HTR1B and HTR2C) in 252 Brazilian male ASD patients of European ancestry. These analyses showed evidence of undertransmission of the HTR1B haplotypes containing alleles -161G and -261A at HTR1B gene to ASD (P=0.003), but no involvement of HTR2C to the predisposition to this disease. Considering the relatively low level of statistical significance and the power of our sample, further studies are required to confirm the association of these serotonin-related genes and ASD.

Clinical pharmacokinetics of lyophilized recombinant human erythropoietin-alpha following single-dose subcutaneous administration in premature newborns.

OBJECTIVE: The aim of this study was to determine pharmacokinetic parameters after subcutaneous administration of a single dose (400 IU/kg) of lyophilized recombinant human erythropoietin-alpha (rhEPOalpha) to preterm newborns. The parameters determined were: maximum concentration (C(max)), time to reach maximum concentration (T(max)), absorption half-life (t(l/2abs)), volume of distribution (Vd), elimination half-life (t(l/2el)), clearance (C(L)), constant of elimination (k(el)) and area under the 0-72 h curve (AUC(0-72). METHODS: The study group comprised 20 premature newborns (eight males and 12 females) delivered in the Teaching Hospital, University of São Paulo. The inclusion criteria were birth weight < 1500 g or gestational age < or = 34 weeks, and clinical and hemodynamic stability. Serum erythropoietin (EPO) concentration was determined before and 1, 4, 6, 12, 24,48 and 72 h after subcutaneous administration of 400 IU/kg rhEPOalpha, and the pharmacokinetic parameters were calculated. RESULTS: There was a significant difference in serum EPO concentration between t72 and t0 (p = 0.001). Mean values (range) of the pharmacokinetic parameters were as follows: C(max), 739.8 (188.0-1390.0) mIU/ml; T(max), 7.7 (4.0-12.0) h; t(l/2abs), 2.9 (0.8-4.8) h, V(d), 0.705 (0.23-1.73) 1/kg; t(l/2e1), 14.9 (8.7-36.1) h; C(L), 0.032 (0.014-0.066) 1/h; k(el), 0.0475 (0.0200-0.0700); and AUC(0-72), 19058.2 (7648.0-34701.5) mIU/ml per h. The Spearman test showed no correlation between the pharmacokinetic parameters analyzed and the characteristics of the population studied. CONCLUSIONS: Studies evaluating the effectiveness of therapy with recombinant human erythropoietin in premature newborns have used various doses, administered at intervals between 24 and 48 h. The kinetics of absorption measured in our study supports the use of 400 IU/kg within an interval of no less than 72 h, together with therapeutic control of the drug and evaluation of the erythropoietic response.

Serum vitamin A levels in patients with ocular lesions attributable to non-complicated malaria in the Brazilian Amazon region.

In order to evaluate the association between serum vitamin A levels and ocular lesions attributable to non-complicated malaria, 200 patients seen consecutively at the Malaria Outpatient Clinic of FUNASA, Manaus, Amazonas, Brazil were included in this study. Ophthalmologic examination consisted of indirect binocular ophthalmoscopy under medicamentous mydriasis, biomicroscopy with a portable slit lamp and measurement of central visual acuity. Vitamin A serum concentration was determined by HPLC, and deficiency was defined as serum values equal to or lower than 0.35 micromol/l. Serum vitamin A values between 0.36 and 0.70 micromol/l were considered as marginal levels. Hypovitaminosis A (

The external calcium-dependent NADPH dehydrogenase from Neurospora crassa mitochondria.

We have inactivated the nuclear gene coding for a putative NAD(P)H dehydrogenase from the inner membrane of Neurospora crassa mitochondria by repeat-induced point mutations. The respiratory rates of mitochondria from the resulting mutant (nde-1) were measured, using NADH or NADPH as substrates under different assay conditions. The results showed that the mutant lacks an external calcium-dependent NADPH dehydrogenase. The observation of NADH and NADPH oxidation by intact mitochondria from the nde-1 mutant suggests the existence of a second external NAD(P)H dehydrogenase. The topology of the NDE1 protein was further studied by protease accessibility, in vitro import experiments, and in silico analysis of the amino acid sequence. Taken together, it appears that most of the NDE1 protein extends into the intermembrane space in a tightly folded conformation and that it remains anchored to the inner mitochondrial membrane by an N-terminal transmembrane domain.

Improving hand hygiene in community healthcare settings: the impact of research and clinical collaboration.

Observation of nursing activities and bacteriological studies undertaken with a sample of nurses employed in a community NHS trust indicated that considerable scope for cross infection existed during domiciliary visits. Poor conditions in patients' homes compromised nurses' ability to perform hand hygiene effectively, increasing risks. A clinical trial indicated that carriage of medically significant bacteria likely to contribute to cross infection could be reduced by applying an antiseptic cream which exhibited residual effectiveness. An audit of hand hygiene throughout the inner city trust indicated the need to pay greater attention to hand hygiene, especially during home nursing visits. The situation was less acute in a rural trust where a second audit was performed for comparative purposes. The motivation of clinical staff to improve hand hygiene precautions was high.

Toxicological evaluation of the hydro-alcohol extract of the dry leaves of Peumus boldus and boldine in rats.

The hydro-alcohol extract of the dry leaves of Peumus boldus and boldine, showed abortive and teratogenic action and changes in the blood levels of bilirubin, cholesterol, glucose, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and urea in rats. The long term administration of the extract and boldine did not cause histological modification during a period of 90 days.

Growth changes induced by gamma radiation on Biomphalaria straminea (Dunker, 1848).

Doses of 60Co gamma radiation with 2.5; 5; 7.5; 10; 15; 20; 25; 30; 35; 40; 45; 50; 55; 60; 80; 160; 320 and 640 Gy were applied to 1, 080 snails Biomphalaria straminea, an intermediate host of Schistosoma mansoni, divided in groups containing 30 mollusks. In addition, 60 non irradiated snails were kept as control. Fifty percent of the population was kept in colonies (allowing cross fertilization) while the other half was maintained in sexual isolation (allowing self fertilization) and during one month their growth was observed through the daily measurement of the shell diameter. Results showed that after 20 Gy doses the growth in shell diameter of irradiated snails was greater than that of the control group after 30 days. At this dose the snail size was the greatest, among all isolated groups. The 80 Gy doses also induced the final shell diameter of isolated snails to be greater then that observed in the control groups. As this effect was most evident among the isolated snails, a possible hormonal role may have been involved in the observed phenomena, which is under investigation with the objective of identifying any future applications that this could have to schistosomiasis control.

Características clínicas de transtorno obsessivo-compulsivo em crianças e adolescentes / Clinical features of obsessive-compulsive disorder in children and adolescents

OBJETIVO: Descrever o quadro clínico, evoluçäo, comorbidade, subtipo de transtorno obsessivo-compulsivo (TOC) e incidencia de patologia familiar em uma amostra clínica de crianças e adolescentes com TOC.CASUISTICA E METODOS: Foram avaliados prospectivamente 40 pacientes menores de 18 anos, do Serviço de Psiquiatria da Infancia e Adolescencia(SEPIA), com TOC, no periodo de junho de 1995 até agosto de 1998, através da entrevista K-SADs, versäo E -IV e escala de sintomas Y-BOCS. Dados de incidencia de patologia familiar foram obtidos por anamnese clínica. Dados de evoluçäo säo referentes ao período em que estiveram em seguimento no SEPIA e, retrospectivamente, desde o início dos sintomas pela história clínica. RESULTADOS: A maioria dos pacientes era do sexo masculino (77,5 por cento), com idade média de início de 10,04 anos. Houve predomínio de subtipo misto de obsessöes e compulsöes, com tipo de sintomas em geral múltiplos e mutáveis ao longo do tempo. Comorbidade foi muito freqüentes nesta amostra (10 por cento). Houve 22,5 por cento de familiares com TOC ou SOC (sintomas obsessivo-compulsivos; subclínico). A evoluçäo foi na maioria crônica flutuante. CONCLUSOES: Trata-se de uma amostra clínica em que os pacientes foram selecionados de uma populaçäo rica em casos psiquiatricos gerais mais graves, por ser um centro de referencia para este atendimento. Portanto há vies quanto ao grau de acometimento e talvez isto tenha influenciado para diferenças em relaçäo a dados de literatura sobre TOC infantil

The 24-kDa iron-sulphur subunit of complex I is required for enzyme activity.

We have cloned the nuclear gene encoding the 24-kDa iron-sulphur subunit of complex I from Neurospora crassa. The gene was inactivated in vivo by repeat-induced point-mutations, and mutant strains lacking the 24-kDa protein were isolated. Mutant nuo24 appears to assemble an almost intact complex I only lacking the 24-kDa subunit. However, we also found reduced levels of the NADH-binding, 51-kDa subunit of the enzyme. Surprisingly, the complex I from the nuo24 strain lacks NADH:ferricyanide reductase activity. In agreement with this, the respiration of intact mitochondria or mitochondrial membranes from the mutant strain is insensitive to rotenone inhibition. These results suggest that the nuo24 complex is not functioning in electron transfer and the 24-kDa protein is absolutely required for complex I activity. This phenotype may explain the findings that the 24-kDa iron-sulphur protein is reduced or absent in human mitochondrial diseases. In addition, selected substitutions of cysteine to alanine residues in the 24-kDa protein suggest that binding of the iron-sulphur centre is a requisite for protein assembly.