Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil / Dieta do cachorro-do-mato (Cerdocyon thous) em duas unidades de conservação da Floresta Amazônica, Brasil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil

Abstract The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Resumo Este estudo objetivou analisar a dieta do cachorro-do-mato, de vida livre, por meio da identificação do conteúdo estomacal de 17 Cerdocyon thous atropelados em duas unidades de conservação da Floresta Amazônica. O conteúdo estomacal foi analisado e os itens alimentares foram quantificados pela frequência de ocorrência (FO) e percentagem de ocorrência (PO). Também foram avaliados os teores de matéria seca (MS), proteína bruta (PB), fibra bruta (FB), extrato etéreo (EE) e, matéria mineral (MM). Foram estimados o extrativo não-nitrogenado (ENN), a energia metabolizável (EM) e a necessidade energética de manutenção. A composição da dieta do cachorro-do-mato apresentou 29 itens alimentares dos diferentes grupos taxonômicos, tendo uma maior diversidade de itens de origem animal (n=22), ainda que a maior FO tenha sido de gramíneas (Poaceae) (41.18%). Dentre os itens de origem animal, 21% eram mamíferos, 18% répteis, 10% anfíbios, 9% invertebrados e 3% aves. Determinou-se um alto teor de FB (62.76%). A média do ENN e da MS foi 5.91% e 141.82 kcal/100g, respectivamente. A média da energia de manutenção foi 447.01 kcal/dia. Estes achados sugerem que o cachorro-do-mato encontrado na Bacia Amazônica é um animal generalista com uma dieta onívora, se alimentando de gramíneas, frutos, insetos, serpentes, anfíbios, aves e pequenos mamíferos, portanto com o mesmo hábito alimentar relatado a esta espécie quando encontrada em outros biomas brasileiros.

Conductive carbon paint: An efficient way to electrodeposit metallic nanowires by using porous membranes.

The present work demonstrates that conductive carbon paint, used for sample preparation in electron microscopy, can be a more straightforward and as-effective substitute for the metallic layer deposition usually used for the electrodeposition of metallic nanowires within porous membranes. AFM images demonstrated the good surface quality of the carbon layer. Raman spectroscopy confirmed the high crystallinity of carbon and high density of π-electrons. The electrical conductivity of the carbon layer was estimated using the linear sweep voltammetry technique. This new cathode was employed to grow continuous (Ni) and composition-modulated (Ni/Cu) nanowires within alumina templates, starting from aqueous solutions of Ni2+ and Cu2+ mixed salts. The obtention of metallic copper and nickel, and their separation can be readily observed by scanning electron microscopy and elemental mapping by EDS.

Diet of crab-eating fox (Cerdocyon thous) in two conservation units of the Amazon rainforest, Brazil.

The present study aimed to evaluate the diet of the free-living crab-eating fox by identifying the stomach contents of the 17 crab-eating foxes (Cerdocyon thous) roadkilled in two conservation units, both located in the Amazon rainforest. The food items were quantified by frequency of occurrence (FO) and percentage of occurrence (PO). The stomach contents were analysed for dry matter (DM), crude protein (CP), crude fibre (CF), ether extract (EE), and mineral matter (MM). Nitrogen-free extractives (NFE), metabolisable energy (ME) values, as well as the energy need for maintenance were estimated. The composition of the diet for the crab-eating fox presented 29 food items from the different taxonomic groups, with a greater diversity of items of animal origin (n=22), although the highest frequency of occurrence was gramineae (Poaceae) (41.18%). Among the items of animal origin, 21% were mammals, 18% reptiles, 10% amphibians, 9% invertebrates and 3% birds. A high content of CF (62.76%) were determined. Nitrogen-free extractive and dry matter averages were 5.91% and 141.82 kcal/100g, respectively. The average maintenance energy was 447.01 kcal/day. These findings suggesting that the crab-eating foxes have a generalist diet with an omnivorous diet in the Amazon basin, feeding on gramineae, fruits, insects, snakes, amphibians, birds and small mammals and have the same feeding habit that present in other Brazilian biomes.

Germination of small tropical seeds has distinct light quality and temperature requirements, depending on microhabitat.

The coexistence of plant species in tropical rainforests is related to specific abiotic resources, varying according to the occurrence microhabitat of each species. Light quality is the main abiotic factor influencing germination of small seeds; however, studies often do not discriminate its effect from that of light irradiance. This study compared specific requirements for seed germination of ten small-seeded species, with restricted occurrence in only one of three contrasting microhabitats: forest understorey, edge of clearings and open areas. Laboratory experiments were carried out to test temperature regime (constant or fluctuating), light quality (R:FR) and light irradiance (PAR), which reproduce high and low conditions commonly found in the microhabitats. Seed germination of all species occurred between 20 and 30 °C, only seeds of open area species were able to germinate at 35 °C and no species required alternating temperatures to germinate. Irrespective of species and microhabitat, a decrease in the R:FR reduced the germination percentage; however, there were differences in the capacity to germinate at low R:FR. The values of R:FR50% were higher for open area and edge species (0.441-0.345) than for understorey species (0.181-0.109), with few exceptions. For all species and most of the tests, germination was not influenced by PAR. Light quality is the most important light signal for germination of small seeds; irradiance has little effect. Our results suggest two distinct patterns of germination for small-seeded species: open area and edge species are light-demanding and require high R:FR to germinate, while understorey species are shade-tolerant and germinate at low R:FR. These differences are responsible for distinct microhabitat occurrence and help to explain the coexistence of species in tropical forests.

Experimental cerebral palsy causes microstructural brain damage in areas associated to motor deficits but no spatial memory impairments in the developing rat.

INTRODUCTION: Cerebral palsy (CP) is the major cause of motor and cognitive impairments during childhood. CP can result from direct or indirect structural injury to the developing brain. In this study, we aimed to describe brain damage and behavioural alterations during early adult life in a CP model using the combination of maternal inflammation, perinatal anoxia and postnatal sensorimotor restriction. METHODS: Pregnant Wistar rats were injected intraperitoneally with 200 µg/kg LPS at embryonic days E18 and E19. Between 3 and 6 h after birth (postnatal day 0 - PND0), pups of both sexes were exposed to anoxia for 20 min. From postnatal day 2 to 21, hindlimbs of animals were immobilized for 16 h daily during their active phase. From PND40, locomotor and cognitive tests were performed using Rota-Rod, Ladder Walking and Morris water Maze. Ex-vivo MRI Diffusion Tensor Imaging (DTI) and Neurite Orientation Dispersion and Density Imaging (NODDI) were used to assess macro and microstructural damage and brain volume alterations induced by the model. Myelination and expression of neuronal, astroglial and microglial markers, as well as apoptotic cell death were evaluated by immunofluorescence. RESULTS: CP animals showed decreased body weight, deficits in gross (rota-rod) and fine (ladder walking) motor tasks compared to Controls. No cognitive impairments were observed. Ex-vivo MRI showed decreased brain volumes and impaired microstructure in the cingulate gyrus and sensory cortex in CP brains. Histological analysis showed increased cell death, astrocytic reactivity and decreased thickness of the corpus callosum and altered myelination in CP animals. Hindlimb primary motor cortex analysis showed increased apoptosis in CP animals. Despite the increase in NeuN and GFAP, no differences between groups were observed as well as no co-localization with the apoptotic marker. However, an increase in Iba-1+ microglia with co-localization to cleaved caspase 3 was observed. CONCLUSION: Our results suggest that experimental CP induces long-term brain microstructural alterations in myelinated structures, cell death in the hindlimb primary motor cortex and locomotor impairments. Such new evidence of brain damage could help to better understand CP pathophysiological mechanisms and guide further research for neuroprotective and neurorehabilitative strategies for CP patients.

Single Layer and Double Layer Duraplasty for the Management of Persistent Cerebrospinal Fluid Rhinorrhea Following Frontal Sinus Trauma: A Retrospective Study.

ABSTRACT: Persistent cerebrospinal fluid (CSF) rhinorrhea is an infrequent complication of frontal sinus trauma. It often require craniotomy for dural repair and obliteration of sinus with well vascularized pericranial flap. The multilayer vascularized techniques have gained popularity over the years owing to low rate of postoperative CSF leaks. The study retrospectively analyzed 25 frontal sinus trauma patients (20 males, 5 females) of mean age 32 ±â€Š10.23 years with persistent CSF rhinorrhea from January 2011 to December 2018. All patients underwent frontal craniotomy and duraplasty was done with pericranial flap alone in 14 patients (Group I) and in combination with fascia lata graft in 11 patients (Group II). The clinical outcomes of single and double layer dural reconstruction techniques in management of CSF leak were analyzed. The size of defects ranged between 3 to 28 mm. Significant association was present between defect size and reconstruction technique with large defects underwent double layer repair. Two patients developed CSF leak following repair with pericranial flap alone. The rate of successful closure of CSF fistula was 100% with double layer reconstruction technique. However, statistically no significant association was noted between postoperative CSF leak and reconstruction technique. There was no evidence of meningitis, subdural hematoma, intracranial abscess, and mucocele formation in all patients during follow-up of 50.7 ±â€Š23.6 months. To conclude, pericranial flap is a reliable reconstruction material for management of post traumatic frontal sinus CSF leaks. Dual flap technique that combine the use of fascia lata provides comprehensive solution of CSF fistula repair.

Previous adaptation triggers distinct molecular pathways and modulates early and long-term neuroprotective effects of pregnancy swimming preventing neonatal hypoxia-ischemia damage in rats.

Neonatal hypoxia-ischemia (HI) is one of the main causes of neurological damage in newborns. Pregnancy swimming (PS) alters brain maturation and has neuroprotective effects following HI; however, variables such as timing play a decisive role in its effects. Prior to mating, we tested if adaptation of female rats to a tank filled with water at 32 °C for 7 days before mating, modulates PS benefits. After mating, rats swam 20 min/day or remained in standard cages. Seven-day-old pups were subjected to HI (right common carotid artery occlusion followed by FiO2 8% for 60 min). Animals were divided into 8 experimental groups, adaptation, swimming and injury. Astrocytic reactivity, apoptosis-related proteins, neurotrophins and cell survival markers expression were assessed in the hippocampus 24 h after HI. From PND45, animals performed behavioral tests followed by histological assessment. Three-way ANOVA showed a significant increase in astrogliosis only in non-adapted HI animals. Swimming decreased apoptotic cell death despite adaptation period in both exercised groups. Cylinder evidenced HI impairments; no effect of swimming or adaptation period were observed. In the open field, only HI animals whose mothers had been adapted had increased locomotion; moreover, swimming reversed HI damage. Hemisphere and hippocampus were preserved only in the HI group whose mothers swam before mating, suggesting a preconditioning effect mediated by the adaptation. In summary, adaptation period plays a major role in the mechanisms involving neuroprotection afforded by PS and needs to be further explored in future studies involving damage to the neonatal brain.

Preventive and therapeutic effects of environmental enrichment in Wistar rats submitted to neonatal hypoxia-ischemia.

Environmental enrichment (EE) at early stages of neurodevelopment attenuates HI-induced behavioral, histological and cellular damage. However, the effects of EE exposure during gestational or early postnatal period and the possible influence of sexual dimorphism on EE protection are not fully understood. Present study evaluated the effects of pre-natal and postnatal EE, as well as their combination, in male and female rats submitted to neonatal HI at postnatal day (PND) 3. Wistar rats were housed in EE or in standard condition (SC) during all pregnancy. At PND1, the litters were randomly allocated to the same prenatal environment during lactation (SC + SC or EE + EE) or housed in a new environment until weaning (SC + EE or EE + SC). Behavioral tasks were performed from PND 60-75. Then, animals were euthanized for biochemical and histological analysis. Prenatal and early postnatal EE alone improved performance of HI males in the Water Maze spatial memory task, while HI females were most benefited from early postnatal stimulation. Moreover, EE attenuated HI-induced lower anxiety-like behavior in rats of both sexes and decreased hyperlocomotion in HI females. Hippocampus tissue preservation and higher VEGF and TrkB levels were observed in all HI groups exposed to EE. Interestingly, HI males exposed to prenatal or postnatal EE alone exhibited higher GFAP levels and additional tissue preservation. Therefore, both prenatal and early postnatal environmental enrichment cause attenuation of HI-induced impairments, revealing their preventive and therapeutic actions, possibly due to VEGF and astrocyte activity; some of these effects are sex-specific.

A participação da família na reabilitação psicossocial do sujeito emsofrimento psíquico / Family involvement in psychosocial rehabilitation of individuals under psychological suffering

Objetivo: conhecer a participação da família na reabilitação psicossocial do sujeito em sofrimento psíquico. Métodos: trata-se de uma pesquisa qualitativa, exploratória e descritiva, realizada em 2012. Os sujeitos do estudo foram seis familiares de usuários de um Centro de Atenção Psicossocial (CAPS). Foi utilizada a análise temática conforme Minayo, que tem como foco encontrar núcleos de sentido na comunicação apresentada pelo material estudado. Resultados: emergiram desse estudo dois eixos temáticos: A inserção do sujeito em sofrimento psíquico na sociedade sob a ótica da família e A participação da família na reabilitação psicossocial do sujeito em sofrimento psíquico. Conclusão: há um certo isolamento do usuário no serviço substitutivo. É importante que os profissionais de saúde invistam na autonomia dos sujeitos, contribuindo para o conhecimento do usuário e sua família acerca dos seus direitos e da proposta da reforma psiquiátrica. Colaborando com o aumento do compromisso da família neste processo.

Objective: to now the family's participation in psychosocial rehabilitation of the individual underpsychological distress. Methods: this is a qualitative, exploratory and descriptive study, held in2012. The study subjects were relatives of individuals with psychological distress, users of aPsychosocial Care Center (CAPS). The thematic analysis according to Minayo was used, whose focusis to find nuclei of meaning in the communication presented by material studied. Results: this studyemerged the themes: The insertion of the subject in psychological distress in society from theperspective of the family and the family's participation in psychosocial rehabilitation of the subject in psychological distress. Conclusions: there’s a certain isolation used in substitute service. It isunderstood that it is important for health professionals bet on education, increasing the user'sknowledge and his family about their rights and psychiatric reform proposal. This may collaboratewith the commitment of the family in this process.

In situ analysis of copper electrodeposition reaction using unilateral NMR sensor.

The uses of high-resolution NMR spectroscopy and imaging (MRI) to study electrochemical reactions in situ have greatly increased in the last decade. However, most of these applications are limited to specialized NMR laboratories and not feasible for routine analysis. Recently we have shown that a bench top, time domain NMR spectrometer can be used to monitor in situ copper electrodeposition reaction and the effect of Lorentz force in the reaction rate. However these spectrometers limit the cell size to the magnet gap and cannot be used with standard electrochemical cells. In this paper we are demonstrating that unilateral NMR sensor (UNMR), which does not limit sample size/volume, can be used to monitor electrodeposition of paramagnetic ions in situ. The copper electrodeposition reaction was monitored remotely and in situ, placing the electrochemical cell on top of the UNMR sensor. The Cu(2+) concentration was measured during three hours of the electrodeposition reactions, by using the transverse relaxation rate (R2) determined with the Carr-Purcell-Meiboom-Gill pulse sequence. The reaction rate increased fourfold when the reaction was performed in the presence of a magnetic field (in situ), in comparison to the reactions in the absence of the magnetic field (ex situ). The increase of reaction rate, in the presence of the UNMR magnet, was related to the magneto hydrodynamic force (FB) and magnetic field gradient force (F∇B). F∇B was calculated to be one order of magnitude stronger than FB. The UNMR sensor has several advantages for in situ measurements when compared to standard NMR spectrometers. It is a low cost, portable, open system, which does not limit sample size/volume and can be easily be adapted to standard electrochemical cells or large industrial reactors.

Knowledge and behavior among health professionals in relation to cadaveric organ donation and transplantation: a questionnaire-based analysis in Portuguese hospitals.

OBJECTIVE: To assess knowledge and behavior relative to cadaveric organ donation and transplantation among physicians and nurses working at nine Portuguese hospitals. METHODS: This prospective study between May and October 2009 included 495 questionnaires that were returned among 840 that were delivered, a 59% response rate. The transplant questionnaire comprised three main sections: section 1, sociodemographic-professional variables of the participants with nine questions; section 2, generic questions on organ donation and transplantation with 12 questions; and section 3 evaluated knowledge with 20 specific questions on organ donation and transplantation. RESULTS: Sixty-one percent of the participants were females, with 40% of the overall cohort between 31 and 40 years old; 62% married; 63% working in the emergency department; and 78% nurses. Although 78% of the participants stated that they had undergone specific training on organ donation and transplantation, 62% felt that they needed more training. The reasons evoked for the low rate of donation included difficulties in initiating the process of organ donation, in diagnosing cerebral death and in obtaining necessary human resources. There were no significant differences in knowledge between physicians and nurses concerning specific organ donation and transplantation issues. Having had specific training on organ donation and transplantation was significantly associated with correct answers to several questions, including those pertaining to knowledge of the national legislation on organ donation and transplantation. CONCLUSIONS: The present study showed a lack of specific knowledge and training on organ donation and transplantation issue. It also suggested that for hospital staff to successfully act as initiators of the organ procurement process, more information and education are necessary. This effort could significantly increase the rate of organ donation and transplantation in Portugal.

Differential expression of alpha-2,3-sialyltransferases and alpha-1,3/4-fucosyltransferases regulates the levels of sialyl Lewis a and sialyl Lewis x in gastrointestinal carcinoma cells.

Sialyl Lewis x and sialyl Lewis a expression depends on sialyltransferases and fucosyltransferases. In this study, we screened for major variations of sialyltransferases and fucosyltransferases involved in the synthesis and regulation of sialyl Lewis x and sialyl Lewis a epitopes in gastrointestinal carcinoma cells. Our results show that expression of ST3Gal IV in several gastrointestinal cell lines is correlated with the expression of sialyl Lewis x at the cell surface. ST3Gal IV overexpressed in the gastric MKN45 cell line, showed exclusive enzymatic activity towards glycoproteins containing terminal Galbeta1-4GlcNAc structure. On the other hand, when ST3Gal III was overexpressed in MKN45, an increase in the expression levels of both sialyl Lewis epitopes was observed. ST3Gal III and ST3Gal IV lead to de novo synthesis of sialyl Lewis x determinant on different molecular weight glycoproteins of MKN45 cells suggesting that each enzyme used different substrates within the available glycoproteome. The final glycosylation step in sialyl Lewis x and sialyl Lewis a biosynthesis in MKN45 cell line was shown to be associated to FUT5, which efficiently fucosylated sialyl Lewis precursors on glycoproteins. Moreover we demonstrate that the expression of sialyl Lewis epitopes in the MKN45 was induced by cell confluence, which can be regarded as a model to study altered glycosylation during tumour progression. This increase was observed together with an increase in mRNA levels of ST3GAL3, FUT5 and FUT6, and a decrease in FUT4 transcript levels in MKN45 confluent cells, suggesting a possible control at the transcriptional level.

Proteolysis in Painho de Portalegre dry fermented sausage in relation to ripening time and salt content.

The effect of salt addition (3% and 6% in the final product) on the shelf-life related physicochemical characteristics and proteolysis profile during the ripening period of a Portuguese dry fermented sausage "Painho de Portalegre", were evaluated. The product with 6% salt concentration had low a(w) and pH values at most ripening periods evaluated, due to the influence of NaCl on the water binding capacity of the protein structure and to the low ammonia accumulation, respectively. Similar changes were observed for total basic volatile nitrogen (TBVN), free amino acid nitrogen (FAAN) and non-protein nitrogen (NPN) fractions in both products. After a clear increase during the first days of the processing phase, all the initial rates slowed down with some fluctuation in FAAN and NPN. In relation to small peptides and free amino acid accumulation, the major differences between the tested formulations were mainly observed on distinct profiles rather than on overall concentrations.

Design, synthesis and activity against Trypanosoma cruzi of azaheterocyclic analogs of megazol.

This study describes the design, synthesis and trypanocidal evaluation of new azaheterocyclic derivatives (4-8). These compounds were designed as megazol (1) analogs based on bioisosterism tools and were synthesized to investigate the possible pharmacophoric contribution of the 1,2,4-triazole nucleus, the position of the heterocyclic nucleus and presence of the nitro group, to the activity against the bloodstream trypomastigote forms of Trypanosoma cruzi. The most potent compound was 6, a nitro derivative obtained by substitution of a thiadiazole by a triazole ring and by moving the nitro group from C-5 position, as in 1, to the C-4 position. Finally, we have used semi-empirical theoretical calculations to discuss the correlation of some stereo electronic properties with biological activity in an attempt to understand the possible mechanism of action of the designed series of compounds.

Differences in peripheral blood lymphocyte phenotypes between Helicobacter pylori-positive children and adults with duodenal ulcer.

The immunological mechanisms involved in the development of duodenal ulcer, especially in childhood, are unclear. Helicobacter pylori-positive children and adults, with and without duodenal ulcer, were therefore compared with respect to CD4(+) T-cells, and CD8(+) T-cells, B-cells and B1a-cells, as well as cell activation (CD4(+)/HLA-DR(+) and CD8(+)/HLA-DR(+)) and co-stimulatory (CD4(+)/CD28(+) and CD8(+)/CD28(+)) markers, in peripheral blood. Children with and without duodenal ulcer differed significantly. In particular, there was a phenotypic change in CD8(+) T-cells from children with ulcer that involved a 200% increase in the number of CD8(+)/HLA-DR(+) cells/mm(3) and a decrease of 34.2% in the number of CD8(+)/CD28(+) cells/mm(3). This phenotype of chronically activated memory CD8(+) T-cells, which has also been observed in patients with AIDS and tuberculosis, is associated with disease severity and progression. A lower frequency of B1a-cells was also observed in the group of children with ulcer. Conversely, no difference between infected adults with and without ulcer was observed, but the percentage of CD4(+)/HLA-DR(+) cells was lower in adults with ulcer, suggesting that a down-regulated immune response may play a role in the development of duodenal ulcer in adults. Gastric inflammation correlated positively with CD4(+) and chronically activated CD4(+) T-cells in children and adults without duodenal ulcer, respectively. These results suggest that there are differences in the immunophenotyping profile between H. pylori-positive children and adults with duodenal ulcer, indicating the possibility of distinct immune mechanisms in the development of the disease according to age.

Phenotypic study of peripheral blood lymphocytes and humoral immune response in Helicobacter pylori infection according to age.

There are differences between children and adults in certain aspects of the Helicobacter pylori (HP) infection, among them the lower titre of IgG antibodies anti-HP in the former group. Thus, we investigated by means of flow cytometry CD4+/CD3+ (CD4+T), CD8+/CD3+ (CD8+T) and CD19+/CD3- (B) cells, activation/co-stimulatory markers (CD4+/HLA-DR+, CD4+/CD28+, CD8+/HLA-DR+ and CD8+/CD28+) and by means of ELISA IgG anti-HP antibodies in the peripheral blood from HP-positive and -negative children and adults. An increased CD4+/CD28+ and CD8+/CD28+ percentage and number of CD4+/CD3+ cells were seen in infected adults. Conversely, no difference was observed between infected and noninfected children, but when they were stratified by age, an increased CD4+/CD28+ cell percentage was seen in the HP-positive group older than 10 years. The mean level of IgG anti-HP was lower in younger infected children, increased with age and correlated with CD4+ cells. Our data suggest that the immune response to HP infection vary according to the age. Low percentage of activated CD4+ cell may contribute to the lower level of serum IgG anti-HP observed in younger infected children. In addition, the CD4+ cell participation during the infection seems to begin after 10 years old, when the immune response becomes similar to that seen in adults.

Effect of the pH of growth on the survival of Lactobacillus delbrueckii subsp. bulgaricus to stress conditions during spray-drying.

AIMS: The aim of this study was to optimize survival of Lactobacillus delbrueckii subsp. bulgaricus during spray-drying and subsequent storage through optimizing the pH of growth conditions. METHODS AND RESULTS: Cell concentrates previously grown without or with pH controlled were spray-dried and stored at 20 degrees C and heat treated at 57 degrees C. Cells grown under noncontrolled pH were more resistant to both drying and heating than cells grown under controlled pH but no significant differences were observed during storage. The intracellular proteins profile of cells grown under both conditions was studied by two-dimensional SDS-polyacrylamide gel electrophoresis. Eight proteins were identified using automated mass spectrometry (MS) and tandem mass spectrometry (MS/MS) data acquisition. Of the identified proteins, only cochaperonin GroES corresponded to a known heat shock protein (HSP). The other proteins identified are proteins involved in glycolysis. For cells grown under noncontrolled pH the expression of the Hsp70, GroES and GroEL, measured by Western blotting, was enhanced. CONCLUSIONS: The higher resistance of cells grown under noncontrolled pH correlates with the enhanced production of heat shock proteins. SIGNIFICANCE AND IMPACT OF THE STUDY: Growth of L. bulgaricus under controlled pH (commonly used by the starter cultures production industry) results in cells more sensitive to stresses frequently encountered by the cells during starter cultures preparation/storage/utilization.

Effect of various growth media upon survival during storage of freeze-dried Enterococcus faecalis and Enterococcus durans.

AIMS: The effects of three different growth media (MRS, M17 and Lee's) on survival during freeze-drying and subsequent storage of six strains of Enterococcus faecalis and two strains of E. durans were investigated. METHODS AND RESULTS: Distinct Enterococcus spp. strains were grown on M17, MRS and Lee's broth, freeze-dried and stored at 20 degrees C in air under darkness. At regular intervals throughout storage, freeze-dried samples were rehydrated and then plated on M17 agar. CONCLUSIONS: A higher survival rate during storage of dried E. durans was obtained when growth occurred in MRS. The same effect was not observed, however, for the majority of E. faecalis strains, which clearly survived better in the dried state when this organism had been grown in M17 or Lee's medium. SIGNIFICANCE AND IMPACT OF STUDY: The survival of the dried Enterococcus spp. tested during storage was shown to be strain-specific and dependent on the growth medium.

Ethical guidance on human embryonic and fetal tissue transplantation: a European overview.

This article presents an overview of regulations, guidelines and societal debates in eight member states of the EC about a) embryonic and fetal tissue transplantation (EFTT), and b) the use of human embryonic stem cells (hES cells) for research into cell therapy, including 'therapeutic' cloning. There appears to be a broad acceptance of EFTT in these countries. In most countries guidance has been developed. There is a 'strong' consensus about some of the central conditions for 'good clinical practice' regarding EFTT. International differences concern, amongst others, some of the informed consent issues involved, and the questions whether an intermediary organisation is necessary, whether the methods of abortion may be influenced by the possible use of EFT, and whether EFTT should only be used for the experimental treatment of rare disorders. The potential use of hES cells for research into cell therapy has given a new impetus to the debate about (human) embryo research. The therapeutic prospects with regard to the retrieval and research use of hES cells appear to function as a catalyst for the introduction of less restrictive regulations concerning research with spare embryos, at least in some European countries. It remains to be seen whether the prospect of treating patients suffering from serious disorders with transplants produced by therapeutic cloning will decrease the societal and moral resistance to allowing the generation of embryos for 'instrumental' use.