Assessment of a tannin-based organic polymer to harvest Chlorella vulgaris biomass from swine wastewater digestate phycoremediation.

This study investigated the efficiency of an organic tannin polymer alone or amended with polyacrylamide to harvest Chlorella vulgaris biomass grown in a laboratory-scale photobioreactor treating swine wastewater digestate. The effect of biomass concentration, tannin (TAN) dosages and changes in pH were evaluated in jar test experiments. Among the TAN concentrations tested (11, 22, 44, 89, 178 mg L(-1)), 11 mg L(-1) showed the highest biomass recovery (97%). The highest coagulation/ flocculation efficiencies were obtained at pH 5 to 7. Flocculation efficiency improved from 50 to 97% concomitant with the increasing biomass concentrations from 45 to 165 mg L(-1), respectively. Recovery efficiencies above 95% were achieved with the same TAN dosage (11 mg L(-1)) irrespective of the concentration of organic carbon present (75 to 300 mg TOC L(-1)). Overall, the results suggest that TAN could become an interesting alternative choice of non-toxic organic polymer for harvesting Chlorella sp. from organic-rich wastewater.

Factors associated with the time taken for diagnosis of amyotrophic lateral sclerosis (ALS) in Brazil. An online population-based inquiry. / Factores asociados al tiempo necesario para el diagnóstico de esclerosis lateral amiotrófica (ELA) en Brasil. Una encuesta poblacional en línea.

OBJECTIVE: This study evaluated factors associated with the time, in months, between the onset of symptoms and the diagnosis (time taken for diagnosis) of ALS for patients in Brazil, in the year 2014. PATIENTS AND METHODS: An electronic questionnaire composed of 38 questions was developed and applied through internet-based social networks of patients. From the 210 replies, 194 were considered (86 from women, 108 from men). Most respondents were 51 to 60 years old. The Mann-Whitney test was used to compare the time taken for diagnosis between the strata of the sample. RESULTS: The mean time taken for diagnosis was 14.21 (±16.87) months. There was a statistically significant difference only for higher education conditions (p = 0.009) and low education status (p = 0.042). There was no statistically significant difference between sexes, bulbar onset, age groups, and the presence of spouse, or 'partnership with ALS patients associations or exchange of experiences'. CONCLUSION: These data suggest that the time taken for diagnosis of ALS is influenced by socioeconomic conditions that promote access to information and/or health services.

TITLE: Factores asociados al tiempo necesario para el diagnóstico de esclerosis lateral amiotrófica (ELA) en Brasil. Una encuesta poblacional en línea.Objetivo. Este estudio evaluó los factores asociados con el tiempo, en meses, entre el inicio de los síntomas y el diagnóstico (tiempo necesario para el diagnóstico) de esclerosis lateral amiotrófica (ELA) de los pacientes en Brasil en 2014. Pacientes y métodos. Se elaboró un cuestionario electrónico compuesto por 38 preguntas y se aplicó a través de redes sociales de pacientes basadas en Internet. De las 210 respuestas, se consideraron 194 (86 de mujeres y 108 de hombres). La mayoría de los encuestados tenía entre 51 y 60 años. Se utilizó la prueba de Mann-Whitney para comparar el tiempo transcurrido hasta el diagnóstico entre los estratos de la muestra. Resultados. El tiempo medio transcurrido hasta el diagnóstico fue de 14,21 (±16,87) meses. Hubo una diferencia estadísticamente significativa sólo para las condiciones de educación superior (p = 0,009) y bajo nivel educativo (p = 0,042). No hubo diferencias estadísticamente significativas entre sexos, inicio bulbar, grupos de edad y presencia de cónyuge, o colaboración con asociaciones de pacientes con ELA o intercambio de experiencias. Conclusión. Estos datos sugieren que el tiempo que se tarda en diagnosticar la ELA está influido por las condiciones socioeconómicas que favorecen el acceso a la información y/o a los servicios sanitarios.

Gene expression and molecular modeling of the HSP104 chaperone of Trypanosoma cruzi.

Heat shock protein (HSP) 104 is a highly conserved molecular chaperone that catalyzes protein unfolding, disaggregation and degradation under stress conditions. We characterized HSP104 gene structure and expression in Trypanosoma cruzi, a protozoan parasite that causes Chagas' disease. The T. cruzi HSP104 is an 869 amino-acid protein encoded by a single-copy gene that has the highest sequence similarity (76%) with that of T. brucei and the lowest (23%) with that of the human protein. HSP104 transcripts were detected at room temperature, and levels increased after incubation at 37° or 40°C. The HSP104 protein was found at low levels in non-heat-shocked cells, and accumulated continuously up to 24 h at elevated temperatures. We developed a predicted structural model of hexameric T. cruzi HSP104, which showed some conserved features.

Cartilage tissue engineering using electrospun PCL nanofiber meshes and MSCs.

Mesenchymal stem cells (MSCs) have been recognized for their ability to differentiate into cells of different tissues such as bone, cartilage, or adipose tissue, and therefore are of great interest for potential therapeutic strategies. Adherent, colony-forming, fibroblastic cells were isolated from human bone marrow aspirates, from patients undergoing knee arthroplasties, and the MSCs phenotype characterized by flow cytometry. Afterward, cells were seeded onto electrospun polycaprolactone nanofiber meshes and cultured in a multichamber flow perfusion bioreactor to determine their ability to produce cartilagineous extracellular matrix. Results indicate that the flow perfusion bioreactor increased the chondrogenic differentiation of hBM-MSCs, as confirmed either by morphological and RT-PCR analysis. Cartilage-related genes such as aggrecan, collagen type II, and Sox9 were expressed. ECM deposition was also detected by histological procedures. Collagen type II was present in the samples, as well as collagen type I. Despite no statistically significant values being obtained for gene expression, the other results support the choice of the bioreactor for this type of culture.

Comparison of bioaugmentation and biostimulation for the enhancement of dense nonaqueous phase liquid source zone bioremediation.

Two 11.7-m(3) experimental controlled release systems (ECRS), packed with sandy model aquifer material and amended with tetrachloroethene (PCE) dense nonaqueous phase liquid (DNAPL) source zone, were operated in parallel with identical flow regimes and electron donor amendments. Hydrogen Releasing Compound (Regenesis Bioremediation Products, Inc., San Clemente, California), and later dissolved lactate, served as electron donors to promote dechlorination. One ECRS was bioaugmented with an anaerobic dechlorinating consortium directly into the source zone, and the other served as a control (biostimulated only) to determine the benefits of bioaugmentation. The presence of halorespiring bacteria in the aquifer matrix before bioaugmentation, shown by nested polymerase chain reaction with phylogenetic primers, suggests that dechlorinating catabolic potential may be somewhat widespread. Results obtained corroborate that source zone reductive dechlorination of PCE is possible at near field scale and that a system bioaugmented with a competent halorespiring consortium can enhance DNAPL dissolution and dechlorination processes at significantly greater rates than in a system that is biostimulated only.

Conditioned medium as a strategy for human stem cells chondrogenic differentiation.

Paracrine signalling from chondrocytes has been reported to increase the synthesis and expression of cartilage extracellular matrix (ECM) by stem cells. The use of conditioned medium obtained from chondrocytes for stimulating stem cells chondrogenic differentiation may be a very interesting alternative for moving into the clinical application of these cells, as chondrocytes could be partially replaced by stem cells for this type of application. In the present study we aimed to achieve chondrogenic differentiation of two different sources of stem cells using conditioned medium, without adding growth factors. We tested both human bone marrow-derived mesenchymal stem cells (hBSMCs) and human Wharton's jelly-derived stem cells (hWJSCs). Conditioned medium obtained from a culture of human articular chondrocytes was used to feed the cells during the experiment. Cultures were performed in previously produced three-dimensional (3D) scaffolds, composed of a blend of 50:50 chitosan:poly(butylene succinate). Both types of stem cells were able to undergo chondrogenic differentiation without the addition of growth factors. Cultures using hWJSCs showed significantly higher GAGs accumulation and expression of cartilage-related genes (aggrecan, Sox9 and collagen type II) when compared to hBMSCs cultures. Conditioned medium obtained from articular chondrocytes induced the chondrogenic differentiation of MSCs and ECM formation. Obtained results showed that this new strategy is very interesting and should be further explored for clinical applications.

Microbiological studies of abscesses complicating Bothrops snakebite in humans: a prospective study.

Forty patients with abscesses at the site of Bothrops bite were evaluated. Ten brought the snake responsible for the bite, Bothrops jararaca in all cases. On admission a sample was collected from each case for aerobic and anaerobic culture, and no abscesses drained spontaneously. None of the patients had received antibacterial therapy at this time. The bacteria most frequently isolated were Morganella morganii (23 patients), Providencia rettgeri (7) Enterobacter sp. (4), Escherichia coli (3), group D streptococcus (11) and Bacteroides sp. (6). All enterobacteria tested were sensitive to chloramphenicol, aminoglycosides and cefotaxime. Group D streptococci were sensitive to chloramphenicol, ampicillin and penicillin G, whereas anaerobic bacteria were sensitive to chloramphenicol and tetracycline. These results suggest that in cases of abscess formation secondary to Bothrops bite, chloramphenicol alone is a good choice, being both safe and cheap.

Preparative purification of tyrosinamide N-linked oligosaccharides.

N-linked oligosaccharides from glycoproteins can be either analyzed on a sub-nanomole scale or preparatively purified on a multi-micromole scale. Each goal necessitates a unique analytical strategy often involving oligosaccharide derivatization to enhance separation and detection. Tyrosinamide-oligosaccharides were developed to facilitate the preparative purification of N-linked oligosaccharides. These have found many uses in oligosaccharide remodeling, in the preparation of neoglycoconjugates, in developing receptor probes, and even as analytical standards in chromatography. This review discusses progress in the preparation of tyrosinamide-oligosaccharides from different glycoproteins and their utility in glycobiology research.

Evidence of central nervous system involvement in Watson syndrome.

In 1967, Watson described 3 families with an autosomal dominant condition characterized by pulmonary valvular stenosis, café-au-lait sports, and short stature. Presumed hamartomatous lesions have been observed in neurofibromatosis type I, but they were not reported to date in Watson syndrome. We report another family with Watson syndrome, in which 1 patient manifested increased intensity T2-weighted lesions on magnetic resonance imaging similar to those occurring in neurofibromatosis type I and possibly hamartomas. This finding demonstrates the overlap between neurofibromatosis type I and Watson syndrome and supports the hypothesis that those conditions are allelic or, less likely, that the gene that determines the Watson phenotype is very closely linked to the neurofibromatosis type I locus.

Structural characterization of Botryosphaeran: a (1-->3;1-->6)-beta-D-glucan produced by the ascomyceteous fungus, Botryosphaeria sp.

The exopolysaccharide, Botryosphaeran, produced by the ligninolytic, ascomyceteous fungus Botryosphaeria sp., was isolated from the extracellular fluid by precipitation with ethanol, and purified by gel permeation chromatography to yield a carbohydrate-rich fraction (96%) composed mainly of glucose (98%). Infra-red and 13C NMR spectroscopy showed that all the glucosidic linkages were in the beta-configuration. Data from methylation analysis and Smith degradation indicated that Botryosphaeran was a (1-->3)-beta-D-glucan with approx 22% side branching at C-6. The products obtained from partial acid hydrolysis demonstrated that the side branches consisted of single (1-->6)-beta-linked glucosyl, and (1-->6)-beta-linked gentiobiosyl residues.

Evaluation of the pharmacokinetic interactions between orally administered trihalomethanes in the rat.

The blood kinetics of trihalomethanes has recently been reported to differ between an oral administration of any single trihalomethane (0.25 mmol/kg) [THMs: chloroform, bromoform, bromodichloromethane (BDCM), dibromochloromethane (DBCM)] and a combined administration of 0.25 mmol/kg of each of the 4 THMs. The significant increase in blood concentrations of THMs could be a consequence of pharmacokinetic interactions between two or more of the THMs present simultaneously. The objective of the present study was to characterize the blood kinetics of THMs following oral administration singly or as binary mixtures in order to assess the relative contribution of each THM to the kinetic interferences observed with the quaternary mixture. A single dose of each THM (0.5 mmol/kg) alone or of a binary mixture containing 0.5 mmol/kg of each THM was administered by gavage to male Sprague-Dawley rats. The venous blood concentrations of unchanged THMs were measured for up to 720 min postadministration by headspace gas chromatography. Results showed that, compared to single administration, each binary mixture caused a significant increase in the blood concentrations of both THMs present and this effect increased with time. The impact, however, was not similar for each mixture, especially during the first hour following administration of the compounds (bromoform and DBCM). Among the four THMs, bromoform and DBCM kinetics appeared to be more sensitive to the mixture effect and to exert the greatest impact on the kinetics of the second THM present in the mixture. Simulation exercises conducted with physiologically based toxicokinetic models suggest metabolic inhibition as the possible mechanism of the interaction between THMs. In conclusion, the results of this study show that, at the dose level investigated, every binary combination of THMs, when orally administered, resulted in a significant modulation of their pharmacokinetics and suggest that this is probably the consequence of a mutual metabolic inhibition between the THMs.

[Bacterial flora of the oral cavity, fangs and venom of Bothrops jararaca: possible source of infection at the site of bite]. / Flora bacteriana da cavidade oral, presas e veneno de Bothrops jararaca: possível fonte de infecção no local da picada.

Culture of fang, fang sheath and venom of fifteen healthy freshly captured Bothrops jararaca were analyzed. The bacteria most frequently encountered were group D streptococci (12 snakes), Enterobacter sp. (6), Providencia rettgeri (6), Providencia sp. (4), Escherichia coli (4), Morganella morganii (3) and Clostridium sp. (5). The bacteria observed are similar to those found in the abscesses from Bothrops bitten patients. Since these snake mouth bacteria may be inoculated during the snake bite, bacterial multiplication and infection may occur under favorable conditions.

[Inhibition of adhesion of the classical enteropathogenic Escherichia coli 0111 ab: H2 to HeLa cells using colostrum and human milk in different experimental conditions]. / Inibição da adesão de Escherichia coli enteropatogênica clássica 0111ab: H2 a células HeLa pelo colostro e leite materno em diferentes condições experimentais.

Adherence of enteropathogenic Escherichia coli (EPEC) to enterocytes with subsequent destruction of microvilli is supposed to be their mechanism of virulence. Adhesion may be studied in vitro systems using HeLa or HEp-2 cells, to which EPEC adhere in a localized pattern. We show here that colostrum and human milk inhibit E. coli 0111ab:H2 adherence to HeLa cells in different experimental conditions. Lactose does not seem to be involved in the in vitro inhibition since no effect was observed when a concentration of 7.5% was used during the test. A bacterial growth curve performed in same conditions of adherence test showed no bacteriostatic effect of human milk. S-IgA and receptor analogues could be responsible for the adherence inhibition observed.

Salt stress response and proline accumulation in Brachiaria humidicola plants with and without mycorrhizal inoculation.

The work was carried out to investigate the effects of different concentrations of NaCl on Brachiaria humidicola Rendle in the presence and absence of Glomus etunicatum Becker and Gerdemann, as well as to evaluate some growth parameters and the accumulation of free proline in the plant leaves. The soil used was a Neosoil Fluvic Eutrophic with pH of 6.5, organic matter, 12.8 g kg-1 and soil texture, sandy loam, in which Brachiaria humidicola Rendle cv. 409 was grown. Five NaCl concentrations were tested 0; 0.22; 1.09; 1.96 and 2.84 g. kg-1 of soil, whose electrical conductivity (EC) were 2.22; 4.00; 8.13; 12.53 and 16.50 dS m-1, respectively. Brachiaria humidicola showed salt tolerance when submitted to an EC of 4 dS m-1. There was a reduction of leaf area, dry matter of shoots and roots for the soil treatments beyond EC at 8 dS m-1. Free proline content in the leaves increased together with the increase in soil salinity (EC at 8 dS m-1) demonstrating that plants submitted to EC of 2 and 4 dS m-1 were less affected by salt stress, and consequently accumulated less proline in the leaves. Root colonization was not affected by the increase of NaCl dosage in the soil.

[Characterization of Rhizobium isolates from acid and alkaline soils in semi-arid regions of Pernambuco]. / Caracterización de aislamientos de rizobios de suelos ácidos y alcalinos en la región semiárida de Pernambuco.

The genetic variety of the Rhizobium isolates from acid and alkaline soils in the semiarid zone of Pernambuco state was evaluated through the use of 17 primers of arbitrary sequence. Amplified products were separated by electrophoresis in agarose gel at 1.4% and visualized by ethidium bromide coloration. The results obtained suggest a high genetic variety of the isolates in relation to the standard strain. Data were analyzed by UPGMA (unweighted pair-group method with arithmetic average), based on Jaccard's coefficient and visualized through dendrograms. The strains isolated from the acid soils were included in one group whereas the strains from alkaline soils were located in other three groups. Meanwhile, one of the groups formed by strain Isol-14, isolated from acid soils is more related to the groups of strains isolated from acid soils than to the remaining groups from alkaline soils.

[Comparison of the subjective global nutrition assessment x objective nutrition evaluation]. / Comparação da avaliação nutricional subjetiva global x avaliação nutricional objetiva.

OBJECTIVE: Nutritional Assessment methods (ONA) are traditionally employed in hospitalized patients (anthropometry, laboratorial exams and immunological tests). The Subjective Global Assessment (SGA) estimates weight loss and changes in dietetic intake as being allegedly efficient for protein-caloric malnutrition diagnosis of the hospitalized adult. PURPOSE: Compare the efficacy of SGA against the traditional ONA. METHODS: The prospective study was done with 100 hospitalized patients from 3/90-5/90 at the Hospital Beneficência Portuguesa de São Paulo. The Detsky et al. questionnaire was applied besides anthropometric measurements (body weight, triceps skinfold thickness-TSF, mirdam circumference MCA) and laboratorial examinations (serun albumin, hemoglobin, peripheral blood lymphocytes) all of them in the first 3 days of admission. SGA always preceded ONA, and was always done by the same observer. RESULTS: There was a significant reduction on the average values of anthropometric and laboratorial measurements with the progressive worsening of nutritional status assessed by SGA. There were significant associations (p < 0.05) between hypoalbuminemia (< 3.5 g/dL) loss of TSF (< 10 mm) and MAC (< 23.3 cm) in those patients classified as moderately and severely malnourished by SGA (14%). Hypoalbuminemia, low TSF and SGA malnutrition were all significantly associated to mortality (p < 0.05). CONCLUSION: SGA in our hospital is a reliable diagnostic method for the diagnosis of malnutrition of hospitalized adult patients and malnutrition of hospitalized adult patients and keeps prognostic association with mortality.

[Inservice education--training for the 4th year nursing students]. / Educaçao em serviço--treinamento para alunos do 4 degrees ano de enfermagem.

This work reports the trainee-nurses contact with recycling of a nursing team at a general hospital through an extension project. It has been observed that the participation in this project reinforced to the trainee-nurses the importance of the nurses educative role and have showed us the necessity of more frequent contacts between trainee-nurses and their educational role, so that, after graduation, they will be able to develop education in service, mainly next to their work-team with security.

Correlating denitrifying catabolic genes with N2O and N2 emissions from swine slurry composting.

This work evaluated N dynamics that occurs over time within swine slurry composting piles. Real-time quantitative PCR (qPCR) analyzes were conducted to estimate concentrations of bacteria community harboring specific catabolic nitrifying-ammonium monooxygenase (amoA), and denitrifying nitrate- (narG), nitrite- (nirS and nirG), nitric oxide- (norB) and nitrous oxide reductases (nosZ) genes. NH3-N, N2O-N, N2-N emissions represented 15.4 ± 1.9%, 5.4 ± 0.9%, and 79.1 ± 2.0% of the total nitrogen losses, respectively. Among the genes tested, temporal distribution of narG, nirS, and nosZ concentration correlated significantly (p<0.05) with the estimated N2 emissions. Denitrifying catabolic gene ratio (cnorB+qnorB)/nosZ ≥ 100 was indicative of N2O emission potential from the compost pile. Considering our current empirical limitations to accurately measure N2 emissions from swine slurry composting at field scale the use of these catabolic genes could represent a promising monitoring tool to aid minimize our uncertainties on biological N mass balances in these systems.

Chondrogenic differentiation of human bone marrow mesenchymal stem cells in chitosan-based scaffolds using a flow-perfusion bioreactor.

Native articular cartilage is subjected to synovial fluid flow during normal joint function. Thus, it is believed that the morphogenesis of articular cartilage may be positively regulated by the application of similar stimulation in vitro. In the present study, the effect of fluid flow over the chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells (hBM-MSCs) was investigated. We intended to find out whether the shear stress caused by perfusion of the medium through the constructs was capable of augmenting the differentiation process. Human BMSCs were isolated from bone marrow aspirates and were characterized by flow cytometry. After expansion, hBM-MSCs were seeded statically onto fibre mesh scaffolds, consisting of a blend of 50:50 chitosan:poly(butylene terephthalate adipate) (CPBTA). Constructs were cultured in a flow-perfusion bioreactor for 28 days, using complete medium for chondrogenesis supplemented by TGFß3. An enhanced ECM deposition and collagen type II production was observed in the bioreactor samples when compared to the static controls. Moreover, it was observed that hBM-MSCs, in static cultures, take longer to differentiate. ECM accumulation in these samples is lower than in the bioreactor sections, and there is a significant difference in the expression of collagen type I. We found that the flow-induced shear stress has a beneficial effect on the chondrogenic differentiation of hMSCs.

Chitosan/polyester-based scaffolds for cartilage tissue engineering: assessment of extracellular matrix formation.

Naturally derived polymers have been extensively used in scaffold production for cartilage tissue engineering. The present work aims to evaluate and characterize extracellular matrix (ECM) formation in two types of chitosan-based scaffolds, using bovine articular chondrocytes (BACs). The influence of these scaffolds' porosity, as well as pore size and geometry, on the formation of cartilagineous tissue was studied. The effect of stirred conditions on ECM formation was also assessed. Chitosan-poly(butylene succinate) (CPBS) scaffolds were produced by compression moulding and salt leaching, using a blend of 50% of each material. Different porosities and pore size structures were obtained. BACs were seeded onto CPBS scaffolds using spinner flasks. Constructs were then transferred to the incubator, where half were cultured under stirred conditions, and the other half under static conditions for 4 weeks. Constructs were characterized by scanning electron microscopy, histology procedures, immunolocalization of collagen type I and collagen type II, and dimethylmethylene blue assay for glycosaminoglycan (GAG) quantification. Both materials showed good affinity for cell attachment. Cells colonized the entire scaffolds and were able to produce ECM. Large pores with random geometry improved proteoglycans and collagen type II production. However, that structure has the opposite effect on GAG production. Stirred culture conditions indicate enhancement of GAG production in both types of scaffold.