Characterization of in vitro effects of patulin on intestinal epithelial and immune cells

The intestinal mucosa is the first biological barrier encountered by natural toxins, and could possibly be exposed to high amounts of dietary mycotoxins. Patulin (PAT), a mycotoxin produced by Penicillium spp. during fruit spoilage, is one of the best known enteropathogenic mycotoxins able to alter functions of the intestine (Maresca et al., 2008). This study evaluated the effects of PAT on barrier function of the gut mucosa utilizing the intestinal epithelial cell model Caco-2, and scrutinized immunomodulatory effects using human peripheral blood mononuclear cells (PBMC) and human blood monocyte-derived dendritic cells (moDCs) as test systems. PAT exposure reduced Caco-2 cell viability at concentrations above 12 mM. As expected, the integrity of a polarized Caco-2 monolayer was affected by PAT exposure, as demonstrated by a decrease in TER values, becoming more pronounced at 50 mM. No effects were detected on the expression levels of the tight junction proteins occludin, claudin-1 and claudin-3 at 50 mM. However, the expression of zonula occludens-1 (ZO-1) and myosin light chain 2 (MLC2) declined. Also, levels of phospho-MLC2 (p-MLC2) increased after 24 h of exposure to 50 mM of PAT. T cell proliferation was highly sensitive to PAT with major effects for concentrations above 10 nM of PAT. The same conditions did not affect the maturation of moDC. PAT causes a reduction in Caco-2 barrier function mainly by perturbation of ZO-1 levels and the phosphorylation of MLC. Low doses of PAT strongly inhibited T cell proliferation induced by a polyclonal activator, but had no effect on the maturation of moDC. These results provide new information that strengthens the concept that the epithelium and immune cells of the intestinal mucosa are important targets for the toxic effects of food contaminants like mycotoxins

Portuguese children exposure to multiple mycotoxins through food consumption: toward a holistic approach for multiple mycotoxins risk assessment

Humans can be exposed to multiple chemicals at once from a variety of sources, and human risk assessment of multiple chemicals poses several challenges to scientists, risk assessors and risk managers. Ingestion of food is considered a major route of exposure to many contaminants, namely mycotoxins, especially for vulnerable population groups, as children. A lack of sufficient data regarding mycotoxins children risk assessment, could contribute to an inaccuracy of the estimated risk. Efforts must be undertaken to develop initiatives that promote a broad overview of multiple mycotoxins risk assessment. The present work, developed within the MYCOMIX project, aims to assess the risk associated to the exposure of Portuguese children (< 3 years old) to multiple mycotoxins through consumption of foods primarily marketed for this age group. A holistic approach was developed applying deterministic and probabilistic tools to the calculation of mycotoxin daily intake values, integrating children food consumption (3-days food diary), mycotoxins occurrence (HPLC-UV, HPLC-FD, LC-MS/MS and GC-MS), bioaccessibility (standardized in vitro digestion model) and toxicological data (in vitro evaluation of cytotoxicity, genotoxicity and intestinal impact). A case study concerning Portuguese children exposure to patulin (PAT) and ochratoxin A (OTA), two mycotoxins co-occurring in processed cereal-based foods (PCBF) marketed in Portugal, was developed. Main results showed that there is low concern from a public health point of view relatively to PAT and OTA Portuguese children exposure through consumption of PCBF, considering the estimated daily intakes of these two mycotoxins (worst case scenarios, 22.930 ng/kg bw/day and 0.402 ng/kg bw/day, for PAT and OTA, respectively), their bioaccessibility and toxicology results. However, the present case study only concerns the risk associated with the consumption of PCBF and child diet include several other foods. The present work underlines the need to adopt a holistic approach for multiple mycotoxins risk assessment integrating data from exposure, bioacessibility and toxicity domains in order to contribute to a more accurate risk assessment.

NIR-spectroscopy for bioprocess monitoring & control

The Ingold port adaption of a free beam NIR spectrometer is tailored for optimal bioprocess monitoring and control. The device shows an excellent signal to noise ratio dedicated to a large free aperture and therefore a large sample volume. This can be seen particularly in the batch trajectories which show a high reproducibility. The robust and compact design withstands rough process environments as well as SIP/CIP cycles. Robust free beam NIR process analyzers are indispensable tools within the PAT/QbD framework for realtime process monitoring and control. They enable multiparametric, non-invasive measurements of analyte concentrations and process trajectories. Free beam NIR spectrometers are an ideal tool to define golden batches and process borders in the sense of QbD. Moreover, sophisticated data analysis both quantitative and MSPC yields directly to a far better process understanding. Information can be provided online in easy to interpret graphs which allow the operator to make fast and knowledge-based decisions. This finally leads to higher stability in process operation, better performance and less failed batches.

Correlaci?n entre la ubicaci?n del centro de gravedad corporal y los cambios posturales en ni?os patinadores

El contenido de este trabajo y en especial el tema, pretende dar un aporte a las investigaciones que se han realizado en el patinaje, con el fin de aumentar el conocimiento cient?fico en los cambios morfol?gicos funcionales que se presentan en los ni?os patinadores debido a su pr?ctica a temprana edad, teniendo como objetivo relacionar la ubicaci?n de su centro de gravedad corporal y la influencia que este tiene en la postura. Esta investigaci?n se har? por medio de la valoraci?n postural bipodal y la ubicaci?n del centro de gravedad corporal por el m?todo de reacci?n en 9 ni?os patinadores de la escuela de patinaje Club Pat?n Sport de la ciudad de Yumbo-Colombia. Para este estudio se necesitara evaluar variables antropom?tricas, posturales, podometricas, podograficas, y motoras, las cuales permitir?n alcanzar el objetivo de esta investigaci?n. El an?lisis de todos los datos recogidos se har? por medio de una estad?stica descriptiva, se comparara las variables de cada ni?o con la valoraci?n obtenida en su postura bipodal, analizando los cambios morfol?gicos y funcionales que se han presentado en ellos. La importancia de esta investigaci?n es poder identificar cu?les han sido los cambios morfol?gicos funcionales que se han presentado en los ni?os evaluados debido a la pr?ctica deportiva del patinaje a temprana edad.

Argamassas de terra e cal - Características e campos de aplicação

As argamassas mistas de cal e terra devem ter resultado da prática profissional quando as condições económicas foram possibilitando a adição de cais aérea às argamassas vernaculares só de terra. Baixas adições de cal aérea a argamassas de terra têm conduzido a drásticas alterações da cor e das resistências mecânicas das argamassas de terra. No entanto, a substituição parcial de massa de cal por terra em argamassas de cal ao traço volumétrico 1:2 (correspondendo a argamassas com traço em massa de 1:8, com substituições de 10% a 50%) resulta em argamassas mais ecológicas e que tecnicamente indiciam ser adequadas para aplicação em rebocos de edifícios antigos e mesmo em construção nova com características compatíveis. Os resultados obtidos foram particularmente interessantes para uma substituição de 10% mas muito há ainda a fazer para um conhecimento aprofundado deste tipo de argamassas mistas de cal e terra, muito utilizadas no passado, com vista à otimização de formulações que possam vir a ser aplicadas correntemente.

Trabajo Final de Grado y Plan de Acción Tutorial en el curriculum del Grado en Enfermería: diseño, desarrollo y evaluación

La acción tutorial (PAT), en los estudios universitarios de Enfermería, se halla en el centro de la acción docente del profesorado. El tutor/a orienta en el proceso de aprendizaje y en la preparación del Trabajo Final de Grado (TFG). En este artículo presentamos los resultados de la investigación realizada con tutores y estudiantes, que tiene por objetivo evaluar el diseño y desarrollo del TFG, los resultados académicos y su vivencia y experiencia. Diseño mixto cuantitativo descriptivo y cualitativo de investigación acción participación. Muestra total de 124 estudiantes de 4º curso de Grado Enfermería y 37 tutores del TFG. Técnicas de recogida de datos: resultados académicos y evaluativos; 3 grupos focales con estudiantes y profesores; 3 entrevistas semiestructuradas abiertas a estudiantes. Los resultados constatan que las tipologías más elegidas fueron revisiones bibliográficas (38,75%) y Planes de cuidados (24,2%). El 63,9 % de la temática escogida es Enfermería Maternal, Pediátrica y Clínica. Estudiantes y tutores viven con dificultad la elaboración del TFG y se requiere una mayor cultura tutorial. Es un proceso complejo de organización y coordinación que, sin embargo, permite un trabajo colaborativo de todo el equipo docente.

Saatana peilissä – 1990-luvun suomalainen saatananpalvontadiskurssi osana folklorea ja kollektiivista mielikuvamaailmaa

Tutkielman aihe on saatananpalvontadiskurssi 1990-luvun Suomessa, sen yhteydet folkloreen ja kollektiiviseen mielikuvamaailmaan (popular imagination), sekä sen rakentuminen ja leviäminen kansanomaisten asiantuntijoiden ja median avulla. Tutkielman ydinkysymykset ovat: mitä evankelikaaliseen aineistoon perustuva saatananpalvontadiskurssi sisältää, miten sitä on rakennettu ja levitetty ja miksi näin on tehty? Tutkimusaineistona ovat Pat Pullingin kirja Noidankehässä (1993), Marja-Liisa Huhdan Saatana kutsuu minua -videokasetti (1994), Riku Rinteen kirjat Syvyyden kuilusta (1994), Pimeys väistyy (1996), Varo Vihollinen! Jumalan pyörremyrsky on tulossa (1999), Keijo Ahorinnan kirja Saatananpalvonnan monet kasvot (1997), sekä lehtiartikkeleita 1990-luvulta. Tutkimuksessa käy ilmi, että evankelikaalisten toimijoiden luomia saatananpalvontatarinoita on levitetty, pidetty ajankohtaisena ja välitetty valtavirtamediaan itseoppineiden asiantuntijoiden toimesta. Kun valtavirtamedia on legitimoinut tarinat, ovat ne palautuneet evankelikaaliseen aineistoon todentuneina. Leviämistä ovat edesauttaneet evankelikaalisen aineiston ja median esittämien tarinoiden synnyttämät huhut, joiden johdosta saatananpalvontadiskurssi on alkanut elää kollektiivisessa mielikuvamaailmassa. Voidakseen levitä ja pysyä ajankohtaisina, tarinoiden tulee sisältää riittävän yksinkertaisia ja voimakkaita aineksia. Saatananpalvontadiskurssissa nämä vaatimukset toteutuvat erityisen selvästi. Saatananpalvoja on hahmona vahvasti kansanperinteen mukainen.

Casas para um planeta pequeno

Dissertação de Mestrado para obtenção do grau de Mestre em Arquitectura, apresentada na Universidade de Lisboa - Faculdade de Arquitectura.

Electrical characterization of organic based transistors: stability issues

An investigation into the stability of metal insulator semiconductor (MIS) transistors based on alpha-sexithiophene is reported. In particular the kinetics of the threshold voltage shift upon application of a gate bias has been determined. The kinetics follow a stretched-hyperbola type behavior, in agreement with the formalism developed to explain metastability in amorphous-silicon thin film transistors. Using this model, quantification of device stability is possible. Temperature-dependent measurements show that there are two processes involved in the threshold voltage shift, one occurring at T approximate to 220 K and the other at T approximate to 300 K. The latter process is found to be sample dependent. This suggests a relation between device stability and alpha-sexithiophene deposition parameters. Copyright (c) 2005 John Wiley A Sons, Ltd.

Large-scale screening of a targeted Enterococcus faecalis mutant library identifies envelope fitness factors

Spread of antibiotic resistance among bacteria responsible for nosocomial and community-acquired infections urges for novel therapeutic or prophylactic targets and for innovative pathogen-specific antibacterial compounds. Major challenges are posed by opportunistic pathogens belonging to the low GC% gram-positive bacteria. Among those, Enterococcus faecalis is a leading cause of hospital-acquired infections associated with life-threatening issues and increased hospital costs. To better understand the molecular properties of enterococci that may be required for virulence, and that may explain the emergence of these bacteria in nosocomial infections, we performed the first large-scale functional analysis of E. faecalis V583, the first vancomycin-resistant isolate from a human bloodstream infection. E. faecalis V583 is within the high-risk clonal complex 2 group, which comprises mostly isolates derived from hospital infections worldwide. We conducted broad-range screenings of candidate genes likely involved in host adaptation (e.g., colonization and/or virulence). For this purpose, a library was constructed of targeted insertion mutations in 177 genes encoding putative surface or stress-response factors. Individual mutants were subsequently tested for their i) resistance to oxidative stress, ii) antibiotic resistance, iii) resistance to opsonophagocytosis, iv) adherence to the human colon carcinoma Caco-2 epithelial cells and v) virulence in a surrogate insect model. Our results identified a number of factors that are involved in the interaction between enterococci and their host environments. Their predicted functions highlight the importance of cell envelope glycopolymers in E. faecalis host adaptation. This study provides a valuable genetic database for understanding the steps leading E. faecalis to opportunistic virulence.

Fluoroquinolone efflux in Streptococcus suis is mediated by SatAB and not by SmrA

Streptococcus suis is an emerging zoonotic pathogen. With the lack of an effective vaccine, antibiotics remain the main tool to fight infections caused by this pathogen. We have previously observed a reserpine-sensitive fluoroquinolone (FQ) efflux phenotype in this species. Here, SatAB and SmrA, two pumps belonging to the ATP binding cassette (ABC) and the major facilitator superfamily (MFS), respectively, have been analyzed in the fluoroquinolone-resistant clinical isolate BB1013. Genes encoding these pumps were overexpressed either constitutively or in the presence of ciprofloxacin in this strain. These genes could not be cloned in plasmids in Escherichia coli despite strong expression repression. Finally, site-directed insertion of smrA and satAB in the amy locus of the Bacillus subtilis chromosome using ligated PCR amplicons allowed for the functional expression and study of both pumps. Results showed that SatAB is a narrow-spectrum fluoroquinolone exporter (norfloxacin and ciprofloxacin), susceptible to reserpine, whereas SmrA was not involved in fluoroquinolone resistance. Chromosomal integration in Bacillus is a novel method for studying efflux pumps from Gram-positive bacteria, which enabled us to demonstrate the possible role of SatAB, and not SmrA, in fluoroquinolone efflux in S. suis.

Haemophilus influenzae clinical isolates with plasmid pB1000 bearing blaROB-1: fitness cost and interspecies dissemination

Plasmid pB1000 is a mobilizable replicon bearing the bla(ROB-1) beta-lactamase gene that we have recently described in Haemophilus parasuis and Pasteurella multocida animal isolates. Here we report the presence of pB1000 and a derivative plasmid, pB1000', in four Haemophilus influenzae clinical isolates of human origin. Pulsed-field gel electrophoresis showed unrelated patterns in all strains, indicating that the existence of pB1000 in H. influenzae isolates is not the consequence of clonal dissemination. The replicon can be transferred both by transformation and by conjugation into H. influenzae, giving rise to recipients resistant to ampicillin and cefaclor (MICs, > or =64 microg/ml). Stability experiments showed that pB1000 is stable in H. influenzae without antimicrobial pressure for at least 60 generations. Competition experiments between isogenic H. influenzae strains with and without pB1000 revealed a competitive disadvantage of 9% per 10 generations for the transformant versus the recipient. The complete nucleotide sequences of nine pB1000 plasmids from human and animal isolates, as well as the epidemiological data, suggest that animal isolates belonging to the Pasteurellaceae act as an antimicrobial resistance reservoir for H. influenzae. Further, since P. multocida is the only member of this family that can colonize both humans and animals, we propose that P. multocida is the vehicle for the transport of pB1000 between animal- and human-adapted members of the Pasteurellaceae.

Multiresistance in Pasteurella multocida is mediated by coexistence of small plasmids

In most gram-negative bacteria, acquired multiresistance is conferred by large plasmids compiling numerous antimicrobial resistance genes. Here, we show an evolutionary alternative strategy used by Pasteurella multocida to become resistant to multiple clinically relevant antibiotics. Thirteen beta-lactam-resistant clinical isolates, concomitantly resistant to tetracyclines and/or streptomycin as well as to sulfonamides, were studied. Pulsed-field gel electrophoresis analysis revealed different profiles among the isolates, showing that clonal dissemination was not the sole event responsible for the spread of multiresistance. Each P. multocida strain carried two or three small plasmids between 4 and 6 kb in size. A direct association between resistance profile and plasmid content was found. Complete nucleotide sequencing of all plasmids revealed seven different replicons, six of them belonging to the ColE1 superfamily. All plasmids carried one, or a maximum of two, antimicrobial resistance determinants. Plasmids pB1000 and pB1002 bore bla(ROB-1), pB1001 carried tet(B), pB1003 and pB1005 carried sul2 and strA, pB1006 harbored tet(O), and p9956 bore the tet(H) gene. All plasmids except pB1002 and pB1006 were successfully transformed into Escherichia coli. pB1000, also involved in beta-lactam resistance in Haemophilus parasuis (A. San Millan et al., Antimicrob. Agents Chemother. 51:2260-2264, 2007), was mobilized in E. coli using the conjugation machinery of an IncP plasmid. Stability experiments proved that pB1000 was stable in P. multocida but highly unstable in E. coli. In conclusion, bla(ROB-1) is responsible for beta-lactam resistance in P. multocida in Spain. Coexistence and the spread of small plasmids are used by P. multocida to become multiresistant.

Physical therapy for urinary incontinence in postmenopausal women with osteoporosis or low bone density : a randomized controlled trial

Objective: To assess the effectiveness of 12 weekly physical therapy sessions for urinary incontinence (UI) compared with a control intervention, for reducing the number of UI episodes measured with the 7-day bladder diary, at 3 months and 1 year postrandomization. Methods: A single parallel-group randomized controlled trial was conducted at one outpatient public health center, in postmenopausal women aged 55 years and over with osteoporosis or low bone density and UI. Women were randomized to physical therapy (PT) for UI or osteoporosis education. The primary outcome measure was number of leakage episodes on the 7-day bladder diary, assessed at baseline, after treatment and at 1 year. The secondary outcome measures included the pad test and disease-specific quality of life and self-efficacy questionnaires assessed at the same timepoints. Results: Forty-eight women participated (24 per group). Two participants dropped out of each group and one participant was deceased before 3-month follow-up. Intention-to-treat analysis was undertaken. At 3 months and 1 year, there was a statistically significant difference in the number of leakage episodes on the 7-day bladder diary (3 mo: P = 0.04; 1 y: P = 0.01) in favor of the PT group. The effect size was 0.34 at 1 year. There were no harms reported. Conclusions: After a 12-week course of PT once per week for UI, PT group participants had a 75% reduction in weekly median number of leakage episodes, whereas the control group's condition had no improvement. At 1 year, the PT group participants maintained this improvement, whereas the control group's incontinence worsened.

Autoinducer-2 plays a crucial role in gut colonization and probiotic functionality of Bifidobacterium breve UCC2003

In the present study we show that luxS of Bifidobacterium breve UCC2003 is involved in the production of the interspecies signaling molecule autoinducer-2 (AI-2), and that this gene is essential for gastrointestinal colonization of a murine host, while it is also involved in providing protection against Salmonella infection in Caenorhabditis elegans. We demonstrate that a B. breve luxS-insertion mutant is significantly more susceptible to iron chelators than the WT strain and that this sensitivity can be partially reverted in the presence of the AI-2 precursor DPD. Furthermore, we show that several genes of an iron starvation-induced gene cluster, which are downregulated in the luxS-insertion mutant and which encodes a presumed iron-uptake system, are transcriptionally upregulated under in vivo conditions. Mutation of two genes of this cluster in B. breve UCC2003 renders the derived mutant strains sensitive to iron chelators while deficient in their ability to confer gut pathogen protection to Salmonella-infected nematodes. Since a functional luxS gene is present in all tested members of the genus Bifidobacterium, we conclude that bifidobacteria operate a LuxS-mediated system for gut colonization and pathogen protection that is correlated with iron acquisition.