Conversion of CD95 (Fas) Type II into Type I signaling by sub-lethal doses of cycloheximide

CD95 (Fas/Apo-1)-mediated apoptosis was shown to occur through two distinct pathways. One involves a direct activation of caspase-3 by large amounts of caspase-8 generated at the DISC (Type I cells). The other is related to the cleavage of Bid by low concentration of caspase-8, leading to the release of cytochrome c from mitochondria and the activation of caspase-3 by the cytochrome c/APAF-1/caspase-9 apoptosome (Type II cells). It is also known that the protein synthesis inhibitor cycloheximide (CHX) sensitizes Type I cells to CD95-mediated apoptosis, but it remains contradictory whether this effect also occurs in Type II cells. Here, we show that sub-lethal doses of CHX render both Type I and Type II cells sensitive to the apoptogenic effect of anti-CD95 antibodies but not to chemotherapeutic drugs. Moreover, Bcl-2-positive Type II cells become strongly sensitive to CD95-mediated apoptosis by the addition of CHX to the cell culture. This is not the result of a restraint of the anti-apoptotic effect of Bcl-2 at the mitochondrial level since CHX-treated Type II cells still retain their resistance to chemotherapeutic drugs. Therefore, CHX treatment is granting the CD95-mediated pathway the ability to bypass the mitochondria requirement to apoptosis, much alike to what is observed in Type I cells.

Culture experiments to evaluate CO2 effects on growth and morphology of the coccolithophore Pleurochrysis cf. pseudoroscoffensis

In the framework of the Italian project CO2 Monitor, two culture experiments were carried out in vertical closed photobioreactors with Pleurochrysis cf. pseudoroscoffensis Gayral & Fresnel 1983, a coccolithophore isolated from the Gulf of Trieste (North Adriatic Sea). The aim of this study was to investigate the effects induced by pH variations due to CO2 emissions on its growth and morphology. Two experiments were carried out with two different CO2 concentrations (1 and 2%). Growth and cell size in light microscopy, morphology and coccolith size in scanning electron microscopy, particulate nitrogen (PN) and particulate inorganic and organic carbon (PIC and POC) content of the coccolithophore were investigated during the light and dark phases. Dissolved inorganic nutrient (nitrate and phosphate) concentrations and pH of the medium and the presence of heterotrophic prokaryotes (HP) were monitored as well.

Analysis of the Pasteurella multocida outer membrane sub-proteome and its response to the in vivo environment of the natural host

This study describes the identification of outer membrane proteins (OMPs) of the bacterial pathogen Pasteurella multocida and an analysis of how the expression of these proteins changes during infection of the natural host. We analysed the sarcosine-insoluble membrane fractions, which are highly enriched for OMPs, from bacteria grown under a range of conditions. Initially, the OMP-containing fractions were resolved by 2-DE and the proteins identified by MALDI-TOF MS. In addition, the OMP-containing fractions were separated by 1-D SDS-PAGE and protein identifications were made using nano LC MS/MS. Using these two methods a total of 35 proteins was identified from samples obtained from organisms grown in rich culture medium. Six of the proteins were identified only by 2-DE MALDI-TOF MS, whilst 17 proteins were identified only by 1-D LC MS/MS. We then analysed the OMPs from P. multocida which had been isolated from the bloodstream of infected chickens (a natural host) or grown in iron-depleted medium. Three proteins were found to be significantly up-regulated during growth in vivo and one of these (Pm0803) was also up-regulated during growth in iron-depleted medium. After bioinformatic analysis of the protein matches, it was predicted that over one third of the combined OMPs predicted by the bioinformatics sub-cellular localisation tools PSORTB and Proteome Analyst, had been identified during this study. This is the first comprehensive proteomic analysis of the P. multocida outer membrane and the first proteomic analysis of how a bacterial pathogen modifies its outer membrane proteome during infection.

Electronic dance music culture and religion: An Overview

This article provides a comprehensive and critical overview of existing research that investigates (directly and indirectly) the religio-spiritual dimensions of electronic dance music culture (EDMC) (from disco, through house, to post-rave forms). Studies of the culture and religion of EDMC are explored under four broad groupings: the cultural religion of EDMC expressed through 'ritual' and 'festal'; subjectivity, corporeality and the phenomenological dance experience (especially 'ecstasy' and 'trance'); the dance community and a sense of belonging (the 'vibe' and 'tribes'); and EDMC as a new 'spirituality of life'. Moving beyond the cultural Marxist approaches of the 1970s, which held youth (sub)cultural expressions as 'ineffectual' and 'tragic', and the postmodernist approaches of the early 1990s, which held rave to be an 'implosion of meaning', recent anthropological and sociological approaches recognise that the various manifestations of this youth cultural phenomenon possess meaning, purpose and significance for participants. Contemporary scholarship thus conveys the presence of religiosity and spirituality within contemporary popular cultural formations. In conclusion, I suggest that this and continuing scholarship can offer useful counterpoint to at least one recent account (of clubbing) that overlooks the significance of EDMC through a restricted and prejudiced apprehension of 'religion'.

Development of a neurotoxicity test-system, using human post-mitotic, astrocytic and neuronal cell lines in co-culture

Astrocytes are essential for neuronal function and survival, so both cell types were included in a human neurotoxicity test-system to assess the protective effects of astrocytes on neurons, compared with a culture of neurons alone. The human NT2.D1 cell line was differentiated to form either a co-culture of post-mitotic NT2.N neuronal (TUJ1, NF68 and NSE positive) and NT2.A astrocytic (GFAP positive) cells (∼2:1 NT2.A:NT2.N), or an NT2.N mono-culture. Cultures were exposed to human toxins, for 4 h at sub-cytotoxic concentrations, in order to compare levels of compromised cell function and thus evidence of an astrocytic protective effect. Functional endpoints examined included assays for cellular energy (ATP) and glutathione (GSH) levels, generation of hydrogen peroxide (H2O2) and caspase-3 activation. Generally, the NT2.N/A co-culture was more resistant to toxicity, maintaining superior ATP and GSH levels and sustaining smaller significant increases in H2O2 levels compared with neurons alone. However, the pure neuronal culture showed a significantly lower level of caspase activation. These data suggest that besides their support for neurons through maintenance of ATP and GSH and control of H2O2 levels, following exposure to some substances, astrocytes may promote an apoptotic mode of cell death. Thus, it appears the use of astrocytes in an in vitro predictive neurotoxicity test-system may be more relevant to human CNS structure and function than neuronal cells alone. © 2007 Elsevier Ltd. All rights reserved.

Brain-derived neurotrophic factor as an indicator of chemical neurotoxicity:an animal-free CNS cell culture model

Recent changes to the legislation on chemicals and cosmetics testing call for a change in the paradigm regarding the current 'whole animal' approach for identifying chemical hazards, including the assessment of potential neurotoxins. Accordingly, since 2004, we have worked on the development of the integrated co-culture of post-mitotic, human-derived neurons and astrocytes (NT2.N/A), for use as an in vitro functional central nervous system (CNS) model. We have used it successfully to investigate indicators of neurotoxicity. For this purpose, we used NT2.N/A cells to examine the effects of acute exposure to a range of test chemicals on the cellular release of brain-derived neurotrophic factor (BDNF). It was demonstrated that the release of this protective neurotrophin into the culture medium (above that of control levels) occurred consistently in response to sub-cytotoxic levels of known neurotoxic, but not non-neurotoxic, chemicals. These increases in BDNF release were quantifiable, statistically significant, and occurred at concentrations below those at which cell death was measureable, which potentially indicates specific neurotoxicity, as opposed to general cytotoxicity. The fact that the BDNF immunoassay is non-invasive, and that NT2.N/A cells retain their functionality for a period of months, may make this system useful for repeated-dose toxicity testing, which is of particular relevance to cosmetics testing without the use of laboratory animals. In addition, the production of NT2.N/A cells without the use of animal products, such as fetal bovine serum, is being explored, to produce a fully-humanised cellular model.

Los números cuentan. Sub-representación de la obra artística de mujeres creadoras en museos y centros de arte contemporáneos

Este artículo cuantifica la presencia de obra artística de mujeres artistas en 21 museos y centros de arte contemporáneo españoles. Los resultados constatan una nítida sub-representación de la obra exhibida, por debajo del 20 por ciento. ¿Por qué sucede esto?, ¿diferencial potencial artístico de mujeres y hombres?, ¿superioridad masculina?, ¿discriminación? o ¿un sistema de arte con sesgo androcéntrico? En estas páginas se discute sobre la presencia de varios factores para explicar la brecha de género y se reclama, de las administraciones públicas y las instituciones de gestión cultural, el cumplimiento de la Ley para la Igualdad para garantizar la paridad.

"Sub Tiberio quies"? Judaea’s Political Situation under the Prefects (6-41 c.e.), between Reality and Ideology

Deeply conflicting views on the political situation of Judaea under the Roman prefects (6-41 c.e.) have been offered. According to some scholars, this was a period of persistent political unrest and agitation, whilst according to a widespread view it was a quiescent period of political calm (reflected in Tacitus’ phrase sub Tiberio quies). The present article critically examines again the main available sources –particularly Josephus, the canonical Gospels and Tacitus– in order to offer a more reliable historical reconstruction. The conclusions drawn by this survey calls into question some widespread and insufficiently nuanced views on the period. This, in turn, allows a reflection on the non-epistemic factors which might contribute to explain the origin of such views.

Global Trade Governance and ASEAN: Studies of Trade-Related Aspects of Labor, Environment and Culture in Southeast Asia

Thesis (Ph.D.)--University of Washington, 2016-08

Risk Factors for Neonatal Sepsis and Method for Reduction of Blood Culture Contamination

Background False-positive blood cultures findings may lead to a falsely increased morbidity and increased hospital costs. Method The survey was conducted as retrospective - prospective study and included 239 preterm infants (born before 37 weeks of gestation) who were treated in Neonatal Intensive Care Unit (NICU) in Institute for Child and Youth Health Care of Vojvodina during one year (January 1st, 2012 to December 31st, 2012). The retrospective part of the study focused on examination of incidence of neonatal sepsis and determination of risk factors. In the prospective part of the study infants were sub-divided into two groups: Group 1- infants hospitalized in NICU during the first 6 months of the study; blood cultures were taken by the ‘’clean technique’’ and checklists for this procedure were not taken. Group 2- neonates hospitalized in NICU during last 6 months of the study; blood cultures were taken by ‘’sterile technique’’ and checklists for this procedure were taken. Results The main risk factors for sepsis were prelabor rupture of membranes, low gestational age, low birth weight, mechanical ventilation, umbilical venous catheter placement, and abdominal drainage. Staphylococcus aureus and coagulase negative Staphylococcus were the most frequently isolated microorganisms in false-positive blood samples. Conclusions Education of employees, use of checklists and sterile sets for blood sampling, permanent control of false positive blood cultures, as well as regular and routine monthly reports are crucial for successful reduction of contamination rates.

Developing reading sub-skills: 15 Lesson Plans Adapted from "The Canterbury Tales”for teenagers

Este trabajo enfatiza el uso de la literatura en Clases EFL. La literatura es usada como una técnica para enseñar las cuatro destrezas básicas: lectura, habilidad de escuchar, escritura y la producción oral. Provee herramientas para mejorar vocabulario, pronunciación y estructuras gramaticales. El leer literatura provee beneficios enlistados en este documento. Además, algunas desventajas también son presentadas. La falta de: capacitación de profesores, interés por parte de los estudiantes y material propio de lectura, hacen de la literatura una tarea difícil de completar. Seleccionar el material correcto para una Clase EFL es el aspecto más importante para generar el interés por la lectura en los estudiantes. En este caso los adolescentes son el objetivo de este estudio. “The Canterbury Tales” por Geoffrey Chaucer, es la obra en la cual este proyecto está basado. La obra literaria de Geoffrey Chaucer “The Canterbury Tales” le da al lector la oportunidad de imaginar una cultura diferente, el comportamiento de las personas, sentimientos, diversión y moralejas con las que las personas se identifican. Quince planificaciones de clase adaptadas de “The Canterbury Tales” es la mejor herramienta para cultivar el interés por la lectura. Las más importantes sub-destrezas son mostradas en estas planificaciones. Cada una de estas sub-destrezas demuestra actividades sugeridas que pueden ser practicadas dentro del aula. La incorporación de la lectura en una Clase EFL facilita el trabajo del profesor con estudiantes quienes encontraran en la lectura facilidad y diversión.

ASSESSING THE DETERMINANTS OF TISSUE CULTURE BANANA ADOPTION IN WESTERN KENYA

In this study cross-section data was used to analyze the effect of farmers’ demographic, socioeconomic and institutional setting, market access and physical attributes on the probability and intensity of tissue culture banana (TCB) adoption. The study was carried out between July 2011 and November 2011. Both descriptive (mean, variance, promotions) and regression analysis were used in the analysis. A double hurdle regression model was fitted on the data. Using multistage sampling technique, four counties and eight sub-locations were randomly selected. Using random sampling technique, three hundred and thirty farmers were selected from a list of banana households in the selected sub-locations. The adoption level of tissue culture banana (TCB) was about 32%. The results also revealed that the likelihood of TCB adoption was significantly influenced by: availability of TCB planting material, proportion of banana income to the total farm income, per capita household expenditure and the location of the farmer in Kisii County; while those that significantly influenced the intensity of TCB adoption were: occupation of farmers, family size, labour source, farm size, soil fertility, availability/access of TCB plantlets to farmers, distance to banana market, use of manure in planting banana, access to agricultural extension services and index of TCB/non-TCB banana cultivar attributes which were scored by farmers. Compared to West Pokot County, farmers located in Bungoma County are more significantly and likely to adopt TCB technology. Therefore, the results of the study suggest that the probability of adoption and intensity of the use of TCB should be enhanced. This can be done by taking cognizance of these variables in order to meet the priority needs of the smallholder farmers who were the target group. This would lead to alleviating banana shortage in the region for enhanced food security. Subsequently, actors along the banana value chain are encouraged to target the intervention strategies based on the identified farmer, farm and institutional characteristics for enhanced impact on food provision. Opening up more TCB multiplication centres in different regions will make farmers access the TCB technology for enhanced impact on the target population.

Up-regulation Of Dnam-1 And Nkp30, Associated With Improvement Of Nk Cells Activation After Long-term Culture Of Mononuclear Cells From Patients With Ovarian Neoplasia.

This study aimed at evaluating the functional activation and activating receptors expression on resting, short- and long-term NK and NK-like T cells from blood of ovarian neoplasia patients. Blood from patients with adnexal benign alterations (n = 10) and ovarian cancer (grade I-IV n = 14) were collected after signed consent. Effector cells activation was evaluated by the expression of the CD107a molecule. Short-term culture was conducted overnight with IL-2 and long-term culture for 21 days, by a method designed to expand CD56(+) lymphocytes. Short-term culture significantly increased NK cells activation compared to resting NK cells (p<0.05), however, the long-term procedure supported an even higher increase (p<0.001). Resting NK-like T cells showed poor activation, which was not altered by the culture procedures. The long-term culture effectively increased the expression of the activating receptors on NK and NK-like T cells, either by increasing the number of cells expressing a given receptor and/or by up-regulating their expression intensity. As a conclusion, the long-term culture system employed, resulted in a high number of functional NK cells. The culture system was particularly efficient on the up-regulation of NKp30 and DNAM-1 receptors on NK cells.

Bartonella Spp. Bacteremia In Blood Donors From Campinas, Brazil.

Bartonella species are blood-borne, re-emerging organisms, capable of causing prolonged infection with diverse disease manifestations, from asymptomatic bacteremia to chronic debilitating disease and death. This pathogen can survive for over a month in stored blood. However, its prevalence among blood donors is unknown, and screening of blood supplies for this pathogen is not routinely performed. We investigated Bartonella spp. prevalence in 500 blood donors from Campinas, Brazil, based on a cross-sectional design. Blood samples were inoculated into an enrichment liquid growth medium and sub-inoculated onto blood agar. Liquid culture samples and Gram-negative isolates were tested using a genus specific ITS PCR with amplicons sequenced for species identification. Bartonella henselae and Bartonella quintana antibodies were assayed by indirect immunofluorescence. B. henselae was isolated from six donors (1.2%). Sixteen donors (3.2%) were Bartonella-PCR positive after culture in liquid or on solid media, with 15 donors infected with B. henselae and one donor infected with Bartonella clarridgeiae. Antibodies against B. henselae or B. quintana were found in 16% and 32% of 500 blood donors, respectively. Serology was not associated with infection, with only three of 16 Bartonella-infected subjects seropositive for B. henselae or B. quintana. Bartonella DNA was present in the bloodstream of approximately one out of 30 donors from a major blood bank in South America. Negative serology does not rule out Bartonella spp. infection in healthy subjects. Using a combination of liquid and solid cultures, PCR, and DNA sequencing, this study documents for the first time that Bartonella spp. bacteremia occurs in asymptomatic blood donors. Our findings support further evaluation of Bartonella spp. transmission which can occur through blood transfusions.

Tissue culture of Cecropia glaziovii Sneth (urticaceae): vegetative micropropagation and plant regeneration from callus

Cecropia glaziovii is a tree with used in Brazilian popular medicine. Methods allowing the clonal propagation of this species are of great interest for superior genotype multiplication and perpetuation. For this reason, we examined the effect of different culture media and different types of explants on adventitious shoot regeneration from callus and buds of C. glaziovii. Leaves, petioles and stipules obtained from aseptically grown seedlings or from pre-sterilized plants were used to initiate cultures. Adventitious shoot regeneration was achieved when apical and axillary buds were inoculated on gelled Murashige & Skoog (MS) medium supplemented with 6-benzylaminopurine alone (BAP) (1.0, 5.0 or 10.0 mg L-1) or combined with -naphthalene acetic acid (NAA) (1.0 or 2.0 mg L-1), after 40 days of culture. Best callus production was obtained after 30 days of petioles' culture on gelled MS medium with 2,4 dichlorophenoxyacetic acid (2,4-D) (5.0 mg L-1) combined with BAP (1.0 mg L-1). Successful shoot regeneration from callus was achieved when MS medium supplemented with zeatin (ZEA) (0.1 mg L-1) alone or combined with 2,4-D (1.0 or 5.0 mg L-1) was inoculated with friable callus obtained from petioles. All shoots were rooted by inoculation on MS medium supplemented with indole-3-acetic acid (IAA) (1.0 mg L-1). Rooted plants transferred to potting soil were successfully established. All in vitro regenerated plantlets showed to be normal, without morphological variations, being also identical to the source plant. Our study has shown that C. glaziovii can be propagated by tissue culture methods, allowing large scale multiplication of superior plants for pharmacological purposes.