Foodborne Cereulide Causes Beta-Cell Dysfunction and Apoptosis

Aims/Hypothesis

To study the effects of cereulide, a food toxin often found at low concentrations in take-away meals, on beta-cell survival and function.

Methods

Cell death was quantified by Hoechst/Propidium Iodide in mouse (MIN6) and rat (INS-1E) beta-cell lines, whole mouse islets and control cell lines (HepG2 and COS-1). Beta-cell function was studied by glucose-stimulated insulin secretion (GSIS). Mechanisms of toxicity were evaluated in MIN6 cells by mRNA profiling, electron microscopy and mitochondrial function tests.

Results

24 h exposure to 5 ng/ml cereulide rendered almost all MIN6, INS-1E and pancreatic islets apoptotic, whereas cell death did not increase in the control cell lines. In MIN6 cells and murine islets, GSIS capacity was lost following 24 h exposure to 0.5 ng/ml cereulide (P<0.05). Cereulide exposure induced markers of mitochondrial stress including Puma (p53 up-regulated modulator of apoptosis, P<0.05) and general pro-apoptotic signals as Chop (CCAAT/-enhancer-binding protein homologous protein). Mitochondria appeared swollen upon transmission electron microscopy, basal respiration rate was reduced by 52% (P<0.05) and reactive oxygen species increased by more than twofold (P<0.05) following 24 h exposure to 0.25 and 0.50 ng/ml cereulide, respectively.

Conclusions/Interpretation

Cereulide causes apoptotic beta-cell death at low concentrations and impairs beta-cell function at even lower concentrations, with mitochondrial dysfunction underlying these defects. Thus, exposure to cereulide even at concentrations too low to cause systemic effects appears deleterious to the beta-cell.     

SYTO-13, a Viability Marker as a New Tool to Monitor In Vitro Pharmacodynamic Parameters of Anti-Pneumocystis Drugs

While Pneumocystis pneumonia (PcP) still impacts the AIDS patients, it has a growing importance in immunosuppressed HIV-negative patients. To determine the anti-Pneumocystis therapeutic efficacy of new compounds, animal and in vitro models have been developed. Indeed, well-designed mouse or rat experimental models of pneumocystosis can be used to describe the in vivo anti-Pneumocystis activity of new drugs. In vitro models, which enable the screening of a large panel of new molecules, have been developed using axenic cultures or co-culture with feeder cells; but no universally accepted standard method is currently available to evaluate anti-Pneumocystis molecules in vitro. Thus, we chose to explore the use of the SYTO-13 dye, as a new indicator of Pneumocystis viability. In the present work, we established the experimental conditions to define the in vitro pharmacodynamic parameters (EC50, Emax) of marketed compounds (trimethoprim/sulfamethoxazole, pentamidine, atovaquone) in order to specifically measure the intrinsic activity of these anti-P. carinii molecules using the SYTO-13 dye for the first time. Co-labelling the fungal organisms with anti-P. carinii specific antibodies enabled the measurement of viability of Pneumocystis organisms while excluding host debris from the analysis. Moreover, contrary to microscopic observation, large numbers of fungal cells can be analyzed by flow cytometry, thus increasing statistical significance and avoiding misreading during fastidious quantitation of stained organisms. In conclusion, the SYTO-13 dye allowed us to show a reproducible dose/effect relationship for the tested anti-Pneumocystis drugs.     

Mechanisms of egg contamination by Salmonella Enteritidis

Salmonella Enteritidis (SE) has been the major cause of the food-borne salmonellosis pandemic in humans over the last 20 years, during which contaminated hen's eggs were the most important vehicle of the infection. Eggs can be contaminated on the outer shell surface and internally. Internal contamination can be the result of penetration through the eggshell or by direct contamination of egg contents before oviposition, originating from infection of the reproductive organs. Once inside the egg, the bacteria need to cope with antimicrobial factors in the albumen and vitelline membrane before migration to the yolk can occur. It would seem that serotype Enteritidis has intrinsic characteristics that allow an epidemiological association with hen eggs that are still undefined. There are indications that SE survives the attacks with the help of antimicrobial molecules during the formation of the egg in the hen's oviduct and inside the egg. This appears to require a unique combination of genes encoding for improved cell wall protection and repairing cellular and molecular damage, among others.     

High diversity of root associated fungi in both alpine and arctic <Emphasis Type="Italic">Dryas octopetala</Emphasis>

Background  

Dryas octopetala is a widespread dwarf shrub in alpine and arctic regions that forms ectomycorrhizal (ECM) symbiotic relationships with fungi. In this study we investigated the fungal communities associated with roots of D. octopetala in alpine sites in Norway and in the High Arctic on Svalbard, where we aimed to reveal whether the fungal diversity and species composition varied across the Alpine and Arctic regions. The internal transcribed spacer (ITS) region of nuclear ribosomal DNA was used to identify the fungal communities from bulk root samples obtained from 24 plants.     

On the evolution of demographic strategies in populations with equilibrium and cyclic densities

Optimal demographic strategies (life histories of l_x-m_x schedules) for populations with stable and cylicly varying densities are predicted on the basis of a density dependent population dynamics model; the cyclic density pattern—often seen in lemmings and voles —is assumed to have relatively slow density increase followed by an abrupt density crash. Analysis shows that low reproductive rates for all ages and a Type I or II survival schedule will be favored by natural selection in stable populations. High reproductive rates for all ages and a Type III survival schedule will be favored by natural selection in cyclic populations. These strategies may be regarded as the endpoints of a continuum. Features predicted to be optimal for opportunistic species are expected to be more pronounced the more hostile the environment is. Since all quantities used in formulating these predictions are operationally defined, the predicted patterns are testable. Predictions derived from the model are compared, in a general way, with available data. This comparison suggests the validity of the theory.     

Relationship Between Pneumocystis carinii Burden and the Degree of Host Immunosuppression in an Airborne Transmission Experimental Model

To quantitatively assess the risk of contamination by Pneumocystis depending on the degree of immunosuppression (ID) of the exposed rat hosts, we developed an animal model, where rats went through different doses of dexamethasone. Then, natural and aerial transmission of Pneumocystis carinii occurred during cohousing of the rats undergoing gradual ID levels (receivers) with nude rats developing pneumocystosis (seeders). Following contact between receiver and seeder rats, the P. carinii burden of receiver rats was determined by toluidine blue ortho staining and by qPCR targeting the dhfr monocopy gene of this fungus. In this rat model, the level of circulating CD4⁺ and CD8⁺ T lymphocytes remained significantly stable and different for each dose of dexamethasone tested, thus reaching the goal of a new stable and gradual ID rat model. In addition, an inverse relationship between the P. carinii burden and the level of circulating CD4⁺ or CD8⁺ T lymphocytes was evidenced. This rat model may be used to study other opportunistic pathogens or even co‐infections in a context of gradual ID.     

Sensitization to apoptosis underlies KrasD12-dependent oncolysis of murine C26 colorectal carcinoma cells by reovirus T3D

Reovirus T3D is an oncolytic agent that preferentially targets tumor cells expressing an activated Ras oncogene. Ras signaling interferes with the cellular stress response that inhibits translation of reovirus RNAs. Murine C26 colorectal carcinoma cells express a mutant KrasD12 gene. Reovirus T3D efficiently kills C26 cells, but not C26 cells in which the KrasD12 mRNA is stably repressed by expression of KrasD12-directed short-hairpin RNAs. Surprisingly, neither reovirus T3D protein synthesis nor T3D virus yields were suppressed by deletion of KrasD12. Rather, reovirus-induced tumor cell apoptosis was completely abrogated as a result of Kras knockdown. We conclude that sensitization of C26 tumor cells to reovirus-induced apoptosis underlies the Ras dependency of reovirus T3D oncolysis.     

Analysis of velocity- and power-load relationships of the free-weight back-squat and hexagonal bar deadlift exercises

The aim of this study was to analyse the load-velocity and load-power relationships in the free-weight back-squat (BSQ) and hexagonal bar deadlift (HBD) exercises. Twenty-five (n = 25) resistance-trained men (age = 23.7 ± 2.8 years) performed a progressive load test at maximal intended velocity to determine their BSQ and HBD one-repetition maximum (1RM). Mean propulsive velocity (MPV) during the concentric phase of the lift was recorded through a linear encoder. Load-velocity and load-power relationships were analysed by fitting linear regression and the second-order polynomial, respectively, to the data. Maximum strength (1RM), MPV (30–80% 1RM), and power output (30–90% 1RM) were higher for HBD compared to BSQ exercise (p < 0.05). A very strong relationship between MPV and relative intensity was found for both BSQ (R² = 0.963) and HBD (R² = 0.967) exercises. The load that maximizes power output (P_max) was 64.6 ± 2.9% (BSQ) and 59.6 ± 1.1% (HBD) 1RM. There was a range of loads at which power output was not different than P_max (BSQ: 40–80% 1RM; HBD: 50–70% 1RM). In conclusion, the load-velocity and load-power relationships might assist strength and conditioning coaches to monitor and prescribe exercise intensity in the BSQ and HBD exercises using the velocity-based training approach.     

The use of real-time monitoring during flywheel resistance training programmes: how can we measure eccentric overload? A systematic review and meta-analysis

This systematic review and meta-analysis aimed to analyse the technologies and main training variables used in the literature to monitor flywheel training devices in real time. In addition, as the main research question, we investigated how eccentric overload can be effectively monitored in relation to the training variable, flywheel shaft type device and the moment of inertia selected. The initial search resulted in 11,621 articles that were filtered to twenty-eight and seventeen articles that met the inclusion criteria for the systematic review and meta-analysis, respectively. The main technologies used included force sensors and rotary/linear encoders, mainly to monitor peak or mean force, power or speed. An eccentric overload was not always achieved using flywheel devices. The eccentric overload measurement was related to the main outcome selected. While mean force (p = 0.011, ES = -0.84) and mean power (p < 0.001, ES = -0.30) favoured the concentric phase, peak power (p < 0.001, ES = 0.78) and peak speed (p < 0.001, ES = 0.37) favoured the eccentric phase. In addition, the lower moments of inertia (i.e., from 0.01 to 0.2 kg·m²) and a cylindrical shaft type (i.e., vs conical pulley) showed higher possibilities to achieve eccentric overload. A wide variety of technologies can be used to monitor flywheel devices, but to achieve eccentric overload, a flywheel cylindrical shaft type with low moments of inertia is advised to be used.