The A1/A2 ß-casein genotype of cows, but not their horn status, influences peptide generation during simulated digestion of milk.

The effect of the horn status of cows on their milk composition and quality is a controversial research topic. In this study, 128 milk samples from 64 horned and 64 disbudded Brown Swiss and Original Braunvieh cows were collected from alpine farms where both horned and disbudded cows were grazing on mountain pastures. The samples were analyzed for their detailed composition and protein digestion in a simulated in vitro digestion (INFOGEST). To exclude probable influences on digestion, the ß-casein genotype with its variants A1 and A2 was also included in the study. The effects of horn status and ß-casein genotype were investigated in linear mixed models, which included additional influencing random factors such as Original Braunvieh blood proportion, stage of lactation, and farm. Horn status did not have any effect on milk composition or digestion. In contrast, milk from A1A1 cows showed a different protein digestion than milk of A1A2 and A2A2 cows in the gastric phase, including smaller amounts of ß-casomorphin(BCM)21-associated peptides and larger amounts of BCM11-associated peptides. Abundances of BCM7 did not differ between ß-casein genotypes. At the end of the intestinal phase, the digested milk of A1A1 and A2A2 b-casein genotypes did not differ.

Validating international CanMEDS-based standards defining education and safe practice of nurse anesthetists.

AIM: To investigate whether the CanMEDS-based International Federation of Nurse Anesthetists' Standards could adequately define the scope of practice and reliably be used to train and evaluate Swiss nurse anesthetists (NAs). BACKGROUND: Although nurse anesthetists represent a majority of the global workforce in anesthesia, policies that define the scope of practice are frequently non-existent. In low- and middle-income countries, the lack of anesthesia providers with adequate training is a major challenge. INTRODUCTION: Despite stringent training requirements, the scope of practice of Swiss nurse anesthetists is actually not defined. Therefore, we surveyed and assessed whether nurse anesthetists felt that the professional competencies outlined in this framework were aligned with their clinical practice. METHODS: A cross-sectional survey investigated Swiss nurse anesthetists' relevance ratings of 76 competencies of the International Federation of Nurse Anesthetists according to their professional practice. Cronbach's alpha coefficients were used to determine the internal consistency of the competencies, as well as factor analyses to assess construct validity of these competencies integrated into the CanMEDS roles model. RESULTS: Participants rated the Standards overall as very relevant with high reliability. Factor analyses provided evidence of construct validity of these. DISCUSSION: The International Federation of Nurse Anesthetists' Standards of Practice provide a highly relevant framework and a valuable set of competencies for the scope of practice of Swiss nurse anesthetists, which enabled translation from global guides to local national standards. CONCLUSION AND IMPLICATION FOR NURSING AND HEALTH POLICY: Adopted by low- and middle-income countries or countries where national standards are non-existent, this survey could introduce national and local policies at minimally acceptable standards of care for nurse anesthetists worldwide. The above standards have the potential to align education, outcomes and assessment of nurse anesthetists with the needs of national healthcare systems.

Correlation between in vitro and in vivo data on food digestion. What can we predict with static in vitro digestion models?

During the last decade, there has been a growing interest in understanding food's digestive fate in order to strengthen the possible effects of food on human health. Ideally, food digestion should be studied in vivo on humans but this is not always ethically and financially possible. Therefore, simple in vitro digestion models mimicking the gastrointestinal tract have been proposed as alternatives to in vivo experiments. Thus, it is no surprise that these models are increasingly used by the scientific community, although their various limitations to fully mirror the complexity of the digestive tract. Therefore, the objective of this article was to call upon the collective experiences of scientists involved in Infogest (an international network on food digestion) to review and reflect on the applications of in vitro digestion models, the parameters assessed in such studies and the physiological relevance of the data generated when compared to in vivo data. The authors provide a comprehensive review in vitro and in vivo digestion studies investigating the digestion of macronutrients (i.e., proteins, lipids, and carbohydrates) as well as studies of the bioaccessibility and bioavailability of micronutrients and phytochemicals. The main conclusion is that evidences show that despite the simplicity of in vitro models they are often very useful in predicting outcomes of the digestion in vivo. However, this has relies on the complexity of in vitro models and their tuning toward answering specific questions related to human digestion physiology, which leaves a vast room for future studies and improvements.

Adjusting surface coverage of Pt nanocatalyst decoration for selectivity control in CMOS-integrated SnO2 thin film gas sensors.

Smart gas-sensor devices are of crucial importance for emerging consumer electronics and Internet-of-Things (IoT) applications, in particular for indoor and outdoor air quality monitoring (e.g., CO2 levels) or for detecting pollutants harmful for human health. Chemoresistive nanosensors based on metal-oxide semiconductors are among the most promising technologies due to their high sensitivity and suitability for scalable low-cost fabrication of miniaturised devices. However, poor selectivity between different target analytes restrains this technology from broader applicability. This is commonly addressed by chemical functionalisation of the sensor surface via catalytic nanoparticles. Yet, while the latter led to significant advances in gas selectivity, nanocatalyst decoration with precise size and coverage control remains challenging. Here, we present CMOS-integrated gas sensors based on tin oxide (SnO2) films deposited by spray pyrolysis technology, which were functionalised with platinum (Pt) nanocatalysts. We deposited size-selected Pt nanoparticles (narrow size distribution around 3 nm) by magnetron-sputtering inert-gas condensation, a technique which enables straightforward surface coverage control. The resulting impact on SnO2 sensor properties for CO and volatile organic compound (VOC) detection via functionalisation was investigated. We identified an upper threshold for nanoparticle deposition time above which increased surface coverage did not result in further CO or VOC sensitivity enhancement. Most importantly, we demonstrate a method to adjust the selectivity between these target gases by simply adjusting the Pt nanoparticle deposition time. Using a simple computational model for nanocatalyst coverage resulting from random gas-phase deposition, we support our findings and discuss the effects of nanoparticle coalescence as well as inter-particle distances on sensor functionalisation.

Static in vitro digestion model adapted to the general older adult population: an INFOGEST international consensus.

Understanding the mechanisms of food digestion is of paramount importance to determine the effect foods have on human health. Significant knowledge on the fate of food during digestion has been generated in healthy adults due to the development of physiologically-relevant in vitro digestion models. However, it appears that the performance of the oro-gastrointestinal tract is affected by ageing and that a model simulating the digestive conditions found in a younger adult (<65 years) is not relevant for an older adult (>65 years). The objectives of the present paper were: (1) to conduct an exhaustive literature search to find data on the physiological parameters of the older adult oro-gastrointestinal tract, (2) to define the parameters of an in vitro digestion model adapted to the older adult. International experts have discussed all the parameters during a dedicated workshop organized within the INFOGEST network. Data on food bolus properties collected in the older adult were gathered, including food particle size found in older adult boluses. In the stomach and small intestine, data suggest that significant physiological changes are observed between younger and older adults. In the latter, the rate of gastric emptying is slowed down, the pH of the stomach content is higher, the amount of secretions and thus the hydrolytic activities of gastric and intestinal digestive enzymes are reduced and the concentration of bile salts lower. The consensus in vitro digestion model of the older adult proposed here will allow significant progress to be made in understanding the fate of food in this specific population, facilitating the development of foods adapted to their nutritional needs. Nevertheless, better foundational data when available and further refinement of the parameters will be needed to implement the proposed model in the future.

Isolated cerebral manifestation of Epstein-Barr virus-associated post-transplant lymphoproliferative disorder after allogeneic hematopoietic stem cell transplantation: a case of clinical and diagnostic challenges.

We present the case of a 49-year-old male patient with Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorder (PTLD) limited to the brain that occurred 6 months after allogeneic hematopoietic stem cell transplantation (HSCT). Clinical symptoms included mental confusion, ataxia, and diplopia. Magnetic resonance imaging (MRI) revealed cerebellar and periventricular lesions consistent with an inflammatory process. Cerebrospinal fluid (CSF) analysis, but not peripheral blood, was positive for EBV-DNA, but no malignant cells were found. Brain biopsy was not feasible because of low platelet counts. As we considered a diagnosis of either EBV-associated encephalitis or PTLD, the patient was treated with rituximab combined with antiviral therapy. However, the cerebral lesions progressed and follow-up CSF testing revealed immunoglobulin H clonality as evidence of a malignant process. Subsequent treatment attempts included 2 donor lymphocyte infusions (DLI). Despite treatment, the patient died from autopsy-proven PTLD within 8 weeks of the onset of symptoms. This case demonstrates the clinical and diagnostic challenges of primary cerebral PTLD in a patient following allogeneic HSCT.

Endoplasmic reticulum stress-induced cell death mediated by the proteasome.

Cells exposed to sustained endoplasmic reticulum (ER) stress undergo programmed cell death and display features typical of apoptosis, such as cysteine aspartyl protease (caspase) activation, cytochrome c release, and DNA fragmentation. Here, we show that the execution of cell death induced by ER stress is mediated via the proteasome. Inhibition of the proteasome by lactacystin prevented ER stress-induced degradation of Bcl-2, release of cytochrome c, processing of effector caspase-3, and exposure of phosphatidylserine. Owing to the ability of lactacystin to inhibit cytochrome c release, we propose that the pro-apoptotic activity of the proteasome lies upstream of mitochondrial activation. Thus, the proteasome serves as a principal mediator of ER stress-induced cell death in this system.

Apoptotic crosstalk between the endoplasmic reticulum and mitochondria controlled by Bcl-2.

Apoptosis involves mitochondrial steps such as the release of the apoptogenic factor cytochrome c which are effectively blocked by Bcl-2. Although Bcl-2 may have a direct action on the mitochondrial membrane, it also resides and functions on the endoplasmic reticulum (ER), and there is increasing evidence for a role of the ER in apoptosis regulation as well. Here we uncover a hitherto unrecognized, apoptotic crosstalk between the ER and mitochondria that is controlled by Bcl-2. After triggering massive ER dilation due to an inhibition of secretion, the drug brefeldin A (BFA) induces the release of cytochrome c from mitochondria in a caspase-8- and Bid-independent manner. This is followed by caspase-3 activation and DNA/nuclear fragmentation. Surprisingly, cytochrome c release by BFA is not only blocked by wild-type Bcl-2 but also by a Bcl-2 variant that is exclusively targeted to the ER (Bcl-2/cb5). Similar findings were obtained with tunicamycin, an agent interfering with N-linked glycosylations in the secretory system. Thus, apoptotic agents perturbing ER functions induce a novel crosstalk between the ER and mitochondria that can be interrupted by ER-based Bcl-2.

Serine proteases mediate apoptosis-like cell death and phagocytosis under caspase-inhibiting conditions.

Effective execution of apoptosis requires the activation of caspases. However, in many cases, broad-range caspase inhibitors such as Z-VAD.fmk do not inhibit cell death because death signaling continues via basal caspase activities or caspase-independent processes. Although death mediators acting under caspase-inhibiting conditions have been identified, it remains unknown whether they trigger a physiologically relevant cell death that shows typical signs of apoptosis, including phosphatidylserine (PS) exposure and the removal of apoptotic cells by phagocytosis. Here we show that cells treated with ER stress drugs or deprived of IL-3 still show hallmarks of apoptosis such as cell shrinkage, membrane blebbing, mitochondrial release of cytochrome c, PS exposure and phagocytosis in the presence of Z-VAD.fmk. Cotreatment of the stressed cells with Z-VAD.fmk and the serine protease inhibitor Pefabloc (AEBSF) inhibited all these events, indicating that serine proteases mediated the apoptosis-like cell death and phagocytosis under these conditions. The serine proteases were found to act upstream of an increase in mitochondrial membrane permeability as opposed to the serine protease Omi/HtrA2 which is released from mitochondria at a later stage. Thus, despite caspase inhibition or basal caspase activities, cells can still be phagocytosed and killed in an apoptosis-like fashion by a serine protease-mediated mechanism that damages the mitochondrial membrane.

A synthetic inhibitor of interleukin-1 beta converting enzyme prevents endotoxin-induced interleukin-1 beta production in vitro and in vivo.

A potent, reversible, tetrapeptide inhibitor of interleukin-1 beta converting enzyme (ICE), L-709,049, has been shown to suppress the in vitro production of mature IL-1 beta. We now report that this inhibitor also effectively suppresses the production of mature IL-1 beta in a murine model of endotoxic shock. Intraperitoneal administration of L-709,049 reduced the elevations of IL-1 beta in the plasma and peritoneal fluid of mice treated with LPS in a dose-related manner (ED50 = 2 +/- 0.9 mg/kg). LPS-induced elevations in IL-1 alpha and IL-6 in these mice were unaffected, indicating that the inhibitor specifically affected IL-1 beta production. Immunoblot analysis of plasma and peritoneal fluid indicated that L-709,049 suppressed the formation of mature IL-1 beta production in vivo. When mouse blood was incubated in vitro with LPS, IL-1 beta was released into the plasma. This assay was used to determine ex vivo the activity of an ICE inhibitor in the blood following its administration to mice. Blood obtained 15 minutes after ip administration of 10 mg/kg of L-709,049 to mice produced 80% less IL-1 beta than control blood, and IL-1 beta production returned to control levels in blood obtained 30 minutes after injection of this inhibitor. In addition, the capacity of the blood plasma obtained from these animals to prevent the cleavage of a synthetic substrate by ICE disappeared within 1 h of ip administration of 50 mg/kg of inhibitor.

Quantitative morphological analysis of spinal motoneurons.

Horseradish peroxidase was injected intracellularly into motoneurons responding to cutaneous stimulation of the central foot pad of the hind limb in cats. Three motoneurons were selected for detailed analysis: two excited by foot pad stimulation, and one postsynaptically inhibited by such stimulation. The overall lengths of the dendritic trees of the 3 cells ranged from 15.2 to 20.4 mm; the total surface areas ranged from 0.161 to 0.185 mm2. One cell had 9 primary dendrites, 39 terminal dendrites, and 69 dendritic branches in all. The second cell had 8 primary dendrites, 44 terminal dendrites, and 80 dendritic branches in all. For the third cell, these numbers were 14, 76 and 136. 75.2% of the total dendritic length of one of the cells was accounted for by branches of 3 of its 9 primary dendrites; for the second, 69.1% by 3 of 8; and for the third cell, 47.9% by 3 of 14. In contrast to the marked disparity in overall length of branches of the various primary dendrites, when the lengths of dendrites were analyzed by order of branching, dendritic branches of orders II-V each included greater than 15% of overall dendritic length. All 3 motoneurons displayed an exponential overall loss of total dendritic width with distance from the cell body, as well as an exponential decrease in Rall's dendritic trunk parameter. Four different patterns of branching were observed. Mean estimated electronic dendritic lengths were 1.3 for two cells, and 1.2 for the third. However, 13% to 28% of the dendrites of the 3 cells terminated more than two space constants from the cell body. In spite of striking differences among these cells in first-order morphology, on many of the detailed measures characterizing the dendritic trees of these motoneurons, they proved to be strikingly similar.

Health behaviours in a Canadian community college sample: prevalence of drug use and interrelationships among behaviours.

This study investigated the prevalence of drug use among a Canadian college sample and the covariation of drug taking and other health-related behaviours. A representative sample of students at a community college in Alberta were interviewed using telephone surveys, mail-in questionnaires and face-to-face interviews. Data was collected on drug, alcohol and caffeine use, cigarette smoking, eating habits, sleep habits and exercise. While use of illicit drugs did not appear to be widespread, alcohol appeared to be a primary substance abuse problem for a minority of subjects. Factor analysis indicated that the various health habits did not form one dimension of health-related behaviours. Four separate factors emerged: abusive drinking, eating habits, a drug use factor (caffeine intake, smoking, cannabis and hallucinogen use), and exercise levels. Findings are discussed in terms of their implications for future research, treatment and intervention.

A standardised static in vitro digestion method suitable for food - an international consensus.

Simulated gastro-intestinal digestion is widely employed in many fields of food and nutritional sciences, as conducting human trials are often costly, resource intensive, and ethically disputable. As a consequence, in vitro alternatives that determine endpoints such as the bioaccessibility of nutrients and non-nutrients or the digestibility of macronutrients (e.g. lipids, proteins and carbohydrates) are used for screening and building new hypotheses. Various digestion models have been proposed, often impeding the possibility to compare results across research teams. For example, a large variety of enzymes from different sources such as of porcine, rabbit or human origin have been used, differing in their activity and characterization. Differences in pH, mineral type, ionic strength and digestion time, which alter enzyme activity and other phenomena, may also considerably alter results. Other parameters such as the presence of phospholipids, individual enzymes such as gastric lipase and digestive emulsifiers vs. their mixtures (e.g. pancreatin and bile salts), and the ratio of food bolus to digestive fluids, have also been discussed at length. In the present consensus paper, within the COST Infogest network, we propose a general standardised and practical static digestion method based on physiologically relevant conditions that can be applied for various endpoints, which may be amended to accommodate further specific requirements. A frameset of parameters including the oral, gastric and small intestinal digestion are outlined and their relevance discussed in relation to available in vivo data and enzymes. This consensus paper will give a detailed protocol and a line-by-line, guidance, recommendations and justifications but also limitation of the proposed model. This harmonised static, in vitro digestion method for food should aid the production of more comparable data in the future.

[Clinical significance of conventional follow-through examinations of the gastrointestinal passage in a university hospital]. / Klinischer Stellenwert der Durchleuchtung der Magen-Darm-Passage (MDP) an einem Universitätsklinikum.

PURPOSE: The follow-through examination (FTE) is still a widely used radiological method. Modern sectional imaging techniques (CT, MRI, sonography) are established routine examinations offering a wider range of information. In this context the study tries to answer the question of the current significance of FTE of the gastrointestinal tract. MATERIALS AND METHODS: We retrospectively analyzed data of 300 patients, who had undergone FTE between 2001 and 2009 in a university hospital. The medical history, current anamnesis and the therapeutic consequences of the examination for each patient were evaluated based on radiological reports and electronic medical files. RESULTS: The most frequent indication to perform the examination was an uncertain gastrointestinal passage or/and the exclusion of stenosis (70%). In 10% of all FTEs there were complications which led to examination abortion in 2% of cases. In patients who underwent surgery of the abdomen, the examination was performed 8 days (median) after surgery. In 35% of these patients, FTE was done within the first 6 days after surgery. 87% of the patients received further diagnostics before getting pharmacotherapy or surgery. None of the analyzed patients had been operated on after an FTE of the abdomen without being investigated by another diagnostic method. The average radiation exposure was 7 mSv. CONCLUSION: Considering the wide availability of modern sectional imaging methods that are usually necessary for taking significant therapeutic steps, the indication for FTE examinations of the gastrointestinal tract should be very restrictive. The relatively high radiation exposure supports this suggestion.

TTP (time to PEAK) and RBV (regional blood volume) as valuable parameters to detect early flap failure.

BACKGROUND: Free flap transplantation is used more and more frequently in order to cover extensive wound defects. The basic prerequisite for successful flap salvage after flap failure is a short time interval from failure until revision. For this reason many different flap monitoring systems have been tested over the last years. OBJECTIVE: The aim of the experiment was to detect critical capillary perfusion using contrast enhanced ultrasound. Quantitative analysis should be performed by a special perfusion software (QONTRAST; Bracco, Italy) appraising digital raw data of contrast enhanced ultrasound (CEUS). Additionally diverse risk factors for free flap transplantation were determined. METHODS: Thirty-one patients were examined after free flap transplantation during the first 72 hours after operation. CEUS was performed with a linear transducer (6-9 MHz, LOGIQ E9/GE) and a bolus injection of 2.4 ml of contrast agent (SonoVue, Bracco, Italy). Operation and examination were performed by either an experienced plastic surgeon or an experienced ultrasound examiner. Depth dependent capillary perfusion was analysed and quantitative perfusions analysis was performed using the perfusions software QONTRAST (Bracco, Italy). Eleven revisions had to be performed: 7 due to haematoma and 4 due to superficial necrosis. RESULTS: Reduced capillary perfusion was seen in all 11 complications using CEUS. Significant difference comparing the no complication and the complication group was observed using TTP (time to PEAK) and RBV (regional blood volume) quantitative analysis. Mean RBV was 922.1 ± 150.9 in the no complication and 303.0 ± 53.9 in the complication group (p = 0.001). Mean TTP was 37.6 ± 3.8 in the no complication and 21.3 ± 3.4 in the complication group (p = 0.006). Tendency to higher complication rate was seen in older male patients with vascular or malignant primary disease. CONCLUSION: In this clinical trial, capillary perfusion after free flap transplantation as well as detection of vascular complications was demonstrated using CEUS. Quantitative perfusions analysis could be performed and flap viability could be assessed easily.

Primary mouse hepatocytes for systems biology approaches: a standardized in vitro system for modelling of signal transduction pathways.

Complex cellular networks regulate regeneration, detoxification and differentiation of hepatocytes. By combining experimental data with mathematical modelling, systems biology holds great promises to elucidate the key regulatory mechanisms involved and predict targets for efficient intervention. For the generation of high-quality quantitative data suitable for mathematical modelling a standardised in vitro system is essential. Therefore the authors developed standard operating procedures for the preparation and cultivation of primary mouse hepatocytes. To reliably monitor the dynamic induction of signalling pathways, the authors established starvation conditions and evaluated the extent of starvation-associated stress by quantifying several metabolic functions of cultured primary hepatocytes, namely activities of glutathione-S-transferase, glutamine synthetase, CYP3A as well as secretion of lactate and urea into the culture medium. Establishment of constant metabolic activities after an initial decrease compared with freshly isolated hepatocytes showed that the cultured hepatocytes achieve a new equilibrium state that was not affected by our starving conditions. To verify the highly reproducible dynamic activation of signalling pathways in the in vitro system, the authors examined the JAK-STAT, SMAD, PI3 kinase, MAP kinase, NF-kappaB and Wnt/beta-catenin signalling pathways. For the induction of gp130, JAK1 and STAT3 phosphorylation IL6 was used, whereas TGFbeta was applied to activate the phosphorylation of SMAD1, SMAD2 and SMAD3. Both Akt/PKB and ERK1/2 phosphorylation were stimulated by the addition of hepatocyte growth factor. The time-dependent induction of a pool of signalling competent beta-catenin was monitored in response to the inhibition of GSK3beta. To analyse whether phosphorylation is actually leading to transcriptional responses, luciferase reporter gene constructs driven by multiple copies of TGFbeta-responsive motives were applied, demonstrating a dose-dependent increase in luciferase activity. Moreover, the induction of apoptosis by the TNF-like cytokine Fas ligand was studied in the in vitro system. Thus, the mouse hepatocyte in vitro system provides an important basis for the generation of high-quality quantitative data under standardised cell culture conditions that is essential to elucidate critical hepatocellular functions by the systems biology approach.

The relationship between kinetics of substrate-limited transitions and steady-state growth in continuous cultures of Aquaspirillum autotrophicum limited by pyruvate.

Heterotrophic growth at steady state and during transient states caused by the sudden change of the concentration of the limiting factor in the feed medium was investigated experimentally for continuous cultures of Aquaspirillum autotrophicum limited by pyruvate. A model for describing the growth at steady state was selected from three unstructured models after statistical tests of the data. This model postulates that the growth yield increases linearly with the growth rate. Growth during transitions where the substrate remained limiting at all times was fitted with first-order kinetics. Theoretical predictions of these kinetics were derived from the unstructured models used to describe steady state. The predicted rate coefficients of the transients were compared to the experimental coefficients. It appeared that the model which best described steady-state growth also provided the best predictions for growth during the transient state. It is a widespread opinion that unstructured models are adequate to describe growth under steady-state conditions but not to predict transitions in continuous culture. However, for the particular case studied here, no higher degree of complexity was required to describe transitions, provided the growth of the culture was always limited by the substrate.

Purification and characterization of the periplasmic domain of EnvZ osmosensor in Escherichia coli.

The EnvZ-OmpR histidyl-aspartyl phosphorelay system in E. coli responds to osmolarity by differentially modulating the expression of the major outer membrane porins OmpF and OmpC. To date, the natural ligand that activates EnvZ, a transmembrane histidine kinase, has not been identified and the role of the periplasmic domain of EnvZ is unclear. We now report on the purification and characterization of the periplasmic domain of EnvZ (Lys48-Arg162) which has been expressed as a soluble protein in fusion with the maltose-binding protein. Overexpression of the fusion protein did not compete for a signal that activates EnvZ. By amylose affinity chromatography and affinity blotting, interacting proteins could not be detected. The periplasmic domain was released by factor Xa and purified to homogeneity. From circular dichroism analysis, the periplasmic domain was estimated to consist of 35% alpha-helices and 16% beta-sheets.