Haloperidol for the Treatment of Delirium in ICU Patients.

BACKGROUND: Haloperidol is frequently used to treat delirium in patients in the intensive care unit (ICU), but evidence of its effect is limited. METHODS: In this multicenter, blinded, placebo-controlled trial, we randomly assigned adult patients with delirium who had been admitted to the ICU for an acute condition to receive intravenous haloperidol (2.5 mg 3 times daily plus 2.5 mg as needed up to a total maximum daily dose of 20 mg) or placebo. Haloperidol or placebo was administered in the ICU for as long as delirium continued and as needed for recurrences. The primary outcome was the number of days alive and out of the hospital at 90 days after randomization. RESULTS: A total of 1000 patients underwent randomization; 510 were assigned to the haloperidol group and 490 to the placebo group. Among these patients, 987 (98.7%) were included in the final analyses (501 in the haloperidol group and 486 in the placebo group). Primary outcome data were available for 963 patients (97.6%). At 90 days, the mean number of days alive and out of the hospital was 35.8 (95% confidence interval [CI], 32.9 to 38.6) in the haloperidol group and 32.9 (95% CI, 29.9 to 35.8) in the placebo group, with an adjusted mean difference of 2.9 days (95% CI, -1.2 to 7.0) (P = 0.22). Mortality at 90 days was 36.3% in the haloperidol group and 43.3% in the placebo group (adjusted absolute difference, -6.9 percentage points [95% CI, -13.0 to -0.6]). Serious adverse reactions occurred in 11 patients in the haloperidol group and in 9 patients in the placebo group. CONCLUSIONS: Among patients in the ICU with delirium, treatment with haloperidol did not lead to a significantly greater number of days alive and out of the hospital at 90 days than placebo. (Funded by Innovation Fund Denmark and others; AID-ICU ClinicalTrials.gov number, NCT03392376; EudraCT number, 2017-003829-15.).

Tailored Polymersomes for Enhanced Oral Drug Delivery: pH-Sensitive Systems for Intestinal Delivery of Immunosuppressants.

Ensuring precise drug release at target sites is crucial for effective treatment. Here, pH-responsive nanoparticles for oral administration of mycophenolate mofetil, an alternative therapy for patients with inflammatory bowel disease unresponsive to conventional treatments is developed. However, its oral administration presents challenges due to its low solubility in the small intestine and high solubility and absorption in the stomach. Therefore, this aim is to design a drug delivery system capable of maintaining drug solubility compared to the free drug while delaying absorption from the stomach to the intestine. Successful synthesis and assembly of a block copolymer incorporating a pH-responsive functional group is achieved. Dynamic light scattering indicated a significant change in hydrodynamic size when the pH exceeded 6.5, confirming successful incorporation of the pH-responsive group. Encapsulation and controlled release of mycophenolate mofetil are efficiently demonstrated, with 90% release observed at intestinal pH. In vitro cell culture studies confirmed biocompatibility, showing no toxicity or adverse effects on Caco-2 cells. In vivo oral rat studies indicated reduced drug absorption in the stomach and enhanced absorption in the small intestine with the developed formulation. This research presents a promising drug delivery system with potential applications in the treatment of inflammatory bowel disease.

Focusing on positive listening experiences improves hearing aid outcomes in first-time hearing aid users: a randomized controlled trial.

OBJECTIVE: The objective of this study was to investigate whether positive focus (PF), an intervention that asks hearing aid users to focus on positive listening experiences, improves hearing aid outcomes for first-time hearing aid users. DESIGN: The participants were randomised into a control or PF group. They were fitted with hearing aids and followed for six months after fitting. The PF group was asked to report positive listening experiences in their daily life via an app. Participants in both groups were periodically prompted by the app to answer questionnaires about hearing aid satisfaction and benefit. Two follow-up visits at approximately one and six months were performed. STUDY SAMPLE: 20 adult first-time hearing aid users in the control and 18 in the PF group. RESULTS: Hearing aid satisfaction and benefit scores were significantly better in the PF group, already at two weeks and throughout the six months. In the PF group, the hearing aid outcomes were positively correlated with the number of submitted positive reports. CONCLUSIONS: These results point to the importance of asking first-time hearing aid users to focus on positive listening experiences and to reflect upon them. This can lead to improved short- and long-term hearing aid outcomes.

Local Delivery of Streptomycin in Microcontainers Facilitates Colonization of Streptomycin-Resistant Escherichia coli in the Rat Colon.

Oral antibiotic treatment is often applied in animal studies in order to allow establishment of an introduced antibiotic-resistant bacterium in the gut. Here, we compared the application of streptomycin dosed orally in microcontainers to dosage through drinking water. The selective effect on a resistant bacterial strain, as well as the effects on fecal, luminal, and mucosal microbiota composition, were investigated. Three groups of rats (n = 10 per group) were orally dosed with microcontainers daily for 3 days. One of these groups (STR-M) received streptomycin-loaded microcontainers designed for release in the distal ileum, while the other two groups (controls [CTR] and STR-W) received empty microcontainers. The STR-W group was additionally dosed with streptomycin through the drinking water. A streptomycin-resistant Escherichia coli strain was orally inoculated into all animals. Three days after inoculation, the resistant E. coli was found only in the cecum and colon of animals receiving streptomycin in microcontainers but in all intestinal compartments of animals receiving streptomycin in the drinking water. 16S rRNA amplicon sequencing revealed significant changes in the fecal microbiota of both groups of streptomycin-treated animals. Investigation of the inner colonic mucus layer by confocal laser scanning microscopy and laser capture microdissection revealed no significant effect of streptomycin treatment on the mucus-inhabiting microbiota or on E. coli encroachment into the inner mucus. Streptomycin-loaded microcontainers thus enhanced proliferation of an introduced streptomycin-resistant E. coli in the cecum and colon without affecting the small intestine environment. While improvements of the drug delivery system are needed to facilitate optimal local concentration and release of streptomycin, the application of microcontainers provides new prospects for antibiotic treatment. IMPORTANCE Delivery of antibiotics in microcontainer devices designed for release at specific sites of the gut represents a novel approach which might reduce the amount of antibiotic needed to obtain a local selective effect. We propose that the application of microcontainers may have the potential to open novel opportunities for antibiotic treatment of humans and animals with fewer side effects on nontarget bacterial populations. In the current study, we therefore elucidated the effects of streptomycin, delivered in microcontainers coated with pH-sensitive lids, on the selective effect on a resistant bacterium, as well as on the surrounding intestinal microbiota in rats.

The moderating role of social network size in the temporal association between formal social participation and mental health: a longitudinal analysis using two consecutive waves of the Survey of Health, Ageing and Retirement in Europe (SHARE).

INTRODUCTION: Previous studies have shown that engaging in formal social participation may protect against declining mental health, but social network size (the number of close social ties a person has) may moderate the relationship. We assessed the potential moderating role of social network size using longitudinal data. METHODS: Nationally representative data from two consecutive waves (2011, 2013) of the SHARE survey were analyzed. The data consisted of 38,300 adults from 13 European countries aged 50 years and older in 2011. Measures pertaining to formal social participation, social network size, quality of life, and depression symptoms were used. Multivariable linear regression models were conducted. RESULTS: The majority of participants (over 70% of the sample) had a social network size of four or less close social ties. We identified significant moderations in both models. Individuals with relatively few close social ties may have benefitted from formal social participation both in terms of reductions in depression symptoms and increases in quality of life, while formal social participation among those with many social ties did not appear to be beneficial, and may even to some extent have been detrimental. CONCLUSIONS: Declines in mental health specifically among those with relatively few close social ties could potentially be prevented through the promotion of formal social participation. It is possible that such strategies could have a greater impact by specifically targeting individuals that are otherwise socially isolated. High levels of formal participation among those with relatively many close social ties may not be pragmatically beneficial.

Development of predictive models evaluating the spoilage-delaying effect of a bioprotective culture on different yeast species in yogurt.

Yeast spoilage of fermented dairy products causes challenges for the dairy industry, including economic losses due to wasted product. Food cultures with bioprotective effects are becoming more widely used to help ensure product quality throughout product shelf life. To assist the dairy industry when evaluating product quality throughout shelf life and the effect of bioprotective cultures, we aimed to build stochastic models that provide reliable predictions of yeast spoilage in yogurt with and without bioprotective culture. Growth characterizations of Debaryomyces hansenii, Yarrowia lipolytica, Saccharomyces cerevisiae, and Kluyveromyces marxianus at storage temperatures of 7, 12, and 16°C during a 30-d storage period were conducted in yogurt with and without a bioprotective culture containing Lacticaseibacillus rhamnosus strains. The kinetic growth parameters were calculated using the Buchanan growth model, and these parameters were used as baseline values in Monte Carlo models to translate the yeast growth into spoilage levels. The models were developed using 100,000 simulations and they predicted yeast spoilage levels in yogurt by the 4 yeast types. Each modeled yogurt batch was set to be contaminated with yeast at a concentration drawn from a normal distribution with a mean of 1 log10 cfu/mL and standard deviation of 1 log10 cfu/mL and stored for 30 d at a temperature drawn from a normal distribution with a mean of 6.1°C and a standard deviation of 2.8°C. Considering a spoilage level of 5 log10 cfu/mL, the predicted number of spoiled samples was reduced 3-fold during the first 10 d and by 2-fold at the end of shelf life when a bioprotective culture was added to the yogurt. The models were evaluated by sensitivity analyses, where the main effect factors were maximum yeast population, storage temperature, and yeast strain. The models were validated by comparing the model output to actual observed spoilage data from a European dairy using the bioprotective culture. When the model prediction, based on a mixture of the 4 specific yeast strains, was compared with spoilage data from the European dairy, the observed effect of bioprotective cultures was considerably higher than predicted, potentially influenced by the presence of contaminating strains more sensitive to a bioprotective culture than those characterized here. The developed Monte Carlo models can predict yeast spoilage levels in yogurt at specific production settings and how this may be affected by various parameters and addition of bioprotective cultures.

Salmonella in eggs: From shopping to consumption-A review providing an evidence-based analysis of risk factors.

Nontyphoidal salmonellae are among the most prevalent foodborne pathogens causing gastrointestinal infections worldwide. A high number of cases and outbreaks of salmonellosis are associated with the consumption of eggs and egg products, and several of these occur at the household level. The aim of the current study is to critically evaluate the current status of knowledge on Salmonella in eggs from a consumer's perspective, analyzing the hazard occurrence and the good practices that should be applied to reduce salmonellosis risk. Following a HACCP (Hazard Analysis and Critical Control Point) based approach, some steps along the food journey were identified as Critical Consumer Handling (CCH)-steps in which consumers, through their behavior or choice, can significantly reduce the level of Salmonella in eggs and egg products. From shopping/collecting to consumption, each of these steps is discussed in this review to provide an evidence-based overview of risk factors of human salmonellosis related to egg consumption. The main message to consumers is to choose Salmonella-free eggs (those that some official entity or producer guarantees that does not contain Salmonella), when available, especially for dishes that are not fully heat treated. Second, as guaranteed Salmonella-free eggs are only available in a few countries, refrigerated storage from the point of collection and proper cooking will significantly reduce the risk of salmonellosis. This will require a revision of the actual recommendations/regulations, as not all ensure that eggs are maintained at temperatures that prevent growth of Salmonella from collection until the time of purchasing.

Cortical Response to Fat Taste.

We sense fat by its texture and smell, but it is still unknown whether we also taste fat despite evidence of both candidate receptors and distinct fat taste sensations. One major reason fat is still not recognized as a basic taste quality is that we first need to demonstrate its underlying neural activity. To investigate such neural fat taste activation, we recorded evoked responses to commercial cow milk products with 0.1%, 4%, and 38 % fat via high-density electroencephalography (EEG) from 24 human participants. The experimental design ensured that the products would only be discriminable via their potential fat taste; all stimuli were carefully controlled for differences in viscosity, lubrication, odor, temperature, and confounding tastes (sweetness, acidity, and "off-taste") and were delivered directly onto the tongue using a set of computer-controlled syringe pumps. Advanced topographical pattern analysis revealed different neural activation to the milk products 85-134 ms after stimulus onset, which, as expected, best discriminated the two milk fat extremes (0.1% and 38% fat). Notably, this time period has previously been shown to also encode basic taste qualities, such as sweet or salty. By adding to the evidence of cortical fat taste processing in response to staple food, our finding not only substantiates that we taste fat but also highlights its potential relevance during our everyday lives with possible large-scale impacts on motivational eating behavior to explain overconsumption of energy-dense foods.

Development and characterization of a PDMS-based masking method for microfabricated Oral drug delivery devices.

With the growing popularity and application of microfabricated devices in oral drug delivery (ODD), masking technologies for drug loading and surface modification become highly relevant. Considering the speed of design and fabrication processes and the necessity for continuous alterations of e.g. the shape and sizes of the devices during the optimization process, there is a need for adaptable, precise and low-cost masking techniques. Here, a novel method is presented for masking ODD microdevices, namely microcontainers, using the physical characteristics of polydimethylsiloxane (PDMS). When compared to a rigid microfabricated shadow mask, used for filling drugs in microcontainers, the PDMS masking technique allows more facile and precise loading of higher quantities of an active compound, without the need of alignment. The method provides flexibility and is adjustable to devices fabricated from different materials with various geometries, topologies and dimensions. This user-friendly flexible masking method overcomes the limitations of other masking techniques and is certainly not limited to ODD and is recommended for a wide range of microdevices.

Surface Stabilization and Dissolution Rate Improvement of Amorphous Compacts with Thin Polymer Coatings: Can We Have It All?

The distinction between surface and bulk crystallization of amorphous pharmaceuticals, as well as the importance of surface crystallization for pharmaceutical performance, is becoming increasingly evident. An emerging strategy in stabilizing the amorphous drug form is to utilize thin coatings at the surface. While the physical stability of systems coated with pharmaceutical polymers has recently been studied, the effect on dissolution performance as a function of storage time, as a further necessary step toward the success of these formulations, has not been previously studied. Furthermore, the effect of coating thickness has not been elucidated. This study investigated the effect of these polymer-coating parameters on the interplay between amorphous surface crystallization and drug dissolution for the first time. The study utilized simple tablet-like coated dosage forms, comprising a continuous amorphous drug core and thin polymer coating (hundreds of nanometers to a micrometer thick). Monitoring included analysis of both the solid-state of the model drug (with SEM, XRD, and ATR FTIR spectroscopy) and dissolution performance (and associated morphology and solid-state changes) after different storage times. Stabilization of the amorphous form (dependent on the coating thickness) and maintenance of early-stage intrinsic dissolution rates characteristic for the unaged amorphous drug were achieved. However, dissolution in the latter stages was likely inhibited by the presence of a polymer at the surface. Overall, this study introduced a versatile coated system for studying the dissolution of thin-coated amorphous dosage forms suitable for different drugs and coating agents. It demonstrated the importance of multiple factors that need to be taken into consideration when aiming to achieve both physical stability and improved release during the shelf life of amorphous formulations.

Cyberbullying and traditional bullying among Nordic adolescents and their impact on life satisfaction.

Aims: The aim of this study was to investigate the prevalence of cybervictimization in the six Nordic countries and to assess its overlap with traditional bullying. A further aim was to examine potential associations between life satisfaction, on the one hand, and traditional bullying and cyberbullying on the other. Methods: Analyses were based on data from the 2013/2014 Health Behaviour in School-aged Children study. It included 32,210 boys and girls, aged 11, 13, and 15, living in the six Nordic countries. Results: The prevalence of cyberbullying by both pictures and by messages was around 2% in all the Nordic countries except Greenland. There it was considerably higher. The prevalence of being bullied in a traditional manner varied widely by country. For boys, this type of bullying was most frequent in the youngest age group and then decreased steadily in the older age groups. Girls were on average more likely to be cyberbullied. Cyberbullying was more common among 13- and 15-year-olds than 11-year-olds. Higher family affluence was unrelated to the risk of cyberbullying. However, it was related to traditional bullying and combined forms of bullying. Compared with intact families, cybervictimization was commoner among single-parent families and stepfamilies. Adjusting for age, gender, family affluence, and family structure, those subjected to cyberbullying had lower life satisfaction than those who were not bullied. Conclusions: We found relatively little overlap between cyberbullying and traditional bullying, indicating that the two may be separate phenomena stemming from different mechanisms, at least in the Nordic context.

Self-efficacy and social competence reduce socioeconomic inequality in emotional symptoms among schoolchildren.

BACKGROUND: Many adolescents experience mental health problems which may have serious consequences for short- and long-term health and wellbeing. This study investigates socioeconomic inequality in emotional symptoms, self-efficacy and social competence. Further, whether self-efficacy and social competence reduce socioeconomic inequalities in emotional symptoms. METHODS: Data stem from the cross-sectional Danish Health Behaviour in School-aged Children Methodology Development Survey 2012. Data were collected among all schoolchildren in grades 5-9 (11-15-year-olds) in 23 public schools in two municipalities. Participation rate was 76.8% (n = 3969). Analyses of the associations between daily emotional symptoms, occupational social class, self-efficacy and social competence were performed through logistic regression analyses using SAS version 9.3. Multilevel logistic regression analyses were used to study effect modification. RESULTS: Schoolchildren from lower socioeconomic positions have higher odds for daily emotional symptoms and lower levels of high self-efficacy and high social competence compared to schoolchildren from higher socioeconomic positions. High self-efficacy and high social competence buffer the association between socioeconomic position and emotional symptoms, i.e. they seem to protect children and adolescents from lower socioeconomic strata against the higher risk of daily emotional symptoms. CONCLUSIONS: High self-efficacy and high social competence buffer the negative effects of low socioeconomic status on emotional symptoms among schoolchildren. Self-efficacy and social competence can be promoted e.g. through school-based initiatives and may be an effective way to improve mental health and reduce socioeconomic inequality in emotional symptoms among children and adolescents.

Predictors of high and low mental well-being and common mental disorders: findings from a Danish population-based study.

BACKGROUND: Mental well-being is fundamental for a good life. Previous literature has examined the predictors of mental disorders and continuous measures of positive mental health. Very few studies have specifically focused on the predictors of different levels of mental well-being, but those that have suggest a different picture. This study aimed to compare socioeconomic and relational/recreational behaviour predictors of different levels of mental well-being as well as common mental disorders (CMDs). METHODS: Data from 3508 adults aged 16+ years old from the Danish Mental Health and Well-Being Survey 2016 were linked to Danish national register-based data. Mental well-being was assessed using the Warwick-Edinburgh Mental Well-being Scale, and information on CMDs was assessed using the Patient Health Questionnaire (PHQ-4). Regression analyses were conducted to estimate the predictors of low and high mental well-being compared to moderate mental well-being and also of CMDs. RESULTS: Lower socioeconomic position (education, income and employment status) was associated with increased odds of low mental well-being and the presence of CMDs, but did not significantly predict high mental well-being. Relational/recreational behaviours (informal and formal social participation, social support and recreational activity) were associated with reduced odds of low mental well-being and CMDs, and also with increased odds of high mental well-being. CONCLUSIONS: Socioeconomic predictors of high mental well-being do not mirror those of low mental well-being and CMDs, whereas relational/recreational predictors of high mental well-being do mirror those of low mental well-being and CMDs. These findings have important implications for public mental health strategies.

Inactivation of salmonella strains in acidified broth and raw egg yolk as a function of pH and acid type.

Raw egg-based dishes present a safety risk when eggs of uncertain Salmonella status are used. Here, the inactivation of four Salmonella serovars at different combinations of acid, pH and temperature were investigated. Strains of egg or broiler-associated Salmonella serovars Enteritidis, Infantis, Typhimurium, and a heat resistant Senftenberg 775W were tested in broth and egg yolk. It was observed that although S. Senftenberg 775W survived better than its mutant lacking the loci of heat resistance, the wild type per se was not acid tolerant. For all strains, egg yolk acidification with vinegar to pH 3.9 and storage at 25 °C or 8 °C resulted in >4Log(cfu/mL) reductions within 2h or 24h, respectively. At pH 4.2, 2-3Log(cfu/mL) reductions were seen within 6h at 25 °C. In contrast, acidification with lemon juice to pH of 3.9 allowed for growth at 25 °C, while a pH of 2.9 ensured >4Log(cfu/mL) reductions within 24h. Egg yolk and acid form the basis for many recipes and with a ratio of 0.82 of vinegar (≥5% acetic acid) to egg yolk or 1.23 of lemon juice to yolk and storage at 25 °C for 2h or 24h, respectively, a high degree of safety can be obtained, if properly chilled raw eggs are used.

Modular, Lightweight, Wireless Potentiostat-on-a-Disc for Electrochemical Detection in Centrifugal Microfluidics.

Interfacing electrochemical sensors in a lab-on-a-disc (LoD) system with a potentiostat is often tedious and challenging. We here present the first multichannel, modular, lightweight, and wirelessly powered, custom-built potentiostat-on-a-disc (PoD) for centrifugal microfluidic applications. The developed potentiostat is in the form factor of a typical digital video disc (DVD) and weighs only 127 g. The design of the potentiostat facilitates easy and robust interfacing with the electrodes in the LoD system, while enabling real-time electrochemical detection during rotation. The device can perform different electroanalytical techniques such as cyclic voltammetry, square wave voltammetry, and amperometry while being controlled by custom-made software. Measurements were conducted with and without rotation using both in-house fabricated and commercial electrodes. The performance of the PoD was in good agreement with the results obtained using a commercial potentiostat with a measured current resolution of 200 pA. As a proof of concept, we performed a real-time release study of an electrochemically active compound from microdevices used for drug delivery.

Characterization of thin gelatin hydrogel membranes with balloon properties for dynamic tissue engineering.

Cell or tissue stretching and strain are present in any in vivo environment, but is difficult to reproduce in vitro. Here, we describe a simple method for casting a thin (about 500 µm) and soft (about 0.3 kPa) hydrogel of gelatin and a method for characterizing the mechanical properties of the hydrogel simply by changing pressure with a water column. The gelatin is crosslinked with mTransglutaminase and the area of the resulting hydrogel can be increased up 13-fold by increasing the radial water pressure. This is far beyond physiological stretches observed in vivo. Actuating the hydrogel with a radial force achieves both information about stiffness, stretchability, and contractability, which are relevant properties for tissue engineering purposes. Cells could be stretched and contracted using the gelatin membrane. Gelatin is a commonly used polymer for hydrogels in tissue engineering, and the discovered reversible stretching is particularly interesting for organ modeling applications.

Increasing prevalence of emotional symptoms in higher socioeconomic strata: Trend study among Danish schoolchildren 1991-2014.

Aims: The aims of this study were: (a) to examine trends in daily emotional symptoms among 11- to 15-year-olds from 1991 to 2014 in Denmark, and (b) to examine trends in social inequality in daily emotional symptoms, that is, whether the differences in prevalence between adolescents with parents of varying occupational social class changed over time. Methods: We combined seven comparable cross-sectional Health Behaviour in School-aged Children surveys (N=31,169). Daily emotional symptoms were measured by the HBSC Symptom Check List and occupational social class (OSC) by students' reports about parents' occupation. We calculated absolute (per cent) differences in emotional symptoms between high and low OSC and relative differences by odds ratio for emotional symptoms by parents' OSC. Results: Eight per cent reported at least one daily emotional symptoms, with an increasing trend from 1991 to 2014 (p<0.001). The prevalence in high, middle and low OSC was 6.2%, 7.4% and 10.6% (p<0.0001). From 1991 to 2014, there was an increase in the prevalence of daily emotional symptoms in high (p<0.0001) and middle (p<0.0001) but not low OSC (p=0.4404). This resulted in a diminishing absolute social inequality in emotional symptoms. The statistical interaction between OSC and survey year was significant (p=0.0023) and suggests a diminishing relative social inequality in emotional symptoms from 1991 to 2014. Conclusions: There was an increasing prevalence of daily emotional symptoms from 1991 to 2014 and a diminishing social inequality in prevalence of daily emotional symptoms in terms of both absolute and relative social inequality.

Cellular Effects and Delivery Propensity of Penetratin Is Influenced by Conjugation to Parathyroid Hormone Fragment 1-34 in Synergy with pH.

The cell-penetrating peptide (CPP) penetratin has demonstrated potential as a carrier for transepithelial delivery of cargo peptides, such as the therapeutically relevant part of parathyroid hormone, i.e., PTH(1-34). The purpose of the present study was to elucidate the relevance of pH for PTH(1-34)-penetratin conjugates and coadministered penetratin with PTH(1-34) regarding transepithelial permeation of PTH(1-34) and cellular effects. Transepithelial permeation was assessed using monolayers of the Caco-2 cell culture model, and effects on Caco-2 cellular viability kinetics were evaluated by using the Real-Time-GLO assay as well as by microscopy following Tryphan blue staining. Morphological Caco-2 cell changes were studied exploiting the impedance-based xCELLigence system as well as optically using the oCelloscope setup. Finally, the effect of pH on the folding propensity of the PTH(1-34)-penetratin conjugate and its ability to disrupt lipid membranes were assessed by circular dichroism (CD) spectroscopy and the calcein release assay, respectively. The transepithelial PTH(1-34) permeation was not pH-dependent when applying the coadministration approach. However, by applying the conjugation approach, the PTH(1-34) permeation was significantly enhanced by lowering the pH from 7.4 to 5 but also associated with a compromised barrier and a lowering of the cellular viability. The negative effects on the cellular viability following cellular incubation with the PTH(1-34)-penetratin conjugate were moreover confirmed during real-time monitoring of the Caco-2 cell viability as well as by enhanced Tryphan blue uptake. In addition, morphological changes were primarily observed for cells incubated with the PTH(1-34)-penetratin conjugate at pH 5, which was moreover demonstrated to have an enhanced membrane permeating effect following lowering of the pH from 7.4 to 5. The latter observation was, however, not a result of better secondary folding propensity at pH 5 when compared to pH 7.4.

Effects of water-absorption and thermal drift on a polymeric photonic crystal slab sensor.

A photonic crystal slab (PCS) sensor is a universal refractive index sensor with possibilities and performance very similar to surface plasmon resonance (SPR), which represents the gold standard of biosensing. Cheap PCS sensors can be made vacuum-free entirely out of polymers, but come with additional challenges, besides those relating to temperature-variations, which must be considered in any refractive index based method: The polymeric waveguide core was found to swell by ∼0.3% as water absorbed into the waveguide core over ∼1.5 h. This was investigated by monitoring the wavelength of resonant reflection during absorption, by monitoring the release of water using ellipsometry, and by rigorous coupled-wave analysis (RCWA). The approach presented here enables monitoring of water uptake and thermal fluctuations, for drift-free, high-performance operation of a polymeric PCS sensor.

Concordance in BRAF V600E status over time in malignant melanoma and corresponding metastases.

AIMS: The aims of the present study were to analyse the usability of an immunohistochemical (IHC) analysis as compared with a frequently used mutation detection analysis, and to examine the extent of intratumour and intertumour heterogeneity of BRAF V600E in primary tumours and their corresponding metastases. In the development of intertumour heterogeneity between the primary tumour and the corresponding metastases, time as a factor was also investigated. METHODS AND RESULTS: In total, 227 samples from 224 melanoma patients were analysed with both the Cobas 4800 BRAF V600 Mutation Test and IHC anti-BRAF V600E staining. In 82 primary tumours and 224 corresponding metastases, the extents of intertumour and intratumour heterogeneity were investigated with IHC staining. In 15 cases, disagreement between IHC analysis and the Cobas test was seen. In all but one of the examined patients, homogeneity between the primary tumour and the corresponding metastasis was found. Except for this one case, no heterogeneity developed over longer periods. CONCLUSION: IHC analysis can be safely used as a BRAF pretreatment screening tool, and no additional test is needed when staining is positive. However, if stains are negative, additional tests are essential for detection of other BRAF mutations. We suggest that using primary melanoma tissues is just as safe as using metastatic tissue for detection of BRAF V600E, as BRAF intertumour heterogeneity is extremely rare. In addition, the time between diagnosis of the primary tumour and diagnosis of the corresponding metastasis seems not to increase the risk of intertumour heterogeneity.