Granulomatous Slack Skin With Lung and Esophagus Involvement: A Case Report and Molecular Analysis.

ABSTRACT: Granulomatous slack skin (GSS) is a rare subtype of mycosis fungoides, and few cases have been known to spread to the blood, lymph nodes, or viscera. We present a case with early dissemination to the lung. A 27-year-old woman, previously healthy, presented with scattered disseminated scaly patches, associated with vulvar and intergluteal firm swelling and groin-skin induration for 1 year. She also reported mild fatigue and breathlessness on moderate exertion. The patient underwent blood tests, skin biopsies, and computed tomography scan. The skin biopsy showed a mildly atypical T-cell lymphoid infiltrate involving the dermis/hypodermis, with focal epidermotropism, associated with a granulomatous infiltrate and elastophagocytosis. The computed tomography scan revealed bilateral ground-glass lung nodular opacities. Positron emission tomography showed an increased signal in the skin and subcutis around the buttocks, inguinal and mediastinal lymph nodes, and lungs. The lung biopsy confirmed a dense T-cell infiltrate with numerous multinucleated giant cells. Subsequently, esophageal involvement was also observed following biopsy. Molecular analyses demonstrated identical T-cell clones in the skin and lung. After 6 cycles of chemotherapy/localized external radiotherapy, the patient had a partial skin response and stable lung disease. A preferred diagnosis of GSS with systemic spread was made based on clinical/histologic/molecular findings, after considering granulomatous mycosis fungoides and peripheral T-cell lymphoma, not otherwise specified. This case highlights the frequent diagnostic difficulty in distinguishing GSS from an inflammatory granulomatous dermatitis. Pulmonary and esophageal involvements are rare in GSS, and the simultaneous presentation of characteristic cutaneous GSS with systemic disease poses an additional classification challenge.

Predictive evolution of metabolic phenotypes using model-designed environments.

Adaptive evolution under controlled laboratory conditions has been highly effective in selecting organisms with beneficial phenotypes such as stress tolerance. The evolution route is particularly attractive when the organisms are either difficult to engineer or the genetic basis of the phenotype is complex. However, many desired traits, like metabolite secretion, have been inaccessible to adaptive selection due to their trade-off with cell growth. Here, we utilize genome-scale metabolic models to design nutrient environments for selecting lineages with enhanced metabolite secretion. To overcome the growth-secretion trade-off, we identify environments wherein growth becomes correlated with a secondary trait termed tacking trait. The latter is selected to be coupled with the desired trait in the application environment where the trait manifestation is required. Thus, adaptive evolution in the model-designed selection environment and subsequent return to the application environment is predicted to enhance the desired trait. We experimentally validate this strategy by evolving Saccharomyces cerevisiae for increased secretion of aroma compounds, and confirm the predicted flux-rerouting using genomic, transcriptomic, and proteomic analyses. Overall, model-designed selection environments open new opportunities for predictive evolution.

Barriers to nurses performing physical assessments in rehabilitation care units: An observational study.

AIMS: To describe the perceived barriers to nurses performing physical assessments of patients in rehabilitation wards. Secondarily, to investigate how sociodemographic and professional characteristics influence the use and frequency of physical assessments by nurses and their perceptions of barriers to their practice. DESIGN: A multicentre, cross-sectional, observational study. METHODS: Data were collected from September to November 2020 among nurses working with inpatients in eight rehabilitation care institutions in French-speaking Switzerland. Instruments included the Barriers to Nurses' use of Physical Assessment Scale. RESULTS: Almost half of the 112 nurses who responded reported performing physical assessments regularly. The predominant perceived barriers to performing physical assessments were 'specialty area', 'lack of nursing role models' and 'lack of time and interruptions'. Greater clinical nursing experience in rehabilitation wards and more senior nurse specialist positions were associated with significantly lower use of physical assessment procedures by nurses. CONCLUSION: The present study revealed heterogeneity in the use of physical assessment by nurses practicing in rehabilitation units and highlighted their perceived barriers to this. IMPACT STATEMENT: Most nurses working in rehabilitation care units did not routinely perform physical assessments as part of their daily clinical practice. These results should raise stakeholders' awareness of this fact. Effective interventions to increase the use of physical assessments in nursing practice are to be recommended, including continuing education or hiring enough highly qualified nurses as role models in wards. This will promote quality of care and patient safety in rehabilitation care units. PUBLIC AND PATIENT ENGAGEMENT AND INVOLVEMENT: There was no patient or public involvement in the present study.

Adaptive laboratory evolution of microbial co-cultures for improved metabolite secretion.

Adaptive laboratory evolution has proven highly effective for obtaining microorganisms with enhanced capabilities. Yet, this method is inherently restricted to the traits that are positively linked to cell fitness, such as nutrient utilization. Here, we introduce coevolution of obligatory mutualistic communities for improving secretion of fitness-costly metabolites through natural selection. In this strategy, metabolic cross-feeding connects secretion of the target metabolite, despite its cost to the secretor, to the survival and proliferation of the entire community. We thus co-evolved wild-type lactic acid bacteria and engineered auxotrophic Saccharomyces cerevisiae in a synthetic growth medium leading to bacterial isolates with enhanced secretion of two B-group vitamins, viz., riboflavin and folate. The increased production was specific to the targeted vitamin, and evident also in milk, a more complex nutrient environment that naturally contains vitamins. Genomic, proteomic and metabolomic analyses of the evolved lactic acid bacteria, in combination with flux balance analysis, showed altered metabolic regulation towards increased supply of the vitamin precursors. Together, our findings demonstrate how microbial metabolism adapts to mutualistic lifestyle through enhanced metabolite exchange.

Model-guided development of an evolutionarily stable yeast chassis.

First-principle metabolic modelling holds potential for designing microbial chassis that are resilient against phenotype reversal due to adaptive mutations. Yet, the theory of model-based chassis design has rarely been put to rigorous experimental test. Here, we report the development of Saccharomyces cerevisiae chassis strains for dicarboxylic acid production using genome-scale metabolic modelling. The chassis strains, albeit geared for higher flux towards succinate, fumarate and malate, do not appreciably secrete these metabolites. As predicted by the model, introducing product-specific TCA cycle disruptions resulted in the secretion of the corresponding acid. Adaptive laboratory evolution further improved production of succinate and fumarate, demonstrating the evolutionary robustness of the engineered cells. In the case of malate, multi-omics analysis revealed a flux bypass at peroxisomal malate dehydrogenase that was missing in the yeast metabolic model. In all three cases, flux balance analysis integrating transcriptomics, proteomics and metabolomics data confirmed the flux re-routing predicted by the model. Taken together, our modelling and experimental results have implications for the computer-aided design of microbial cell factories.

Cyber-harassment victimization of Portuguese adolescents: A lifestyle-routine activities theory approach.

Cyber-harassment refers to a pattern of repeated, intentional, and unwanted annoyances, impositions, threats, or other aggravating actions mediated by Information and Communication Technologies. Globally, substantial proportions of adolescents experience cyber-harassment, resulting in a host of deleterious health-related consequences. This study tested the empirical utility of the Lifestyle-Routine Activities Theory in accounting for adolescent cyber-victimization. A representative sample of 627 adolescents, aged 12-16, enrolled in schools from northern Portugal and Azores, were surveyed. Consistent with the theoretical assumptions, older adolescents, who used tablets, published information on social network profiles, added unknown people as friends, met face-to-face with them, and had parents with less knowledge of their online contacts were at increased risk of being cyber-victimized. Theoretical and practical implications are elaborated.

Collaborative medication management for older adults after hospital discharge: a qualitative descriptive study.

BACKGROUND: Safe medication management for older adults after hospital discharge requires a well-coordinated, interprofessional, patient-centered approach. This study aimed to describe the perceived needs for collaborative medication management for older adults taking several different medications at home after hospital discharge. METHODS: A qualitative descriptive study was conducted using semi-structured interviews with older adults (n = 28), informal (n = 17), and professional caregivers (n = 13). RESULTS: Findings revealed four main needs: older adults and informal caregivers' perceived needs for greater involvement in discharge planning; older adults' perceived needs to be informed, listened to, and to be actively involved in decision-making; informal caregivers' perceived needs for help in supporting and coordinating medication management; and older adults' and informal and professional caregivers' perceived needs for better communication and coordination between professional caregivers. CONCLUSION: This study revealed two underutilized pathways towards improving collaborative medication management: medication follow-up involving a community healthcare professional taking an overarching responsibility and empowering older adults and their informal caregivers in medication management after hospital discharge.

Very Late Onset of Obsessive-Compulsive Disorder: Case Report and Review of Published Cases in Those More Than 60 Years Old.

ABSTRACT: It is widely agreed that obsessive-compulsive disorder (OCD) is less common among the elderly. However, several studies suggest that a third peak of OCD onset may occur after the age of 65. The onset of OCD in the elderly is unusual and mostly related to nonpsychiatric diagnoses. Nonetheless, some reports have documented late-onset OCD in older adults with no detection of cerebral abnormalities. Such differences in age of onset may be associated with phenotypical differences in disease severity, comorbidity, and treatment response across patients. In this report, we describe the case of late-onset OCD in an 80-year old man with no specific focal brain structural abnormality. The report could improve awareness of the disorder in the elderly and contribute to a better identification of clinical characteristics and additional risk factors of OCD.

Integration of family caregivers in delirium prevention care for hospitalized older adults: A case study analysis.

AIM: To understand how family caregivers of older adults hospitalized for orthopaedic surgery are integrated by nurses in delirium prevention care. DESIGN: Multiple case study. METHODS: The sample consisted of eight cases. Each case comprised an older adult, a family caregiver, and a nurse. Data were collected from September 2017 - April 2018 through various instruments, including semi-structured interviews and family caregiver logs. Within- and across-case analyses were conducted, based on the model of The Care Partner Engagement developed by Hill, Yevchak, Gilmore-Bykovskyi, & Kolanowski (Geriatric Nursing, 35, 2014, 272). RESULTS: Two themes emerged: (a) family caregivers were engaged in caring for the older adults during their hospital stay, though they had differences in terms of views and needs; and (b) family caregivers communicated with nurses but nurses did not recognize their role and did not integrate them much in care. CONCLUSION: The presence and availability of family caregivers, their sense of responsibility towards the hospitalized older adults, and their positive effects on them suggest that family caregivers could be integrated more systematically in a care partnership with nurses. Poor integration of family caregivers in delirium prevention care shows that nurse delirium prevention competencies and their relational skills for communicating effectively with family caregivers need to be developed further. IMPACT: Integrating family caregivers in delirium prevention care for older adults is a challenge for nurses. Family caregivers are engaged during the hospitalization of older adults, though differences and problems exist between the two groups. While there is communication between patients, family caregivers, and nurses, nurses do not recognize the role of family caregivers and hardly integrated them in the delirium prevention care of hospitalized older adults. Nurses must adopt a patient- and family-centred approach. Care and training facilities must make resources available to implement this approach in nursing practice.

Beliefs and implementation of evidence-based practice among nurses in the nursing homes of a Swiss canton: An observational cross-sectional study.

AIMS AND OBJECTIVES: Examine beliefs about EBP and its level of implementation among nurses working in nursing homes in a bilingual canton of Switzerland and explore associations between these aspects and nurses' sociodemographic and professional characteristics. BACKGROUND: Although evidence-based practice (EBP) is recognised as an effective strategy for improving the quality and safety of care, little is known about its use in nursing homes. Nurses' beliefs about EBP and their implementation of it in Switzerland's nursing homes have never been explored. DESIGN: An observational cross-sectional study. METHODS: Beliefs about and implementation of EBP were evaluated using validated French- and German-language versions of the EBP Beliefs Scale and the EBP Implementation Scale, developed by Melnyk and Fineout-Overholt (Melnyk, Fineout-Overholt, & Mays, 2008, Worldviews on Evidence-Based Nursing, 5, 208). The STROBE checklist for cross-sectional studies was used in reporting this study. RESULTS: The participation rate was 40.6% (N = 194). Most participants stated that they had some knowledge of EBP and held favourable beliefs about it. Nevertheless, 37.1% of participants found the concept complicated and 36.1% found it time-consuming. Participants were more likely to implement stages in the EBP process linked to direct clinical practice rather than those which required scientific knowledge and skills. CONCLUSION: Most participants had favourable beliefs about EBP, but the level of implementation of EBP among nurses in their daily clinical practice was sub-optimal. RELEVANCE TO CLINICAL PRACTICE: A greater emphasis should be put on fostering the use of EBP among nurses working in nursing homes. This could be achieved via training and the development of individual, institutional and contextual strategies promoting the integration of EBP in clinical settings.

Identification of hospitalized patients with community-acquired infection in whom treatment guidelines do not apply: a validated model.

OBJECTIVES: To develop and validate a clinical model to identify patients admitted to hospital with community-acquired infection (CAI) caused by pathogens resistant to antimicrobials recommended in current CAI treatment guidelines. METHODS: International prospective cohort study of consecutive patients admitted with bacterial infection. Logistic regression was used to associate risk factors with infection by a resistant organism. The final model was validated in an independent cohort. RESULTS: There were 527 patients in the derivation and 89 in the validation cohort. Independent risk factors identified were: atherosclerosis with functional impairment (Karnofsky index <70) [adjusted OR (aOR) (95% CI) = 2.19 (1.41-3.40)]; previous invasive procedures [adjusted OR (95% CI) = 1.98 (1.28-3.05)]; previous colonization with an MDR organism (MDRO) [aOR (95% CI) = 2.67 (1.48-4.81)]; and previous antimicrobial therapy [aOR (95% CI) = 2.81 (1.81-4.38)]. The area under the receiver operating characteristics (AU-ROC) curve (95% CI) for the final model was 0.75 (0.70-0.79). For a predicted probability ≥22% the sensitivity of the model was 82%, with a negative predictive value of 85%. In the validation cohort the sensitivity of the model was 96%. Using this model, unnecessary broad-spectrum therapy would be recommended in 30% of cases whereas undertreatment would occur in only 6% of cases. CONCLUSIONS: For patients hospitalized with CAI and none of the following risk factors: atherosclerosis with functional impairment; previous invasive procedures; antimicrobial therapy; or MDRO colonization, CAI guidelines can safely be applied. Whereas, for those with some of these risk factors, particularly if more than one, alternative antimicrobial regimens should be considered.

Perceptions About Technologies That Help Community-Dwelling Older Adults Remain at Home: Qualitative Study.

BACKGROUND: The population of Europe is aging rapidly. Most community-dwelling older adults (CDOAs) want to remain in their homes, particularly those experiencing functional decline. Politicians and academics repeatedly praise technological instruments for being the preferred solution for helping older adults with deteriorating health to remain at home. OBJECTIVE: This study aimed to understand the perceptions of CDOAs and their informal caregivers (ICs) and professional caregivers (PCs) about technologies that can help keep older adults at home. METHODS: This qualitative study used personal interviews, focus groups, and photo-elicitation interviews to better understand the perceptions of a convenience sample of 68 CDOAs, 21 ICs, and 32 PCs. RESULTS: A fraction of CDOAs did not perceive technological instruments to be a very useful means of helping them remain at home. However, the ICs and PCs were more positive. The CDOAs preferred and were more willing to adopt technologies related to their mobility and safety and those that would help slow down their cognitive decline. The ICs preferred technological aids that assist in the activities of daily living as well as safety-related technologies for detecting falls and helping to locate disoriented older adults. The PCs preferred integrated communication and information systems to improve collaboration between all stakeholders, housing equipped with technologies to manage complex care, high-performance ancillary equipment to transfer people with reduced mobility, and surveillance systems to ensure safety at home. CONCLUSIONS: Although our study reports that CDOAs have limited interest in innovative technologies to help them remain at home, their technological skills will undoubtedly improve in the future, as will those of ICs and PCs. Technological tools will play an increasingly important role in home health care.

Demographic and Clinical Characteristics of Regular GHB-Users with and without GHB-Induced Comas.

BACKGROUND: Gamma hydroxybutyric acid (GHB) has been used recreationally for nearly three decades and its chronic use is frequently associated with serious adverse events including GHB-intoxication with GHB-induced comas. Moreover, despite its low prevalence, the number of individuals with GHB-use disorders is steadily increasing. However, the risk-factors associated with chronic GHB-use or the development of a GHB-use disorders remain poorly understood. Purpose: This study aims to profile two types of GHB-users, those with and those without GHB-induced comas. Methods: We included 27 GHB users with ≥4 GHB-induced comas (GHB-Coma), 27 GHB users without a coma (GHB-NoComa), and 27 polydrug users who never used GHB (No-GHB). Participants completed self-reported questionnaires in order to assess their demographic and clinical features, and their use profile of GHB and other drugs. Results: The typical GHB user in our sample was young, single, living alone, well-educated, and a student. The GHB-Coma group had lower self-control and reported higher negative affect than the GHB-NoComa group. GHB-Coma participants were heavier GHB users and mostly used GHB alone at home, whereas the GHB-NoComa group mostly used GHB with friends and in nightclubs. Remarkably, the majority of participants were not concerned about potential neurocognitive impairments induced by GHB-intoxication and/or GHB-induced comas. Conclusion: In this assessment, different profiles for recreational users with and without GHB-induced comas were well expressed. Their description contributes to a better understanding of the risk factors associated with recreational GHB-use, GHB-induced coma, and the development of GHB-use disorders.

Recreational use of GHB is associated with alterations of resting state functional connectivity of the central executive and default mode networks.

Gamma-hydroxybutyrate acid (GHB) is a recreational drug with a high addictive potential. Severe side effects such as GHB-induced coma are common and linked to increased emergency room attendances. Task-based functional-imaging studies have revealed an association between the regular use of GHB and multiple GHB-induced comas, and altered neurocognitive function. However the effects of multiple GHB-induced comas and regular GHB-use on intrinsic brain connectivity during rest remain unknown. The study population consisted of 23 GHB-users with ≥4 GHB-induced comas (GHB-Coma), 22 GHB-users who never experienced a GHB-induced coma (GHB-NoComa) and 24 polydrug users who never used GHB (No-GHB). Resting-state scans were collected to assess resting-state functional-connectivity within and between the default mode network (DMN), the bilateral central executive network (CEN) and the salience network (SN). The GHB-NoComa group showed decreased rsFC of the right CEN with a region in the anterior cingulate cortex (pFWE = 0.048) and decreased rsFC between the right CEN and the DMN (pFWE = 0.048) when compared with the No-GHB group. These results suggest that regular GHB-use is associated with decreased rsFC within the right CEN and between the right CEN and the DMN. The presence of multiple GHB-induced comas is not associated with (additional) alterations in rsFC.

Influence of Gamma-Hydroxybutyric Acid-Use and Gamma-Hydroxybutyric Acid-Induced Coma on Affect and the Affective Network.

BACKGROUND: Gamma-hydroxybutyric acid (GHB) is a drug of abuse associated with increased emergency room attendances, due to GHB-induced comas. Withdrawal from GHB often increases social anxiety and is linked to alterations in emotion processing. However, little is known about the effects of GHB-use and GHB-induced comas on affect regulation in humans. OBJECTIVES: We aimed to assess the effect of GHB-use and GHB-induced comas on the affective network. METHOD: We recruited 27 GHB users with ≥4 GHB-induced comas (GHB-Coma), 27 GHB users without a GHB-induced coma (GHB-NoComa), and 27 polydrug users who never used GHB (No-GHB). Participants completed self-report questionnaires assessing negative affect (depression, anxiety and stress) and performed an emotional face matching task during functional magnetic resonance imaging to probe activity of the amygdala and the hippocampus. RESULTS: The GHB-Coma group reported higher levels of depression, anxiety, and stress; showed decreased activity of the hippocampus; and increased functional connectivity of the left hippocampus with the left fusiform gyrus and a cluster on the left temporal-parietal-occipital junction, when compared with the 2 other groups. The GHB-NoComa group showed decreased functional connectivity of the left hippocampus with the amygdala in comparison with the No-GHB group. CONCLUSIONS: GHB-use but in particular GHB-induced comas, are associated with altered emotion identification and hippocampal functioning. Awareness campaigns are required to raise consciousness about the adverse effects of GHB-induced comas on affect regulation, despite the absence of subjective side effects.

Understanding the integration of family caregivers in delirium prevention care for hospitalized older adults: A case study protocol.

AIM: To understand family caregiver involvement in delirium prevention care for older adults hospitalized for orthopaedic surgery hospitals and family caregiver integration by nurses. DESIGN: Multiple-case study. METHODS: The model of Care Partner Engagement was selected as theoretical framework. Eight cases will comprise an older adult hospitalized a family caregiver and a ward nurse. They will be recruited with a non-probability sampling on two orthopaedic surgery wards in two hospitals. Semi-structured interviews with participants will be audiotaped. Sociodemographic data will be collected. These data, researcher field notes and interview transcripts will be subjected to within- and across-case thematic analysis. Regional ethics committee approved the study protocol in August 2018. DISCUSSION: The study will allow surgical nursing teams to gain a better understanding of the issues and possibilities regarding family caregiver integration in delirium prevention care for older adults.

Laboratory evolution reveals regulatory and metabolic trade-offs of glycerol utilization in Saccharomyces cerevisiae.

Most microbial species, including model eukaryote Saccharomyces cerevisiae, possess genetic capability to utilize many alternative nutrient sources. Yet, it remains an open question whether these manifest into assimilatory phenotypes. Despite possessing all necessary pathways, S. cerevisiae grows poorly or not at all when glycerol is the sole carbon source. Here we discover, through multiple evolved lineages, genetic determinants underlying glycerol catabolism and the associated fitness trade-offs. Most evolved lineages adapted through mutations in the HOG pathway, but showed hampered osmotolerance. In the other lineages, we find that only three mutations cause the improved phenotype. One of these contributes counter-intuitively by decoupling the TCA cycle from oxidative phosphorylation, and thereby hampers ethanol utilization. Transcriptomics, proteomics and metabolomics analysis of the re-engineered strains affirmed the causality of the three mutations at molecular level. Introduction of these mutations resulted in improved glycerol utilization also in industrial strains. Our findings not only have a direct relevance for improving glycerol-based bioprocesses, but also illustrate how a metabolic pathway can remain unexploited due to fitness trade-offs in other, ecologically important, traits.

Beliefs and implementation of evidence-based practice among community health nurses: A cross-sectional descriptive study.

AIMS AND OBJECTIVES: To describe beliefs about evidence-based practice and record levels of implementation among community health nurses working independently and in community healthcare centres in the canton of Valais, Switzerland. BACKGROUND: In many settings, evidence-based practice is considered a key means of delivering better and secure health care. However, there is a paucity of published studies on the implementation of evidence-based practice in community health care. DESIGN: Cross-sectional descriptive study (n = 100). METHODS: Beliefs about evidence-based practice and levels of implementation were measured using validated scales developed by Melnyk et al. (Worldviews on Evidence-Based Nursing, 5, 2008, 208). Information on respondents' sociodemographic and professional characteristics was collected. Data were analysed using descriptive and inferential statistics. RESULTS: The final response rate was 32.3% (n = 100). More than half of respondents had previously heard about evidence-based practice; most believed in the value of using evidence to guide their practice and were prepared to improve their skills to be able to do so. However, the rate of implementation of evidence-based practice in daily practice in the 8 weeks before the survey was poor. Statistically significant positive associations were found between beliefs about evidence-based practice and how respondents had heard about it and between implementation rates and whether they had heard about evidence-based practice and how they had done so. Evidence-based practices requiring scientific knowledge and skills were implemented less frequently. Greater professional community healthcare experience and management roles did not increase implementation of evidence-based practice. CONCLUSIONS: The systematic implementation of evidence-based practice by community health nurses working independently and in healthcare centres in Valais was rare, despite their positive beliefs about it. RELEVANCE TO CLINICAL PRACTICE: These results revealed the level of implementation of evidence-based practice by nurses in community healthcare settings in Valais. Further research is required to better understand their needs and expectations and to develop suitable strategies that will allow the integration of evidence-based practice into nurses' daily practice.

Covalent immobilisation of antibodies in Teflon-FEP microfluidic devices for the sensitive quantification of clinically relevant protein biomarkers.

This study reports for the first time the sensitive colorimetric and fluorescence detection of clinically relevant protein biomarkers by sandwich immunoassays using the covalent immobilisation of antibodies onto the fluoropolymer surface inside Teflon®-FEP microfluidic devices. Teflon®-FEP has outstanding optical transparency ideal for high-sensitivity colorimetric and fluorescence bioassays, however this thermoplastic is regarded as chemically inert and very hydrophobic. Covalent immobilisation can offer benefits over passive adsorption to plastic surfaces by allowing better control over antibody density, orientation and analyte binding capacity, and so we tested a range of different and novel covalent immobilisation strategies. We first functionalised the inner surface of a 10-bore, 200 µm internal diameter FEP microcapillary film with high-molecular weight polyvinyl alcohol (PVOH) without changing the outstanding optical transparency of the device delivered by the matched refractive index of FEP and water. Glutaraldehyde immobilisation was compared with the use of photoactivated linkers and NHS-ester crosslinkers for covalently immobilising capture antibodies onto PVOH. Three clinically relevant sandwich ELISAs were tested against the cytokine IL-1ß, the myocardial infarct marker cardiac troponin I (cTnI), and the chronic heart failure marker brain natriuretic peptide (BNP). Overall, glutaraldehyde immobilisation was effective for BNP assays, but yielded unacceptable background for IL-1ß and cTnI assays caused by direct binding of the biotinylated detection antibody to the modified PVOH surface. We found NHS-ester groups reacted with APTES-treated PVOH coated fluoropolymers. This facilitated a novel method for capture antibody immobilisation onto fluoropolymer devices using a bifunctional NHS-maleimide crosslinker. The density of covalently immobilised capture antibodies achieved using PVOH/APTES/NHS/maleimide approached levels seen with passive adsorption, and sensitive and quantitative assay performance was achieved using this method. Overall, the PVOH coating provided an excellent surface for controlled covalent antibody immobilisation onto Teflon®-FEP for performing high-sensitivity immunoassays.

Glucose intolerance after chronic stress is related with downregulated PPAR-γ in adipose tissue.

BACKGROUND: Chronic stress is associated with increased risk of glucose intolerance and cardiovascular diseases, albeit through undefined mechanisms. With the aim of gaining insights into the latter, this study examined the metabolic profile of young adult male rats that were exposed to chronic unpredictable stress. METHODS: Young adult male rats were submitted to 4 weeks of chronic unpredictable stress and allowed to recover for 5 weeks. An extensive analysis including of morphologic, biochemical and molecular parameters was carried out both after chronic unpredictable stress and after recovery from stress. RESULTS: After 28 days of chronic unpredictable stress (CUS) the animals submitted to this protocol displayed less weight gain than control animals. After 5 weeks of recovery the weight gain rebounded to similar values of controls. In addition, following CUS, fasting insulin levels were increased and were accompanied by signs of impaired glucose tolerance and elevated serum corticosteroid levels. This biochemical profile persisted into the post-stress recovery period, despite the restoration of baseline corticosteroid levels. The mRNA expression levels of peroxisome proliferator-activated receptor (PPAR)-γ and lipocalin-2 in white adipose tissue were, respectively, down- and up-regulated. CONCLUSIONS: Reduction of PPAR-γ expression and generation of a pro-inflammatory environment by increased lipocalin-2 expression in white adipose tissue may contribute to stress-induced glucose intolerance.