Reliable assessment of carbon black nanomaterial of a variety of cell culture media for in vitro toxicity assays by asymmetrical flow field-flow fractionation.

Carbon black nanomaterial (CB-NM), as an industrial product with a large number of applications, poses a high risk of exposure, and its impact on health needs to be assessed. The most common testing platform for engineered (E)NMs is in vitro toxicity assessment, which requires prior ENM dispersion, stabilization, and characterization in cell culture media. Here, asymmetric flow field-flow fractionation (AF4) coupled to UV-Vis and dynamic light scattering (DLS) detectors in series was used for the study of CB dispersions in cell culture media, optimizing instrumental variables and working conditions. It was possible to disperse CB in a non-ionic surfactant aqueous solution due to the steric effect provided by surfactant molecules attached on the CB surface which prevented agglomeration. The protection provided by the surfactant or by culture media alone was insufficient to ensure good dispersion stability needed for carrying out in vitro toxicity studies. On the other hand, cell culture media in combination with the surfactant improved dispersion stability considerably, enabling the generation of shorter particles and a more favourable zeta potential magnitude, leading to greater stability due to electrostatic repulsion. It was demonstrated that the presence of amino acids in the culture media improved the monodisperse nature and stability of the CB dispersions, and resulted in a turn towards more negative zeta potential values when the pH was above the amino acid isoelectric point (IEP). Culture media used in real cell culture scenarios were also tested, and in vitro toxicity assays were developed optimizing the compatible amount of surfactant.

Biotechnological development of Trichoderma-based formulations for biological control.

Trichoderma spp. are a genus of well-known fungi that promote healthy growth and modulate different functions in plants, as well as protect against various plant pathogens. The application of Trichoderma and its propagules as a biological control method can therefore help to reduce the use of chemical pesticides and fertilizers in agriculture. This review critically discusses and analyzes groundbreaking innovations over the past few decades of biotechnological approaches to prepare active formulations containing Trichoderma. The use of various carrier substances is covered, emphasizing their effects on enhancing the shelf life, viability, and efficacy of the final product formulation. Furthermore, the use of processing techniques such as freeze drying, fluidized bed drying, and spray drying are highlighted, enabling the development of stable, light-weight formulations. Finally, promising microencapsulation techniques for maximizing the performance of Trichoderma spp. during application processes are discussed, leading to the next-generation of multi-functional biological control formulations. KEY POINTS: • The development of carrier substances to encapsulate Trichoderma propagules is highlighted. • Advances in biotechnological processes to prepare Trichoderma-containing formulations are critically discussed. • Current challenges and future outlook of Trichoderma-based formulations in the context of biological control are presented.

Biopolymer-based emulsions for the stabilization of Trichoderma atrobrunneum conidia for biological control.

Trichoderma spp. are ubiquitous soil-borne fungi that are widely used in biological control to promote and regulate healthy plant growth, as well as protect against plant pathogens. However, as with many biological materials, the relative instability of Trichoderma propagules limits its practical use in industrial applications. Therefore, there has been significant research interest in developing novel formulations with various carrier substances that are compatible with these fungal propagules and can enhance the shelf-life and overall efficacy of the Trichoderma. To this end, herein, we investigate the use of a variety of biopolymers and nanoparticles for the stabilization of Trichoderma atrobrunneum T720 conidia for biological control. The best-performing agents-agar and cellulose nanocrystals (CNC)-were then used in the preparation of oil-in-water emulsions to encapsulate conidia of T720. Emulsion properties including oil type, oil:water ratio, and biopolymer/particle concentration were investigated with respect to emulsion stability, droplet size, and viability of T720 conidia over time. Overall, agar-based formulations yielded highly stable emulsions with small droplet sizes, showing no evidence of drastic creaming, or phase separation after 1 month of storage. Moreover, agar-based formulations were able to maintain ~ 100% conidial viability of T720 after 3 months of storage, and over 70% viability after 6 months. We anticipate that the results demonstrated herein will lead to a new generation of significantly improved formulations for practical biological control applications. KEY POINTS: • Various biopolymers were evaluated for improving the stability of Trichoderma conidia • Oil in water emulsions was prepared using cellulose nanocrystals and agar as interface stabilizers • Agar-based emulsions showed ~ 100% viability for encapsulated conidia after 3 months of storage.

Determination of antioxidant activity by in situ synthesis of AgNPs using in-tube SPME coupled on-line to capillary liquid chromatography.

A chromatographic system based on in-tube SPME coupled to capillary LC-DAD has been used to study the synthesis of silver nanoparticles using polyphenols in different scenarios: excess of the reducing agent or of the silver salt, addition of the cationic surfactants, and thermal synthesis. The optimized synthesis conditions allowed to quantify the polyphenols used as reducing agents, such as Trolox and chlorogenic acid. Two chromatographic peaks with different absorption spectrum were monitored during the syntheses. Depending on the molar relationship, a linear relation between the area of the chromatographic peaks and the concentration of the silver or polyphenol was established. For stabilization of silver nanoparticles, different cationic surfactants were used allowing to evaluate the role of anion (chloride and bromide) and of the alkyl chain. The proposed methodology can be used to determine chlorogenic acid up to 3 mM with a detection limit of 34 µM at λ= 400 nm. Chlorogenic acid was determined in dietary products with successful results. Precision (RSD=10%) and recovery (97-100%) were also satisfactory.

Scientific evidence on the efficacy of effervescent tablets for cleaning removable prostheses. A systematic review and meta-analysis.

STATEMENT OF PROBLEM: Scientific evidence to determine the optimal method of cleaning and disinfecting removable prostheses is lacking. PURPOSE: The purpose of this systematic review and meta-analysis was to evaluate the effectiveness of effervescent tablets in the cleaning and sanitizing of removable prostheses compared with other chemical and physical methods by assessing the reduction of biofilm, microbial levels, and material stability. MATERIAL AND METHODS: A systematic literature search and meta-analysis was conducted in August 2021 in the MEDLINE/PubMed, Cochrane, Embase, Scopus, and Web of Science databases. Randomized and nonrandomized controlled clinical trials published in English were included without publication year limits. A total of 23 studies were included in the systematic review and 6 in the meta-analysis, which had been registered in the international prospective register of systematic reviews (PROSPERO) database (CRD42021274019). The Cochrane Collaboration tool was used to assess the risk of bias of randomized clinical trials. The physiotherapy evidence database (PEDro) scale was used to analyze the internal validity of clinical trials by assessing the quality of the data obtained. The studies included in the meta-analysis were combined by using a random-effects model with the inverse variance method. Publication bias was analyzed by using the Duvall and Tweedie trim-and-fill method. RESULTS: With regard to biofilm reduction, the standardized mean difference estimated with the 4 studies combined in the meta-analysis was P=.012: mean difference=-1.92; 95% confidence interval=-3.45 to -0.38, indicating a "large" effect of the combination of brushing and effervescent tablet versus brushing alone. To estimate the reduction in the total bacteria levels in the 3 combined studies, a "large" effect size was obtained for the combination of brushing and using an effervescent tablet versus brushing alone, P<.001: mean difference=-4.43; 95% confidence interval=-8.29 to -0.55. Finally, when the 3 studies were combined to assess the reduction of Candida or fungal infection, a "moderate" effect size was obtained for the combination of brushing combined with the use of an effervescent tablet, P<.001: mean difference=-0.78; 95% confidence interval=-1.19 to -0.37. CONCLUSIONS: The combination of brushing and the use of effervescent tablets versus brushing alone had a significantly higher effect on reducing biofilm and bacterial counts and a moderate effect on reducing Candida. Regarding color and dimensional stability, few studies were found, with the results depending on the concentration of the product and the immersion time of the device.

Correction to: Effectiveness and safety of vitamin K antagonists and new anticoagulants in the prevention of thromboembolism in atrial fibrillation in older adults - a systematic review of reviews and the development of recommendations to reduce inappropriate prescribing.

CORRECTION: After publication of the original article [1] it was found that author Marc Krause's name had been spelt incorrectly. In the original article it is presented as Mark Krause, rather than Marc Krause. The revised spelling has been included in the author list for this Correction.

Peptide Metal-Organic Frameworks for Enantioselective Separation of Chiral Drugs.

We report the use of a chiral Cu(II) 3D metal-organic framework (MOF) based on the tripeptide Gly-l-His-Gly (GHG) for the enantioselective separation of metamphetamine and ephedrine. Monte Carlo simulations suggest that chiral recognition is linked to preferential binding of one of the enantiomers as a result of either stronger or additional H-bonds with the framework that lead to energetically more stable diastereomeric adducts. Solid-phase extraction of a racemic mixture by using Cu(GHG) as the extractive phase permits isolating >50% of the (+)-ephedrine enantiomer as target compound in only 4 min. To our knowledge, this represents the first example of a MOF capable of separating chiral polar drugs.

Intrahepatic angiopoietin-2 correlates with chronic hepatitis C progression and is induced in hepatitis C virus replicon systems.

BACKGROUND & AIMS: Chronic hepatitis C (CHC) is a major cause of cirrhosis and hepatocellular carcinoma and angiogenesis is closely related to the pathogenesis and progression of different chronic liver diseases (CLD). Thus, the intrahepatic expression of angiopoietins 1 and 2 (Ang1 and Ang2), as relevant mediators of pathological angiogenesis in several CLD, was investigated. In addition, the differential influence of structural and non-structural genomic regions of HCV on the expression of angiopoietins and the possible signalling involved were studied. METHODS: Ang1 and Ang2 expression was evaluated by western blotting and enzyme-linked immunosorbent assay (ELISA) in liver homogenates of CHC patients (n=47) and uninfected subjects (n=8). Their association with disease progression (according to METAVIR classification) was assessed by Spearman's correlation. Statistical differences among the expression of angiopoietins at different CHC stages were calculated by Mann-Whitney U-test. Finally, the in vitro expression of Angiopoietins in HCV replicons (complete or non-structural subgenomic) and the main signalling pathways involved were also examined. RESULTS: Ang2 levels were significantly higher in the liver of CHC patients compared to controls and significantly correlated with inflammation and fibrosis. Accordingly, an increased expression of Ang2 was found in all HCV replicons tested. Interestingly, the inhibition of MEK and PI3K signalling pathways exerted differential effects on Ang2 expression concerning to the genomic region of HCV. CONCLUSIONS: Hepatitis C virus induces Ang2 expression in hepatocytes through different signalling routes which may lead to the disregulation of vascular homeostasis in the liver. Thus, pharmacologic intervention on Ang2 signalling might constitute an important therapeutic tool.

A set of systematic reviews to help reduce inappropriate prescribing to older people: study protocol.

BACKGROUND: Multimorbidity and polypharmacy are common in older people. Assessment tools or lists of criteria aimed at supporting prescription decisions for older people exist, but have often been based on expert opinion with insufficient consideration of the evidence available. The present paper describes the methods we are using to systematically review the existing evidence on the efficacy and safety of the most commonly prescribed drugs for older people in the management of their chronic medical conditions and to develop recommendations to reduce inappropriate prescriptions for incorporation into the Comprehensive Medication Review (CMR) tool developed by the PRIMA-eDS European project. METHODS: We selected the 20 most relevant drugs/drug classes in terms of prescription volumes and risk of hospitalisation for older people and the most relevant indications for the most common chronic conditions among older people and a total of 35 distinct drug-indication pairs were chosen. Based on clinical considerations we collapsed some indications together, reducing the 35 pairs to a final total of 22 separate systematic reviews (SR). A common methodology will be used for each individual SR, based on the methodological manuals of the Cochrane collaboration and the PRISMA statement for reporting systematic reviews. Our search strategy will have a staged approach where we initially search for systematic reviews and meta-analyses, but if relevant reviews are not found, then search for individual studies (controlled intervention and observational studies). Our pilot work and initial scoping of the literature suggested that very few, relevant individual trials or existing systematic reviews have researched or reported exclusively on older people. Therefore, sufficient data might not be available to perform meta-analysis but we will provide a narrative synthesis describing characteristics and findings of included studies. The collected evidence will be used to construct recommendations on when not to use or to discontinue a drug, or when to reduce its dose. Recommendations will be developed in team meetings using the GRADE methodology to reflect the strength of the recommendation and the quality of the evidence. Recommendations will be built into the CMR tool. DISCUSSION: This protocol has been prepared for a series of systematic reviews which will provide research-based evidence to develop recommendations to reduce inappropriate polypharmacy in older people as part of the CMR tool of the PRIMA-eDS project.

Effectiveness and patient safety of platelet aggregation inhibitors in the prevention of cardiovascular disease and ischemic stroke in older adults - a systematic review.

BACKGROUND: Platelet aggregation inhibitors (PAI) are among the most frequently prescribed drugs in older people, though evidence about risks and benefits of their use in older adults is scarce. The objectives of this systematic review are firstly to identify the risks and benefits of their use in the prevention and treatment of vascular events in older adults, and secondly to develop recommendations on discontinuing PAI in this population if risks outweigh benefits. METHODS: Staged systematic review consisting of three searches. Searches 1 and 2 identified systematic reviews and meta-analyses. Search 3 included controlled intervention and observational studies from review-articles not included in searches 1 and 2. All articles were assessed by two independent reviewers regarding the type of study, age of participants, type of intervention, and clinically relevant outcomes. After data extraction and quality appraisal we developed recommendations to stop the prescribing of specific drugs in older adults following the Grading of Recommendations Assessment Development and Evaluation (GRADE) methodology. RESULTS: Overall, 2385 records were screened leading to an inclusion of 35 articles reporting on 22 systematic reviews and meta-analyses, 11 randomised controlled trials, and two observational studies. Mean ages ranged from 57.0 to 84.6 years. Ten studies included a subgroup analysis by age. Overall, based on the evaluated evidence, three recommendations were formulated. First, the use of acetylsalicylic acid (ASA) for primary prevention of cardiovascular disease (CVD) in older people cannot be recommended due to an uncertainty in the risk-benefit ratio (weak recommendation; low quality of evidence). Secondly, the combination of ASA and clopidogrel in patients without specific indications should be avoided (strong recommendation; moderate quality of evidence). Lastly, to improve the effectiveness and reduce the risks of stroke prevention therapy in older people with atrial fibrillation (AF) and a CHA2DS2-VASc score of ≥ 2, the use of ASA for the primary prevention of stroke should be discontinued in preference for the use of oral anticoagulants (weak recommendation; low quality of evidence). CONCLUSIONS: The use of ASA for the primary prevention of CVD and the combination therapy of ASA and clopidogrel for the secondary prevention of vascular events in older people may not be justified. The use of oral anticoagulants instead of ASA in older people with atrial fibrillation may be recommended. Further high quality studies with older adults are needed.

Efficacy and safety of metformin in the management of type 2 diabetes mellitus in older adults: a systematic review for the development of recommendations to reduce potentially inappropriate prescribing.

BACKGROUND: Metformin is usually prescribed as first line therapy for type 2 diabetes mellitus (DM2). However, the benefits and risks of metformin may be different for older people. This systematic review examined the available evidence on the safety and efficacy of metformin in the management of DM2 in older adults. The findings were used to develop recommendations for the electronic decision support tool of the European project PRIMA-eDS. METHODS: The systematic review followed a staged approach, initially searching for systematic reviews and meta-analyses first, and then individual studies when prior searches were inconclusive. The target population was older people (≥65 years old) with DM2. Studies were included if they reported safety or efficacy outcomes with metformin (alone or in combination) for the management of DM2 compared to placebo, usual or no treatment, or other antidiabetics. Using the evidence identified, recommendations were developed using GRADE methodology. RESULTS: Fifteen studies were included (4 intervention and 11 observational studies). In ten studies at least 80% of participants were 65 years or older and 5 studies reported subgroup analyses by age. Comorbidities were reported by 9 studies, cognitive status was reported by 4 studies and functional status by 1 study. In general, metformin showed similar or better safety and efficacy than other specific or non-specific active treatments. However, these findings were mainly based on retrospective observational studies. Four recommendations were developed suggesting to discontinue the use of metformin for the management of DM2 in older adults with risk factors such as age > 80, gastrointestinal complaints during the last year and/or GFR ≤60 ml/min. CONCLUSIONS: On the evidence available, the safety and efficacy profiles of metformin appear to be better, and certainly no worse, than other treatments for the management of DM2 in older adults. However, the quality and quantity of the evidence is low, with scarce data on adverse events such as gastrointestinal complaints or renal failure. Further studies are needed to more reliably assess the benefits and risks of metformin in very old (>80), cognitively and functionally impaired older people.

Thiazides in the management of hypertension in older adults - a systematic review.

BACKGROUND: Thiazides are commonly prescribed to older people for the management of hypertension. The objective of this study was to identify the evidence on the risks and benefits of their use among adults aged ≥65 years and to develop recommendations to reduce potentially inappropriate use. METHODS: Systematic review (SR) of the literature covering six databases. We applied a staged search approach, where each search was undertaken only if the previous one did not yield high quality results. Searches 1 and 2 identified relevant SRs and meta-analyses published up to December 2015 from all databases. Search 3 identified additional individual interventional studies (IS) and observational studies (OS) not identified by the preceding searches. We included all studies evaluating the effect of thiazides on patient-relevant outcomes in the management of hypertension with a sufficient number of participants aged ≥65 years or a subgroup analysis based on age. Two independent reviewers extracted data and carried out quality appraisal. Recommendations were developed using the GRADE methodology. RESULTS: Searches 1 to 3 were performed. We included 34 articles reporting on 12 IS and 4 OS. Mean ages ranged from 59 to 83.8 years. Four studies had performed a subgroup analysis by age. Information on comorbidity, polypharmacy and frailty of the participants was scarce or not available. The IS compared thiazides to placebo or other antihypertensive drugs and evaluated cardiovascular endpoints or all-cause-mortality as primary outcomes. The OS investigated the association between thiazide use and the risk of gout, fractures and adverse effects. Our results suggest that thiazides are efficacious in preventing cardiovascular events for this population group. Low-dose regimens of thiazides may be safer than high-dose (low quality of evidence), and a history of gout may increase the risk of adverse events (low quality of evidence). Three recommendations were developed. CONCLUSIONS: The use of low dose treatment with thiazides for the management of hypertension in adults aged 65 and older seems justified, unless a history of gout is present. The quality of the evidence is low and studies rarely describe characteristics of the participants such as polypharmacy and frailty. Further good quality studies are needed.

Effectiveness and safety of beta blockers in the management of hypertension in older adults: a systematic review to help reduce inappropriate prescribing.

BACKGROUND: The benefit from a blood pressure lowering therapy with beta blockers may not outweigh its risks, especially in older populations. The aim of this study was to look for evidence on risks and benefits of beta blockers in older adults and to use this evidence to develop recommendations for the electronic decision support tool of the PRIMA-eDS project. METHODS: Systematic review of the literature using a stage approach with searches for systematic reviews and meta-analyses first, and individual studies only if the previous searches are inconclusive. The target population were older adults (≥65 years old) with hypertension. We included studies reporting on the effectiveness and/or safety of beta blockers on clinically relevant endpoints (e.g. mortality, cardiovascular events, and stroke) in the management of hypertension. The recommendations were developed according to the GRADE methodology. RESULTS: Fifteen studies were included, comprising one meta-analysis, four randomized controlled trials, six secondary analyses of randomized controlled trials and four observational studies. Seven studies involved only older adults and eight studies reported subgroup analyses by age. With regard to a composite endpoint (death, stroke or myocardial infarction) beta blockers were associated with a higher risk of events then were other antihypertensive agents. Further, beta blockers showed no benefit compared to other antihypertensive agents or placebo regarding mortality. They appear to be less effective than other antihypertensive agents in reducing cardiovascular events. Contradictory results were found regarding the effect of beta blockers on stroke. None of the studies explored the effect on quality of life, hospitalisation, functional impairment/status, safety endpoints or renal failure. CONCLUSION: The quality of current evidence to interpret the benefits of beta blockers in hypertension is rather weak. It cannot be recommended to use beta blockers in older adults as first line agent for hypertension.

Effectiveness and safety of dipeptidyl peptidase 4 inhibitors in the management of type 2 diabetes in older adults: a systematic review and development of recommendations to reduce inappropriate prescribing.

BACKGROUND: Preventable drug-related hospital admissions can be associated with drugs used in diabetes and the benefits of strict diabetes control may not outweigh the risks, especially in older populations. The aim of this study was to look for evidence on risks and benefits of DPP-4 inhibitors in older adults and to use this evidence to develop recommendations for the electronic decision support tool of the PRIMA-eDS project. METHODS: Systematic review using a staged approach which searches for systematic reviews and meta-analyses first, then individual studies only if prior searches were inconclusive. The target population were older people (≥65 years old) with type 2 diabetes. We included studies reporting on the efficacy and/or safety of DPP-4 inhibitors for the management of type 2 diabetes. Studies were included irrespective of DPP-4 inhibitors prescribed as monotherapy or in combination with any other drug for the treatment of type 2 diabetes. The target intervention was DPP-4 inhibitors compared to placebo, no treatment, other drugs to treat type 2 diabetes or a non-pharmacological intervention. RESULTS: Thirty studies (reported in 33 publications) were included: 1 meta-analysis, 17 intervention studies and 12 observational studies. Sixteen studies were focused on older adults and 14 studies reported subgroup analyses in participants ≥65, ≥70, or ≥75 years. Comorbidities were reported by 26 studies and frailty or functional status by one study. There were conflicting findings regarding the effectiveness of DPP-4 inhibitors in older adults. In general, DPP-4 inhibitors showed similar or better safety than placebo and other antidiabetic drugs. However, these safety data are mainly based on short-term outcomes like hypoglycaemia in studies with HbA1c control levels recommended for younger people. One recommendation was developed advising clinicians to reconsider the use of DPP-4 inhibitors for the management of type 2 diabetes in older adults with HbA1c <8.5% because of scarce data on clinically relevant benefits of their use. Twenty-two of the included studies were funded by pharmaceutical companies and authored or co-authored by employees of the sponsor. CONCLUSIONS: Other than the surrogate endpoint of improved glycaemic control, data on clinically relevant benefits of DPP-4 inhibitors in the treatment of type 2 diabetes mellitus in older adults is scarce. DPP-4 inhibitors might have a lower risk of hypoglycaemia compared to other antidiabetic drugs but data show conflicting findings for long-term benefits. Further studies are needed that evaluate the risks and benefits of DPP-4 inhibitors for the management of type 2 diabetes mellitus in older adults, using clinically relevant outcomes and including representative samples of older adults with information on their frailty status and comorbidities. Studies are also needed that are independent of pharmaceutical company involvement.

New Tools for Characterizing Metallic Nanoparticles: AgNPs, A Case Study.

Currently, transmission electron microscopy (TEM) is the main technique for estimating the sizes of spherical nanoparticles (NPs) and through them, their concentrations. This paper demonstrates for the first time that C18 reversed-phase capillary liquid chromatography (Cap-LC) coupled to diode array detection (DAD) has the potential to estimate mean concentrations of silver nanoparticles (AgNPs) and thereby determine their average size. Direct injection of the sample without previous extraction or separation steps is carried out. Only a unique standard with a known AgNP size is needed for the calibration. In a first approach, the new method has been tested over silver nanoparticles, produced using different methods of synthesis, and their water dilutions. Good results were achieved: relative errors ranged up to 5% compared with TEM. Also stability and functionality-related NP properties, as well as nonspherical AgNPs, can be studied using this method. Moreover, by coupling online in-tube solid-phase microextraction (IT-SPME) to Cap-LC-DAD, the effect of the dilution can be studied as particles distribute by polarity in two groups, a distribution that responds to average particle size of not only AgNPs, but also gold nanoparticles (AuNPs). In such a distribution, the average particle size is correlated with the peak area ratio. Additionally, besides higher sensitivity and concentration-dependent signals, IT-SPME-Cap-LC responds to changes in the particle's hydrodynamic diameter allowing, for instance, the detection of cationic surfactants. Size-exclusion and hydrophobic effects are the mechanisms involved to explain this behavior.

The individual contribution and relative importance of self-management and quality of care on glycaemic control in type 2 diabetes.

OBJECTIVE: To evaluate the relative importance of selfmanagement (SM) and quality of care (QoC) inpredicting glycaemic control in patients with type 2 diabetes. MATERIALS AND METHODS: A longitudinal cohort study was conducted in 204 adults diagnosed with type 2 diabetes. Self-management and quality of care were measured at baseline. HbA1c was measured at baseline and at six-month follow-up. RESULTS: None of the measures of self-management were significantly associated with HbA1c.Treatment intensification (TI) (a proxy for quality of care) resulted in lower HbA1c at follow-up. Other variables were associated with HbA1c at follow-up: HbA1c at baseline, age, diabetes duration, and combination of oral glucose-lowering medications. An exploratory analysis showed that patients who did not receive treatment intensification but performed more self-management behaviours had lower HbA1c levels at follow-up. CONCLUSION: Treatment intensification might be more important for glycaemic control than self-management but the interaction between treatment intensification and self-management needs further research.

Hepatitis C virus variants resistant to macrocyclic NS3-4A inhibitors subvert IFN-ß induction by efficient MAVS cleavage.

BACKGROUND & AIMS: The hepatitis C virus (HCV) NS3-4A protease is essential for the HCV life cycle and a prime target of antiviral treatment strategies. Protease inhibitors, however, are limited by emergence of resistance-associated amino acid variants (RAVs). The capacity to cleave and inactivate mitochondrial antiviral-signaling protein (MAVS) in the RIG-I-signaling pathway is a cardinal feature of NS3-4A, by which HCV blocks induction of interferon-(IFN)-ß, thereby promoting viral persistence. Here, we aimed to investigate the impact of NS3-4A RAVs on MAVS cleavage. METHODS: The impact of NS3-4A RAVs on MAVS cleavage was assessed using immunoblot analyses, luciferase reporter assays and molecular dynamics simulations to study the underlying molecular principles. IFN-ß was quantified in serum from patients with different NS3-4A RAVs. RESULTS: We show that macrocyclic NS3-4A RAVS with substitutions at residue D168 of the protease result in an increased capacity of NS3-4A to cleave MAVS and suppress IFN-ß induction compared with a comprehensive panel of RAVs and wild type HCV. Mechanistically, we show the reconstitution of a tight network of electrostatic interactions between protease and the peptide substrate that allows much stronger binding of MAVS to D168 RAVs than to the wild-type protease. Accordingly, we could show IFN-ß serum levels to be lower in patients with treatment failure due to the selection of D168 variants compared to R155 RAVs. CONCLUSIONS: Our data constitutes a proof of concept that the selection of RAVs against specific classes of direct antivirals can lead to the predominance of viral variants with possibly adverse pathogenic characteristics.

Analysis of polar triazines and degradation products in waters by in-tube solid-phase microextraction and capillary chromatography: an environmentally friendly method.

This paper describes a new method for the determination of polar triazines, including some degradation products, which combines online in-tube solid-phase microextraction (IT-SPME) and capillary liquid chromatography with UV-diode array detection (DAD). Different extractive coatings have been evaluated for IT-SPME, a capillary column with a polydimethylsiloxane (PDMS) coating, and the same coating modified with carboxylated single-wall carbon nanotubes (c-SWCNTs) and carboxylated multiwall carbon nanotubes (c-MWCNTs). On the basis of the results obtained, a new method is presented for the identification and determination of triazines in water samples. A careful selection of the eluent composition provides the required selectivity and sensitivity for the quantification of the target analytes, even those highly polar (log K ow ≤ 2.3). The proposed conditions have been successfully used for the quantitation of the analytes in the 0.25-50.0 µg L(-1) range. The limits of detection (LODs) are in the 0.02-0.1 µg L(-1) range, and the intraday and interday relative standard deviation (RSD) coefficients are ≤9 and ≤17 %, respectively. The reliability of the described method has been tested by analyzing several real water samples. The proposed method can be considered an environmentally friendly and cost-effective alternative for routine monitoring of triazines and their degradation products in waters.

The relationship between quality of care and self-management in patients with type 2 diabetes: a cross-sectional survey in primary care in Mexico.

BACKGROUND: Achieving glycaemic control in diabetes reduces complications and improves outcomes. Glycaemic control requires both good quality clinical care and effective self-management support. However, the relationship between these factors in routine clinical practice is unclear. AIMS: To evaluate baseline levels of self-management and clinical quality of care in patients with type 2 diabetes in primary care in Mexico and to explore relationships between measures of self-management and clinical quality of care. METHODS: The sample consisted of adults (N=205) diagnosed with type 2 diabetes for over a year and registered at one of five practices in the Mexican Institute of Social Security in Aguascalientes. Self-management and quality of care were measured using medical record review and interviews, including validated measures of diabetes knowledge, self-care behaviours, self-efficacy, treatment intensification, continuity of care, doctor-patient communication, and patient satisfaction with diabetes care. HbA1c and cholesterol tests were taken. RESULTS: There were few associations between measures of self-management, and between measures of the quality of clinical care. 'Strong' knowledge about medical prescription was associated with higher diabetes knowledge (OR = 1.2, 95% CI 1.08 to 1.32). Diabetes self-efficacy was associated with self-care behaviours (OR = 1.51, 95% CI 1.26 to 1.81). Patient-doctor communication was associated with continuity of care (Chi-squared = 11.03, P <0.05), with patient satisfaction (ß = 6.17, 95% CI 4.47 to 7.93) and with diabetes self-efficacy (ß = 0.70, 95% CI 0.19 to 1.20, P <0.01). Patient satisfaction was associated with continuity of care (F = 7.82, P <0.001). CONCLUSION: The associations between measures of self-management and quality of care were modest. Patients who were achieving high levels of one aspect of care were not necessarily receiving high levels of the other. This indicates that different factors are likely to be driving each aspect of care and highlights the importance of measuring their relative importance.

Supplementation with a Symbiotic Induced Neuroprotection and Improved Memory in Rats with Ischemic Stroke.

After an ischemic stroke, various harmful mechanisms contribute to tissue damage, including the inflammatory response. The increase in pro-inflammatory cytokines has been related to greater damage to the neural tissue and the promotion of neurological alterations, including cognitive impairment. Recent research has shown that the use of prebiotics and/or probiotics counteracts inflammation and improves cognitive function through the production of growth factors, such as brain-derived neurotrophic factor (BDNF), by reducing inflammatory molecules. Therefore, in this study, the effect of the symbiotic inulin and Enterococcus faecium on neuroprotection and memory improvement was evaluated in a rat model of transient middle cerebral artery occlusion (tMCAO). In order to accomplish this, the animals were subjected to ischemia; the experimental group was supplemented with the symbiotic and the control group with the vehicle. The neurological deficit as well as spatial and working memory were evaluated using the Zea Longa scale, Morris water maze, and the eight-arm maze tests, respectively. Infarct size, the levels of BDNF, and tumor necrosis factor-alpha (TNF-α) were also assessed. The results show that supplementation with the symbiotic significantly diminished the neurological deficit and infarct size, improved memory and learning, increased BDNF expression, and reduced TNF-α production. These findings provide new evidence about the therapeutic use of symbiotics for ischemic stroke and open up the possibilities for the design of further studies.