Long-term urological outcome of cloaca patients with multidisciplinary management.

PURPOSE: Urological management of Cloacal Malformation (CM) focuses on preserving renal function and continence. Study aim was to analyze urinary and intestinal outcomes in CM patients, considering the length of common channel (CC) and presence of occult spinal dysraphism (OSD). METHODS: Retrospective review of CM treated at our institution by a multidisciplinary team from 1999 to 2020. Patients with follow-up < 2.5 years were excluded. Length of CC, renal function, urinary and bowel outcomes, presence of associated anomalies (especially OSD) were evaluated. RESULTS: Twenty patients were included, median age at follow-up: 8 years (4-15). A long CC > 3 cm was described in 11 (55%). Chronic kidney disease was found in 3 patients. Urinary continence was achieved in 8/20 patients, dryness (with intermittent catheterization) in 9/20. Fecal continence was obtained in 3/20, cleanliness in 14 (under bowel regimen). OSD was present in 10 patients (higher prevalence in long-CC, 73%). Among OSD, 1 patient reached fecal continence, 7 were clean; 2 achieved urinary continence, while 6 were dry. CONCLUSIONS: Length of CC and OSD may affect urinary and fecal continence. An early counseling can improve outcome at long-term follow-up. Multidisciplinary management with patient centralization in high grade institutions is recommended to achieve better results.

Inactivation Mechanism of the Fatty Acid Amide Hydrolase Inhibitor BIA 10-2474.

BIA 10-2474 is a time-dependent inhibitor of fatty acid amide hydrolase (FAAH) that was under clinical development for the treatment of neurological conditions when the program was terminated after one subject died and four were hospitalized with neurological symptoms during a first-in-human clinical study. The present work describes the mechanism of FAAH inhibition by BIA 10-2474 as a target-specific covalent inhibition, supported by quantum mechanics and molecular modelling studies. The inhibitor incorporates a weakly reactive electrophile which, upon specific binding to the enzyme's active site, is positioned to react readily with the catalytic residues. The reactivity is enhanced on-site by the increased molarity at the reaction site and by specific inductive interactions with FAAH. In the second stage, the inhibitor reacts with the enzyme's catalytic nucleophile to form a covalent enzyme-inhibitor adduct. The hydrolysis of this adduct is shown to be unlikely under physiological conditions, therefore leading to irreversible inactivation of FAAH. The results also reveal the important role played by FAAH Thr236 in the reaction with BIA 10-2474, which is specific to FAAH and is not present in other serine hydrolases. It forms a hydrogen bond with the imidazole nitrogen of the inhibitor and helps lowering the activation free energy of the first step of the reaction, by pre-orienting and stabilizing the inhibitor in a near-reactive configuration. In the second step, Thr236 can also serve as a mechanistic alternative to protonate the leaving group.

Continuing invariant solutions towards the turbulent flow.

A new mathematical framework is proposed for characterizing the coherent motion of fluctuations around a mean turbulent channel flow. We search for statistically invariant coherent solutions of the unsteady Reynolds-averaged Navier-Stokes equations written in a perturbative form with respect to the turbulent mean flow, using a suitable approximation of the Reynolds stress tensor. This is achieved by setting up a continuation procedure of known solutions of the perturbative Navier-Stokes equations, based on the continuous increase of the turbulent eddy viscosity towards its turbulent value. The recovered solutions, being sustained only in the presence of the Reynolds stress tensor, are representative of the statistically coherent motion of turbulent flows. For small friction Reynolds number and/or domain size, the statistically invariant motion is almost identical to the corresponding invariant solution of the Navier-Stokes equations. Whereas, for sufficiently large friction number and/or domain size, it considerably departs from the starting invariant solution of the Navier-Stokes equations, presenting spatial structures, main wavelengths and scaling very close to those characterizing both large- and small-scale motion of turbulent channel flows. This article is part of the theme issue 'Mathematical problems in physical fluid dynamics (part 2)'.

Occurrence, distribution and dynamics of perchlorate in soil, water, fertilizers, vegetables and fruits and associated human exposure in Chile.

Perchlorate (ClO4-) has been identified as a persistent environmental contaminant of concern. Perchlorate exposure is a potential health concern because it interferes with hormone production by thyroid gland. Food (fruits and vegetables) and drinking water are an important source of human exposure to perchlorate. However, little is known about the occurrence of perchlorate in Chile. The purpose of this study was to determine the occurrence of perchlorate in 145 samples (soil, drinking water, surface water, groundwater, fertilizers, fruits and vegetables) collected across Chile and estimate associated exposure to consumers. Our results show that perchlorate was detected in soil (median: 22.2 ng g-1), drinking water (median: 3.0 ng mL-1), fruits (median: 0.91 ng g-1 fresh weight [FW]), lettuce (median: 5.0 ng g-1 FW) and chard (median: 4.15 ng g-1 FW). Interestingly, perchlorate concentrations detected in drinking water from three regions (Serena, Copiapo and Illapel) exceeded the USEPA interim drinking water health advisory level of 15 ng mL-1. Median concentrations of perchlorate in non-nitrogenous fertilizers (3.1 mg kg-1) were higher than those in nitrogenous fertilizers (1.3 mg kg-1). Estimated daily intake (EDI) of perchlorate via drinking water was lower than the USEPA's reference dose (7000 ng kg-1 bw day-1). The EDI of perchlorate via vegetables (chard and lettuce) produced in northern Chile was three times higher than those produced in other regions. The results of this study provide information about perchlorate sources in Chile, which will be helpful in modifying current regulations.

Consensus statement from the international consensus meeting on post-traumatic cranioplasty.

BACKGROUND: Due to the lack of high-quality evidence which has hindered the development of evidence-based guidelines, there is a need to provide general guidance on cranioplasty (CP) following traumatic brain injury (TBI), as well as identify areas of ongoing uncertainty via a consensus-based approach. METHODS: The international consensus meeting on post-traumatic CP was held during the International Conference on Recent Advances in Neurotraumatology (ICRAN), in Naples, Italy, in June 2018. This meeting was endorsed by the Neurotrauma Committee of the World Federation of Neurosurgical Societies (WFNS), the NIHR Global Health Research Group on Neurotrauma, and several other neurotrauma organizations. Discussions and voting were organized around 5 pre-specified themes: (1) indications and technique, (2) materials, (3) timing, (4) hydrocephalus, and (5) paediatric CP. RESULTS: The participants discussed published evidence on each topic and proposed consensus statements, which were subject to ratification using anonymous real-time voting. Statements required an agreement threshold of more than 70% for inclusion in the final recommendations. CONCLUSIONS: This document is the first set of practical consensus-based clinical recommendations on post-traumatic CP, focusing on timing, materials, complications, and surgical procedures. Future research directions are also presented.

How to dissect the plasticity of antigen-specific immune response: a tissue perspective.

Generation of antigen-specific humoral responses following vaccination or infection requires the maturation and function of highly specialized immune cells in secondary lymphoid organs (SLO), such as lymph nodes or tonsils. Factors that orchestrate the dynamics of these cells are still poorly understood. Currently, experimental approaches that enable a detailed description of the function of the immune system in SLO have been mainly developed and optimized in animal models. Conversely, methodological approaches in humans are mainly based on the use of blood-associated material because of the challenging access to tissues. Indeed, only few studies in humans were able to provide a discrete description of the complex network of cytokines, chemokines and lymphocytes acting in tissues after antigenic challenge. Furthermore, even fewer data are currently available on the interaction occurring within the complex micro-architecture of the SLO. This information is crucial in order to design particular vaccination strategies, especially for patients affected by chronic and immune compromising medical conditions who are under-vaccinated or who respond poorly to immunizations. Analysis of immune cells in different human tissues by high-throughput technologies, able to obtain data ranging from gene signature to protein expression and cell phenotypes, is needed to dissect the peculiarity of each immune cell in a definite human tissue. The main aim of this review is to provide an in-depth description of the current available methodologies, proven evidence and future perspectives in the analysis of immune mechanisms following immunization or infections in SLO.

[Prevalence of transmitted drug resistance in HIV-infected treatment-naive patients in Chile]. / Prevalencia de resistencia transmitida a drogas antirretrovirales en pacientes con infección por VIH en Chile (2014-2018).

BACKGROUND: Transmitted drug resistance (TDR) occurs in patients with HIV infection who are not exposed to antiretroviral drugs but who are infected with a virus with mutations associated with resistance. AIM: To determine the prevalence of TDR and characterize HIV reverse transcriptase and protease mutation patterns. MATERIAL AND METHODS: HIV infected antiretroviral treatment-naive patients treated in three centers between 2014 and 2018 were studied. A genotyping study was carried out. The HIVdb Program (Stanford University) and the World Health Organization (WHO) TDR surveillance mutation list were used to register resistance-associated mutations. RESULTS: We enrolled 220 patients aged a median of 29 (interquartile range (IQR) 24-34) years, 99% men. Median CD4 count was 365 cells/µL (IQR 250-499 cells/µL) and median viral load was 39.150 copies/mL (IQR 9,270 -120,000). The overall prevalence of RTD was 10.45% (95% CI 6.7-15.2, N = 23/220). The higher frequency of TDR was against non-nucleoside reverse transcriptase inhibitors, reaching 9.0% (95% CI 5.6-13.6), followed by nucleoside reverse transcriptase inhibitors reaching 1.8% (95% CI 0.49-4.5) and protease inhibitors reaching 0.45% (95% CI 0.01-2.5). The mutations in reverse transcriptase were M41L, L210W, D67N, K70E, M184V, K103N (6.36%, 95% CI 3.5-10.4), G190A, E138A, K101E, and I84V in protease. CONCLUSIONS: These results should prompt a change in recommendations for starting antiretoviral therapy, especially in first-line regimens that include non-nucleoside reverse transcriptase inhibitors.

Chemical and ecotoxicological effects of the use of drinking-water treatment residuals for the remediation of soils degraded by mining activities.

The aim of this study was to evaluate the use of drinking-water treatment residuals (DWTR) in the amendment of a soil affected by mining activities (Aljustrel mine, Portuguese sector of the Iberian Pyrite Belt), considering the effects on its chemical, biochemical and ecotoxicological characteristics. The DWTR had neutral characteristics (pH 6.7) and an organic matter (OM) content of 575 g kg-1 dry matter (DM), which makes them a potential amendment for the remediation of mine degraded soils, as they may correct soil acidity and reduce the extractable metal fraction. An incubation assay, with soil and DWTR, with or without lime, was carried out to test the doses to be used in the assisted-phytostabilization experiment. Based on the results obtained, the doses of DWTR used were the equivalent to 48, 96, and 144 t DM ha-1, with and without lime application (CaCO3 11 t DM ha-1). Agrostis tenuis Sibth was used as the test plant. Some amendments doses were able to improve soil characteristics (pH and OM content), to decrease metal extractability by 0.01 M CaCl2 (especially for Cu and Zn), and to allow plant growth, that did not occur in the non-amended soil. Copper, Pb and Zn concentrations in the plant material were lower than the maximum tolerable level for cattle feed, used as an indicator of risk of entry of those metals into the human food chain. The simultaneous application of DWTR (96 and 144 t ha-1), with lime, allowed a reduction in the mine soil ecotoxicity, as evaluated by some lethal and sub-lethal bioassays, including luminescence inhibition of Vibrio fischeri, Daphnia magna acute immobilization test, mortality of Thamnocephalus platyurus, and 72-h growth inhibition of the green microalgae Pseudokirchneriella subcapitata. However, DWTR were unable to increase soil microbial activity, evaluated by dehydrogenase activity, an important soil-health indicator. Also, OM content and NKjeldahl, concentrations increased slightly but remained low or very low (P and K extractable concentrations were not affected). In general, the bioassays highlighted a decrease in soil ecotoxicity with the presence of lime and DWTR (144 t DM ha-1). In conclusion, DWTR are recommended to amend acidic soils, with high concentrations of trace elements, but an additional application of organic or mineral fertilizers should be considered.

Association of the Familial Coexistence of Child Stunting and Maternal Overweight with Indigenous Women in Guatemala.

Objectives This study investigated the association of the familial coexistence of child stunting and maternal overweight with indigenous women in Guatemala. Methods We selected 2388 child-mother pairs from the data set of the Living Standards Measurement Study conducted in Guatemala in 2000. This study examined the association between maternal and household characteristics and the nutritional status of children aged 6-60 months and mothers aged 18-49 years by using multivariable logistic regression models. Results Compared with non-indigenous households, a significantly higher percentage of indigenous households exhibited stunted child and overweight mother (SCOM) pairs (15.9 vs. 22.2%). Compared with normal-weight mothers, overweight mothers were less likely to have stunted children [adjusted odds ratio (AOR) 0.66, 95% confidence interval (CI) 0.50-0.88]. However, compared with mothers who were not short and overweight, short and overweight mothers were significantly more likely to have stunted children (AOR 1.80, 95% CI 1.19-2.73) and were more likely to be indigenous women living in urban areas (AOR 3.01, 95% CI 1.19-7.60) or rural areas (AOR 3.02, 95% CI 1.28-7.14). The order of observed prevalence of SCOM pairs in different types of households was as follows: urban indigenous (25.0%), rural indigenous (21.2%), rural non-indigenous (19.8%), and urban non-indigenous households (10.7%). Conclusions for Practice Urban indigenous households were more likely to have SCOM pairs. This study provided useful information for identifying the most vulnerable groups and areas with a high prevalence of the familial coexistence of child stunting and maternal overweight.

A review of perchlorate (ClO4-) occurrence in fruits and vegetables.

Since the 1990s, a large number of studies around the world have reported the presence of perchlorate in different types of environmental matrices. In view of their inherent characteristics, such as high solubility, mobility, persistence, and low affinity for the surface of soil, perchlorates are mobilized through the water-soil system and accumulate in edible plant species of high human consumption. However, the ingestion of food products containing perchlorate represents a potential health risk to people due to their adverse effects on thyroid, hormone, and neuronal development, mainly in infants and fetuses. At present, research has been centered on determining sources, fates, and remediation methods and not on its real extension in vegetables under farming conditions. This review presents a comprehensive overview and update of the frequent detection of perchlorate in fruits and vegetables produced and marketed around the world. Additionally, the impact of fertilizer on the potential addition of perchlorate to soil and its mobility in the water-soil-plant system is discussed. This review is organized into the following sections: sources of perchlorate, mobility in the water-soil system, presence in fruits and vegetables in different countries, international regulations, and toxicological studies. Finally, recommendations for future studies concerning perchlorate in fruits and vegetables are presented.

Antibodies to human serum amyloid P component eliminate visceral amyloid deposits.

Accumulation of amyloid fibrils in the viscera and connective tissues causes systemic amyloidosis, which is responsible for about one in a thousand deaths in developed countries. Localized amyloid can also have serious consequences; for example, cerebral amyloid angiopathy is an important cause of haemorrhagic stroke. The clinical presentations of amyloidosis are extremely diverse and the diagnosis is rarely made before significant organ damage is present. There is therefore a major unmet need for therapy that safely promotes the clearance of established amyloid deposits. Over 20 different amyloid fibril proteins are responsible for different forms of clinically significant amyloidosis and treatments that substantially reduce the abundance of the respective amyloid fibril precursor proteins can arrest amyloid accumulation. Unfortunately, control of fibril-protein production is not possible in some forms of amyloidosis and in others it is often slow and hazardous. There is no therapy that directly targets amyloid deposits for enhanced clearance. However, all amyloid deposits contain the normal, non-fibrillar plasma glycoprotein, serum amyloid P component (SAP). Here we show that administration of anti-human-SAP antibodies to mice with amyloid deposits containing human SAP triggers a potent, complement-dependent, macrophage-derived giant cell reaction that swiftly removes massive visceral amyloid deposits without adverse effects. Anti-SAP-antibody treatment is clinically feasible because circulating human SAP can be depleted in patients by the bis-d-proline compound CPHPC, thereby enabling injected anti-SAP antibodies to reach residual SAP in the amyloid deposits. The unprecedented capacity of this novel combined therapy to eliminate amyloid deposits should be applicable to all forms of systemic and local amyloidosis.

Pathogenetic mechanisms of amyloid A amyloidosis.

Systemic amyloid A (AA) amyloidosis is a serious complication of chronic inflammation. Serum AA protein (SAA), an acute phase plasma protein, is deposited extracellularly as insoluble amyloid fibrils that damage tissue structure and function. Clinical AA amyloidosis is typically preceded by many years of active inflammation before presenting, most commonly with renal involvement. Using dose-dependent, doxycycline-inducible transgenic expression of SAA in mice, we show that AA amyloid deposition can occur independently of inflammation and that the time before amyloid deposition is determined by the circulating SAA concentration. High level SAA expression induced amyloidosis in all mice after a short, slightly variable delay. SAA was rapidly incorporated into amyloid, acutely reducing circulating SAA concentrations by up to 90%. Prolonged modest SAA overexpression occasionally produced amyloidosis after long delays and primed most mice for explosive amyloidosis when SAA production subsequently increased. Endogenous priming and bulk amyloid deposition are thus separable events, each sensitive to plasma SAA concentration. Amyloid deposits slowly regressed with restoration of normal SAA production after doxycycline withdrawal. Reinduction of SAA overproduction revealed that, following amyloid regression, all mice were primed, especially for rapid glomerular amyloid deposition leading to renal failure, closely resembling the rapid onset of renal failure in clinical AA amyloidosis following acute exacerbation of inflammation. Clinical AA amyloidosis rarely involves the heart, but amyloidotic SAA transgenic mice consistently had minor cardiac amyloid deposits, enabling us to extend to the heart the demonstrable efficacy of our unique antibody therapy for elimination of visceral amyloid.

Hereditary systemic amyloidosis due to Asp76Asn variant ß2-microglobulin.

We describe a kindred with slowly progressive gastrointestinal symptoms and autonomic neuropathy caused by autosomal dominant, hereditary systemic amyloidosis. The amyloid consists of Asp76Asn variant ß(2)-microglobulin. Unlike patients with dialysis-related amyloidosis caused by sustained high plasma concentrations of wild-type ß(2)-microglobulin, the affected members of this kindred had normal renal function and normal circulating ß(2)-microglobulin values. The Asp76Asn ß(2)-microglobulin variant was thermodynamically unstable and remarkably fibrillogenic in vitro under physiological conditions. Previous studies of ß(2)-microglobulin aggregation have not shown such amyloidogenicity for single-residue substitutions. Comprehensive biophysical characterization of the ß(2)-microglobulin variant, including its 1.40-Å, three-dimensional structure, should allow further elucidation of fibrillogenesis and protein misfolding.

Salivary Urease and ADS Enzymatic Activity as Endogenous Protection against Dental Caries in Children.

UNLABELLED: The aim of this cross sectional study was to evaluate the ureolytic and arginolytic activities of saliva in children and associate them with their caries status. STUDY DESIGN: 65, 8 year old children, were randomly selected. The ureolytic and arginolytic activity of non stimulated saliva was studied and associated with DMFT and dmft index. Saliva of children were sampled under fasting conditions; Children refrained from any oral hygiene procedures during the 12 hours that preceded the sample collection. Caries activity was scored and divided in 3 groups: Group A: Index zero: without lesions; Group B: Moderate Index: 1 to 3 enamel caries lesions; and Group C: High Index: more than 4 dentin caries lesions. RESULTS: DMFT scores were moderate: 0.4(±0.79) and dmft: 2.78(±2.45). Results expressed in µmol/min/mg/protein, for urease activity were statistically significant (p=0.048): Group A= 0.69 (±0.7); Group B= 0.45 (±0.43); and Group C= 0.39 (±0.55). The arginine deiminase activity was not statistically significant (p=0.16): Group A= 2.53 (±1.42), Group B= 2.31 (±1.57) and Group C= 1.97 (±2.0). CONCLUSION: Higher levels of ureolytic (statistically significant) and arginolytic activity (trend) in saliva were associated with lower DMFT/dmft scores in 8 year old children. There was a higher production of ammonia from the arginine deiminase system than the urease enzyme in saliva (p>0.05).

Perchlorate levels in soil and waters from the Atacama Desert.

Perchlorate is an anion that originates as a contaminant in ground and surface waters. The presence of perchlorate in soil and water samples from northern Chile (Atacama Desert) was investigated by ion chromatography-electrospray mass spectrometry. Results indicated that perchlorate was found in five of seven soils (cultivated and uncultivated) ranging from 290 ± 1 to 2,565 ± 2 µg/kg. The greatest concentration of perchlorate was detected in Humberstone soil (2,565 ± 2 µg/kg) associated with nitrate deposits. Perchlorate levels in Chilean soils are greater than those reported for uncultivated soils in the United States. Perchlorate was also found in superficial running water ranging from 744 ± 0.01 to 1,480 ± 0.02 µg/L. Perchlorate water concentration is 30-60 times greater than levels established by the United States Environmental Protection Agency (24.5 µg/L) for drinking.

Integrated environmental assessment of freshwater sediments: a chemical and ecotoxicological approach at the Alqueva reservoir.

In order to study the pollution of an aquatic ecosystem, it is necessary to analyze not only the levels of chemical pollutants in water, but also those accumulated in the sediment matrix, as well as to assess its ecotoxicological status. The Alqueva reservoir, the largest artificial lake in Europe, was chosen as case study as it constitutes the most important water supply source in southern Portugal. It is located in the Guadiana River Basin, in a semi-arid region with high levels of water scarcity and where agriculture is one of the main activities. The evaluation of sediments comprised: (1) physical and chemical analysis (grain size, pH, organic matter, nitrogen, phosphorus); (2) potentially toxic trace elements (Cu, As, Pb, Cr, Cd, Zn and Ni); and (3) ecotoxicological evaluation with Vibrio fischeri, Thamnocephalus platyurus, Daphnia magna, and Heterocypris incongruens. Total trace element concentrations indicated that As, Cd, and Pb surpassed the Canadian levels for the protection of aquatic life, in most of Alqueva's sites. The results of the toxicity assessment showed that some locations induced acute and chronic toxicity in the species used. Further, the H. incongruens was the most sensitive species as far as the contamination found in the sediment is concerned, followed by the bacteria V. fischeri. This integrative approach, together with the water column quality assessment, allowed a comprehensive evaluation of the environmental quality of this strongly modified water body and will allow the implementation of remediation strategies to obtain a good ecological potential as proposed in the Water Framework Directive.

Exploring nasopharyngeal microbiota profile in children affected by SARS-CoV-2 infection.

The relationship between COVID-19 and nasopharyngeal (NP) microbiota has been investigated mainly in the adult population. We explored the NP profile of children affected by COVID-19, compared to healthy controls (CTRLs). NP swabs of children with COVID-19, collected between March and September 2020, were investigated at the admission (T0), 72 h to 7 days (T1), and at the discharge (T2) of the patients. NP microbiota was analyzed by 16S rRNA targeted-metagenomics. Data from sequencing were investigated by QIIME 2.0 and PICRUSt 2. Multiple machine learning (ML) models were exploited to classify patients compared to CTRLs. The NP microbiota of COVID-19 patients (N = 71) was characterized by reduction of α-diversity compared to CTRLs (N = 59). The NP microbiota of COVID-19 cohort appeared significantly enriched in Streptococcus, Haemophilus, Staphylococcus, Veillonella, Enterococcus, Neisseria, Moraxella, Enterobacteriaceae, Gemella, Bacillus, and reduced in Faecalibacterium, Akkermansia, Blautia, Bifidobacterium, Ruminococcus, and Bacteroides, compared to CTRLs (FDR < 0.001). Exploiting ML models, Enterococcus, Pseudomonas, Streptococcus, Capnocytopagha, Tepidiphilus, Porphyromonas, Staphylococcus, and Veillonella resulted as NP microbiota biomarkers, in COVID-19 patients. No statistically significant differences were found comparing the NP microbiota profile of COVID-19 patients during the time-points or grouping patients on the basis of high, medium, and low viral load (VL). This evidence provides specific pathobiont signatures of the NP microbiota in pediatric COVID-19 patients, and the reduction of anaerobic protective commensals. Our data suggest that the NP microbiota may have a specific disease-related signature since infection onset without changes during disease progression, regardless of the SARS-CoV-2 VL. IMPORTANCE: Since the beginning of pandemic, we know that children are less susceptible to severe COVID-19 disease. A potential role of the nasopharyngeal (NP) microbiota has been hypothesized but to date, most of the studies have been focused on adults. We studied the NP microbiota modifications in children affected by SARS-CoV-2 infection showing a specific NP microbiome profile, mainly composed by pathobionts and almost missing protective anaerobic commensals. Moreover, in our study, specific microbial signatures appear since the first days of infection independently from SARS-CoV-2 viral load.

Premature ageing of the immune system relates to increased anti-lymphocyte antibodies (ALA) after an immunization in HIV-1-infected and kidney-transplanted patients.

Low-affinity immunoglobulin (Ig)G with potential autoreactivity to lymphocytes and hypergammaglobulinaemia have been described previously in HIV-1-infected patients. Whether such antibodies increase after challenging the immune system, for example with an immunization, is not known. In the present study, the modulation of antibodies with low affinity and potential autoreactivity was evaluated after 2012-13 seasonal flu vaccination with a simple empirical laboratory test measuring the titres of anti-lymphocyte antibodies (ALA) in two different models of secondary immunodeficiency: HIV-1 vertically infected patients (HIV) and patients treated with immunosuppressive therapies after kidney transplantation (KT) compared to healthy individuals (HC). In parallel, the activation status of B cells and their degree of immune senescence was evaluated by measuring the B cell interleukin (IL)-21R expression/plasma IL-21 levels and the frequencies of mature-activated (MA) and double-negative (DN) B cells. A significant increase of ALA titres was observed after vaccination in HIV and KT but not in HC, and this correlated directly with the frequencies of both MA and DN and inversely with the B cell IL-21R expression. This suggests that the quality of an immune response triggered by flu vaccination in HIV and KT may depend upon the activation status of B cells and on their degree of immune senescence. Further investigations are needed to verify whether high frequencies of MA and DN may also relate to increase autoimmunity after immunization in high-risk populations.

Opicapone: a short lived and very long acting novel catechol-O-methyltransferase inhibitor following multiple dose administration in healthy subjects.

AIMS: The aim of this study was to assess the tolerability, pharmacokinetics and inhibitory effect on erythrocyte soluble catechol-O-methyltransferase (S-COMT) activity following repeated doses of opicapone. METHODS: This randomized, placebo-controlled, double-blind study enrolled healthy male subjects who received either once daily placebo or opicapone 5, 10, 20 or 30 mg for 8 days. RESULTS: Opicapone was well tolerated. Its systemic exposure increased in an approximately dose-proportional manner with an apparent terminal half-life of 1.0 to 1.4 h. Sulphation was the main metabolic pathway. Opicapone metabolites recovered in urine accounted for less than 3% of the amount of opicapone administered suggesting that bile is likely the main route of excretion. Maximum S-COMT inhibition (Emax ) ranged from 69.9% to 98.0% following the last dose of opicapone. The opicapone-induced S-COMT inhibition showed a half-life in excess of 100 h, which was dose-independent and much longer than plasma drug exposure. Such a half-life translates into a putative underlying rate constant that is comparable with the estimated dissociation rate constant of the COMT-opicapone complex. CONCLUSION: Despite its short elimination half-life, opicapone markedly and sustainably inhibited erythrocyte S-COMT activity making it suitable for a once daily regimen.

Trapping of palindromic ligands within native transthyretin prevents amyloid formation.

Transthyretin (TTR) amyloidosis is a fatal disease for which new therapeutic approaches are urgently needed. We have designed two palindromic ligands, 2,2'-(4,4'-(heptane-1,7-diylbis(oxy))bis(3,5-dichloro-4,1-phenylene)) bis(azanediyl)dibenzoic acid (mds84) and 2,2'-(4,4'-(undecane-1,11-diylbis(oxy))bis(3,5-dichloro-4,1-phenylene)) bis(azanediyl)dibenzoic acid (4ajm15), that are rapidly bound by native wild-type TTR in whole serum and even more avidly by amyloidogenic TTR variants. One to one stoichiometry, demonstrable in solution and by MS, was confirmed by X-ray crystallographic analysis showing simultaneous occupation of both T4 binding sites in each tetrameric TTR molecule by the pair of ligand head groups. Ligand binding by native TTR was irreversible under physiological conditions, and it stabilized the tetrameric assembly and inhibited amyloidogenic aggregation more potently than other known ligands. These superstabilizers are orally bioavailable and exhibit low inhibitory activity against cyclooxygenase (COX). They offer a promising platform for development of drugs to treat and prevent TTR amyloidosis.