Mutations in PIK3R1 can lead to APDS2, SHORT syndrome or a combination of the two.

Mutations in PIK3R1 gene have been associated to two different conditions: a primary immunodeficiency, called APDS2, of recent description and SHORT syndrome. 47 patients with APDS2 have been reported to date, only one of them sharing both PIK3R1-related phenotypes. Here we describe two more patients affected by APDS2 and SHORT syndrome, which highlights that this association may not be so infrequent. We recommend that patients with mutations in PIK3R1 gene should be assessed by both clinical immunologists and clinical geneticists.

Serum properdin consumption as a biomarker of C5 convertase dysregulation in C3 glomerulopathy.

Properdin (P) stabilizes the alternative pathway (AP) convertases, being the only known positive regulator of the complement system. In addition, P is a pattern recognition molecule able to initiate directly the AP on non-self surfaces. Although P deficiencies have long been known to be associated with Neisseria infections and P is often found deposited at sites of AP activation and tissue injury, the potential role of P in the pathogenesis of complement dysregulation-associated disorders has not been studied extensively. Serum P levels were measured in 49 patients with histological and clinical evidence of C3 glomerulopathy (C3G). Patients were divided into two groups according to the presence or absence of C3 nephritic factor (C3NeF), an autoantibody that stabilizes the AP C3 convertase. The presence of this autoantibody results in a significant reduction in circulating C3 (P < 0·001) and C5 levels (P < 0·05), but does not alter factor B, P and sC5b-9 levels. Interestingly, in our cohort, serum P levels were low in 17 of the 32 C3NeF-negative patients. This group exhibited significant reduction of C3 (P < 0·001) and C5 (P < 0·001) and increase of sC5b-9 (P < 0·001) plasma levels compared to the control group. Also, P consumption was correlated significantly with C3 (r = 0·798, P = 0·0001), C5 (r = 0·806, P < 0·0001), sC5b-9 (r = -0·683, P = 0·043) and a higher degree of proteinuria (r = -0·862, P = 0·013). These results illustrate further the heterogeneity among C3G patients and suggest that P serum levels could be a reliable clinical biomarker to identify patients with underlying surface AP C5 convertase dysregulation.

Sports injuries in Brazilian blind footballers.

The purpose of this study was to assess the characteristics and prevalence of sports-related injuries in visually disabled athletes of the Brazilian football 5-a-side team. The participants were 13 male athletes, all classified as B1 visual class, members of the Brazilian team, who played in five consecutive international competitions. Data were collected using the Brazilian Paralympic Committee and the Brazilian Confederation of Sports for the Blind report form. From the total of 13 athletes, 11 succumbed to some form of injury during the 5 competitions, which incorporated 23 matches, representing a prevalence of 84.6%. A total of 35 sports injuries were recorded, giving a clinical incidence of 2.7 injuries per athlete and an injury risk of 0.85 and an incidence rate of 0.12 injuries per match. Traumatic injuries (80%) were more common than overuse injuries (20%) (p<0.05). The highest distribution of injury was in the lower limbs (80%), followed by the head (8.6%), spine (5.7%) and upper limbs (5.7%). The body regions most affected were the knee (28.6%), feet (17.1%), ankle (11.4%) and thigh (11.4%). Contusions (31.4%), sprains (25.7%) and tendinopathy (8.6%) were the most frequent diagnoses. This is the first study to describe the nature and prevalence of sports-related injuries in 5-a-side football in blind athletes. The results are important in guiding strategies to inform the implementation of preventive pathways and provide a strong rationale for the compulsory use of additional protective equipment.

Performance of a single stage vertical flow constructed wetland system treating raw domestic sewage in Brazil.

This study investigated the behaviour of two intermittently fed vertical flow constructed wetlands (one planted with Tifton 85 and the other unplanted) working in parallel, treating raw municipal sewage in Brazil for a population equivalent around 100 inhabitants. Based on a monitoring programme of over 2 years, the following items were evaluated: influence of batch frequency and the presence of Tifton 85 on the wetlands performance in terms of several physico-chemical and biological constituents. The unit with plants performed better than the one without, indicating a positive influence of the presence of plants. More attachment by total and volatile solids and larger amount of bacteria involved in the nitrogen cycle were observed in the planted filter medium, which can explain its higher nitrification and solids removal. The application of a smaller influent volume with a higher batch frequency improved the performance of both units. No signs of medium clogging have been observed in both units. The system simplicity and the good removal efficiency of organic matter, suspended solids, ammonia and helminth eggs indicate its high applicability in small communities in developing countries such as Brazil.

The hyperbenthic environment: A forgotten habitat for plastic pollution.

This study investigates the abundances and composition of microplastics (MP) among the shallow layers of a coastal Mediterranean Marine Protected Area (Cabrera MPA), seafloor sediments, hyperbenthic environment, and the water column. The mid waters samples were collected mid-way between the sea surface and the seafloor and hyperbenthic samples at the water layer adjacent to the seafloor. Sampling was carried out on patchiness seafloor of Posidonia oceanica meadows. The seafloor sediments showed a mean abundance of 378,769.20 ± 508,109.11 MPs/m3, three orders of magnitude higher than the hyperbenthic (209.17 ± 117.07 MPs/m3), and the mid waters layer (106.48 ± 107.17 MPs/m3). An increasing vertical gradient in MP abundances, mainly composed of fibers was observed. Fibers were made-up mainly of polystyrene (PS, 25 %), expanded polystyrene (EPS, 18 %) and cellulose acetate (CA, 16 %). The results stress the need to increase efforts to find solutions to mitigate fiber pollution in the marine environment.

Breaking the paradigm: Marine sediments hold two-fold microplastics than sea surface waters and are dominated by fibers.

We conducted one of the first studies to integrate the quantification and characterization of microplastics (MPs), including fibers, in different habitats (sea surface, seafloor and beach sediments) of a coastal Mediterranean marine protected area, analyzing their ingestion in several marine species. The objectives of the study were to evaluate the distribution of MPs according to shape and polymer, to assess the contribution of fibers to local plastic pollution and to evaluate their ingestion in fish and invertebrates species that inhabit the study area (Pagrus pagrus, Serranus scriba, Spondyliosoma cantharus, Diplodus vulgaris, Oblada melanura, Holothuria forskalii, Holothuria tubularis, Holothuria polis, Arbacia lixula, Paracentrotus lividus, Modiolus barbatus, Mytilus galloprovincialis and Arca noae). A total of 111 environmental samples were analyzed. The mean abundance of MPs (excluding fibers) quantified in beach sediments (13,418.86 ± 28,787.99 MPs/m2) was two orders of magnitude higher than that found in seafloor sediments (76.92 ± 108.84 MPs/m2), which in turn was two orders of magnitude higher than sea surface samples (0.17 ± 0.39 MPs/m2). The fibers were the most abundant shape of MPs identified in all habitats. Variability in MPs ingestion was detected between species, with ingestion rates ranging from 43 % to 100 % for general MPs and ranging from 7 % to 100 % for fibers. The highest ingestion was observed in Holoturians, representing suitable bioindicators for plastic pollution. The composition of the polymer varies weakly depending on habitats and biota, but the result is strongly correlated with the morphology of the plastic. Fibers were mainly composed of cellulose acetate (29 %), styrofoam of polystyrene (18 %), and filaments, films and fragments of polyethylene and polypropylene. The results highlighted the need to expand integrated approaches to effectively study marine plastic pollution and to undertake efficient actions to limit the input of plastics, particularly fibers, into the marine environment.

Research-based flow cytometry assays for pathogenic assessment in the human B-cell biology of gene variants revealed in the diagnosis of inborn errors of immunity: a Bruton's tyrosine kinase case-study.

Introduction: Inborn errors of immunity (IEI) are an expanding group of rare diseases whose field has been boosted by next-generation sequencing (NGS), revealing several new entities, accelerating routine diagnoses, expanding the number of atypical presentations and generating uncertainties regarding the pathogenic relevance of several novel variants. Methods: Research laboratories that diagnose and provide support for IEI require accurate, reproducible and sustainable phenotypic, cellular and molecular functional assays to explore the pathogenic consequences of human leukocyte gene variants and contribute to their assessment. We have implemented a set of advanced flow cytometry-based assays to better dissect human B-cell biology in a translational research laboratory. We illustrate the utility of these techniques for the in-depth characterization of a novel (c.1685G>A, p.R562Q) de novo gene variant predicted as probably pathogenic but with no previous insights into the protein and cellular effects, located in the tyrosine kinase domain of the Bruton's tyrosine kinase (BTK) gene, in an apparently healthy 14-year-old male patient referred to our clinic for an incidental finding of low immunoglobulin (Ig) M levels with no history of recurrent infections. Results and discussion: A phenotypic analysis of bone marrow (BM) revealed a slightly high percentage of pre-B-I subset in BM, with no blockage at this stage, as typically observed in classical X-linked agammaglobulinemia (XLA) patients. The phenotypic analysis in peripheral blood also revealed reduced absolute numbers of B cells, all pre-germinal center maturation stages, together with reduced but detectable numbers of different memory and plasma cell isotypes. The R562Q variant allows Btk expression and normal activation of anti-IgM-induced phosphorylation of Y551 but diminished autophosphorylation at Y223 after anti IgM and CXCL12 stimulation. Lastly, we explored the potential impact of the variant protein for downstream Btk signaling in B cells. Within the canonical nuclear factor kappa B (NF-κB) activation pathway, normal IκBα degradation occurs after CD40L stimulation in patient and control cells. In contrast, disturbed IκBα degradation and reduced calcium ion (Ca2+) influx occurs on anti-IgM stimulation in the patient's B cells, suggesting an enzymatic impairment of the mutated tyrosine kinase domain.

Decrease in the expression of N-methyl-D-aspartate receptors in the nucleus tractus solitarii induces antinociception and increases blood pressure.

N-methyl-D-aspartate receptors (NMDAR) have a role in cardiovascular control at the nucleus tractus solitarii (NTS), eliciting increases or decreases in blood pressure (BP), depending on the area injected with the agonists. In spite of the association between cardiovascular control and pain modulation, the effects of manipulating NMDAR in pain responses have never been evaluated. In this study, we decreased the expression of NMDAR in the NTS using gene transfer to target receptor subunits and evaluate long-term effects. Seven days after the injection of lentiviral vectors containing the NR1a subunit cDNA of NMDAR, in antisense orientation, into the intermediate NTS of Wistar rats, BP was measured, and the formalin test of nociception was performed. The antisense vector induced a decrease of NR1 expression in the NTS and elicited BP rises and hypoalgesia. Antisense vectors inhibited formalin-evoked c-Fos expression in the spinal cord, indicating decreased nociceptive activity of spinal neurons. Using a time-course approach, we verified that the onset of both the increases in BP and the hypoalgesia was at 4 days after vector injection into the NTS. The injection of NMDA into the NTS reversed the effects of antisense vectors in pain behavioral responses and spinal neuronal activation and decreased BP and heart rate. The present study shows that the NR1 subunit of the NMDAR at the NTS is critical in the regulation of tonic cardiovascular and nociceptive control and shows an involvement of the nucleus in the modulation of sustained pain.

Methodological guidelines for preparing a structured therapeutic education program: From design to evaluation.

BACKGROUND AND OBJECTIVE: Therapeutic patient education (TPE) is effective and essential in the context of the growing prevalence of chronic diseases in which tools are needed for planning structured programs. The objective of this project was to develop guidelines for designing and assessing a TPE program. METHODS: 1) We assembled a multidisciplinary group of 8 leaders in TPE, chronicity, quality and safety from the hospital and the university. 2) We conducted an exhaustive review of the scientific literature on the planning of TPE programs directed at chronically ill patients, their relatives and caregivers. 3) The final text underwent comments and suggestions by participants from the hospital and primary care centre during a course on information and TPE methodology. The recommendations were unanimously agreed upon by the writing group. RESULTS: We obtained a standardised work procedure targeted at professionals involved in planning TPE programs, based on international recommendations. The document is structured into sections: a) Definition of the health problem and analysis of the situation; b) Program structure (human resources and materials); objectives (health-related, behaviour-related and educational) and methodology; c) Path the patient and family/caregiver follows in the program; and d) Assessment and indicators. Assessment of the procedure, within the framework of the methodology courses, was favourable. CONCLUSIONS: The methodology provided by this document serves as an instrument for the standardised and systematic planning of educational programs and unifies the criteria in their drafting. However, the document needs to be adapted to the condition and population to which each program is addressed.

Giardia duodenalis: kinetics of cyst elimination and the systemic humoral and intestinal secretory immune responses in gerbils (Meriones unguiculatus) experimentally infected.

This study aimed to determine the pre-patent period and to evaluate the kinetics of cyst elimination and the systemic humoral (IgA, IgG(1), IgG(2a), IgM, IgE) and intestinal secretory (IgA) immune responses in gerbils (Meriones unguiculatus) experimentally innoculated with different doses of Giardia duodenalis trophozoites. Forty-eight animals aged 6-8 weeks were used, equally distributed among six groups, five groups innoculated with different doses of trophozoites (10(1), 10(2), 10(3), 10(4), 10(5)) and one control (non-infected) group. Coproparasitological examinations were carried out daily up to 91 days after inoculation (d.a.i.) to determine the pre-patent period and the kinetics of cyst elimination. Blood and stool samples were weekly collected for antibody assays. The pre-patent period was observed from the 9 d.a.i. onwards, with intermittent elimination of variable quantities of cysts up to 27 d.a.i.. All infected gerbils, irrespective of the dose received, were able to mount systemic humoral immune responses as evidenced by specific IgM titers from 7 to 28 d.a.i., corresponding to the peak of cyst elimination, followed by high and persistent IgG1 titers. Intestinal secretory responses were also seen with two peaks of fecal IgA titers, corresponding to IgM and IgG1 response peaks, respectively. In conclusion, systemic and intestinal humoral immune responses were related to the control of giardiasis in this experimental model.

Microinjection of angiotensin II in the caudal ventrolateral medulla induces hyperalgesia.

Nociceptive transmission from the spinal cord is controlled by supraspinal pain modulating systems that include the caudal ventrolateral medulla (CVLM). The neuropeptide angiotensin II (Ang II) has multiple effects in the CNS and at the medulla oblongata. Here we evaluated the expression of angiotensin type 1 (AT(1)) receptors in spinally-projecting CVLM neurons, and tested the effect of direct application of exogenous Ang II in the CVLM on nociceptive behaviors. Although AT(1)-immunoreactive neurons occurred in the CVLM, only 3% of AT(1)-positive neurons were found to project to the dorsal horn, using double-immunodetection of the retrograde tracer cholera toxin subunit B. In behavioral studies, administration of Ang II (100 pmol) in the CVLM gave rise to hyperalgesia in both the tail-flick and formalin tests. This hyperalgesia was significantly attenuated by local administration of the AT(1) antagonist losartan. The present study demonstrates that Ang II can act on AT(1) receptors in the CVLM to modulate nociception. The effect on spinal nociceptive processing is likely indirect, since few AT(1)-expressing CVLM neurons were found to project to the spinal cord. The renin-angiotensin system may also play a role in other supraspinal areas implicated in pain modulation.

Interferences in the transverse profile of a toluene beam induced by a resonant RF electric field.

In this work, the interaction of a supersonic beam of toluene diluted in He with a resonant oscillating RF field is investigated both experimental and theoretically. It is shown how the resonant field induces a peak structure in the transverse beam profile which can be explained by the onset of molecular interferences. Specifically, the interaction of a toluene beam of 0.12 eV of translational energy with a resonant RF field of 1.12 kV/m amplitude, and -610 kV/m(2) of gradient at the horizontal plane, during 160 micros produces a series of maxima in the transverse beam profile. The observed structure was satisfactorily reproduced by a quantum interference model based on the interaction of two coherent superpositions induced by the resonant RF field. It appears the present experimental technique could be useful to investigate the spectroscopy and dynamical behavior of coherent beams of polar molecules.

Comparative analysis of cell phenotypes in different severe clinical forms of Chagas' disease.

The understanding of the role of the immune response in the development of gastrointestinal and cardio-digestive (CD) forms of Chagas disease has received little attention. In this paper, the commitment of each leukocyte population of peripheral blood to the production of IFN-gamma, TNF-alpha, IL-12, IL-4, IL-5 and IL-10 was studied in patients with the CD form of Chagas disease. The data show that cells from patients with the CD form of the disease have distinct cytokine profiles when compared with the other clinical forms of Chagas disease and suggest that eosinophils are the major source of cytokine production in this clinical entity. The data presented in this paper demonstrate that patients with CD form can be distinguished from patients with gastrointestinal or cardiac forms of the disease by the distinct cytokine profile of peripheral blood cells.

Laboratory guidelines for the diagnosis and follow-up of patients with monoclonal gammopathies.

We present guidelines from the Immunochemistry group of the Spanish Society for Immunology that are designed to provide a practical tool for the diagnosis and follow-up of monoclonal gammopathies. We review the clinical and analytical features of various monoclonal gammopathies, international consensus guidelines and techniques used to detect and follow-up monoclonal components.

Efficacy of silver diamine fluoride for caries reduction in primary teeth and first permanent molars of schoolchildren: 36-month clinical trial.

We hypothesized that the six-monthly application of silver diamine fluoride (SDF) can arrest the development of caries in the deciduous dentition of six-year-old schoolchildren and prevent caries in their first permanent molars. A prospective controlled clinical trial was conducted on the efficacy of a 38% SDF solution for caries reduction. Four hundred and twenty-five six-year-old children were divided into two groups: One group received SDF solution in primary canines and molars and first permanent molars every 6 mos for 36 mos. The second group served as controls. The 36-month follow-up was completed by 373 children. The mean number of new decayed surfaces appearing in primary teeth during the study was 0.29 in the SDF group vs. 1.43 in controls. The mean of new decayed surfaces in first permanent molars was 0.37 in the SDF group vs. 1.06 in controls. The SDF solution was found to be effective for caries reduction in primary teeth and first permanent molars in schoolchildren.

Anti-inflammatory action of glucagon in rats.

1 Subcutaneous administration of glucagon (1 and 0.5 mg/kg) 30 min before the injection of carrageenin or dextran into the rat's paw reduced oedema and the local exudation of Evans blue previously given intravenously. 2 The effect persisted after removal of the adrenal medulla but not after adrenalectomy. 3 When glucagon (1 mg/kg, s.c.) was given daily after a local reaction to Freund's adjuvant injected into the paw had developed, a decrease in the reaction was observed up to 12 days. Blood sugar levels remained within the normal range. 4 Glucagon may exert an anti-inflammatory effect through the release of adrenal corticosteroids and thus help modulate inflammatory reactions.

Cellular immune responses of chagasic patients to antigens derived from different Trypanosoma cruzi strains and clones.

The reactivity of peripheral blood mononuclear cells (PBMN) from 62 chagasic patients to antigens prepared with different Trypanosoma cruzi strains and clones belonging to different zymodemes was evaluated by the incorporation of 3H-thymidine into DNA. Standardization of experimental conditions was carried out by establishing the proper antigen concentration (15-20 micrograms protein), the adequate period of time (5-6 days) and the best cell concentration (300,000/well). Individual analysis of 62 patients showed 2 distinct patterns of cellular response. One group of patients (32%) had low cellular responses to all antigens tested while the remaining patients had high response to at least 1 of the antigens. No relationship of the immune responsiveness to the patients' clinical forms could be established. In addition, the PBMN response to different strain and clone antigens was not statistically significant. Thus, it appears that the cellular response induced by any particular clone or strain represents an expression of the stimulation of their common antigenic make-up.

A comparative study of postsynaptic alpha2-adrenoceptors of the dog mesenteric and rat femoral veins.

The aim of the present study was to compare the subtype of postjunctional alpha2-adrenoceptors of canine mesenteric and rat femoral veins. To check whether the presence of alpha1-adrenoceptors in both tissues might interfere with alpha2-adrenoceptor-mediated effects, the experiments were carried out under two experimental conditions: without and with alpha1-adrenoceptor blockade. The selective and irreversible alpha1-adrenoceptor antagonist phenoxybenzamine (30 nM) was used to eliminate alpha1-adrenoceptors. The pA2 values for the antagonism exerted by eight alpha-adrenoceptor antagonists (rauwolscine, yohimbine, RX821002, WB4101, idazoxan, phentolamine, spiroxatrine and prazosin) against the highly selective alpha2-adrenoceptor agonist UK-14,304 were determined under these two experimental conditions and correlated with pKi values of the same antagonists at cloned human alpha2A-, alpha2B- and alpha2C-adrenoceptors expressed in Chinese hamster lung cells and at the alpha2D-adrenoceptors in the rat submaxillary gland or the bovine pineal gland. In most of the experiments carried out in the canine mesenteric vein, the concentration-response curves for UK-14,304 were biphasic; the first phase was antagonized by low concentrations (2-50 nM) of the antagonists used except prazosin, while the second one was antagonized by 30 nM of either prazosin or phenoxybenzamine. In the rat femoral vein, the concentration-response curves to UK-14,304 were monophasic. In either tissue, the pA2 values obtained in untreated preparations and in preparations pretreated with phenoxybenzamine were not significantly different, showing that effects resulting from the activation of alpha1-adrenoceptors can be avoided simply by using low concentrations of the highly selective alpha2-adrenoceptor agonist UK-14,304. The correlation between pA2 and pKi values showed that, while in the canine mesenteric vein the postjunctional alpha2-adrenoceptors resemble alpha2A-adrenoceptors more closely, in the rat femoral vein they are more closely related to alpha2D-adrenoceptors. In either species, therefore, they belong to the genetic alpha2A/D type of alpha2-adrenoceptor.

Cytokine profiles during experimental Chagas' disease.

People infected with Trypanosoma cruzi remain so for life, yet only 30-40% of these individuals develop characteristic chagasic cardiomyopathies. Similarly, when infected with the Brazilian strain of T. cruzi, DBA/2 mice develop severe cardiac damage while B10.D2 mice do not. To better understand the immunological parameters that may be involved in the disease process, we have used this murine model (DBA/2 vs B10.D2) and compared the changes in cytokine production during the course of infection with T cruzi. Concanavalin A (Con A) stimulation of spleen cells harvested during the acute phase (day 30) resulted in similarly high levels of IFN-gamma in both mouse strains. However, the amount of IFN-gamma in supernatants from cultures of B10.D2 spleen cells initiated during the chronic phase (day 72) was at subacute levels, whereas secretion by chronic DBA/2 spleen cells remained high. In addition, Con A-stimulated spleen cells from acute DBA/2 mice produced approximately twice as much IL-10 and significantly more IL-4 than cells from B10.D2 mice. IL-4 secretion remained low by cells from chronic B10.D2 mice, but when using cells from chronic DBA/2 mice, levels continued to increase beyond the already high levels secreted by cells harvested during the acute phase. Proliferative responses to Con A stimulation by spleen cells from DBA/2 mice were significantly higher than those from B10.D2 mice in both the acute and chronic phases. These data suggest that enhanced responses in DBA/2 mice, which may be related to a higher parasite burden, a lack of down-regulation, and/or the onset of autoimmune phenomena, correlate with the more severe cardiomyopathy seen in pathopermissive mice.

Losartan and atenolol on hypertension induced by adenosine receptor blockade.

1. The prolonged infusion of 1,3-dipropyl-8-sulfophenylxanthine (DPSPX), a non-selective antagonist of adenosine receptors, induces hypertension, an increase in plasma renin activity and morphological cardiovascular changes. 2. The aim of this work was to evaluate the effects of losartan, a selective AT1 receptor antagonist, and atenolol, a beta-adrenoceptor antagonist, on DPSPX-induced hypertension. 3. Male Wistar rats (250-300 g, n = 4-6) were treated for 1 or 4 weeks with: saline i.p.; DPSPX (90 microg kg(-1) h(-1)) i.p.; losartan (15 mg kg(-1) day(-1)) p.o.; atenolol (25 mg kg(-1) day(-1)) p.o.; DPSPX (90 microg kg(-1) h(-1)) i.p. + losartan (15 mg kg(-1) day(-1)) p.o.; DPSPX (90 microg kg(-1) h(-1)) i.p. + atenolol (25 mg kg(-1) day(-1)) p.o. Blood pressure was measured by the 'tail-cuff' method in conscious animals. Fragments of the mesenteric and tail arteries were processed for morphological study and the mean diameter of the vascular smooth muscle cells was determined. 4. DPSPX increased blood pressure. Losartan and atenolol prevented this rise but had no effect on blood pressure of control rats. DPSPX-treated groups showed hypertrophy of the vascular smooth muscle cells and proliferation of subintimal cells. Losartan but not atenolol prevented these changes. Losartan had no effect on the vascular morphology of control rats, while treatment with atenolol for 4 weeks induced hypertrophy of the vascular smooth muscle cells. 5. Both losartan and atenolol counteract the development of DPSPX-induced hypertension but only losartan prevents the alterations in vascular morphology.