Pan-Enteric Capsule Endoscopy: Current Applications and Future Perspectives.

Background: The role of capsule endoscopy in the evaluation of the small bowel is well established, and current guidelines position it as a first-line test in a variety of clinical scenarios. The advent of double-headed capsules further enabled the endoscopic assessment of colonic mucosa and the opportunity for a one-step noninvasive examination of the entire bowel (pan-enteric capsule endoscopy [PCE]). Summary: We reviewed the technical procedure and preparation of patients for PCE, as well as its current clinical applications and future perspectives. In non-stricturing and non-penetrating Crohn's disease affecting the small bowel and colon, PCE monitors disease activity by assessing mucosal healing, a major treatment outcome, with a higher diagnostic yield than cross-sectional imaging or conventional colonoscopy. Also in ulcerative colitis, double-headed capsules have been used to monitor disease activity noninvasively. Currently, validated scoring systems have been specifically devised for these double-headed capsules and permit a standardized assessment of the inflammatory burden. In suspected mid-lower digestive bleeding, some exploratory studies have demonstrated the feasibility and high diagnostic yield of PCE, which may work as a filter indicating which patients may benefit of further invasive procedures, namely, for planned hemostatic procedures. The possibility of using PCE is also discussed in the context of polyposis syndromes with simultaneous involvement of the small intestine and colon. Key Messages: PCE is a feasible, effective, and safe diagnostic procedure to evaluate the small bowel and colon. It has been increasingly explored in the setting of inflammatory bowel diseases and, more recently, in suspected mid-lower digestive bleeding. PCE is expected to reduce the demand for invasive procedures and expand the scope of noninvasive intestinal evaluation in the coming future.

Introdução: O papel da endoscopia por cápsula na avaliação do intestino delgado encontra-se bem estabelecido, e as orientações atuais posicionam-na como um teste de primeira linha numa variedade de cenários clínicos. O advento das cápsulas de dupla câmara permitiu expandir a sua aplicação para a avaliação endoscópica da mucosa do cólon, oferecendo a oportunidade de um exame não invasivo de todo o intestino (endoscopia pan-entérica por cápsula, PCE). Sumário: Procedemos a uma revisão de vários aspectos do procedimento e preparação dos doentes para a PCE, bem como as aplicações clínicas atuais e as perspetivas futuras das cápsulas de dupla câmara. Na doença de Crohn não estenosante e não penetrante localizada ao intestino delgado e cólon, a PCE permite monitorizar a atividade da doença e avaliar a cicatrização da mucosa, um indicador importante da eficácia da terapêutica, com um rendimento de diagnóstico superior aos métodos convencionais, nomeadamente os exames imagiológicos ou a colonoscopia invasiva. Também na colite ulcerosa, as cápsulas de dupla câmara têm sido utilizadas para monitorizar a atividade da doença de forma não invasiva. Existem índices endoscópicos validados e especificamente concebidos para as cápsulas de dupla câmara, que permitem uma avaliação sistematizada e quantificação objetiva da atividade inflamatória. Na suspeita de hemorragia digestiva média ou baixa, alguns estudos exploratórios demonstraram a aplicabilidade e o elevado rendimento diagnóstico da PCE, podendo funcionar como um filtro de modo a permitir indicar quais os doentes que mais irão beneficiar de um procedimento invasivo subsequente, nomeadamente para a realização de procedimentos hemostáticos dirigidos. A possibilidade de utilização da PCE é também discutida no contexto das síndromes de polipose com envolvimento simultâneo do intestino delgado e do cólon. Mensagens-chave: A PCE é um procedimento diagnóstico eficaz e seguro para avaliar diretamente a mucosa do intestino delgado e cólon. A sua aplicação tem vindo a expandir-se no contexto das Doenças Inflamatórias Intestinais e, mais recentemente, na suspeita de hemorragia digestiva média ou baixa. Existe a expectativa de que no futuro próximo possamos assistir a uma redução substancial da demanda por procedimentos endoscópicos invasivos, face à utilização crescente da PCE enquanto método de diagnóstico pan-intestinal não invasivo.

Tight control using fecal calprotectin and early disease intervention increase the rates of transmural remission in Crohn's disease.

BACKGROUND: Increasing evidence supports the use of transmural remission as a treatment target in Crohn's disease (CD), but it is seldom achieved in clinical practice. Tight monitoring of inflammation using fecal calprotectin with reactive treatment escalation may potentially improve these results. AIMS: To evaluate if treatment escalation based on fecal calprotectin can improve the rates of transmural remission in CD. The influence of the timing of intervention on this strategy was also evaluated. METHODS: Retrospective cohort study including 256 CD patients with 2 consecutive assessments by MRI-enterography and colonoscopy and with regular monitoring using fecal calprotectin. For each occurrence of an elevated fecal calprotectin (≥250 µg/g), we evaluated whether a reactive adjustment of medical treatment was performed. The ratio of treatment escalation/elevated fecal calprotectin was correlated with the chances of reaching transmural remission. Early disease was defined as disease duration <18 months without previous exposure to immunomodulators and biologics. RESULTS: After a median follow-up of 2 years (IQR 1-4), 61 patients (23.8%) reached transmural remission. Ratios of escalation ≥50% resulted in higher rates of transmural remission (34.2% vs. 15.1%, p < 0.001). The effect was more pronounced in patients with early disease (50.0% vs. 12.0%, p = 0.003). In multivariate analysis, a treatment escalation ratio ≥50% (OR 3.46, 95% CI 1.67-7.17, p = 0.001) and early disease intervention (OR 3.24, 95% CI 1.12-9.34, p = 0.030) were independent predictors of achieving transmural remission. CONCLUSION: Tight-monitoring and reactive treatment escalation increase the rates of transmural remission in CD. Intervention in early disease further improves these results.

Seasonal and inter-seasonal RSV activity in the European Region during the COVID-19 pandemic from autumn 2020 to summer 2022.

Background: The emergence of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in early 2020 and subsequent implementation of public health and social measures (PHSM) disrupted the epidemiology of respiratory viruses. This work describes the epidemiology of respiratory syncytial virus (RSV) observed during two winter seasons (weeks 40-20) and inter-seasonal periods (weeks 21-39) during the pandemic between October 2020 and September 2022. Methods: Using data submitted to The European Surveillance System (TESSy) by countries or territories in the World Health Organization (WHO) European Region between weeks 40/2020 and 39/2022, we aggregated country-specific weekly RSV counts of sentinel, non-sentinel and Severe Acute Respiratory Infection (SARI) surveillance specimens and calculated percentage positivity. Results for both 2020/21 and 2021/22 seasons and inter-seasons were compared with pre-pandemic 2016/17 to 2019/20 seasons and inter-seasons. Results: Although more specimens were tested than in pre-COVID-19 pandemic seasons, very few RSV detections were reported during the 2020/21 season in all surveillance systems. During the 2021 inter-season, a gradual increase in detections was observed in all systems. In 2021/22, all systems saw early peaks of RSV infection, and during the 2022 inter-seasonal period, patterns of detections were closer to those seen before the COVID-19 pandemic. Conclusion: RSV surveillance continued throughout the COVID-19 pandemic, with an initial reduction in transmission, followed by very high and out-of-season RSV circulation (summer 2021) and then an early start of the 2021/22 season. As of the 2022/23 season, RSV circulation had not yet normalised.

Impact of inbreeding and genetic parameter estimates for seminal traits in Lusitano horses.

The objectives of this study were to establish baseline information for seminal traits in Lusitano stallions, to assess the impact of inbreeding, interval between collections and age on semen quality during the breeding and non-breeding seasons, and to estimate the corresponding genetic parameters. A total of 2129 ejaculates by 146 Lusitano stallions used for artificial insemination, obtained from four equine reproduction centers distributed throughout Portugal, over a period of 14 years (2008-2021), were included in the study. The seminal traits analyzed, and the corresponding means and standard deviations, were gel-free volume (56.95 ± 28.76 mL), concentration (186.48 ± 104.68 × 106), motility (64.1 ± 16.9%), total number of spermatozoa (TNS) (9.271 ± 4.956 × 109) and total number of motile spermatozoa per ejaculate (TNMS) (5.897 ± 3.587 × 109). These results are in the normal range of values described for other breeds. In the stallions analyzed, the mean value for the inbreeding coefficient was 7.93 ± 5.29%, and for age it was 12.70 ± 6.83 years. A significant decline in sperm concentration, motility, TNS, and TNMS was observed as inbreeding increased. The season also influenced sperm concentration, motility, TNS and TNMS, with the highest values observed during the breeding season. When considering the impact of age on Lusitano seminal parameters, results showed a nonlinear relationship, with a positive effect until 18 years of age for volume, motility, TNS and TNMS and a negative effect after this age, with a slow decrease. However, age had a markedly negative effect on sperm concentration. The interval between semen collections only affected (P < 0.05) sperm motility, with a regression coefficient of +1.89 ± 2.17% per additional day. Genetic parameters were estimated with an Animal Model, and the estimated heritability (repeatability) was 0.27 (0.35) for volume, 0.02 (0.38) for sperm concentration, 0.24 (0.44) for motility, 0.29 (0.39) for TNS and 0.41 (0.41) for TNMS. These results suggest that it is possible to improve semen quality by selection and that the properties of semen produced by a stallion tend to remain consistent throughout its lifetime. Furthermore, the impact of inbreeding should be taken into consideration when selecting Lusitano stallions for fertility.

Swiss public health measures associated with reduced SARS-CoV-2 transmission using genome data.

Genome sequences from evolving infectious pathogens allow quantification of case introductions and local transmission dynamics. We sequenced 11,357 severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) genomes from Switzerland in 2020-the sixth largest effort globally. Using a representative subset of these data, we estimated viral introductions to Switzerland and their persistence over the course of 2020. We contrasted these estimates with simple null models representing the absence of certain public health measures. We show that Switzerland's border closures decoupled case introductions from incidence in neighboring countries. Under a simple model, we estimate an 86 to 98% reduction in introductions during Switzerland's strictest border closures. Furthermore, the Swiss 2020 partial lockdown roughly halved the time for sampled introductions to die out. Last, we quantified local transmission dynamics once introductions into Switzerland occurred using a phylodynamic model. We found that transmission slowed 35 to 63% upon outbreak detection in summer 2020 but not in fall. This finding may indicate successful contact tracing over summer before overburdening in fall. The study highlights the added value of genome sequencing data for understanding transmission dynamics.

Influence of Spinal Shock on the Neurorehabilitation of ANNPE Dogs.

Acute noncompressive nucleus pulposus extrusion (ANNPE) is related to contusive spinal cord injuries, and dogs usually appear to be exercising vigorously at the time of onset. ANNPE has a characteristic peracute onset of clinical signs during exercise or following trauma, with non-progressive signs during the first 24 h and possibly signs of spinal shock. The main aim was to assess if the presence of spinal shock affects the neurorehabilitation outcomes of ANNPE dogs. This prospective controlled cohort clinical study was conducted at the Arrábida Rehabilitation Center. All of the dogs had T3−L3 injuries and were paraplegic/monoplegic with/without nociception, the study group (n = 14) included dogs with ANNPE spinal shock dogs, and the control group (n = 19) included ANNPE dogs without spinal shock. The study group was also evaluated using a new scale­the Spinal Shock Scale (SSS)­and both groups were under the same intensive neurorehabilitation protocol. Spinal shock was a negative factor for a successful outcome within less time. SSS scores > 4 required additional hospitalization days. The protocol was safe, tolerable, and feasible and accomplished 32% ambulation within 7 days, 29% in 14 days, and 29% in 30 days. The results were better than those obtained in previous studies­94% at 60 days­and 75% of the dogs without nociception recovered ambulation. Long-term follows-ups carried out 4 years later revealed a positive evolution.

Proactive infliximab is more effective than vedolizumab in inducing fecal calprotectin remission in inflammatory bowel disease.

BACKGROUND: Current evidence suggests vedolizumab (VDZ) may be as effective as Infliximab (IFX) in inflammatory bowel disease. It is unknown if proactive therapeutic drug monitoring (PTDM) of IFX may improve these results. METHODS: Case-control study including consecutive patients with primary response to conventional IFX (n = 70), proactive IFX (n = 148), and VDZ (n = 95). PTDM was performed at week 14 and every other infusion, aiming at a trough level between 5 and 10 µg/ml. The primary outcome was fecal calprotectin (Fc) remission (<250 µg/g) at 1 year of treatment. Secondary outcomes included Fc remission at week 14 (proactive IFX/VDZ), clinical remission, treatment discontinuation, hospitalization, and surgery at 1-year of follow-up. RESULTS: Proactive IFX was superior to conventional IFX and VDZ in inducing Fc remission at 1-year (69.4% vs 47.1% vs 37.9%, p = .003 and p < .001). Results remained significant in biologic naïve patients (70.8% vs 44.4% vs 51.4%, p = .001 and p = .043) but comparisons between conventional IFX and VDZ were not significant (p = .265 and p = .664). In multivariate analysis correcting for prior biologic exposure, proactive IFX was more effective than conventional IFX (OR 2.480 95%CI [1.367-4.499], p = .003) and VDZ (OR 3.467 95%CI [1.578-7.617], p = .002) in inducing Fc remission. Amongst secondary outcomes, only clinical remission was significant between proactive IFX and VDZ in the overall cohort (80.4% vs 55.8%, p < .001) and in biologic naïve patients (80.2% vs 62.9%, p = .043). Fc remission at 1-year was associated with better results in most secondary outcomes. CONCLUSION: Proactive IFX was superior to VDZ in inducing Fc remission at 1-year, which was associated with improved clinical outcomes.SUMMARYCurrent evidence suggests that vedolizumab may be as effective as Infliximab in the treatment of patients with inflammatory bowel disease.There have been no studies comparing vedolizumab with proactively optimized Infliximab based on trough levels.We confirm that conventional IFX is as effective as vedolizumab but proactive IFX appears superior to vedolizumab in inducing fecal calprotectin remission.Fecal calprotectin remission associates with better clinical outcomes.

The Ant-like Tachydromia Complex in the Iberian Peninsula-Insights from Habitat Suitability Modelling for the Conservation of an Endemism (Diptera: Hybotidae).

Ant-like flies comprise nine Iberian endemic species of flightless Tachydromia. Severe knowledge gaps on distribution and ecological requirements hinder conservation assessments. Species distribution models were applied to unveil habitat suitability and to provide guidelines for future studies. An ensemble modeling approach combining ten different techniques was implemented with the biomod2 package. Occurrence data was partitioned into six sets, including two multi-species groups and four species. The most relevant drivers of habitat suitability are climate-related, followed by forest type and structure, according to well-defined biogeographic gradients. T. lusitanica and T. ebejeri are adapted to mild temperatures and high-humidity environments. Their distribution is connected to the Temperate-Eurosiberian life zone. T. semiaptera and T. iberica are adapted to progressively drier and hotter central and southern parts of the Iberian Peninsula, connected to transitional Temperate-submediterranean areas. Ant-like fly' ranges overlap with deciduous/marcescent oak species, acting as suitable indicators of their presence in Iberia. Southern marcescent forests emerge as "islands" with particular interest for future prospections. Ant-like flies are threatened by several factors such as climate change and habitat destruction, including urbanization and forest fires. This study provides vital tools to better assess the ant-like flies' conservation status and to manage their habitat.

αIIbß3 variants in ten families with autosomal dominant macrothrombocytopenia: Expanding the mutational and clinical spectrum.

BACKGROUND: Rare pathogenic variants in either the ITGA2B or ITGB3 genes have been linked to autosomal dominant macrothrombocytopenia associated with abnormal platelet production and function, deserving the designation of Glanzmann Thrombasthenia-Like Syndrome (GTLS) or ITGA2B/ITGB3-related thrombocytopenia. OBJECTIVES: To describe a series of patients with familial macrothrombocytopenia and decreased expression of αIIbß3 integrin due to defects in the ITGA2B or ITGB3 genes. METHODS: We reviewed the clinical and laboratory records of 10 Portuguese families with GTLS (33 patients and 11 unaffected relatives), including the functional and genetic defects. RESULTS: Patients had absent to moderate bleeding, macrothrombocytopenia, low αIIbß3 expression, impaired platelet aggregation/ATP release to physiological agonists and low expression of activation-induced binding sites on αIIbß3 (PAC-1) and receptor-induced binding sites on its ligand (bound fibrinogen), upon stimulation with TRAP-6 and ADP. Evidence for constitutive αIIbß3 activation, occurred in 2 out of 9 patients from 8 families studied, but also in 2 out of 12 healthy controls. We identified 7 missense variants: 3 in ITGA2B (5 families), and 4 in ITGB3 (5 families). Three variants (αIIb: p.Arg1026Trp and p.Arg1026Gln and ß3: p.Asp749His) were previously reported. The remaining (αIIb: p.Gly1007Val and ß3: p.Thr746Pro, p.His748Pro and p.Arg760Cys) are new, expanding the αIIbß3 defects associated with GTLS. The integration of the clinical and laboratory data allowed the identification of two GTLS subgroups, with distinct disease severity. CONCLUSIONS: Previously reported ITGA2B and ITGB3 variants related to thrombocytopenia were clustered in a confined region of the membrane-proximal cytoplasmic domains, the inner membrane clasp. For the first time, variants are reported at the outer membrane clasp, at the transmembrane domain of αIIb, and at the membrane distal cytoplasmic domains of ß3. This is the largest single-center series of inherited macrothrombocytopenia associated with αIIbß3 variants published to date.

An Online Influenza Surveillance System for Primary Care Workers in Switzerland: Observational Prospective Pilot Study.

BACKGROUND: A better understanding of the influenza epidemiology among primary care workers could guide future recommendations to prevent transmission in primary care practices. Therefore, we designed a pilot study to assess the feasibility of using a work-based online influenza surveillance system among primary care workers. Such an approach is of particular relevance in the context of the coronavirus disease (COVID-19) pandemic, as its findings could apply to other infectious diseases with similar mechanisms of transmission. OBJECTIVE: This study aims to determine the feasibility of using a work-based online influenza surveillance system for primary care workers in Switzerland. METHODS: Physicians and staff of one walk-in clinic and two selected primary care practices were enrolled in this observational prospective pilot study during the 2017-2018 influenza season. They were invited to record symptoms of influenza-like illness in a weekly online survey sent by email and to self-collect a nasopharyngeal swab in case any symptoms were recorded. Samples were tested by real-time polymerase chain reaction for influenza A, influenza B, and a panel of respiratory pathogens. RESULTS: Among 67 eligible staff members, 58% (n=39) consented to the study and 53% (n=36) provided data. From the time all participants were included, the weekly survey response rate stayed close to 100% until the end of the study. Of 79 symptomatic episodes (mean 2.2 episodes per participant), 10 episodes in 7 participants fitted the definition of an influenza-like illness case (attack rate: 7/36, 19%). One swab tested positive for influenza A H1N1 (attack rate: 3%, 95% CI 0%-18%). Swabbing was considered relatively easy. CONCLUSIONS: A work-based online influenza surveillance system is feasible for use among primary care workers. This promising methodology could be broadly used in future studies to improve the understanding of influenza epidemiology and other diseases such as COVID-19. This could prove to be highly useful in primary care settings and guide future recommendations to prevent transmission. A larger study will also help to assess asymptomatic infections.

Proactive Infliximab Drug Monitoring Is Superior to Conventional Management in Inflammatory Bowel Disease.

BACKGROUND: Increasing evidence supports the use of reactive therapeutic drug monitoring (TDM) in Crohn's disease (CD) and ulcerative colitis (UC) following secondary loss of response. It is still unknown if proactive TDM can improve clinical outcomes. METHODS: Consecutive patients completing infliximab (IFX) induction therapy were prospectively allocated into a proactive TDM protocol (pTDM). Before the fourth infusion and every 2 infusions, IFX trough levels and antidrug antibodies were measured using a drug-sensitive assay (Theradiag, Lisa Tracker). Treatment was proactively escalated aiming at an IFX trough level between 3 and 7 ug/mL (CD) and 5 and 10 ug/mL (UC). A retrospective cohort treated with IFX but without TDM served as the reference group. End points included the need for surgery, hospitalization, treatment discontinuation, and mucosal healing at 2 years of follow-up. RESULTS: Two hundred five patients were included, 56 in the proactive regimen. Treatment escalation was more common in pTDM patients (76.8% vs 25.5%; P < 0.001), who also required less surgery (8.9% vs 20.8%; P = 0.032) and presented higher rates of mucosal healing (73.2% vs 38.9%; P < 0.0001). Proactive TDM significantly decreased the odds of reaching any unfavorable outcome (odds ratio, 0.358; 95% confidence interval, 0.188-0.683; P = 0.002). CONCLUSIONS: Proactive TDM is associated with fewer surgeries and higher rates of mucosal healing than conventional non-TDM-based management.

Genetic and Environmental Factors Influencing Gestation Length in Lusitano Horses.

Breeding and parturition records collected over a period of 35 years in the Alter Real stud of Lusitano horses were used to calculate gestation length (GL). The 1027 gestations by 209 mares mated to 60 stallions had a mean GL of 338.1 ± 9.26 days. The mixed model analysis of variance indicated that the sex of the foal and inbreeding of the dam and foal had no significant effect on GL (P > .05). On the other hand, GL increased linearly with mare age, with an estimated regression coefficient of 0.155 ± 0.069 days/year (P < .05). Year and conception month affected GL (P < .05), with longer gestations observed when breeding occurred between January and April, followed by a decline of about 5 and 10 days in GL of mares bred in May and in June-July, respectively. Important differences were observed between stallions (P < .05), with most stallions resulting in a distribution of GL in an interval of ±5 days. The inclusion of the mare as a random effect in the mixed model resulted in an estimated repeatability of GL of 0.427, indicating that mares tend to be regular in having long or short gestations across their lifetime. Variance components estimated in an Animal Model resulted in heritability estimates of 0.39 for maternal genetic effects and 0.19 for direct genetic effects, with no association between the two components. Overall, the mare seems to have the major genetic influence on GL in Lusitano horses, but environmental factors, such as month of conception and also the age of the mare, have a noticeable effect.

Antigen Extraction and B Cell Activation Enable Identification of Rare Membrane Antigen Specific Human B Cells.

Determining antigen specificity is vital for understanding B cell biology and for producing human monoclonal antibodies. We describe here a powerful method for identifying B cells that recognize membrane antigens expressed on cells. The technique depends on two characteristics of the interaction between a B cell and an antigen-expressing cell: antigen-receptor-mediated extraction of antigen from the membrane of the target cell, and B cell activation. We developed the method using influenza hemagglutinin as a model viral membrane antigen, and tested it using acetylcholine receptor (AChR) as a model membrane autoantigen. The technique involves co-culturing B cells with adherent, bioorthogonally labeled cells expressing GFP-tagged antigen, and sorting GFP-capturing, newly activated B cells. Hemagglutinin-specific B cells isolated this way from vaccinated human donors expressed elevated CD20, CD27, CD71, and CD11c, and reduced CD21, and their secreted antibodies blocked hemagglutination and neutralized viral infection. Antibodies cloned from AChR-capturing B cells derived from patients with myasthenia gravis bound specifically to the receptor on cell membrane. The approach is sensitive enough to detect antigen-specific B cells at steady state, and can be adapted for any membrane antigen.

Predicting the Course of Disease in Hospitalized Patients With Acute Severe Ulcerative Colitis.

BACKGROUND: Up to one-third of patients with acute severe ulcerative colitis (ASUC) will fail intravenous steroid (IVS) treatment, requiring rescue therapy with cyclosporin (Cys), infliximab (IFX), or colectomy. Although several scores for predicting response to IVS exist, formal comparison is lacking. METHODS: We performed a single-center retrospective analysis including 489 patients with ulcerative colitis. In patients with ASUC, the Mayo endoscopic subscore and the Oxford, Edinburgh, and Lindgren scores were assessed. Outcomes included IVS failure, need for rescue medical therapy, and surgery. RESULTS: One hundred twelve patients presented with ASUC. Forty-two percent showed an incomplete or absent response to IVS, 28.6% received rescue therapy (22 with IFX, 10 with Cys, and 1 with sequential treatment), and 26.8% required surgery. The Lindgren score showed the highest performance in predicting IVS failure (are under the curve [AUC], 0.856; 95% confidence interval [CI], 0.784-0.928), need for medical rescue therapy (AUC, 0.826; 95% CI, 0.749-0.902), and surgery (AUC, 0.836; 95% CI, 0.712-0.960; all P < 0.01). CONCLUSIONS: In our series, the Lindgren score was superior to the Mayo, Oxford, and Edinburgh scores in predicting major clinical outcomes in ASUC.

Molecular pathway of influenza pan-neuraminidase inhibitor resistance in an immunocompromised patient.

BACKGROUND: Neuraminidase (NA) inhibitors (NAIs), including oseltamivir and zanamivir, play an important therapeutic role against influenza infections in immunocompromised patients. In such settings, however, NAI therapy may lead to the emergence of resistance involving mutations within the influenza surface genes. The aim of this study was to investigate the evolution of NA and haemagglutinin (HA) genes of influenza A(H1N1)pdm09 virus in an immunocompromised patient receiving oseltamivir then zanamivir therapies. METHODS: Nasopharyngeal swab (NPS) samples were collected between 27 January 2018 and 11 April 2018 from a haematopoietic stem cell transplant recipient. These include 10 samples collected either pre-therapy, during oseltamivir and zanamivir treatment as well as after therapy. The A(H1N1)pdm09 HA/NA genes were sequenced. The H275Y NA substitution was quantified by droplet digital RT-PCR assay. A(H1N1)pdm09 recombinant viruses containing HA mutations were tested by HA elution experiments to investigate in vitro binding properties. RESULTS: Oseltamivir rapidly induced the H275Y NA mutation which constituted 98.33% of the viral population after 15 days of oseltamivir treatment. The related HA gene contained S135A and P183S substitutions within the receptor-binding site. After a switch to zanamivir, 275H/Y and 119E/G/D mixed populations were detected. In the last samples, the double H275Y-E119G NA variant dominated with S135A and P183S HA substitutions. CONCLUSIONS: This report confirms that oseltamivir can rapidly induce the emergence of the H275Y substitution in A(H1N1)pdm09 viruses and subsequent switch to zanamivir can lead to additional substitutions at codon E119 resulting in multi-drug resistance. Such data additionally suggest a potential compensatory role for HA substitutions near the receptor binding site.

Stress e engagement na profissão de enfermagem: Análise de dois contextos internacionais / Stress and engagement in the nursing profession: Analysis of two international contexts / Estrés y engagement en la profesión de enfermería: Análisis de dos contextos internacionales

CONTEXTO: As mudanças constantes nas condições de trabalho, têm conduzido ao aparecimento de riscos psicossociais que conjugados com alterações organizacionais, de gestão de recursos humanos e das cargas horárias, podem levar ao aumento dos níveis de stress e ao comprometimento da saúde física e mental dos trabalhadores. OBJETIVO(S): Objetivou-se caraterizar duas amostras de enfermeiros (espanhóis e portugueses) e analisar as dimensões do stress e engagement. METODOLOGIA: Estudo comparativo e correlacional. Amostra não probabilística por conveniência, 504 enfermeiros portugueses (65.8%) e 363 espanhóis (34.2%) inscritos nas respetivas Ordens. Utilizou-se um questionário online, composto pela escala de stress, a “Nursing Stress Scale”, medindo sete dimensões de stress, e a escala de motivação, a “Utrecht Work Engagement Scale” (UWES), medindo três dimensões de motivação. RESULTADOS: Os enfermeiros portugueses apresentam pontuações médias superiores no Stress Total (2.2962±.43731) do que os espanhóis (2.2518±.46651) e nas dimensões “Morte e Morrer” e “Falta de Apoio”, essas diferenças são estatisticamente significativas (p=.015 e p<.001 respetivamente), sendo que no engagement os espanhóis apresentam resultados consistentemente superiores e estatisticamente significativos (p<.001) nas três dimensões. A variável profissional que mais dimensões do stress e do engagement influencia, é o tipo de horário, sendo o horário rotativo mais indutor de stress e menos motivador do que o horário fixo, sendo essas diferenças estatisticamente significativas (p≤.001) para o Stress Total e as três dimensões da UWES. CONCLUSÕES: Existem diferenças a nível do stress e motivação entre os enfermeiros portugueses e espanhóis.

BACKGROUND: Constant changes in working conditions have led to the emergence of psychosocial risks that, combined with organizational changes, human resources management and working hours, can lead to an increase in stress levels and compromise the physical and mental health of workers. AIM: To characterize two samples of nurses (Spanish and Portuguese) and analyse the dimensions of stress and engagement. METHODS: Comparative and correlational study. Non-probabilistic sample for convenience, 504 Portuguese nurses (65.8%) and 363 Spaniards (34.2%) enrolled in the respective Nurses Associations. An online questionnaire was used, consisting of the stress scale, the Nursing Stress Scale, measuring seven dimensions of stress, and the Utrecht Work Engagement Scale (UWES), measuring three dimensions of motivation. RESULTS: Portuguese nurses have higher mean scores in Total Stress (2.2962 ± .43731) than the Spanish (2.2518 ± .46651) and in the "Death and Dying" and "Lack of Support" dimensions, those differences are statistically significant (p =. 015 and p <.001 respectively), and in engagement, the Spaniards presented consistently higher and statistically significant scores (p <.001) in the three dimensions. The professional variable that more dimensions of stress and engagement influences, is the type of schedule, with rotating hours inducing more stress and less motivating than fixed hours, these differences being statistically significant (p≤.001) for Total Stress and the three dimensions of UWES. CONCLUSIONS: There are differences in stress and motivation among Portuguese and Spanish nurses.

CONTEXTO: Los cambios constantes en las condiciones de trabajo, han conducido a la aparición de riesgos psicosociales que, conjugados con alteraciones organizacionales, de gestión de recursos humanos y de las cargas horarias, pueden conducir al aumento de los niveles de estrés y al comprometimiento de la salud física y mental trabajadores. OBJETIVO(S): El objetivo fue caracterizar dos muestras de enfermeras (España y Portugal) y el análisis de las dimensiones de estrés y de compromiso. METODOLOGÍA: Estudio comparativo y correlacional. La muestra no probabilística de conveniencia, portugués 504 enfermeras (65,8%) y 363 españoles (34,2%) inscritos en los respectivos Órdenes. Se utilizó un cuestionario online, compuesto por la escala de estrés, la "Nursing Stress Scale", midiendo siete dimensiones de estrés, y la escala de motivación, la "Utrecht Work Engagement Scale" (UWES), midiendo tres dimensiones de motivación. RESULTADOS: Las enfermeras portuguesas tienen mayores puntuaciones medias en estrés total (2,2962 ± 0,43731) que la española (2,2518 ± 0,46651) y dimensiones "muerte y el morir" y "falta de apoyo", estas diferencias son estadísticamente significativas (p=.015 e p<.001 respetivamente) y en el compromiso los españoles presentan resultados consistentemente superiores y estadísticamente significativos (p <.001) en las tres dimensiones. La variable profesional que más dimensiones del estrés y del compromiso influye, es el tipo de horario, siendo el horario rotativo más inductor de estrés y menos motivador que el horario fijo, siendo esas diferencias estadísticamente significativas (p≤.001) para el Estrés Total y las tres dimensiones de la UWES. CONCLUSIONES: Existen diferencias en el estrés y la motivación entre los enfermeros de Portugal y España.

Influenza A Virus Genetic Tools: From Clinical Sample to Molecular Clone.

Implementation of reverse genetics for influenza A virus, that is, the DNA-based generation of infectious viral particles in cell culture, opened new avenues to investigate the function of viral proteins and their interplay with host factors on a molecular level. This powerful technique allows the introduction, depletion, or manipulation of any given sequence in the viral genome, as long as it gives rise to replicating virus progeny. Reverse genetics can be used to generate targeted reassortant viruses by mixing segments of different viral strains, thus providing insight into phenotypes of potentially pandemic viruses arising from natural reassortment. It was further instrumental for the development of novel vaccine strategies, allowing rapid and targeted exchange of viral surface antigens on a well-replicating genetic backbone of cell culture-adapted or cold-adapted/attenuated viral strains. Establishment of reverse genetics and rescue of molecular clones of influenza A virus have been extensively described before. Here we give a detailed stand-alone protocol encompassing clinical sampling of influenza A virus specimens and subsequent plasmid-based genetics to rescue, manipulate, and confirm a fully infectious molecular clone. This protocol is based on the combined techniques and experience of a number of influenza laboratories, which are credited and referenced whenever appropriate.

Systemic sclerosis and sarcoidosis: a rare case of chronic intestinal pseudo-obstruction

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Systemic sclerosis and sarcoidosis: a rare case of chronic intestinal pseudo-obstruction.

The coexistence of systemic sclerosis (SSc) and sarcoidosis is an extremely rare phenomenon; some studies question its existence. We report the case of a male with a diagnosis of sarcoidosis that was admitted due to abdominal distension and pain. After a thorough investigation, he was diagnosed with severe chronic intestinal pseudo-obstruction as a manifestation of SSc.

Clinical performance of an infliximab rapid quantification assay.

BACKGROUND: Therapeutic drug monitoring (TDM)-based algorithms can be used to guide infliximab (IFX) adjustments in inflammatory bowel disease (IBD) patients. This study aimed to explore a rapid IFX-quantification test from a clinical perspective. METHODS: This manuscript describes a prospective cohort study involving 110 ulcerative colitis (UC) patients on the maintenance phase of IFX. IFX trough levels were quantified using a rapid quantification assay and a commonly-used reference kit. RESULTS: Irrespective of the assay used to measure IFX, its through levels were statistically different between patients with and without endoscopic remission (Mayo endoscopic score = 0), as well as between patients stratified by their faecal calprotectin (FC) levels. Despite the fact that the two methods correlated well with each other [Spearman's rank correlation coefficient = 0.843, p < 0.001; intraclass correlation coefficients = 0.857, 95% confidence interval (CI): 0.791-0.903], there was a discernible systematic variation; values obtained with the reference kit were on average 2.62 units higher than those obtained with the rapid assay. Notwithstanding, 3 µg/ml was shown to be an acceptable cut-off to assess endoscopic status and inflammatory burden levels using both assays. The percentage of patients that had a positive outcome when the IFX concentration measured by the rapid assay ranked above 3 µg/ml was 88% both for a Mayo endoscopic score ⩽ 1 and for an FC concentration <250 µg/g. CONCLUSIONS: Based on this study, we concluded that using the rapid IFX assessment system with a 3 µg/ml threshold is a reliable alternative to the time-consuming enzyme-linked immunosorbent assays in patients on the maintenance phase of IFX.