Pseudomonas aeruginosa isolates from cystic fibrosis patients induce neutrophil extracellular traps with different morphologies that could correlate with their disease severity.

Cystic fibrosis (CF) is a disease characterized by bacterial chronic infection of the respiratory tract and inflammation, which leads to a progressive decrease in lung function. Pseudomonas aeruginosa is commonly isolated from the sputum of patients and their presence is associated with a predominant airway inflammation with neutrophils, causing chronic colonization and higher mortality rates. Neutrophil extracellular traps (NETs) have been observed in response against Pseudomonas, however, these cannot eliminate the pathogen from the respiratory tract, so one possibility is that the bacteria could promote their production to use them as a scaffold to colonize the lungs and as a nutrient source, however, their overproduction could also lead to increased damage to the lungs. In this work, we evaluated NETs formation by Pseudomonas clinical isolates obtained from CF patients and found that these induced NETs formation with globular or spread morphologies, of note, we found that there is a trend by which the spread forms were induced mainly by isolates obtained from patients with severe disease, whereas, the globular morphologies were observed for isolates obtained from patients with mild/moderate disease. Finally, we screened for bacterial molecules implicated in NETs formation and found that Exotoxin S, pyocin S2 and pyoverdine could participate in the process.

Cirrhosis, high age and high body mass index are risk factors for persisting advanced fibrosis after sustained virological response in chronic hepatitis C.

We aimed to assess fibrosis with liver stiffness measurement long-term after sustained virological response of chronic hepatitis C and to identify risk factors associated with persisting fibrosis. In this cross-sectional study, patients with chronic hepatitis C and pretreatment advanced fibrosis or cirrhosis treated successfully at Karolinska University Hospital with an interferon-containing regimen underwent liver stiffness measurement with FibroScan. The impact of potential risk factors for persisting fibrosis was estimated. We included 269 patients with a median follow-up time of 7.7 years (range 0-20), 84 with a follow-up time of ≥10 years. Patients with pretreatment cirrhosis had a significantly higher median liver stiffness level (8.5 kPa 95% CI 7-9.1) at follow-up, than patients with advanced fibrosis (6 kPa 95% CI 5.5-6.4). A majority improved their fibrosis stage after sustained virological response, but 24% had persisting advanced fibrosis with a liver stiffness level of ≥ 9.5 kPa. Among patients with pretreatment cirrhosis, the proportion with persisting advanced fibrosis diminished with longer follow-up time, from 48% after <5 years to 21% after >10 years. The main risk factors for persisting advanced fibrosis were pretreatment cirrhosis, high age and body mass index. In conclusion, fibrosis improves substantially during long-term follow-up after sustained virological response in hepatitis C patients with pretreatment advanced liver fibrosis. Lifestyle intervention to decrease weight in obese persons and treatment before establishment of cirrhosis should therefore be recommended to avoid persistence of advanced fibrosis after virological cure.

Long-term liver-related morbidity and mortality related to chronic hepatitis C virus infection in Swedish patients with inherited bleeding disorders.

INTRODUCTION: Hepatitis C virus (HCV) infection is common in patients with inherited bleeding disorders treated with clotting factor concentrates prior to the introduction of viral inactivation of these products. The long-term consequences of hepatitis C infection in Swedish patients are not fully understood. AIM: To examine the impact of HCV infection on liver-related morbidity and mortality in Swedish patients with inherited bleeding disorders. METHODS: We retrospectively collected data on 183 patients with inherited bleeding disorders infected with HCV who attended the Coagulation Unit at Karolinska University Hospital, Sweden. Data regarding end-stage liver disease (ESLD), defined as presence of ascites, encephalopathy, variceal bleeding, hepatocellular carcinoma or liver-related death, were collected from the patient records and the national registers. RESULTS: The median follow-up time was 35.9 years (IQR 29.0-41.2). A total of 41% had achieved sustained virological response (SVR) after treatment. In total, 14.2% developed ESLD at the median age of 52.6 years (IQR 46.5-64.7). Nineteen (35.8%) of all deaths were due to liver-related causes. Co-infection with human immunodeficiency virus (HIV), older age at time of infection and severe form of bleeding disorder was associated with higher risk of developing ESLD, while SVR was a strong protective factor. CONCLUSIONS: This study demonstrated that liver-related morbidity and mortality was significant in patients with bleeding disorders and HCV infection in Sweden. Patients with HCV-infection should be candidates for treatment with the new highly effective antiviral drugs, since SVR proved to be a strong protective factor.

Long-term follow-up of successful hepatitis C virus therapy: waning immune responses and disappearance of liver disease are consistent with cure.

BACKGROUND: A sustained viral response (SVR) after interferon-based therapy of chronic hepatitis C virus (HCV) infection is regarded to represent a cure. Previous studies have used different markers to clarify whether an SVR truly represents a cure, but no study has combined a clinical work-up with highly sensitive HCV RNA detection, and the determination of immune responses. AIM: To determine clinical, histological, virological and immunological markers 5-20 years after SVR. METHODS: In 54 patients, liver biochemistry, histology and elastography were evaluated. Liver biopsies, plasma and peripheral blood mononuclear cells (PBMCs) were tested for minute amounts of HCV RNA. HCV-specific T-cell responses were monitored by ELISpot and pentamer staining, and humoral responses by measuring HCV nonstructural (NS)3-specific antibodies and virus neutralisation. RESULTS: Liver disease regressed significantly in all patients, and 51 were HCV RNA-negative in all tissues tested. There was an inverse association between liver disease, HCV-specific T-cell responses and HCV antibody levels with time from SVR, supporting that the virus had been cleared. The three patients, who all lacked signs of liver disease, had HCV RNA in PBMCs 5-9 years after SVR. All three had HCV-specific T cells and NS3 antibodies, but no cross-neutralising antibodies. CONCLUSIONS: Our combined data confirm that a SVR corresponds to a long-term clinical cure. The waning immune responses support the disappearance of the antigenic stimulus. Transient HCV RNA traces may be detected in some patients up to 9 years after SVR, but no marker associates this with an increased risk for liver disease.

Strategies to manage hepatitis C virus (HCV) disease burden.

The number of hepatitis C virus (HCV) infections is projected to decline while those with advanced liver disease will increase. A modeling approach was used to forecast two treatment scenarios: (i) the impact of increased treatment efficacy while keeping the number of treated patients constant and (ii) increasing efficacy and treatment rate. This analysis suggests that successful diagnosis and treatment of a small proportion of patients can contribute significantly to the reduction of disease burden in the countries studied. The largest reduction in HCV-related morbidity and mortality occurs when increased treatment is combined with higher efficacy therapies, generally in combination with increased diagnosis. With a treatment rate of approximately 10%, this analysis suggests it is possible to achieve elimination of HCV (defined as a >90% decline in total infections by 2030). However, for most countries presented, this will require a 3-5 fold increase in diagnosis and/or treatment. Thus, building the public health and clinical provider capacity for improved diagnosis and treatment will be critical.

The present and future disease burden of hepatitis C virus (HCV) infection with today's treatment paradigm.

The disease burden of hepatitis C virus (HCV) is expected to increase as the infected population ages. A modelling approach was used to estimate the total number of viremic infections, diagnosed, treated and new infections in 2013. In addition, the model was used to estimate the change in the total number of HCV infections, the disease progression and mortality in 2013-2030. Finally, expert panel consensus was used to capture current treatment practices in each country. Using today's treatment paradigm, the total number of HCV infections is projected to decline or remain flat in all countries studied. However, in the same time period, the number of individuals with late-stage liver disease is projected to increase. This study concluded that the current treatment rate and efficacy are not sufficient to manage the disease burden of HCV. Thus, alternative strategies are required to keep the number of HCV individuals with advanced liver disease and liver-related deaths from increasing.

Proyecto de mejora en prevención de infecciones del tracto urinario asociadas a sondaje vesical / Improvement in urinary tract infections rates in a department of internal medicine

Objetivo. Valorar el impacto de una intervención desarrollada en el servicio de Medicina Interna del Hospital Universitario Insular de Gran Canaria para prevenir las infecciones del tracto urinario asociadas a sondaje vesical (ITU-SV). Material y métodos. El proyecto se desarrolló en 3 fases de vigilancia prospectivas de 2 meses de duración cada una entre 2009 y 2011. Durante las fases 1 y 2 se siguieron los criterios diagnósticos de los Centers for Disease Control and Prevention (CDC) de 2004 para la clasificación de las infecciones y se obtuvieron los indicadores de resultado. En la fase 3 se emplearon los criterios de los CDC de 2009 y se obtuvieron tanto indicadores de resultado como de proceso. Se comparó la incidencia acumulada de ITU-SV de las 3 fases mediante el test de tendencia ji-cuadrado. Resultados. El número total de días de sondaje vesical (SV) disminuyó desde la primera a la tercera fase, así como la incidencia acumulada de ITU-SV. Sin embargo, en la segunda fase la media de días de SV aumentó, volviendo a disminuir en la tercera fase. Se realizó una intervención con envío de recordatorios a través de la historia clínica electrónica durante la tercera fase, en la que se logró disminuir las tasas de ITU-SV y reducir la duración media de SV. Conclusiones. Las tasas de ITU-SV podrían ser minimizadas mediante un enfoque multidisciplinar que incluya tanto formación a los sanitarios implicados como vigilancia, retroalimentación y uso específico de recordatorios electrónicos(AU)

Objective. To evaluate the impact on the urinary tract infections (UTI) rates of an intervention implemented in the Department of Internal Medicine of the Hospital Universitario Insular de Gran Canaria. Material and methods. Infection control practitioners implemented a three phase project, each lasting two months, focusing on surveillance and feed-back, between 2009 and 2011. During phases 1 and 2, the 2004 Centers for Disease Control and Prevention (CDC)–diagnostic criteria for nosocomial infections were followed, and only rates of infections were calculated. For phase 3, the criteria published in 2009 were used, and rates of infections plus processes rates were obtained. The cumulative incidence of UTI in the three periods was compared using a chi-square for trends test. Results. The total number of catheter days, as well as the cumulative incidence of UTI dropped from phase 1 to 3. Nevertheless, in phase 2 the mean urinary catheter days increased. We detected a decrease in the UTI rates and urinary catheter days mean after introducing an electronic reminder in the patient electronic charts. Conclusions. A multidisciplinary approach, including surveillance, reminders, and feed-back, has proved useful in controlling UTI rates in our hospital(AU)

[Improvement in urinary tract infections rates in a department of internal medicine]. / Proyecto de mejora en prevención de infecciones del tracto urinario asociadas a sondaje vesical.

OBJECTIVE: To evaluate the impact on the urinary tract infections (UTI) rates of an intervention implemented in the Department of Internal Medicine of the Hospital Universitario Insular de Gran Canaria. MATERIAL AND METHODS: Infection control practitioners implemented a three phase project, each lasting two months, focusing on surveillance and feed-back, between 2009 and 2011. During phases 1 and 2, the 2004 Centers for Disease Control and Prevention (CDC)-diagnostic criteria for nosocomial infections were followed, and only rates of infections were calculated. For phase 3, the criteria published in 2009 were used, and rates of infections plus processes rates were obtained. The cumulative incidence of UTI in the three periods was compared using a chi-square for trends test. RESULTS: The total number of catheter days, as well as the cumulative incidence of UTI dropped from phase 1 to 3. Nevertheless, in phase 2 the mean urinary catheter days increased. We detected a decrease in the UTI rates and urinary catheter days mean after introducing an electronic reminder in the patient electronic charts. CONCLUSIONS: A multidisciplinary approach, including surveillance, reminders, and feed-back, has proved useful in controlling UTI rates in our hospital.

The burden of hepatitis C in Sweden: a national study of inpatient care.

The spread of hepatitis C virus (HCV) in Sweden in the 1970s indicated that serious liver complications (SLC) would increase in the 2000s. The aim of this study was to analyse the burden of HCV-associated inpatient care in Sweden, to demonstrate the changes over time and to compare the findings with a noninfected population. The HCV-cohort (n: 43,000) was identified from the national surveillance database 1990-2006, and then linked to national registers to produce an age-, sex-, and region-matched noninfected comparison population (n: 215,000) and to obtain information on demographics, cancers, inpatient care and prescriptions. Cox regression was used to estimate the likelihood (hazard ratios) for admission to hospital in the HCV compared with the noninfected cohort. The hazard ratios were 4.03 (95% CI: 3.98-4.08) for all care, 77.52 (71.02-84.60) for liver-related care and 40.74 (30.58-54.27) for liver cancer care. The admission rate in the HCV-cohort compared with the noninfected cohort, the rate ratio (age- and sex-adjusted) for all inpatient care was 5.91 (95% CI: 5.87-5.94), and the rate ratio for liver-related care was 70.05 (66.06-74.28). In the HCV-cohort, 45% of all episodes were for psychiatric, mostly drug-related, care. Inpatient care for SLC increased in the 2000s. To conclude, drug-related care was common in the HCV-infected cohort, the demand for liver-related care was very high, and SLC increased notably in the 2000s, indicating that the burden of inpatient care from serious liver disease in HCV-infected individuals in Sweden is an increasing problem.

Cleavage of the IPS-1/Cardif/MAVS/VISA does not inhibit T cell-mediated elimination of hepatitis C virus non-structural 3/4A-expressing hepatocytes.

BACKGROUND: Hepatitis C virus (HCV) effectively establishes persistent infection in human livers. The non-structural (NS) 3/4A complex participates in this process by cleavage of interferon beta (IFN beta) promoter stimulator-1 (IPS-1; also termed Cardif/MAVS/VISA), which inhibits responses to double stranded (ds) RNA. However, it is not known whether this effect extends beyond innate responses. AIMS: To test if HCV NS3/4A affects innate and adaptive immune responses in vivo. METHODS: NS3 levels were semi-quantified in human liver biopsies, transfected cells, and in transgenic (Tg) mouse livers by western blot. The effect of NS3/4A on dsRNA-mediated signalling and on the integrity of IPS-1 was analysed using in vitro translation, transfected cells and Tg mice. Cytotoxic T cell (CTL)-mediated clearance of transient firefly luciferase (FLuc)- and/or NS3/4A-Tg hepatocytes was determined using in vivo imaging and western blot. RESULTS: NS3 protein levels were in a comparable range (0.1-49 microg/g tissue) in infected human livers and Tg mouse livers. Importantly, these levels of NS3/4A reduced murine innate responses to synthetic dsRNA in vivo, supporting the possibility that this occurs also in infected humans. The likely explanation for this was the NS3/4A-mediated cleavage of mouse IPS-1, albeit less efficiently than human IPS-1. Despite this, FLuc- and/or NS3/4A-expressing murine hepatocytes were effectively eliminated by hepatic CTLs, utilising the classical molecules for virus-infected cell lysis, including CD8, IFN gamma, perforin and FasL. CONCLUSIONS: Although HCV NS3/4A inhibits the innate immunity, this does not prevent CTL-mediated clearance of NS3/4A-expressing hepatocytes in vivo. Thus, other HCV proteins are most likely responsible for interfering with the adaptive immunity.

Detection and characterization of focal liver lesions with contrast-enhanced ultrasonography in patients with hepatitis C-induced liver cirrhosis.

BACKGROUND: Hepatitis C virus (HCV)-associated liver cirrhosis provides a major preneoplastic condition for hepatocellular carcinoma (HCC). Ultrasonography (US) is usually used for screening of HCC, but needs improvement. PURPOSE: To assess whether use of a second-generation ultrasound contrast agent can improve characterization of focal liver lesions and detection of HCC in HCV-infected patients with liver cirrhosis. MATERIAL AND METHODS: In total, 96 US studies in 49 HCV-infected patients with liver cirrhosis were performed. The patients were first examined with a baseline US. After this, a diagnostic decision was made and recorded. The patients were then re-examined with contrast-enhanced ultrasound (CEUS), and the diagnostic triage was repeated. The patients were followed up for at least 1 year. RESULTS: On baseline US, indeterminate focal lesions were found in 27 examinations. After CEUS, a confident diagnosis of HCC was made in eight of these examinations. In an additional eight US examinations, diagnosis of regenerative/dysplastic noduli was established. In one patient with no detectable focal lesion at baseline examination, an indeterminate malignant lesion was detected with CEUS. This lesion was further investigated with computed tomography and diagnosed as HCC. CONCLUSION: Our study indicates that the use of CEUS significantly improves diagnostic confidence. CEUS improves the detection of HCC in patients with HCV-induced liver cirrhosis. Also, CEUS makes it possible to rule out malignancy in many cases where baseline US shows indeterminate focal lesions. In low-endemic countries, the use of CEUS in screening for HCC may be considered.

Fascitis necrotizante en recien nacidos / Necrotizing fascitis in newborns

La fascitis Necrotizante (FN) es una emergencia quirúrgica, resultado de la infección de los tejidos subcutáneos y de la fascia superficial, por una gran variedad de bacterias. En esta etapa neonatal, esta afección puede alcanzar una mortalidad mayor al 70 por ciento. El éxito del tratamiento requiere un preciso diagnóstico y precoz y agresivo desbridamiento de los tejidos afectados, la cobertura por vía parenteral de antibióticos de amplio espectro y un soporte adecuado en cuidados intensivos. Reportamos un caso de FN en recién nacidos de sexo masculino, en quien la enfermedad se desencadenó probablemente luego de una onfalitis. Este reporte ilustra la naturaleza devastadora de este tipo de infección, sin embargo, con tratamiento agresivo y precoz es posible lograr resultados satisfactorios.

Plasma levels of soluble CD27: a simple marker to monitor immune activation during potent antiretroviral therapy in HIV-1-infected subjects.

Plasma levels of soluble CD27 (sCD27) are elevated in diseases characterized by T cell activation and are used as a marker of immune activation. We assessed the usefulness of determining plasma sCD27 as a marker for monitoring immune activation in HIV-1-infected patients treated with highly active antiretroviral therapy (HAART). A first cross-sectional examination of 68 HIV-1-infected and 18 normal subjects showed high levels of sCD27 in HIV-1 infection; plasma sCD27 was correlated to HIV-1 viraemia and inversely correlated to CD4+ T cell count. Twenty-six HIV-1-infected patients undergoing HAART were studied at baseline and after 6, 12, 18 and 24 months of therapy. Seven additional patients under HAART were analysed at baseline, during and after interruption of therapy. In the total population, HAART induced a significant and progressive reduction, but not a normalization, of plasma levels of sCD27 after 24 months. A full normalization of plasma sCD27 was observed in the virological responders (undetectable HIV-1 RNA at months 18 and 24) and also in patients with moderate immunodeficiency at baseline (CD4+ T cell count >200 cells/mm3). Changes in plasma neopterin paralleled the changes in sCD27 but only baseline sCD27 levels were predictive of a greater increase in CD4+ T cell count during the follow-up. Discontinuation of therapy resulted in a rapid increase of sCD27 plasma levels associated with viraemia rebound and drop in CD4+ T cell count. Our findings suggest that plasma sCD27 may represent an alternative and simple marker to monitor immune activation during potent antiretroviral therapy. HIV-1-induced immune activation can be normalized by HAART in successfully treated patients where the disease is not advanced.

Human immunodeficiency virus type 1 variability and long-term non-progression.

A high heterogeneity is found in the HIV-1 genome in vivo, not only between individuals, but also within a single individual. Different types of genetic heterogeneity of HIV-1 can be analyzed: the extension and the evolution of the viral quasispecies in blood, the variation between the virus obtained from different body compartment, the differences between isolates from diverse individuals and between HIV-1 subtypes. The virus population during primary HIV-1 infection is generally homogeneous and the intrahost viral evolution is thought to be forced (in absence of antiviral therapy) by the immune system pressure and is generally related to the length of the immunocompetent period. A group of 12 Italian and Swedish well characterized HIV-1 infected long-term nonprogressors (LNTP) have been analyzed for the viral heterogeneity, calculated in the nef gene and in the long terminal repeat (LTR). The intra-sample variations in LTNP were found comparable with those from 8 progressor patients, while a lower inter-individual diversity was observed in the former. In one LTNP the viral evolution during a four-years period was extremely low suggesting that other factors than the host immune pressure may be involved in modulating the intra- and inter-sample HIV-1 diversity.

[Autoimmune hepatitis with prolonged IGM antibodies to HAV. Are serological markers reliable in ruling out a diagnosis of autoimmune hepatitis?]. / Hepatitis autoinmune con positividad mantenida al IgM VHA. Son los marcadores serológicos fiables para descartar el diagnóstico de la hepatitis autoinmune?

We present the case of a 44-year-old woman who was referred to us for jaundice, choluria and acholia. Review of the patient's clinical history revealed that tests for hepatitis A virus (HAV) IgM antibody had been positive 4 and 2 years previously and continued to be so. The patient showed a score of 16 in the International Autoimmune Hepatitis Group's system when the HAV marker was not taken into account and a score of 13 when this marker was taken into account. The persistence of this viral marker for so many years in such patients is probably associated with polyclonal hypergammaglobulinemia. Consequently, we believe that the inclusion of this antibody as a negative datum in the diagnosis of autoimmune virus should be viewed with caution.

Diferencias en ansiedad, depresión, estrés y apoyo social entre sujetos hipertensos y normotensos / Differences in anxiety, depression, stress and social support between hypertensive an normotensive subjects

La hipertensión arterial (HTA) es uno de los principales factores de riesgo de las enfermedades cardiovasculares (ECV). Existe una fuerte evidencia de la relación entre la depresión, la ansiedad, el estrés y el déficit de apoyo social y las ECV, e indicios de la influencia de dichas variables psicológicas sobre la HTA. Nuestro objetivo ha sido continuar ahondando en las supuestas relaciones entre dichas variables y la HTA. Método: se estudiaron las diferencias en ansiedad, depresión, nivel de estrés y nivel de apoyo social, entre dos grupos, 73 hipertensos diagnosticados y 73 controles sanos sociodemográficamente equivalentes. Resultados: se encontraron diferencias significativas en ansiedad y depresión entre ambos grupos, siendo más elevadas en el grupo de hipertensos. No se constataron diferencias significativas en nivel de estrés y en nivel de apoyo socia entre ambos grupos (AU)

High plasma levels of soluble fas in HIV type 1-infected subjects are not normalized during highly active antiretroviral therapy.

Plasma levels of soluble Fas (sFas) are elevated in human immunodeficiency virus type 1 (HIV-1) infection, indicating dysregulation of the Fas apoptosis pathway and chronic immune activation. We performed a retrospective study to investigate the effects of HAART on plasma levels of sFas. A cross-sectional study of 27 drug-naive infected subjects and 49 patients under antiretroviral treatment showed that plasma levels of sFas were higher in HIV-1-infected subjects than in 52 HIV-1-negative controls, independently of the treatment status. In a longitudinal study of 69 patients undergoing HAART, we observed a minimal, but significant decrease in sFas plasma levels after 1 year of therapy. Levels of sFas, however, remained still higher than physiologic values. Patients undergoing HAART were further classified as nonresponders or responders on the basis of viremia suppression; no significant changes in plasma levels of sFas were observed between the two groups. These findings show that 1 year of HAART has a minor effect on the sFas levels in plasma. Long-term HAART may be required to normalize the dysregulation of the Fas apoptotic pathway and the persistent immune activation initiated by HIV-1.

Proviral HIV-1 dynamics and evolution in patients receiving efficient long-term antiretroviral combination therapy.

OBJECTIVES: Different experimental approaches have shown that, despite plasma viral loads under the threshold of detection, HIV-1 frequently continues to replicate in patients receiving potent antiretroviral therapy. However, whether this low-grade viral replication is sufficient for the generation of new major quasispecies has not been studied. Thus, in order to evaluate the extent of variation in the major proviral HIV-1 population, we monitored proviral DNA sequences in such patients over a time period of up to 30 months. METHODS: DNA was extracted from peripheral blood mononuclear cells (PBMC) and the V3 region was amplified by nested polymerase chain reaction (PCR) and directly sequenced. Additionally, both HIV-1 RNA and DNA levels and CD4+ T-lymphocyte counts were monitored. RESULTS: Analysing the V3 gene sequences of 17 patients, we observed a sequence evolution in nine patients. Interestingly, the majority of these changes (77%) occurred in the first interval following the initiation of therapy and despite signs of ongoing replication the proviral DNA levels continued to decrease in all patients. CONCLUSIONS: Our data suggest that, although available data report that HIV-1 continues to replicate in patients with undetectable viraemia, the extent of viral replication in many of these patients is not sufficient to result in changes in the major viral population.