Cases Of Severe Disease Research Articles

Disease severity is associated with differential gene expression at the early and late phases of infection in nonhuman primates infected with different H5N1 highly pathogenic avian influenza viruses.

Occasional transmission of highly pathogenic avian H5N1 influenza viruses to humans causes severe pneumonia with high mortality. To better understand the mechanisms via which H5N1 viruses induce severe disease in humans, we infected cynomolgus macaques with six different H5N1 strains isolated from human patients and compared their pathogenicity and the global host responses to the virus infection. Although all H5N1 viruses replicated in the respiratory tract, there was substantial heterogeneity in their replicative ability and in the disease severity induced, which ranged from asymptomatic to fatal. A comparison of global gene expression between severe and mild disease cases indicated that interferon-induced upregulation of genes related to innate immunity, apoptosis, and antigen processing/presentation in the early phase of infection was limited in severe disease cases, although interferon expression was upregulated in both severe and mild cases. Furthermore, coexpression analysis of microarray data, which reveals the dynamics of host responses during the infection, demonstrated that the limited expression of these genes early in infection led to a failure to suppress virus replication and to the hyperinduction of genes related to immunity, inflammation, coagulation, and homeostasis in the late phase of infection, resulting in a more severe disease. Our data suggest that the attenuated interferon-induced activation of innate immunity, apoptosis, and antigen presentation in the early phase of H5N1 virus infection leads to subsequent severe disease outcome. Highly pathogenic avian H5N1 influenza viruses sometimes transmit to humans and cause severe pneumonia with ca. 60% lethality. The continued circulation of these viruses poses a pandemic threat; however, their pathogenesis in mammals is not fully understood. We, therefore, investigated the pathogenicity of six H5N1 viruses and compared the host responses of cynomolgus macaques to the virus infection. We identified differences in the viral replicative ability of and in disease severity caused by these H5N1 viruses. A comparison of global host responses between severe and mild disease cases identified the limited upregulation of interferon-stimulated genes early in infection in severe cases. The dynamics of the host responses indicated that the limited response early in infection failed to suppress virus replication and led to hyperinduction of pathological condition-related genes late in infection. These findings provide insight into the pathogenesis of H5N1 viruses in mammals.

Role of iNOS-NO-cGMP signaling in modulation of inflammatory and myelination processes.

Nitric oxide (NO) is the main activator of the soluble guanylate cyclase (sGC)-guanosine 3'5' cyclic monophosphate (cGMP) pathway. The level of cGMP is regulated by phosphodiesterases (PDEs), which break down cGMP. It has been reported that levels of NO in the central nervous system (CNS) can greatly increase during demyelination and/or neuroinflammation. Controversially, in demyelination models, mice without iNOS may develop more severe cases of disease. Furthermore, cGMP accumulation caused by PDE inhibitors has an anti-inflammatory/neuroprotective effect in MS-models. The role of the NO-cGMP pathway in the nervous tissue is, therefore, complex and not fully understood. The aim of the present study was to contribute to existing knowledge of the role of this pathway in the CNS. Wild type (WT - C57BL/6) and iNOS(-/-) animals were treated with sildenafil (25mg/kg) for 8 weeks. Control animals were not treated. VCAM and ICAM (adhesion proteins), GFAP and Iba-1 (astrocyte and microglia markers, respectively), PKG (cGMP-dependent protein kinase), sGC, eNOS (constitutive endothelial NO sinthase) and GSTpi (a marker of mature oligodendrocytes) were evaluated in the cerebellum using immunohistochemistry or western blotting. Myelin was assessed by luxol fast blue staining and electron transmission microscopy. Treatment with sildenafil reduced ICAM and VCAM levels (anti-inflammatory effect) and increased GFAP and Iba-1 expression (clearance phenotype) in WT animals. The expression of VCAM, ICAM, GFAP, PKG and sGC was lower in iNOS(-/-) mice than in WT control animals. The treatment of iNOS(-/-) animals with sildenafil resulted in an increase of all proteins (pro-inflammatory effect). There was overexpression of eNOS in untreated iNOS(-/-) mice. The myelin structure of iNOS(-/-) animals was damaged in comparison with WT control. Sildenafil increased GSTpi and resulted in an improved myelin structure in iNOS(-/-) mice. In conclusion, NO-cGMP signaling plays a role in the regulation of inflammation and myelination processes. The accumulation of cGMP produced opposite effects in WT and iNOS(-/-) mice. This can be explained by the overexpression of eNOS in iNOS(-/-) mice, unbalancing cGMP signaling, or cGMP has a dual role in inflammation. Drugs that modulate the NO-sGC-cGMP pathway may be clinically beneficial in the treatment of neuroinflammatory/demyelinating disorders, but further studies of the regulation of this pathway are required.

Changed terms for drug payment influenced GPs’ diagnoses and prescribing practice for inhaled corticosteroids

Background: Inhaled glucocorticosteroids (ICS) are first-line anti-inflammatory treatment in asthma, but not in chronic obstructive pulmonary disease (COPD). To restrict ICS use in COPD to cases of severe disease, new terms for reimbursement of drug costs were introduced in Norway in 2006, requiring a diagnosis of COPD to be verified by spirometry.Objectives: To describe how GPs’ diagnoses and treatment of patients who used ICS before 2006 changed after a reassessment of the patients that included spirometry.Methods: From the shared electronic patient record system in one group practice, patients ≥ 50 years prescribed ICS (including in combination with long-acting beta2-agonists) during the previous year were identified and invited to a tailored consultation including spirometry to assure the quality of diagnosis and treatment. GPs’ diagnoses and ICS prescribing patterns after this reassessment were recorded, retrospectively.Results: Of 164 patients identified, 112 were included. Post-bronchodilator spirometry showed airflow limitation indicating COPD in 55 patients. Of the 57 remaining patients, five had a positive reversibility test. The number of patients diagnosed with asthma increased (from 25 to 62) after the reassessment. A diagnosis of COPD was also more frequently used, whereas fewer patients had other pulmonary diagnoses. ICS was discontinued in 31 patients; 20 with mild to moderate COPD and 11 with normal spirometry.Conclusion: Altered reimbursement terms for ICS changed GPs’ diagnostic practice in a way that made the diagnoses better fit with the treatment given, but over-diagnosis of asthma could not be excluded. Spirometry was useful for identifying ICS overuse.

Variability in disease burden and management of rheumatic fever and rheumatic heart disease in two regions of tropical Australia

Acute rheumatic fever (ARF) and rheumatic heart disease (RHD) contribute to Aboriginal Australian and Torres Strait Islander health disadvantage. At the time of this study, specialist ARF/RHD care in the Kimberley region of Western Australia was delivered by a broad range of providers. In contrast, in Far North Queensland (FNQ), a single-provider model was used as part of a coordinated RHD control programme. To review ARF/RHD management in the Kimberley and FNQ to ascertain whether differing models of service delivery are associated with different disease burden and patient care. An audit of ARF/RHD management. Classification and clinical management data were abstracted from health records, specialist letters, echocardiograms and regional registers using a standardised data collection tool. Four hundred and seven patients were identified, with 99% being Aboriginal and/or Torres Strait Islanders. ARF without RHD was seen in 0.4% of Aboriginal and/or Torres Strait Islander residents and RHD in 1.1%. The prevalence of RHD was similar in both regions but with more severe disease in the Kimberley. More FNQ RHD patients had specialist review within recommended time frames (67% vs 45%, χ(2) , P < 0.001). Of patients recommended benzathine penicillin secondary prophylaxis, 17.7% received ≥80% of scheduled doses in the preceding 12 months. Prescription and delivery of secondary prophylaxis was greater in FNQ. FNQ's single-provider model of specialist care and centralised RHD control programme were associated with improved patient care and may partly account for the fewer cases of severe disease and reduced surgical procedures and other interventions observed in this region.

Counting the Cost of Meningococcal Disease

Meningococcal disease can result in severe disabling sequelae, but there is no published information about the lifelong rehabilitation costs of patients with severe outcome in the UK. As cost-effectiveness studies play a crucial role in determining whether immunization programmes will be implemented, it is important to identify these costs. The aim of the study was to estimate lifelong rehabilitation costs associated with severe cases of meningococcal disease and to present these costs in a format appropriate for use in a cost-effectiveness analysis. Two severe scenarios of meningococcal disease with major sequelae were developed; one that presented acutely as meningitis and the other as septicemia. Scenarios were based on systematic reviews of the literature describing the sequelae of meningococcal disease, dialogue with Meningitis Research Foundation members who have experience of the disease, and discussions with pediatricians who have been responsible for managing children with this disease over many years. The two scenarios were devised to represent cases typical of the severe end of the spectrum. To obtain a comprehensive list of the health, educational and other resources used by survivors during and since their acute illness, families of individuals with sequelae similar to those in each of our scenarios were interviewed. Relevant academics and professionals in health, social care and education were consulted in order to ensure that our scenarios accurately represented the treatment and support that individuals with such sequelae might realistically receive from the National Health Service (NHS), the local authority and Personal Social Services (PSS). The majority of costs were derived from English Department of Health reference costs and unit costs of health and social care reflecting values for the financial year 2008-2009 indexed to 2010-2011. Costs were based on a life expectancy of 70 years in each scenario and are presented at a discount rate of 3.5 % for the first 30 years and 3 % thereafter, as recommended by the UK treasury (non-discounted costs are also presented for comparison). Costs are presented from both an NHS/PSS perspective and a government perspective. This study has revealed that severe cases of disease that result in long-term sequelae can result in costs to the NHS/PSS of around £160,000-£200,000 in the first year alone. Over a lifetime of 70 years, discounted costs from an NHS/PSS perspective ranged from approximately £590,000 to £1,090,000 (£1,250,000-£3,320,000 undiscounted) and discounted costs from a government perspective ranged from £1,360,000 to £1,720,000 (£3,030,000-£4,470,000 undiscounted). This study fills a gap in the UK literature and produces estimates that can be used in cost-effectiveness analysis to better represent the cost of illness at the severe end of the spectrum. Costs from a government perspective highlight the wider impacts of this disease, which is important for clinical decision makers, and budgetary and service planners to be aware of when making decisions about the benefits of implementing public health interventions such as immunization programmes.

Glycobiology and the Paediatric Eye in Health and Disease

Inflammatory bowel disease, especially Crohn disease, had increased in the last five decades. We analyzed all the patients of our unit diagnosed with the disease between 2001-2012 but we just considered the patients who respected the Porto criteria to define IBD at presentation. We diagnosed 51 children and adolescents with the disease. Crohn disease was responsible for 62.7% of cases, ulcerative colitis for 31.4% and indeterminate colitis for 5.9%. Considering the 4 different periods of time (2001-2003/2004-2006/2007-2009 and 2010-2012), we observed and increase in the number of cases, but a decrease in the time from onset of symptoms to diagnosis. Our medical approach with tumor necrosis factor antagonists was reserved for severe disease cases and to spare patients from adverse effects of purine analogues and corticosteroids. Mucosal healing and remission of the disease was achieved in all of the patients under biologic therapy and no serious or life-threating event was reported with their use.

Inflammatory Responses Associated with the Induction of Cerebral Malaria: Lessons from Experimental Murine Models

Inflammatory Responses Associated with the Induction of Cerebral Malaria: Lessons from Experimental Murine Models

Molecular Characterization and Phylogenetic Analysis of the Hemagglutinin 1 Protein of Human Influenza A Virus Subtype H1N1 Circulating in Kenya During 2007-2008

Among influenza viruses, type A viruses exhibit the greatest genetic diversity, infect the widest range of host species, and cause the vast majority of cases of severe disease in humans, including cases during the great pandemics. The hemagglutinin 1 (HA1) domain of the HA protein contains the highest concentration of epitopes and, correspondingly, experiences the most intense positive selection pressure. We sought to isolate and genetically characterize influenza A virus subtype H1N1 (A[H1N1]) circulating in Kenya during 2007-2008, using the HA1 protein. Nasopharyngeal swab specimens were collected from patients aged ≥ 2 months who presented to 8 healthcare facilities in Kenya with influenza-like illness. We tested specimens for seasonal influenza A viruses, using real-time reverse-transcription polymerase chain reaction (RT-PCR). Viruses were subtyped using subtype-specific primers. Specimens positive for seasonal A(H1N1) were inoculated onto Madin-Darby canine kidney cells for virus isolation. Viral RNAs were extracted from isolates, and the HA1 gene was amplified by RT-PCR, followed by nucleotide sequencing. Nucleotide sequences were assembled using BioEdit and translated into amino acid codes, using DS Gene, version 1.5. Multiple sequence alignments were performed using MUSCLE, version 3.6, and phylogenetic analysis was performed using MrBayes software. We found that, similar to A/Brisbane/59/2007 (H1N1)-like virus, which was included in the southern hemisphere vaccine for the 2009 influenza season, all 2007 Kenyan viruses had D39N, R77K, T132V, K149R, and E277K amino acid substitutions, compared with A/Solomon Islands/3/2006 (H1N1)-like virus, a component of the southern hemisphere vaccine for the 2008 influenza season. However, the majority of 2008 viruses from Kenya also had R192K and R226Q substitutions, compared with A/Solomon Islands/3/2006 (H1N1)-like virus. These 2 changes occurred at the receptor binding site. The majority of the 2008 Kenyan isolates contained N187S, G189N, and A193T mutations, which differed from A/Brisbane/59/2007 (H1N1)-like virus. The A193T substitution is involved in binding the sialic acid receptor. Phylogenetically, the 2008 Kenyan isolates grouped into 2 clusters. The main cluster contained viruses with N187S and A193T changes; residue 187 is involved in receptor binding, whereas residue 193 is at antigenic site Sb. Overall, the major genetic variations that occurred in seasonal A(H1) viruses either affected receptor binding or altered epitopes at the immunodominant sites. These genetic variations in seasonal A(H1N1) isolated in Kenya during 2007-2008 highlight the importance of continuing surveillance and characterization of emerging drift variants of influenza virus in Africa.

Easy Diagnosis of Invasive Pneumococcal Disease

To the Editor: Invasive pneumococcal disease (IPD) causes many cases of severe disease and death among children <5 years of age, mostly in developing countries (1,2). Before conjugate vaccines can be introduced in developing countries, information about disease epidemiology is urgently needed. The lack of laboratories equipped to perform pneumococcal serotyping leads to the need to send isolates to reference laboratories. Good sample preservation is necessary to prevent samples from arriving at the laboratory in poor condition. We evaluated the usefulness of multiplex real-time PCR from strains and blood samples kept at room temperature on dried blood spot (DBS) filter paper for detecting and serotyping Streptococcus pneumoniae. DBS screening is a reliable method that requires only a small amount of blood; it is used for the diagnosis of several human diseases (3,4). To validate the technique, we selected 15 pneumococcus clinical isolates representing 15 serotypes (1, 5, 19A, 19F, 14, 3, 7F, 4, 6A, 6B, 8, 9N, 18C, 23A, 23F) obtained during 2009 from patients at Hospital Sant Joan de Deu, in Barcelona. These isolates, used as controls, had been serotyped by quellung reaction at the Instituto de Salud Carlos III, Majadahonda-Madrid, Spain. These strains were cultured overnight at 35°C in 5% carbon dioxide on Columbia agar plates with 5% sheep blood (bioMerieux SA, Marcy l’Etoile, France). A suspension of each strain was adjusted to match a 0.5 McFarland standard (equivalent to 108 colony-forming units (CFU)/mL). Stock solutions of pneumococcus culture for each previously identified serotype were injected into blood previously extracted from 2 healthy volunteers. Serial dilutions of 100,000 CFU/mL to 1,000 CFU/mL (1,000 to 10 CFU equivalents/PCR) were performed. A total of 100 µL of blood was applied to DBS filter paper, and another 100 µL was used for DNA extraction from fresh blood. All DBS samples were air dried for 1 week. The procedure was also performed on negative control blood samples. DNA was extracted from DBS and fresh blood samples by using the NucliSense easyMAG automated extraction platform (bioMerieux, Boxtel, the Netherlands) according to the manufacturer’s instructions. DNA detection of the pneumolysin (ply) gene by real-time PCR was performed according to a published assay (5). In addition, we performed a multiplex real-time PCR for molecular serotype detection of serotypes 1, 3, 5, 4, 6A, 6B, 7FA, 8, 9VANL, 14, 15BC, 18CB, 19A, 19FBC, 23F, 23A and the conserved capsular gene wzg as described by Tarrago et al. (6). DNA extracts were amplified with the Applied Biosystems 7300 Real-time PCR System (Applied Biosystems, Foster City, CA, USA). Negative results were defined as those with cycle threshold >40. To evaluate the reliability obtained with this in vivo approach, we performed identification and serotyping of S. pneumoniae in 25 DBS samples from 25 children at Saint John of God Hospital in Mabesseneh-Lunsar, Sierra Leone. This hospital does not perform blood cultures. IPD was confirmed when DNA of a pneumolysin (ply) gene and an additional capsular gene of S. pneumoniae were detected by multiplex real-time PCR of DBS samples. Detection of ply, wzg, and the specific gene for molecular serotype showed that both fresh blood and DBS samples yielded correctly positive results from the 10-fold serial dilutions analyzed (Table). With respect to the 25 (11 female and 14 male) patients from Sierra Leone who had suspected IPD, the median age was 25.71 months (range 15 days to 96 months); all had a diagnosis of fever without apparent source, and 16 also had malaria. Of these 25 children, DBS samples from 15 (60%) yielded a positive result for the ply and wzg genes, so they were considered confirmed episodes of IPD. A serotype included in 13-valent conjugate vaccine was detected in 6 (40%) of 15 positive samples: serotypes 3, 7FA, 19A, 6A, 6B, and 9VNL (1 sample each). In the remaining 9 samples, the results for ply gene and wzg gene were positive, but none of the 24 tested serotypes was detected. Table Sensitivity of real-time PCR for detecting Streptococcus pneumoniae ply or wzg genes or a specific gene for molecular serotype from fresh or dried blood spot samples* This preliminary study enabled us to demonstrate that DBS screening is a reliable and easy method for diagnosing IPD and also for epidemiologic surveillance of the more frequent serotypes. The main limitation of our study is the small number of DBS samples sent from Saint John of God Hospital in Sierra Leone. In conclusion, the DBS technique enables reproducible transport of samples for identification and serotyping of S. pneumoniae by multiplex PCR. The use of DBS on filter paper is an attractive alternative method for storing samples at room temperature and easily transporting them. Additional studies, including evaluation of the relative sensitivity of this method compared to direct culture, are necessary.

Analysis of 5′ Nontranslated Region of Hepatitis A Viral RNA Genotype I from South Korea: Comparison with Disease Severities

The aim of the study was to analyze genotype I hepatitis A virus (HAV) 5′ nontranslated region (NTR) sequences from a recent outbreak in South Korea and compare them with reported sequences from Japan. We collected a total of 54 acute hepatitis A patients' sera from HAV genotype I [27 severe disease (prothrombin time INR≥1.50) and 27 mild hepatitis (prothrombin time INR <1.00)], performed nested RT-PCR of 5′ NTR of HAV directly sequenced from PCR products (∼300 bp), and compared them with each other. We could detect HAV 5′NTR sequences in 19 of the 54 (35.1%) cases [12 of 27 severe cases (44.4%) and 7 of 27 self-limited cases (25.9%)], all of which were subgenotype IA. Sequence analysis revealed that sequences of severe disease had 93.6%–99.0% homology and of self-limited disease 94.3%–98.6% homology, compared to subgenotype IA HAV GBM wild-type IA sequence. In this study, confirmation of the 5′NTR sequence differences between severe disease and mild disease was not carried out. Comparison with Japanese HAV A10 revealed 222C to G or T substitution in 8/12 cases of severe disease and 222C to G or T and 392G to A substitutions in 5/7 and 4/7 cases of mild disease, respectively, although the nucleotide sequences in this study showed high homology (93.6%–100%). In conclusion, HAV 5′NTR subgenotype IA from Korea had relatively high homology to Japanese sequences previously reported from Japan, and this region would be considered one of the antiviral targets. Further studies will be needed.

Influenza A virus transmission: contributing factors and clinical implications

Efficient human-to-human transmission is a necessary property for the generation of a pandemic influenza virus. To date, only influenza A viruses within the H1-H3 subtypes have achieved this capacity. However, sporadic cases of severe disease in individuals following infection with avian influenza A viruses over the past decade, and the emergence of a pandemic H1N1 swine-origin virus in 2009, underscore the need to better understand how influenza viruses acquire the ability to transmit efficiently. In this review, we discuss the biological constraints and molecular features known to affect virus transmissibility to and among humans. Factors influencing the behaviour of aerosols in the environment are described, and the mammalian models used to study virus transmission are presented. Recent progress in understanding the molecular determinants that confer efficient transmission has identified crucial roles for the haemagglutinin and polymerase proteins; nevertheless, influenza virus transmission remains a polygenic trait that is not completely understood. The clinical implications of this research, including methods currently under investigation to mitigate influenza virus human-to-human transmission, are discussed. A better understanding of the viral determinants necessary for efficient transmission will allow us to identify avian influenza viruses with pandemic potential.

Virulence-Associated Substitution D222G in the Hemagglutinin of 2009 Pandemic Influenza A(H1N1) Virus Affects Receptor Binding

The clinical impact of the 2009 pandemic influenza A(H1N1) virus (pdmH1N1) has been relatively low. However, amino acid substitution D222G in the hemagglutinin of pdmH1N1 has been associated with cases of severe disease and fatalities. D222G was introduced in a prototype pdmH1N1 by reverse genetics, and the effect on virus receptor binding, replication, antigenic properties, and pathogenesis and transmission in animal models was investigated. pdmH1N1 with D222G caused ocular disease in mice without further indications of enhanced virulence in mice and ferrets. pdmH1N1 with D222G retained transmissibility via aerosols or respiratory droplets in ferrets and guinea pigs. The virus displayed changes in attachment to human respiratory tissues in vitro, in particular increased binding to macrophages and type II pneumocytes in the alveoli and to tracheal and bronchial submucosal glands. Virus attachment studies further indicated that pdmH1N1 with D222G acquired dual receptor specificity for complex α2,3- and α2,6-linked sialic acids. Molecular dynamics modeling of the hemagglutinin structure provided an explanation for the retention of α2,6 binding. Altered receptor specificity of the virus with D222G thus affected interaction with cells of the human lower respiratory tract, possibly explaining the observed association with enhanced disease in humans.

Introduction of Virulence Markers in PB2 of Pandemic Swine-Origin Influenza Virus Does Not Result in Enhanced Virulence or Transmission

In the first 6 months of the H1N1 swine-origin influenza virus (S-OIV) pandemic, the vast majority of infections were relatively mild. It has been postulated that mutations in the viral genome could result in more virulent viruses, leading to a more severe pandemic. Mutations E627K and D701N in the PB2 protein have previously been identified as determinants of avian and pandemic influenza virus virulence in mammals. These mutations were absent in S-OIVs detected early in the 2009 pandemic. Here, using reverse genetics, mutations E627K, D701N, and E677G were introduced into the prototype S-OIV A/Netherlands/602/2009, and their effects on virus replication, virulence, and transmission were investigated. Mutations E627K and D701N caused increased reporter gene expression driven by the S-OIV polymerase complex. None of the three mutations affected virus replication in vitro. The mutations had no major impact on virus replication in the respiratory tracts of mice and ferrets or on pathogenesis. All three mutant viruses were transmitted via aerosols or respiratory droplets in ferrets. Thus, the impact of key known virulence markers in PB2 in the context of current S-OIVs was surprisingly small. This study does not exclude the possibility of emergence of S-OIVs with other virulence-associated mutations in the future. We conclude that surveillance studies aimed at detecting S-OIVs with increased virulence or transmission should not rely solely on virulence markers identified in the past but should include detailed characterization of virus phenotypes, guided by genetic signatures of viruses detected in severe cases of disease in humans.

Comparative Quality of Laparoscopic and Open Cholecystectomy in the Elderly Using Propensity Score Matching Analysis

The safety of laparoscopic cholecystectomy (LC) in patients ≥65 years of age requires further investigation of postoperative outcomes before it becomes more widely accepted as a safe technique. The advantages of using LC versus open cholecystectomy (OC) in elderly patients were analyzed using propensity score matching. The demographics, cholecystitis severity, comorbidities, complications, and admission and discharge Barthel Index (BI) scores of patients with benign gallbladder diseases were analyzed. Outcomes were analyzed by age, length of stay (LOS), total charges (TCs), BI improvement, and postoperative complications. OC, which was indicated in severe disease cases, increased hospital resource use and caused more complications than LC, but did not improve BI. Advanced age and OC resulted in greater LOS and TCs and was the best indicator of BI deterioration. Whenever possible, surgeons should use LC in elderly patients to minimize postoperative complications and allow them to regain a good quality of life.

Simplified Early Predictors of Severe Acute Pancreatitis: A Prospective Study

BackgroundTo propose simple tests for the prediction of severe acute pancreatitis (SAP), which are accurate and could be performed at emergency departments and outpatient clinics.MethodsA prospective study was performed on 149 patients admitted with acute pancreatitis. Body mass index (BMI), plain chest radiograph, blood biochemical data were obtained at the time of admission; white cell, lymphocyte and platelet counts, hematocrit level, prothrombin time, PaO2, creatinine, calcium, blood sugar, total protein, aspartate aminotransferase, total bilirubin, amylase, lipase and C-reaction protein were determined. Patients were graded into severe and mild acute pancreatitis based on CT Balthazar grading system.ResultsTwenty-seven patients were diagnosed to have SAP and 122 patients considered mild acute pancreatitis. Comparing parameters between both groups; significant factors (P < 0.05) were blood sugar level, haematocrit level, BMI and presence of pleural effusion in chest X-ray. The hematocrit at admission and at approximately 24 hours was significantly higher among patients with SAP. Twenty-two of 27 cases of severe disease and only 10 of 122 cases of mild acute pancreatitis diagnosed to have pleural effusion (P < 0.001).ConclusionBMI, blood glucose ≥ 190 mg/dL, hematocrit level ≥ 43 % and pleural effusion detected by plain chest radiograph are simple tests and provide significant predictive power for clinical decision-making.

Swine Flu (H1N1/09)

The most recent epidemiological weekly reports from the Robert Koch Institute show that the swine flu epidemic is now spreading with increasing speed in Germany too. The number of new cases in the country is more than doubling every week—in some states (Lander) it is even faster. This has become clear not just from the statutory notification data, but also from the data emanating from the Sentinel system run by the Working Group on Influenza (Arbeitsgruppe Influenza). The following facts are particularly notable: First, the influenza activity is beginning earlier than we are used to seeing with seasonal flu. Second, in the laboratory-confirmed cases the patients are infected almost exclusively with the novel influenza A (H1N1), not with seasonal influenza strains. Third, in by far the larger number of cases now, infection is autochthonous. In addition, the numbers of patients requiring hospitalization and who have a severe illness ending in death are also increasing. This development indicates the start of a qualitative change in the course of the epidemic in Germany, just as was seen much earlier in other countries, such as the USA, the UK, and France. The article by Winzer et al. is the first report on retrospectively evaluated data from a large clinical cohort of patients infected with the novel influenza A (H1N1) in Germany, starting from the beginning of the pandemic. The authors give an exemplary description of the beginning and course of the first, non-seasonal months of the epidemic, in which the infection was reported first in travelers from South and North America, then in people returning from holidays in Spain. They also describe the implementation and success of rigorous containment measures, including in the initial phase, especially in Dusseldorf, inpatient admission under quarantine conditions. In addition to epidemiological data, comprehensive information about the manifestation, symptoms, laboratory test parameters, course, and treatment were obtained. Many infected individuals remained without symptoms, and most of those who became ill became only mildly so. No cases of severe disease requiring intensive care were observed. A publication by Kumar et al. (1) gives a very different picture. This gives the results of a multicenter observational study of a Canadian cohort over the same time period. Those affected were primarily young, female, and of native ethnicity (Indian, Eskimo), without serious underlying disease. A history of lung disease and smoking and clinical findings of hypertension, diabetes, and obesity were the main comorbidities. About one-third of the 168 patients who were critically ill and in intensive care required special ventilation regimens, in some cases even extracorporeal membrane oxygenation; 49 (17.3%) died. Of 50 children, 4 died (8%). A substantial number of patients suffered bacterial complications, usually due to S. aureus or S. pneumoniae. Compared to seasonal flu, notable findings were gastrointestinal symptoms, dyspnea, and purulent sputum. The Canadian authors particularly emphasized the medical-logistical and social-ethical implications: even before the normal flu season, the intensive care facilities were nearly completely taken up by patients infected with the novel influenza A (H1N1). An increased wave of infections such as one might expect at this time of year would—or indeed will—pose a challenge to the (limited) high-tech intensive care facilities in our healthcare system too, and one such as it has never seen before. So far Germany has succeeded, by a variety of hygiene measures, to keep the wave of the epidemic fairly “flat.” The aim of these efforts was to keep a lid on any epidemic/pandemic until a vaccine was available. The article in this issue by Mikolajczyk et al. on the modelling of infectious diseases attempts to explain the principles upon which calculations are made about the courses of epidemics. The contribution does not contain any specific prediction about the novel influenza A (H1N1), but the basic principles of course apply to any influenza epidemic.

The Oxazolidinone Derivative Locostatin Induces Cytokine Appeasement

Damaging inflammation arising from autoimmune pathology and septic responses results in severe cases of disease. In both instances, anti-inflammatory compounds are used to limit the excessive or deregulated cytokine responses. We used a model of robust T cell stimulation to identify new proteins involved in triggering a cytokine storm. A comparative proteomic mining approach revealed the differential mapping of Raf kinase inhibitory protein after T cell recall in vivo. Treatment with locostatin, an Raf kinase inhibitory protein inhibitor, induced T cell anergy by blocking cytokine production after Ag recall. This was associated with a reduction in Erk phosphorylation. Importantly, in vivo treatment with locostatin profoundly inhibited TNF-alpha production upon triggering the Ag-specific T cells. This effect was not limited to a murine model because locostatin efficiently inhibited cytokine secretion by human lymphocytes. Therefore, locostatin should be a useful therapeutic to control inflammation, sepsis, and autoimmune diseases.

Malaria is associated with poor school performance in an endemic area of the Brazilian Amazon

BackgroundApproximately 40% of the world's population is at risk for malaria. In highly endemic tropical areas, malaria is a major cause of morbidity and mortality during infancy. There is a complex interrelationship between malaria, malnutrition and intestinal helminths, and this may impair cognitive development in children. The aim of this study was to determine the relationship between malaria and school performance in children living in an endemic area where Plasmodium vivax is the species responsible for most of the cases.MethodsThe study was conducted in the Municipality of Careiro, Amazonas, Brazil, with five to14 year-old children, studying the first eight grades of public school, during the year 2008. After an initial active case detection, during nine months of follow-up, passive malaria cases detection was instituted, through a thick blood smear performed in every child with fever. School performance was evaluated by the final notes in Mathematics and Portuguese Language. Performance was considered poor when either of the final notes in these disciplines was below the 50th percentile for the respective class and grade.ResultsThe total number of students followed-up in the cohort was 198. Malarial attacks were reported in 70 (35.4%) of these students, with no cases of severe disease. Plasmodium vivax was detected in 69.2% of the attacks, Plasmodium falciparum in 25.5% and both species in 5.3%. In the multivariate analysis, adjusting for age, mother's education, time living in the study area and school absenteeism, presenting with at least one episode of malaria independently predicted a poor performance at school [OR = 1.91 (1.04-3.54); p = 0.039].ConclusionNon-severe malaria compromises the school performance of children even during a nine-month follow-up, potentially contributing to the maintenance of underdevelopment in countries endemic for malaria. This is the first evidence of such impact in Latin America, where P. vivax is responsible for the majority of the cases.

Genetic Characterization of Venezuelan Equine Encephalitis Virus from Bolivia, Ecuador and Peru: Identification of a New Subtype ID Lineage

Venezuelan equine encephalitis virus (VEEV) has been responsible for hundreds of thousands of human and equine cases of severe disease in the Americas. A passive surveillance study was conducted in Peru, Bolivia and Ecuador to determine the arboviral etiology of febrile illness. Patients with suspected viral-associated, acute, undifferentiated febrile illness of <7 days duration were enrolled in the study and blood samples were obtained from each patient and assayed by virus isolation. Demographic and clinical information from each patient was also obtained at the time of voluntary enrollment. In 2005–2007, cases of Venezuelan equine encephalitis (VEE) were diagnosed for the first time in residents of Bolivia; the patients did not report traveling, suggesting endemic circulation of VEEV in Bolivia. In 2001 and 2003, VEE cases were also identified in Ecuador. Since 1993, VEEV has been continuously isolated from patients in Loreto, Peru, and more recently (2005), in Madre de Dios, Peru. We performed phylogenetic analyses with VEEV from Bolivia, Ecuador and Peru and compared their relationships to strains from other parts of South America. We found that VEEV subtype ID Panama/Peru genotype is the predominant one circulating in Peru. We also demonstrated that VEEV subtype ID strains circulating in Ecuador belong to the Colombia/Venezuela genotype and VEEV from Madre de Dios, Peru and Cochabamba, Bolivia belong to a new ID genotype. In summary, we identified a new major lineage of enzootic VEEV subtype ID, information that could aid in the understanding of the emergence and evolution of VEEV in South America.

The practitioners guide for dealing with the novel Influenza A, H1N1 pandemic

AbstractWithin three months of its discovery, the new Influenza A (Him) swineflu strain has spread to such an extent that a pandemic has been declared by the World Health Organization (WHO). Although most cases seem to be mild, cases of severe disease have also been reported and by 6 July 2009, 94 912 cases and 429 deaths were reported worldwide. At this point the WHO concluded that further spread within and to new countries is inevitable and sustained community transmission will make it impossible to confirm all cases by laboratory testing. In South Africa the 100 case mark was reached on 16 July 2009 and the laboratory testing strategy was modified. All cases of suspected swine flu will no longer be tested by the National Institute for Communicable Diseases (NICD) although continued monitoring of cases of severe or fatal respiratory illness will continue. This also places the responsibility on the health care provider to manage mild cases, treat moderate to severe cases and request confirmatory diagnost...