Evaluación preliminar del impacto en México de la influenza AH1N1: documento elaborado por el equipo conjunto CEPAL

Incluye Bibliografía

Caracterização dos genes codificadores da hemaglutinina e PB2 do vírus Influenza A (H1N1) pandêmico isolado na mesorregião metropolitana de Belém

A recente pandemia de gripe de 2009/2010 causada pelo vírus A (H1N1) pandêmico mostrou um perfil de gravidade diferente da gripe sazonal, pois um percentual considerável de casos graves e fatais ocorreu em indivíduos adultos jovens, sem comorbidade. A virulência dos vírus Influenza A (H1N1) pandêmico resulta de interações protéicas complexas e depende essencialmente de alguns genes virais. O objetivo deste estudo foi caracterizar os genes codificadores da hemaglutinina (H1) e polimerase básica 2 (PB2) do vírus Influenza A (H1N1) pandêmico mediante a obtenção de cepas provenientes de pacientes com gripe procedente da mesorregião metropolitana de Belém-PA. O tamanho amostral foi constituído de 87 amostras aleatórias de ambos os sexos de 0 a 96 anos, com síndrome respiratória aguda grave (SRAG) sem nenhuma comorbidade relatada, no período de maio de 2009 a agosto de 2010. As amostras foram isoladas em cultura de célula MDCK e analisadas por técnicas de biologia molecular que compreenderam três etapas principais: a) extração do RNA viral (RNAv) a partir do sobrenadante celular; b) amplificação do RNAv pela técnica de Reação em Cadeia mediada pela Polimerase precedida de Transcrição Reversa (RT-PCR); c) sequenciamento completo dos genes codificadores da H1 e PB2. Das 87 cepas amplificadas pelo RT-PCR, em 82 tornou-se possível a obtenção e análise de sequências para o gene HA, enquanto que de 81 amostras virais obteve-se sequências para o gene PB2. A análise comparativa das sequências obtidas com a sequência da cepa vacinal (A/California/07/2009(H1N1)) revelou substituições aminoacídicas na HA (P83S; D97N; S203T; D222G; Q293H e I321V) e na PB2 (K340N; K526R e M631L), no entanto sem associação a hospitalização. Ao nível de substituição na HA, a D97N isolada ou associada com a S203T, foi detectada com mais frequência na primeira onda. Já ao nível da PB2 a substituição K526R foi mais encontrada em cepas que circularam na primeira onda, enquanto que, a M631L foi mais evidenciada na segunda. A substituição D222G na HA só foi encontrada em casos de óbitos. Por fim, observou-se uma tendência de alterações nos sítios antigênicos da HA. Sendo assim, a contínua vigilância genética e antigênica do vírus Influenza A (H1N1) pdm em circulação, bem como o compartilhamento de informações é de extrema importância para a melhor recomendação possível para os vírus que entram na composição vacinal evitando assim maior risco de epidemias severas no futuro.

Perfil genotípico de resistência do vírus Influenza A (H1N1) pandêmico aos inibidores da neuraminidase em pacientes procedentes da mesorregião de Belém no período de maio de 2009 a maio de 2012

O vírus Influenza é o responsável pela gripe, uma doença que ocasiona milhões de mortes e hospitalizações todos os anos. Nas infecções severas, especialmente em pessoas com risco para complicações, os antivirais tornam-se os principais meios para o manejo clínico, merecendo especial destaque os inibidores da neuraminidase (INAs). De fato, na pandemia de 2009 a Organização Mundial da Saúde (OMS) recomendou o uso do oseltamivir para o tratamento dos doentes. Porém, devido à evolução genética viral, surgiram cepas com mutações no gene codificador da neuraminidase (NA) responsáveis por substituições aminoacídicas que levam à resistência aos fármacos INAs. Assim, a OMS passou a recomendar a vigilância de resistência genotípica para os vírus Influenza. Este trabalho teve como objetivos verificar a ocorrência de mutações no gene codificador da NA dos vírus Influenza A (H1N1) pandêmico que possam estar relacionadas à resistência aos INAs em cepas circulantes na mesorregião metropolitana de Belém no período de maio de 2009 a maio de 2012 e analisar, através da modelagem de proteínas, as substituições aminoacídicas da NA que possam estar influenciando na conformação protéica. Durante o período de estudo, foram recebidas no Laboratório de Vírus Respiratórios 2619 amostras clínicas de pacientes que apresentavam sinais e sintomas de infecção respiratória aguda com até cinco dias de evolução. Para a detecção do genoma viral foi feita a extração do RNA viral, seguida de RT-PCR em tempo real utilizando marcadores específicos para Influenza A H1N1pdm, resultando em 744 (28,4%) positivas. Parte das amostras positivas foram então inoculadas em células MDCK. Para as amostras isoladas em cultura de células, foi feita uma nova extração do RNA viral seguida de uma RT-PCR e semi-nested (PCR) utilizando iniciadores específicos para o gene NA, e posterior análise em sequenciador automático ABI Prism 3130xl (Applied Biosystems). A modelagem molecular da NA foi realizada através dos softwares SWISS-MODEL, MODELLER 9.10, PROCHECK, VERIFY3D e PYMOL. A análise parcial das sequências da neuraminidase nas amostras sequenciadas mostrou que não houve a circulação de cepas de vírus H1N1pdm com a mutação H275Y, a principal envolvida na resistência ao oseltamivir. Porém, em duas amostras foi identificada a substituição D199N que já foi relatada em vários estudos mostrando uma possível associação com o aumento da resistência ao oseltamivir. As amostras de 2012 apresentaram duas substituições (V241I e N369K) que estão relacionadas com um possível papel na compensação dos efeitos negativos causados pela mutação H275Y. A modelagem molecular mostrou que na mutação D199N houve uma alteração na estrutura da proteína NA próxima ao sítio de ligação ao antiviral. A análise filogenética revelou que as amostras de 2012 formaram um cluster isolado, demonstrando uma variação muito mais temporal do que geográfica. Este representa o primeiro estudo de resistência dos vírus Influenza H1N1pdm na mesorregião metropolitana de Belém, representando um importante instrumento para que os profissionais de saúde adotem estratégias mais eficazes no manejo da doença e no desenvolvimento de novos fármacos anti-influenza.

Immunogenicity and Reactogenicity of 2009 Influenza A (H1N1) Inactivated Monovalent Non-Adjuvanted Vaccine in Elderly and Immunocompromised Patients

Background Immunosuppressed individuals present serious morbidity and mortality from influenza, therefore it is important to understand the safety and immunogenicity of influenza vaccination among them. Methods This multicenter cohort study evaluated the immunogenicity and reactogenicity of an inactivated, monovalent, non-adjuvanted pandemic (H1N1) 2009 vaccine among the elderly, HIV-infected, rheumatoid arthritis (RA), cancer, kidney transplant, and juvenile idiopathic arthritis (JIA) patients. Participants were included during routine clinical visits, and vaccinated according to conventional influenza vaccination schedules. Antibody response was measured by the hemagglutination-inhibition assay, before and 21 days after vaccination. Results 319 patients with cancer, 260 with RA, 256 HIV-infected, 149 elderly individuals, 85 kidney transplant recipients, and 83 with JIA were included. The proportions of seroprotection, seroconversion, and the geometric mean titer ratios postvaccination were, respectively: 37.6%, 31.8%, and 3.2 among kidney transplant recipients, 61.5%, 53.1%, and 7.5 among RA patients, 63.1%, 55.7%, and 5.7 among the elderly, 59.0%, 54.7%, and 5.9 among HIV-infected patients, 52.4%, 49.2%, and 5.3 among cancer patients, 85.5%, 78.3%, and 16.5 among JIA patients. The vaccine was well tolerated, with no reported severe adverse events. Conclusions The vaccine was safe among all groups, with an acceptable immunogenicity among the elderly and JIA patients, however new vaccination strategies should be explored to improve the immune response of immunocompromised adult patients. (ClinicalTrials.gov, NCT01218685)

Influenza A(H1N1) infection and severe cardiac dysfunction in adults: A case series

BACKGROUND: While viral myocarditis and heart failure are recognized and feared complications of seasonal influenza A infection, only limited information is available for 2009 influenza A(H1N1)-induced heart failure. METHODS AND MAIN FINDINGS: This case series summarizes the disease course of four patients with 2009 influenza A(H1N1) infection who were treated at our institution from November 2009 until September 2010. All patients presented with severe cardiac dysfunction (acute heart failure, cardiogenic shock or cardiac arrest due to ventricular fibrillation) as the leading symptom of influenza A(H1N1) infection. Two patients most likely had pre-existent cardiac pathologies, and three required catecholamine therapy to maintain hemodynamic function. Except for one patient who died before influenza A(H1N1) infection had been diagnosed, all patients received antiviral therapy with oseltamivir and supportive critical care. Acute respiratory distress syndrome due to influenza A(H1N1) infection developed in one patient. Heart function normalized in two of the three surviving patients but remained impaired in the other one at hospital discharge. CONCLUSIONS: Influenza A(H1N1) infection may be associated with severe cardiac dysfunction which can even be the leading clinical symptom at presentation. During an influenza pandemic, a thorough history may reveal flu-like symptoms and should indicate testing for H1N1 infection also in critically ill patients with acute heart failure.

The development of poultry farms risk assessment tool for avian influenza in Imo State, Nigeria

Background: Nigeria was one of the 13 countries where avian influenza outbreak in poultry farms was reported during the 2006 avian influenza pandemic threat and was also the first country in Africa to report the presence of H5N1influenza among its poultry population. There are multiple hypotheses on how the avian influenza outbreak of 2006 was introduced to Nigeria, but the consensus is that once introduced, poultry farms and their workers were responsible for 70% of the spread of avian influenza virus to other poultry farms and the population. ^ The spread of avian influenza has been attributed to lack of compliance by poultry farms and their workers with poultry farm biosecurity measures. When poultry farms fail to adhere to biosecurity measures and there is an outbreak of infectious diseases like in 2006, epidemiological investigations usually assess poultry farm biosecurity—often with the aid of a questionnaire. Despite the importance of questionnaires in determining farm compliance with biosecurity measures, there have been few efforts to determine the validity of questionnaires designed to assess poultry farms risk factors. Hence, this study developed and validated a tool (questionnaire) that can be used for poultry farm risk stratification in Imo State, Nigeria. ^ Methods: Risk domains were generated using literature and recommendations from agricultural organizations and the Nigeria government for poultry farms. The risk domains were then used to develop a questionnaire. Both the risk domain and questionnaire were verified and modified by a group of five experts with a research interest in Nigeria's poultry industry and/or avian influenza prevention. Once a consensus was reached by the experts, the questionnaire was distributed to 30 selected poultry farms in Imo State, Nigeria that participated in this study. Survey responses were received for all the 30 poultry farms that were selected. The same poultry farms were visited one week after they completed the questionnaires for on-site observation. Agreement among survey and observation results were analyzed using a kappa test and rated as poor, fair, moderate, substantial, or nearly perfect; and internal consistency of the survey was also computed. ^ Result: Out of the 43 items on the questionnaire, 32 items were validated by this study. The agreement between the survey result and onsite observation was analyzed using kappa test and ranged from poor to nearly perfect. Most poultry farms had their best agreements in the contact section of the survey. The least agreement was noted in the farm management section of the survey. Thirty-two questions on the survey had a coefficient alpha > 0.70, which is a robust internal consistency for the survey. ^ Conclusion: This study developed 14 risk domains for poultry farms in Nigeria and validated 32 items from the original questionnaire that contained 43 items. The validated items can be used to determine the risk of introduction and spread of avian influenza virus in poultry farms in Imo State, Nigeria. After further validations in other states, regions and poultry farm sectors in Nigeria; this risk assessment tool can then be used to determine the risk profile of poultry farms across Nigeria.^

Influenza A virus ribonucleoproteins modulate host recycling by competing with Rab11 effectors

Influenza A virus assembly is an unclear process, whereby individual virion components form an infectious particle. The segmented nature of the influenza A genome imposes a problem to assembly because it requires packaging of eight distinct RNA particles (vRNPs). It also allows genome mixing from distinct parental strains, events associated with influenza pandemic outbreaks. It is important to public health to understand how segmented genomes assemble, a process that is dependent on the transport of components to assembly sites. Previously, it has been shown that vRNPs are carried by recycling endosome vesicles, resulting in a change of Rab11 distribution. Here, we describe that vRNP binding to recycling endosomes impairs recycling endosome function, by competing for Rab11 binding with family-interacting proteins, and that there is a causal relationship between Rab11 ability to recruit family-interacting proteins and Rab11 redistribution. This competition reduces recycling sorting at an unclear step, resulting in clustering of single- and double-membraned vesicles. These morphological changes in Rab11 membranes are indicative of alterations in protein and lipid homeostasis during infection. Vesicular clustering creates hotspots of the vRNPs that need to interact to form an infectious particle.

Monitoring influenza vaccine effectiveness using the national influenza surveillance system

Background: Flu vaccine composition is reformulated on a yearly basis. As such, the vaccine effectiveness (VE) from previous seasons cannot be considered for subsequent years, and it is necessary to monitor the VE for each season. This study (MonitorEVA- monitoring vaccine effectiveness) intends to evaluate the feasibility of using the national influenza surveillance system (NISS) for monitoring the influenza VE. Material and methods: Data was collected within NISS during 2004 to 2014 seasons. We used a case-control design where laboratory confirmed incident influenza like illness (ILI) patients (cases) were compared to controls (ILI influenza negative). Eligible individuals consisted on all aged individuals that consult a general practitioner or emergency room with ILI symptoms with a swab collected within seven days of symptoms onset. VE was estimated as 1- odds ratio of being vaccinated in cases versus controls adjusted for age and month of onset by logistic regression. Sensitivity analyses were conducted to test possible effect of assumptions on vaccination status, ILI definition and timing of swabs (<3 days after onset). Results: During the 2004-2014 period, a total of 5302 ILI patients were collected but 798 ILI were excluded for not complying with inclusion criteria. After data restriction the sample size in both groups was higher than 148 individuals/ season; minimum sample size needed to detect a VE of at least 50% considering a level of significance of 5% and 80% power. Crude VE point estimates were under 45% in 2004/05, 2005/06, 2011/12 and 2013/14 season; between 50%-70% in 2006/07, 2008/09 and 2010/11 seasons, and above 70% in 2007/08 and 2012/13 season. From season 2006/07 to 2013/14, all crude VE estimates were statistically significant. After adjustment for age group and month of onset, the VE point estimates decreased and only 2008/09, 2012/13 and 2013/14 seasons were significant. Discussion and Conclusions: MonitorEVA was able to provide VE estimates for all seasons, including the pandemic, indicating if the VE was higher than 70% and less than 50%. When comparing with other observational studies, MonitorEVA estimates were comparable but less precise and VE estimates were in accordance with the antigenic match of the circulating virus/ vaccine strains. Given the sensitivity results, we propose a MonitorEVA based on: a) Vaccination status defined independently of number of days between vaccination and symptoms onset; b) use of all ILI data independent of the definition; c) stratification of VE according to time between onset and swab (< 3 and ≥3 days).

Augmentation de l'immunogénicité de la nucléoprotéine de l'influenza par multimérisation in vitro

Les travaux effectués au cours de ce mémoire ont permis de développer une alternative aux vaccins présentement utilisés contre le virus de l’influenza. Nous avons utilisé la nucléoprotéine (NP) de l’influenza comme base vaccinale puisque cette protéine est conservée chez les souches d’influenza A et qu’elle possède un potentiel de protection croisée. Nous avons montré que la multimérisation de la NP grâce à un gabarit d’ARN permet d’augmenter son immunogenicité. Cette multimérisation en pseudo-nucléoparticule virale (NLP) a augmenté la réponse humorale et cellulaire spécifique à NP et l’ajout d’un adjuvant (PAL) a permis d’amplifier davantage la réponse humorale contre NP. Une dose du vaccin candidat NLP-PAL n’a pas réussi à protéger des souris contre une infection létale avec une souche homotypique d’influenza. Cependant, des résultats avec un régime de deux immunisations montrent des résultats encourageants qui permettent d’espérer une protection envers une infection virale.

Cochrane review summary for cancer nursing : influenza vaccination in children having chemotherapy for cancer

Background: Children having chemotherapy for cancer are prone to developing influenza infections. Influenza virus infection may lead to hospitalization/prolonged hospitalization, interruption of treatment, and other severe adverse outcomes such as death. Although clinical guidelines recommend children who are being treated for cancer be vaccinated against influenza, evidence supporting this recommendation is unclear.--------- Objectives: The objectives of this review were to (1) assess the efficacy of influenza vaccination in stimulating immunologic response in children with cancer receiving chemotherapy, compared with other control groups; (2) assess the efficacy of influenza vaccination in preventing influenza infection; and (3) establish any adverse effects associated with influenza vaccines in children with cancer.

An examination of the predictors of blood donors' intentions to donate during two phases of an avian influenza outbreak

BACKGROUND: Data from prior health scares suggest that an avian influenza outbreak will impact on people’s intention to donate blood; however research exploring this is scarce. Using an augmented theory of planned behavior (TPB), incorporating threat perceptions alongside the rational decision-making components of the TPB, the current study sought to identify predictors of blood donors’ intentions to donate during two phases of an avian influenza outbreak. STUDY DESIGN AND METHODS: Blood donors (N = 172) completed an on-line survey assessing the standard TPB predictors as well as measures of threat perceptions from the health belief model (HBM; i.e., perceived susceptibility and severity). Path analyses examined the utility of the augmented TPB to predict donors’ intentions to donate during a low- and high-risk phase of an avian influenza outbreak. RESULTS: In both phases, the model provided a good fit to the data explaining 69% (low risk) and 72% (high risk) of the variance in intentions. Attitude, subjective norm, and perceived susceptibility significantly predicted donor intentions in both phases. Within the low-risk phase, gender was an additional significant predictor of intention, while in the high-risk phase, perceived behavioral control was significantly related to intentions. CONCLUSION: An augmented TPB model can be used to predict donors’ intentions to donate blood in a low-risk and a high-risk phase of an outbreak of avian influenza. As such, the results provide important insights into donors’ decision-making that can be used by blood agencies to maintain the blood supply in the context of an avian influenza outbreak.

Detection of bacteriophage and respiratory viruses in droplets

Influenza is a widespread disease occurring in seasonal epidemics, and each year is responsible for up to 500,000 deaths worldwide. Influenza can develop into strains which cause severe symptoms and high mortality rates, and could potentially reach pandemic status if the virus’ properties allow easy transmission. Influenza is transmissible via contact with the virus, either directly (infected people) or indirectly (contaminated objects); via reception of large droplets over short distances (one metre or less); or through inhalation of aerosols containing the virus expelled by infected individuals during respiratory activities, that can remain suspended in the air and travel distances of more than one metre (the aerosol route). Aerosol transmission of viruses involves three stages: production of the droplets containing viruses; transport of the droplets and ability of a virus to remain intact and infectious; and reception of the droplets (via inhalation). Our understanding of the transmission of influenza viruses via the aerosol route is poor, and thus our ability to prevent a widespread outbreak is limited. This study explored the fate of viruses in droplets by investigating the effects of some physical factors on the recovery of both a bacteriophage model and influenza virus. Experiments simulating respiratory droplets were carried out using different types of droplets, generated from a commonly used water-like matrix, and also from an ‘artificial mucous’ matrix which was used to more closely resemble respiratory fluids. To detect viruses in droplets, we used the traditional plaque assay techniques, and also a sensitive, quantitative PCR assay specifically developed for this study. Our results showed that the artificial mucous suspension enhanced the recovery of infectious bacteriophage. We were able to report detection limits of infectious bacteriophage (no bacteriophage was detected by the plaque assay when aerosolised from a suspension of 103 PFU/mL, for three of the four droplet types tested), and that bacteriophage could remain infectious in suspended droplets for up to 20 minutes. We also showed that the nested real-time PCR assay was able to detect the presence of bacteriophage RNA where the plaque assay could not detect any intact particles. Finally, when applying knowledge from the bacteriophage experiments, we reported the quantitative recoveries of influenza viruses in droplets, which were more consistent and stable than we had anticipated. Influenza viruses can be detected up to 20 minutes (after aerosolisation) in suspended aerosols and possibly beyond. It also was detectable from nebulising suspensions with relatively low concentrations of viruses.

Epidemiologic modeling with FluSurge for pandemic (H1N1) 2009 outbreak, Queensland, Australia

At the beginning of the pandemic (H1N1) 2009 outbreak, we estimated the potential surge in demand for hospital-based services in 4 Health Service Districts of Queensland, Australia, using the FluSurge model. Modifications to the model were made on the basis of emergent evidence and results provided to local hospitals to inform resource planning for the forthcoming pandemic. To evaluate the fit of the model, a comparison between the model's predictions and actual hospitalizations was made. In early 2010, a Web-based survey was undertaken to evaluate the model's usefulness. Predictions based on modified assumptions arising from the new pandemic gained better fit than results from the default model. The survey identified that the modeling support was helpful and useful to service planning for local hospitals. Our research illustrates an integrated framework involving post hoc comparison and evaluation for implementing epidemiologic modeling in response to a public health emergency.

Documentary as history, memory, dream and film

A century ago, as the Western world embarked on a period of traumatic change, the visual realism of photography and documentary film brought print and radio news to life. The vision that these new mediums threw into stark relief was one of intense social and political upheaval: the birth of modernity fired and tempered in the crucible of the Great War. As millions died in this fiery chamber and the influenza pandemic that followed, lines of empires staggered to their fall, and new geo-political boundaries were scored in the raw, red flesh of Europe. The decade of 1910 to 1919 also heralded a prolific period of artistic experimentation. It marked the beginning of the social and artistic age of modernity and, with it, the nascent beginnings of a new art form: film. We still live in the shadow of this violent, traumatic and fertile age; haunted by the ghosts of Flanders and Gallipoli and its ripples of innovation and creativity. Something happened here, but to understand how and why is not easy; for the documentary images we carry with us in our collective cultural memory have become what Baudrillard refers to as simulacra. Detached from their referents, they have become referents themselves, to underscore other, grand narratives in television and Hollywood films. The personal histories of the individuals they represent so graphically–and their hope, love and loss–are folded into a national story that serves, like war memorials and national holidays, to buttress social myths and values. And, as filmic images cross-pollinate, with each iteration offering a new catharsis, events that must have been terrifying or wondrous are abstracted. In this paper we first discuss this transformation through reference to theories of documentary and memory–this will form a conceptual framework for a subsequent discussion of the short film Anmer. Produced by the first author in 2010, Anmer is a visual essay on documentary, simulacra and the symbolic narratives of history. Its form, structure and aesthetic speak of the confluence of documentary, history, memory and dream. Located in the first decade of the twentieth century, its non-linear narratives of personal tragedy and poetic dreamscapes are an evocative reminder of the distance between intimate experience, grand narratives, and the mythologies of popular films. This transformation of documentary sources not only played out in the processes of the film’s production, but also came to form its theme.