Dengue virus (DENV), which consists of four serotypes (DENV1-4), infects over 400 million people annually. Previous studies have indicated most human monoclonal antibodies (HMAbs) from dengue patients are cross-reactive and poorly neutralizing. Rare neutralizing HMAbs are usually serotype-specific and bind to quaternary structure-dependent epitopes. We determined the structure of DENV1 complexed with Fab fragments of a highly potent HMAb 1F4 to 6 Å resolution by cryo-EM. Although HMAb 1F4 appeared to bind to virus and not E proteins in ELISAs in the previous study, our structure showed that the epitope is located within an envelope (E) protein monomer, and not across neighboring E proteins. The Fab molecules bind to domain I (DI), and DI-DII hinge of the E protein. We also showed that HMAb 1F4 can neutralize DENV at different stages of viral entry in a cell type and receptor dependent manner. The structure reveals the mechanism by which this potent and specific antibody blocks viral infection.Subject Categories Microbiology, Virology & Host Pathogen Interaction; Immunology Flaviviridae, is transmitted to humans by Aedes mosquitoes. Other flaviviruses that are important human pathogens include West Nile virus (WNV), yellow fever virus, Japanese encephalitis virus, and tick-borne encephalitis virus. DENV targets susceptible populations residing in tropical and sub-tropical regions of the globe. An estimated 400 million people worldwide are infected with DENV annually, leading to approximately 100 million cases of dengue and 21 000 deaths , a member of the family et al, et al, DENV consists of four distinct serotypes (DENV1–4), and the amino acid sequence variation of the polyprotein between serotypes is about 25–40%  and envelope (E) proteins  showed that the most potent neutralizing Abs are serotype-specific and bind to DIII  compared with full-length IgG  demonstrated that it is a highly potent Ab with a NeutIn mice that had been given 20 μg of HMAb 1F4 prior to infection with a sub-lethal dose of DENV1, a significant reduction in the viral genomic RNA copy number as compared to the isotype control was observed in serum and bone marrow  as when added after attachment to cells (FRNT50 of 0.37 μg/ml)   . A micrograph of an untreated control DENV1 (PVP159) , which was grown at 28°C in C6/36 cells and kept at 4°C, showed that the sample consisted of mainly smooth mature virus particles, with about 15% spiky immature particles and a few broken particles. DENV2 particles have been shown to exhibit a change from a smooth to a bumpy surface when incubated at 37°C for 30 min   , Eo strand (136–138), part of the EoFo loop, Fo strand (155–165), Go strand, the following GoHo loop (170–177), and kl loop (272–276)  in an asymmetric unit Fig A and B. Doa loop 6–52, Eo Doa loop 6–52, Eo Fab 1F4 is unable to bind to the E proteins near the 3-fold vertices (molecule C) Fig A. Comparin vitro . Similar interactions between Fab and amino residues on β-strands Fo and Go and the loop FoGo were also found in the complex structure of Fab 14c10  of DC-SIGN complexed with DENV, showed that one CRD binds across two glycosylation sites at position 67 from two neighboring E proteins  was obtained from the EDEN patient cohort  was administered intravenously (i.v.) in a total volume of 100 μl. Three days post-infection, mice were sacrificed. Serum was obtained from whole blood by centrifugation and stored at −80°C. Bone marrow cells were processed as previously described . AG129 mice , as per the manufacturer's instructions. Similarly, RNA was extracted from the 1 ml bone marrow collection using an RNeasy Mini kit (Qiagen). The ABI Prism Sequence Detection System 7300 was used to perform quantitative RT-PCR (qRT-PCR). Viral load was determined using previously published primers and probe sequences , resulting in a pixel size of 1.81 Å per pixel, at an underfocus range of 1–4 μm. Images showing drift and strong astigmatism were discarded, and 752 images were selected for further processing.DENV1 was mixed with Fab 1F4 in a molar ratio of one Fab molecule to every E protein and then incubated at one of two different temperatures (4 or 37°C), for 30 min followed by another 2 h at 4°C. The corresponding controls (DENV1 without Ab at 4 or 37°C) were also included. A 2.5 μl sample was pipetted onto ultra-thin carbon-coated lacey carbon grids (Ted Pella), and then blotted with filter paper for 2 s and flash-frozen in liquid ethane using the FEI Vitrobot Mark IV plunger. The frozen grid was kept at liquid nitrogen temperature. The virus particles were imaged using a 300-kV FEI Titan Krios electron microscope at a nominal magnification of 47 000 with an electron dose of 17.5 e/Ået al, et al, et al, Reconstruction of the DENV1-Fab complex particles was initiated by manually picking the spiky-looking unbroken particles using the program e2boxer. A total of 10 270 particles were selected and the contrast transfer function parameters were determined by using e2ctf. Following the class averaging, an initial model was generated based on a set of class-averages using e2initialmodel. The programs e2boxer, e2ctf and e2initialmodel are available in EMAN2 software package . As for the Fab molecules, a homology model was built for Fab 1F4 by using Swiss-model server (Arnold http://helixweb.nih.gov/structbio/basic.html) for calculating the contact area between two molecules, Multalin  and a two-tailed Wilcoxon rank-sum test. Analysis of attachment assays were conducted using a 2-way repeated measures analysis of variance (RM ANOVA).Dengue is a tropical disease that affects hundreds of millions of people annually and poses an economic burden in many tropical countries. There are four dengue serotypes (DENV1-4). Infection with virus from one serotype in a primary infection may potentially prime the individual to develop a more severe disease when infected with virus from another DENV serotype in a secondary infection. This is likely caused in part by antibody-dependent enhancement of infection of virus into monocyte/macrophage cells via Fcγ receptors, facilitated by the binding of non-neutralizing antibodies to virus. This phenomenon complicates the development of vaccines and therapeutics. One strategy to produce a safe dengue antibody therapeutic is to identify four potently neutralizing antibodies, each specific to a DENV serotype, and to then combine them into a cocktail. We therefore conducted structural studies to understand how a potent human antibody binds and neutralizes DENV1.in vitro and in an AG129 mouse model. We determined the structure of DENV1 complexed with Fab 1F4 to a resolution of 6 Å by using cryo-electron microscopy (cryo-EM). The structure showed that the antibody binds to domain (D) I, and the DI-DII hinge region on an envelope protein monomer. Previous studies on HMAb 1F4 had demonstrated that it only binds to intact virus and not to recombinant envelope (rE) protein. Comparison of cryo-EM structures of virus E proteins to rE crystal structures showed that the E proteins on the virus had a conserved DI-DII hinge angle, whereas the hinge angle on the rE proteins is highly variable. As the DI-DII hinge forms part of the HMAb 1F4 epitope, we propose that HMAb 1F4 may be very sensitive to the conformation of this region. We also determined the mechanisms of neutralization of HMAb 1F4 in different cell lines. In Vero cells, the antibody prevents virus infection at a post-attachment step, whereas in DC-SIGN-expressing U937 cells, the HMAb can also prevent virus attachment. Using the cryoEM structure of 1F4 complexed with DENV, we discuss how the antibody could neutralize these steps of the virus infection.Human monoclonal antibody (HMAb) 1F4 was shown to be highly neutralizing We have identified and characterized an antibody that could potentially be used as a DENV1 therapeutic. The results also contribute significantly to vaccine design. Firstly, by comparing the HMAb 1F4-DENV1 structure to another potent HMAb 14c10-DENV1 structure, we observed an overlap at the DI-DII hinge, suggesting that this region is likely to be one of the principal determinants in eliciting type-specific neutralizing antibodies in humans. The inclusion of this region is thus important for the development of an effective vaccine. In addition, binding of HMAb 1F4 to E protein monomer is likely to be very sensitive to the hinge conformation. This conformation is held in proper configuration on the virus by the interactions between E ectodomains and also the E ectodomains with the E and M stem region. This has important implications for the development of an effective rE protein-based vaccine.


The continuous growth of malware presents a problem for internet computing due to increasingly sophisticated techniques for disguising malicious code through mutation and the time required to identify signatures for use by antiviral software systems (AVS). Malware modelling has focused primarily on semantics due to the intended actions and behaviours of viral and worm code. The aim of this paper is to evaluate a static structure approach to malware modelling using the growing malware signature databases now available. We show that, if malware signatures are represented as artificial protein sequences, it is possible to apply standard sequence alignment techniques in bioinformatics to improve accuracy of distinguishing between worm and virus signatures. Moreover, aligned signature sequences can be mined through traditional data mining techniques to extract metasignatures that help to distinguish between viral and worm signatures. All bioinformatics and data mining analysis were performed on publicly available tools and Weka.  If users do not have confidence that their machines will not be attacked when connected to the internet, major areas of computing will be constrained due to fear of denial of service and massive data fraud . SymanteResearch has continued in static structure checking algorithms –24 despiAntiviral signatures can be calculated from a pattern of operations in the malware code or can represent the encryption algorithm used to hide the virus or worm. Signatures were originally and continue to be identified and calculated by human experts and are typically a sequence of hexadecimal numbers intended to uniquely identify viruses and worms. Automatic generation of signatures for new malware continues to be a difficult problem . Such siAVS scanners use a dictionary or library of signatures in a variety of different ways. For instance, for simple polymorphic malware detection, the hexadecimal representation of a signature can be used to match against incoming network packets containing bytes also represented in hexadecimal. This allows the AVS to check for contiguous similarities between parts of the signature and packet contents. For metamorphic and more complex polymorphic malware detection, increasingly sophisticated techniques must be used that allow for contiguous parts of the signature to be detected noncontiguously across different packets . SignatuSequence analysis is used in biology to understand the relationship between two or more sequences  of genetic information, such as DNA or amino acids. There are databases of genetic information which are processed by string alignment algorithms to better understand the relationship between species and also determine the location of specific genes. In particular, sequence analysis and alignment can be used to identify conserved regions or motifs (regions of similarity) in biological data that identify common genes and shared ancestry as well as common structure and function of amino acid sequences . One advSequence alignment techniques are not confined to biological sequences, however, and there have been applications of sequence alignment in linguistics  and markSubsequent work  reportedThe aim of this paper is to significantly extend the work started in  and to eThere are several tools for alignment and many algorithms used in the study of biosequence analysis. In general, an alignment is an adjustment of a sequence in relation to other sequences. The aim is to arrange two  or more  possibly variable length sequences of DNA or protein in such a way that regions of similarity across sequences (rows of a matrix) fall in the same successive columns of the matrix, where such similarity signifies functional, structural, or evolutionary commonality. Global alignment tries to align every item in every sequence and tends to work best when the sequences are of roughly similar length, such as the Needleman-Wunsch technique . Local avirus.1C.Tanga.a computer virus signature has the hexadecimal coding 8e5ef1aec91259d70c5e62 and the worm Bat.Agent.bo, the hexadecimal coding fb56373bde3881741. The hexadecimal code for 60 viruses belonging to 12 families and 60 worms belonging to 13 families were downloaded from VX Heavens ..[61][b2]b..2....2..” “..1..b3..4..0..1..c..1..”; Worm signature if “[2a]..3..2....6..a..88..1..” “..0..2..8[30]e..7..0..f..[6e]f[0a][3c]..” Virus signature if “R2:[67]..[25][5c]..f..2..1....6..1..” “..e..4..9[ad]e..8..b..” Virus signature if “..[32]..8..[b3]..8..” Worm signature if “R3:..11..4..”; Worm signature if “[90]..0..a..1..[60]....1..7..” Virus signature if “R4: Virus signature if “..2..1..2..”R5:[6e]8c5..757...”; Worm signature if “..3..” Virus signature if “.Converting these metasignatures back into hexadecimal patterns produces :[6e]8c5..757...”) in particular contains a number of contiguous hexadecimal characters (no gaps) that could be useful for future AVS to help distinguish viral malware from nonmalware. Across the three machine learning algorithms, R5 was best for accuracy and specificity , and R3 for sensitivity (0.978). When R1 was used with 60 virus and 60 worm signatures, the metasignatures “..1..b3..4..0..1..1(/c)..1..” for virus and “..0..2..83(/0)e.. 7..0..f..6(/c)fa(/0)3(/c)..” were reported . The resThe results indicate that aligning computer virus and worm signatures using multiple alignment techniques leads to improved classification accuracy using the techniques described in this paper. While the differences in representation are reflected to some extent in classification accuracy after alignment, there is a difference when PRISM is used, with R1 producing more informative metasignatures for both virus and worm. The method of converting malware hexadecimal signatures to residue representation has been clearly demonstrated to affect learning and the motifs extracted. More work is required to determine the tradeoff between representations and richness or usefulness of motifs extracted. Converting the hexadecimal signatures of viruses and worms to amino acids and then rational numbers between 0 to 1 has also been shown to be effective for perceptron learning. Naive Bayes for separating worm from virus signatures after alignment has also been shown to be the most accurate. However, extracting the knowledge contained in Naive Bayes is not easy and symbolic rule extraction techniques are to be preferred when trying to generate malware signatures for scanning files and network packets directly. The derivation of metasignatures provides a new way to look at viral and worm signatures at a “motif” level. These motifs represent common subpatterns among the signatures after initial alignment of virus and worm signatures separately  and then together . The machine learning task was to separate worm signatures from virus signatures using their hexadecimal form and different amino acid representations, rather than distinguish malware from nonmalware. To test for malware versus nonmalware classification will require mapping the metasignatures back to malware op code, and this is not possible for the signatures used in this research. Nevertheless, we have shown how the growing databanks of malware signatures can be mined for interesting signature information, even if the relationship back to op code is lost or not available. More work is required, however, to identify the most effective alignment algorithms, substitution matrices, and representations for rich and informative metasignatures extraction.


Bolomys lasiurus and Oligoryzomys nigripes are rodent reservoirs of Araraquara-like and Juquitiba-like hantaviruses, which cause HPS in Brazil. Bolomys lasiurus (hairy-tailed Bolo mouse) and the Juquitiba-like virus is associated with Oligoryzomys nigripes (black-footed pigmy rice rat).We describe the genetic analysis of samples from hantavirus pulmonary syndrome (HPS) patients from southern and southeastern states of Brazil and rodents captured at the presumed site of infection of these patients. A total of 65 samples that were antibody-positive for Sin Nombre or Laguna Negra virus by enzyme-linked immunosorbent assay were processed by nested reverse transcription–polymerase chain reaction (RT-PCR) by using several primer combinations in the M and S genome segments. PCR products were amplified and sequenced from samples from 11 HPS patient and 7 rodent samples. Phylogenetic analysis of nucleotide sequence differences showed the cocirculation of Araraquara and Juquitiba-like viruses, previously characterized from humans. Our genetic data indicate that Araraquara virus is associated with  Bunyaviridae ; 10 samples from São Paulo ; 7 samples from Paraná ; 7 samples from Santa Catarina ; 10 samples from Rio Grande do Sul . Of seven samples from HPS patients from Santa Catarina, five samples were from a family cluster reported by Seara  set in rural or sylvan environments, around the presumed sites of HPS infection. The rodents were processed in the field; biologic samples  were obtained according to established biosafety guidelines (Sigmodontinae) were studied by nested RT-PCR: 3 samples were from Uberlândia (2 Bolomys lasiurus) and Uberaba (1 B. lasiurus), in Minas Gerais; 13 samples were from Araraquara (1 B. lasiurus), Batatais (2 B. lasiurus), Franca , Cassia dos Coqueiros (1 Oximycterus rutilans), Cravinhos (1 B. lasiurus), Fartura (1 Akodon sp.), Mariápolis (2 B. lasiurus), Nuporanga (1 B. lasiurus and 1 Oligoryzomys nigripes), and São Carlos (1 B. lasiurus), in São Paulo; 3 samples from General Carneiro (2 O. nigripes and 1 Akodon sp.) in Paraná; 3 samples from Seara (2 O. nigripes and 1 Bolomys sp.) in Santa Catarina; 3 samples from Marcelino Ramos (1 O. nigripes and 1 Akodon sp.) and São Lourenço do Sul (1 Akodon sp.), in Rio Grande do Sul.A total of 25 rodent samples  Kit  as described elsewhere  or GFXTM PCR DNA and Gel Band Purification Kit . The nucleotide sequence of these products was determined on an ABI PRISM 377 Genetic Analyzer  using the dydeoxy cycle sequencing technique (Bolomys lasiurus) from the state of São Paulo, and 1 HPS case-patient and one rodent (B. lasiurus) from Minas Gerais. Pairwise comparisons of the sequences of ARA virus strains from HPS patients and B. lasiurus showed an 85.1%-99.7% nt and 95%-99% amino acid (aa) identity. The viral sequence from the HPS patient Hu237251 from Patrocínio, Minas Gerais, the northernmost location included in this study, was most divergent from the other members of this group (16.7%). The second hantavirus genetic lineage identified was closely related to JUQ virus. Samples from seven HPS patients and three Oligoryzomys nigripes from the states of Rio Grande do Sul, Santa Catarina, Paraná, and São Paulo fall into this group (PCR products of the expected size were amplified from 11 of 40 HPS patient samples  and 7 ofis group .O. nigripes sequences (distance 0.273). The distance observed between these two subclades is intermediate between in-group and between group distances , showed 85.7%-97.7% nt identity . The maxistances .O. nigripes samples of this group showed 86.6%-99.7 % nt identity. Sequence comparison with the corresponding to ARA group showed 20.8% nt and 4.9% amino acid (aa) divergence. Genetic distances between the sequences studied are shown in Comparison of the 303-nt G2-encoding region sequences derived from HPS-patient and B. lasiurus sequences with ARA virus showed a 96.8%-nt and 99.3%-aa identity for the 1,239-nt G2-M piece, and 98.8% nt and 100 % aa identity for the S segment piece. When RNA viral sequences from B. lasiurus were compared with those from O. nigripes, 77.8% nt and 93.4% aa identity for the 1,239-nt G2-M piece, and 84.1% nt and 98.8% aa identity for the S segment piece were observed.A longer fragment of 1,239 nt of the G2 encoding region of the M segment , as wellB. lasiurus fell into the ARA virus group, whereas all samples from O. nigripes fell into the JUQ-like genetic group. The nodes separating these groups had high bootstraps values ; however, the exact branching order among the Brazilian and the other hantaviruses cannot be resolved by the present phylogenetic analysis. The phylogenetic tree, based on the analysis of the 1,239-nt M segment sequence differences, showed the same topology of the 303-nt sequence, data-based tree. Both maximum parsimony  from Akodon azarae, although with a low bootstrap support (66%). The JUQ-like virus from O. nigripes 193576 is ambiguously placed within the South American viruses, together with CAS virus from Brazil and the rest of the Oligoryzomys-borne Argentinean hantaviruses , as well as LN virus from Paraguay. A slight difference in the topology of this 1,239-nt database tree was observed when neighbor-joining analysis was used; the grouping of this Oligoryzomys-borne Argentinean hantavirus clustered in a subclade with a bootstrap support of 79% (data not shown).Within this clade the ARA virus forms a subclade with other two akodontine-borne Argentinean hantaviruses, Maciel (MAC) from ,,Eleven (27.5%) of 40 human samples and 7 (28.0%) of 25 rodent samples studied tested positive for hantavirus by RT-PCR. Previous studies have characterized three different hantavirus genetic lineages associated with HPS cases in Brazil: CAS, ARA, and JUQ . The two subclades observed in The data we report on the phylogenetic analysis of viral M and S genome segment fragments, from HPS patients and rodent samples from different locations of southern and southeastern Brazil, showed the circulation of two distinct hantaviruses, closely related to ARA and JUQ viruses, previously characterized only from humans. Our data on the phylogenetic analysis of a 303-nt G2 encoding region of the M genome segment represents the first genetic evidence of the role of B. lasiurus) and HPS patient samples showed an identity of up to 99.7% at the nucleotide level, while within the JUQ-like virus group, the comparison between O. nigripes and HPS patient virus strains showed an identity of up to 97.7%. However, the phylogenetic relationship of JUQ-like hantavirus to other members of the South American hantavirus lineages, determined on the nucleotide sequences of the 303-nt sequence, as well as on a longer fragment of a 1,239-nt piece of the M segment performed for one representative strain derived from one O. nigripes, systematically failed to resolve the branching order.Comparison of the sequences of ARA virus strains obtained from rodents  and sugar cane cultures. These rodents are aggressive and usually dominate the areas they infest  and showed the highest prevalence of antibodies to hantavirus (11%). Akodon spp. and O. nigripes were the two most abundant among those rodents captured in the transition area between cerrado and the Atlantic rainforest, but the highest prevalence of antibodies to hantavirus was found in O. nigripes specimens (8%) (B. lasiurus and O. nigripes in the transmission of ARA and JUQ-like viruses, respectively, to humans.Among the rodents captured in the cerrado,


There are few health promotion tools for urban Inuit, and there is a specific dearth of evaluations on such tools.The current study used a community-specific approach in the evaluation of a health promotion tool, based on an urban Inuit community's preferences of health knowledge sources and distribution strategies. In partnership with the Tungasuvvingat Inuit Family Health Team in Ottawa, a CD-Rom was developed featuring an Inuk Elder presenting prenatal health messages in both Inuktitut and English. Also, relevant evaluation materials were developed.Using a mixed methods approach, 40 participants completed interviews prior to viewing the CD-Rom and participated in a focus group at follow-up. Questionnaires were also completed pre- and post-viewing to assess changes between expectations and reactions in order to document acceptability.Significant increases were found on satisfaction, acceptability of medium and relevance of content ratings. Qualitative findings also included (a) interest, uncertainty and conditional interest prior to viewing; and (b) positive evaluations of the CD-Rom.This suggests that CD-Rom technology has the potential for health promotion for urban Inuit, and the community-specific evaluation approach yielded useful information. Health promotion programmes represent a promising approach to improving the health of Inuit, as messages would be spread effectively among community members, given the strong kinship ties in Inuit communities . AlthougThis project began with a detailed consultation process with the Inuit community of Ottawa focused on health information sources and distribution strategies . ThroughAnother feature identified was the importance of Elders as sources and distributors of health information. It has been found that Elders could play an important role in public education about teenage pregnancy . In InuiIn collaboration with Tungasuvvingat Inuit Family Health Team (TIFHT), a CD-Rom was developed showing 2 messages about pregnancy and family health presented by an Inuk Elder in Inuktitut. The use of oral and visual media in the CD-Rom represented a match to Inuit learning styles  and prevThe goal of the present study was to evaluate the acceptability of this guided CD-Rom as a health promotion tool for urban Inuit in Ottawa. Together, the selected medium, source and content were expected to make the CD-Rom an acceptable health promotion resource as it was judged to best represent the articulated patterns and preferences of the community. Given such a match, the acceptability of the tool would be evidenced if after viewing the CD-Rom individuals would (a) share the information with others; (b) consider the medium acceptable; (c) evaluate the source positively; and (d) consider the content helpful.This project followed a community-based participatory approach to research and the research team comprised both academic and community researchers. In brief, the project arouse out of long-standing partnership between the community partner and the second author, initially, and the later inclusion of the first author as a student researcher. For this project, the academic researchers and community researchers undertook all aspects of this research together in a collaborative manner and jointly shared decision-making. A more detailed description of the development of the partnership with TIFHT, the participating community, appears elsewhere ,8,9. TheIndividuals were recruited by staff from the Tungasuvvingat Inuit Family Resource Centre (TIFRC) and for the TIFHT in Ottawa, Canada. The Ottawa Inuit community is the largest Inuit community outside of Nunavut, with a population estimate of 725 Inuit residing in the Ottawa-Gatineau area  at the tThe community research team members, who were Inuit community members and front-line health workers, initially identified potential participants. Identified individuals were provided with a description of the study and were then invited to participate if they were interested . Forty adults were recruited to participate in the study . This sample included 9 males and 31 females. One participant did not complete the initial interview and another participant did not complete the initial questionnaire. Four participants did not complete the post-CD-Rom evaluation. Three of the 4 were no longer interested in participating in the study, and 1 returned to live in the north.Planning for a Healthy Family, contained 2 segments, each with a teaching and text, and a set of summary slides containing major themes and illustrations (see The CD-Rom was developed in consultation with 2 Elders (from the north) and the academic and community research team. Video segments were selected from a series of brief teachings provided by an Elder on midwifery and prenatal care, based on her >60 years of practice as a midwife. The 2 selected teachings focused on (a) pregnancy and labour and (b) how to support mothers during pregnancy. The Elder believed that it would be of benefit to most members of the Inuit community in Ottawa, whether they were expectant parents themselves or grandparents. The criteria for message selection included clarity and coherence, cultural relevance and consistency with current recommendations regarding the clinical management of pregnancy with respect to physical activity . The CD-The academic research team, in consultation with the community research team, developed evaluation materials. Open-ended questions were direct and succinct (e.g. “Tell me what it will be ….” vs. “What do you think it will be ….”). In the questionnaire, comparison questions were avoided as they would have required changing the scale anchors, and this was believed by the community research team to create confusion for respondents. For scale anchors, 5 pie charts were used with 3 anchors . No mid point anchors were used to denote “2” and “4” because in Inuktitut, ratings such as “a little” and “a bit” do not exist. See Pre) and the second to assess reactions to viewing the CD-Rom (Post). Using a pre–post design, the goal was to assess whether post-ratings would be higher than expectations, which would also account for any existing high expectations that could influence results if post-only data had been used.The questionnaire items assessed 4 construct areas (a) satisfaction; (b) medium; (c) source; and (d) content. The constructs (b–d) map onto the 3 articulated preferences of the community. Two sets of questionnaires were developed see ; the firFor qualitative data collection, interviews were conducted prior to viewing the CD-Rom, and a focus group was conducted at follow-up (approximately 3 months after viewing the CD-Rom). The interview items were used to explore participants’ expectations about the CD-Rom (e.g. “Tell me what you think it will be like to use the CD-Rom”) and participants’ experience with the CD-Rom (e.g. “How was the CD-Rom different than what you expected?”) and evaluations of features of the CD-Rom (e.g. “What did you like the most about the CD-Rom?”). Questions at the focus group examined how the CD-Rom as a health promotion tool compared to other tools and explored suggestions for changes . All materials were translated into Inuktitut prior to commencing the data collection. In addition, an interpreter was used to translate questions during the focus group.Post), participants viewed the CD-Rom with the assistance of a community facilitator. Afterwards, all participants completed a questionnaire. A final focus group was conducted 3 months following the viewing of the CD-Rom, where a subsample of the participants (n=20) participated in a focus group (Follow-up).Interested individuals were identified by community research team members and were then provided with a description of the study. If they were interested, participants were presented with a consent form in English and were provided with a description of the study and the consent process in Inuktitut. At the first data collection point, participants answered the interview questions regarding their expectations of using the CD-Rom and completed a questionnaire. About 1 month later . Twenty-seven individuals completed the interview and questionnaire in Inuktitut and 13 completed them in English.Interviews were all recorded, transcribed and translated for coding purposes. Data analysis used a mixed iterative editorial and organisational approach . This ap2=0.44, but no language by time interaction, F=1.37, ns, partial η2=0.12. Inspection of the specific dependent variables showed a significant language effect for item 11 , F=27.64, p<0.01, partial η2=0.30, and for item 12 (CD-Rom like reading a brochure), F=34.91, p<0.001, partial η2=0.38. For both items 11 and 12, ratings completed by Inuktitut-speakers  were higher than ratings completed by English-speakers . Therefore, Inuktitut-speakers expected and found their interactions with the CD-Rom to be higher in similarity to both talking to someone and reading a brochure, compared to English-speakers. No other MANOVA was significant. As the language by time interaction was not significant, it was not necessary to control for language in the quantitative analyses assessing changes in ratings.In order to assess for any differences in ratings as a result of language, mixed multivariate analysis of variance (MANOVAs) were performed, with time and outcome category as within subjects variables and language as a between subjects variable. Four MANOVAs were performed, 1 for each outcome category: (a) satisfaction ; (b) medium ; (c) content ; and (d) source (items 8 and 9). The MANOVA for medium revealed a significant main effect for language, based on Wilks’ criteria, F=7.71, p<0.01, partial ηMultivariate analyses of variance (MANOVAs) were performed to examine differences between pre- and post-ratings. As there were multiple items measuring the same construct, a MANOVA was selected to assess differences between pre and post for each of the 4 constructs. Four MANOVAs were performed, with time and outcome category as independent variables, 1 for each outcome category: (a) satisfaction ; (b) medium ; (c) content ; and (d) source (items 8 and 9). Descriptive statistics are presented in 2=0.34. Inspection of the specific dependent variables revealed a significant effect for time for item 2 (recommend to someone), F=5.09, p<0.05, partial η2=0.15. Ratings of likelihood to recommend the CD-Rom were higher after having viewed it =4.71, p<0.01, partial ηd it see .2=0.28. Inspection of the specific dependent variables revealed a significant effect for time for item 11 , F=11.73, p<0.01, partial η2=0.27. Ratings of similarity to talking to someone were higher after having viewed it =3.84, p<0.05, partial ηd it see .2=0.22. Inspection of the specific dependent variables revealed a significant effect for time for item 5 (message clarity), F=7.85, p<0.01, partial η2=0.21. Ratings of how much sense the message made were higher after having viewed it =2.67, p<0.10, partial ηd it see .2=0.03.The overall multivariate test for a time effect on the 2 source-dependent variables, based on Wilks’ criteria, was not significant, F=1.00, p>0.10, partial ηinterest in the CD-Rom, including the technological aspects of the tool, the cultural content (language and Elder), as well as the information contained in the tool. They also spoke about their intent to use the information and the CD-Rom, and some requested to have a copy of the CD-Rom at the end of the study. One participant commented, “I know I'm going to want one1, I know it's going to help a lot of people”.To supplement and clarify the quantitative results, the interviews conducted prior to viewing the CD-Rom were coded for themes. Three main themes emerged were interest, uncertainty and conditional interest. Individuals expressed an uncertainty about the CD-Rom. Some expressed concern specifically about technology . Others mentioned concern about the content, for instance expressing concern about the ability to understand the message and language. Some individuals expressed an interest in the CD-Rom and its information if a certain condition was met; for example, someone commented it would be good “If I'm doing it with someone”. Other criteria included ease of technology, clarity of content and cultural-appropriateness of message.Individuals expressed a general I found <the Elder> to be, her voice is very soothing she is very low key, it's very easy to listen to her, and her, she has a very benign, not a preachy kind of demeanor, she's passing on information, and that, there just something very easy to listen.Qualitative results of the focus group focused on the preferred aspects of the CD-Rom and suggestions for future work. When community members were asked which feature they preferred the most about the CD-Rom, they were divided between “All” and “the Elder”. One person described the effect of the Elder's voice.She like <s> the idea of talking about pregnancy from a wholistic point of view with the help of the mother and the help of the father and that is something that we need talk more <about> as a family value.Some people commented about liking the traditional information and the fact that the CD-Rom was set up with 2 focused messages. The wholistic nature of the messages was again mentioned.The visuals good too because even when you have it in English you can still read her body language.Others made comments about the visual information. For instance, 1 participant stated:When asked what they would change about the CD-Rom, community members stated that the message was too short. Individuals were divided about whether there should be a few long messages or whether there should be many short messages. One person commented about the English translation: “As an urban Inuit I thought the English translation was a bit repetitive, so maybe have it ah, more edited a bit on the translation”. There was a request for additional topics, akin to a series that the Elder could talk about. There were also a few comments about enhancing the technology by allowing for a full screen viewing and mouse-over pop-up features for the graphics.When participants were asked about how the CD-Rom compared to other ways of sharing information, 1 individual commented that “It's not just something you can just pick up and throw away”. One comment was made that it was not as good as face-to-face, but it was better than paper-based information. There was a discussion around accessibility, given that a computer is needed. However, others pointed out that a computer was available at the various community based services.Few health promotion tools have been evaluated for use with Inuit, and none that we are aware of have been developed and evaluated for urban Inuit specifically. This study evaluated the acceptability of a guided CD-Rom as a health promotion tool for urban Inuit in Ottawa. Overall, the quantitative analyses largely supported predictions regarding the feasibility of such a tool. Although initial expectations were highly positive, overall increases in satisfaction ratings were observed, in particular when assessing the likelihood to recommend the tool to another. This is particularly meaningful as it suggests that this tool represents a viable medium for information sharing. It is also very encouraging that even though the CD-Rom focused specifically on pregnancy and prenatal care, a broad range of community members would recommend the tool to others. This might be because prenatal teachings are important to all members of the community, regardless of their stage of life. In addition, the messages provided participants the opportunity to gain more knowledge about traditional Inuit health. As 1 person commented, “I'm half Inuit and I don't know a lot of our traditional ways so this will give me a good idea of the traditional ways”. Furthermore, given that the Inuk Elder was from the north, this provided community members with an opportunity to learn new Inuktitut words. Given that the Inuktitut language has changed over the years, it is extremely important that traditional Inuktitut words survive . The CD-Ratings of similarity between CD-Rom and engaging in a conversation with someone increased from pre to post. This is significant because face-to-face communication has been identified as 1 of the key methods of sharing health information in the urban Inuit community. Furthermore, community members have previously commented about the difficulty in accessing Elders  in theirWith respect to the technology, it appears that the CD-Rom was easy to navigate. However, it is possible that having a community facilitator present during the viewing of the CD-Rom was responsible for the acceptance, as it is unclear if individuals would have regarded the CD-Rom as easy to use if they had been using a computer on their own, potentially in their home. Also, there remains the issue of accessibility to a computer with which to view the CD-Rom. The follow-up focus group captured the concern that some community members felt about being able to access a computer. For this study, community members viewed the CD-Rom on 1 of 2 office computers that are normally reserved for employees. Alternatively, if DVD technology was used, it is possible that there would be greater accessibility.Overall, the evaluation of the CD-Rom was positive and indicates that CD-Rom technology has strong potential as a medium for health information sharing in this urban Inuit community. Additional research is needed to assess the impact that the information has on participants’ experiences with pregnancy and prenatal care, and whether attitudes or behaviours changed as a result of having viewed the CD-Rom. The technology itself represents a practical and cost-effective way to connect community members in urban areas with Elders in the north. During the course of this research project, community members expressed a desire for additional health promotion materials, and several participants articulated an interest in materials about parenting young children.


Proc. Natl. Acad. Sci. USA 2010, 107, 19508–19513 [Cafeteria roenbergensis producing a lytic infection.Viruses with genomes larger than 300 kb and up to 1.2 Mb, which encode hundreds of proteins, are being discovered and characterized with increasing frequency. Most, but not all, of these large viruses (often referred to as giruses) infect protists that live in aqueous environments. Bioinformatic analyses of metagenomes of aqueous samples indicate that large DNA viruses are quite common in nature and await discovery. One issue that is perhaps not appreciated by the virology community is that large viruses, even those classified in the same family, can differ significantly in morphology, lifestyle, and gene complement. This brief commentary, which will mention some of these unique properties, was stimulated by the characterization of the newest member of this club, virus CroV (Fischer, M.G.; Allen, M.J.; Wilson, W.H.; Suttle, C.A. Giant virus with a remarkable complement of genes infects marine zooplankton.  However, large viruses with huge dsDNA genomes that encode hundreds of proteins are being discovered with increasing frequency. These large viruses have also been referred to as giruses in order to emphasize their unique properties . MimivirCafeteria roenbergensis [A recent report describes the newest girus, a lytic virus (named CroV) that infects the marine microzooplankton ergensis . CroV haergensis ,9.Mycoplasma genitalium, encodes ∼470 CDSs [Carsonella ruddii [Hodgkinia cicadicola [To place the size of these large viruses into perspective, the smallest free-living bacterium, 470 CDSs . Althoug470 CDSs ,12, somea ruddii  and Hodgcadicola  have genParamecium bursaria chlorella virus (PBCV-1) [Except for bacteriophage G ,16 and t(PBCV-1) , giruses(PBCV-1) . Althoug(PBCV-1) .2.Cafeteria roenbergensis indicates that giruses infect protists besides amoeba and algae.Metagenomic studies indicate that giruses are common in nature and wait to be discovered. For example, one study used the Sorcerer II Global Ocean metagenome to determine the frequency that DNA polymerase fragments could be assigned to virus groups. The results indicated that Mimiviruses were second in abundance to bacteriophages . Another3.2 fixed on the planet is by photosynthetic microorganisms, including cyanobacteria and microalgae (collectively referred to as phytoplankton). Current estimates are that at any one time ∼20% of phytoplankton cells are infected by viruses, including viruses that qualify as giruses. Additional components of this aquatic foodweb are microzooplankton that graze on these microorganisms, referred to as protistan grazers. CroV is the first virus to be characterized that infects a protistan grazer.In addition to the interesting biology associated with giruses that is briefly mentioned below, giruses along with smaller viruses play major roles in the ecology of aqueous environments, which is only now becoming apparent. Viruses are the most abundant biological entities on earth and are major drivers of nutrient and energy cycles on the planet (for detailed discussions see –24). MorEmiliania huxleyi, is one of the most abundant and widely distributed photosynthetic unicellular eukaryotes in the oceans. Coccolithophores produce skeletons of minute calcite platelets ; consequently, they are major contributors to the oceanic carbon cycle and thus to the flux of CO2 between the atmosphere and oceans. E. huxleyi cells periodically form huge blooms covering wide coastal and mid-oceanic areas at high latitudes in both the northern and southern hemispheres. Large viruses that infect E. huxleyi (named EhV viruses) are largely responsible for the termination of these blooms . This tE. huxleyi blooms also results in the release of dimethylsulfoniopropionate (DMSP) from the dying alga, which is cleaved by DMSP lyases; DMSP lyases are common in marine microorganisms. The cleavage products are acrylic acid and dimethylsulfide; dimethylsulfide is released into the atmosphere inducing cloud formation and rain. Thus, EhV infection of its host plays a significant role in climate conditions .E. huxleyi life cycle. The haploid state is resistant to EhV [One interesting feature of EhV is that it only infects the diploid phase of the t to EhV .4.Mimiviridae, Phycodnaviridae, Poxviridae, Asfarviridae, Iridoviridae, and Ascoviridae probably have a common evolutionary ancestor and are referred to as nucleocytoplasmic large DNA viruses (NCLDVs) [Mimiviridae family because 32% or the CroVs are Mimivirus homologs [Viruses classified in virus families (NCLDVs) –29. Rece(NCLDVs) . The newhomologs .Comparative analysis of 45 NCLDVs identified five common genes in all the viruses and 177 additional genes that are shared by at least two of these virus families . The fivet al. [Although common ancestry of NCLDVs is generally accepted, there is disagreement on the size and morphology of its ancestor and how it evolved into the different virus families. Like cellular organisms, gene and genome duplication contributed to the large genome of Mimivirus and maybe other giruses . A maximet al.  proposedThere are also disagreements on the origin of the NCLDVs. For example, some researchers have suggested that NCLDVs should be considered the fourth kingdom of life ,35, otheSulfolobus solfataricus. This structural similarity suggests that these three viruses might have an evolutionary connection to NCLDVs, despite the lack of amino acid sequence similarity among their MCPs (for a detailed discussion see [Contributing to the discussion about NCLDV evolution is the discovery that the structure of the chlorella virus PBCV-1 major capsid protein (MCP) resembles MCPs from some smaller dsDNA viruses with hosts in all three domains of life, including human adenoviruses, bacteriophage PRD1, and a virus infecting an archaeon, sion see ,40). If vice versa [Another interesting hypothesis is that a primitive NCLDV gave rise to the eukaryotic nucleus or ce versa ,42. Takece versa .Detailed reviews by Forterre  and Koon5.i.e., the viral factory [The discovery and characterization of giruses has revived a discussion of whether viruses should be considered to be living organisms, e.g., . One per factory ), which  factory ,46.These ideas have stimulated a lively discussion as to whether the tree of life should include viruses  The DNA polymerase from three putative phycodnaviruses, CdV01, PpV01, and PoV01 , is moreAdding to the classification issue is the proposal that the taxonomic status of viruses should be elevated to the same level as cells by dividing the biological world into two classes of organisms, those encoding capsids, and those encoding ribosomes .7.One issue that is perhaps not appreciated by the virology community is that these large viruses, even members within the same family, can differ in morphology see  and lifeIn contrast, the entire particle of another phycodnavirus, EhV, which has an external membrane, enters the host intact via either endocytosis or an envelope fusion mechanism with the host plasma membrane and then rapidly disassembles . In the ii) All NCLDVs are assembled in “virus factories” located in the cytoplasm. However, the role of the nucleus in the replication of NCLDVs varies. For example, Mimivirus, like the poxviruses, appears to carry out its entire life cycle in the cytoplasm . The intMimivirus has provided another surprise for virologists; all known icosahedral viruses with a unique vertex package their DNA at the same vertex that releases DNA, e.g., tailed bacteriophage. However, Mimivirus is reported to package its DNA through a face-centered aperture rather than the vertex-centered stargate structure that is involved in DNA release . If thisiii) Exit of newly formed viruses from the cell can also differ among giruses. The phycodnavirus EhV buds from its host , whereasEctocapus siliculosus; EsV has a lysogenic life cycle [Finally, it should be noted that most girus genomes do not integrate into their host genomes. The one exception is the phycodnavirus EsV, which infects the filamentous marine brown alga fe cycle .8.Not surprisingly, giruses encode an amazing array of proteins and even metabolic pathways, as well as properties that are typically the function of the host. Of course, the function of more than 50% of girus CDSs are unknown. Because of space limitations, only a few unusual virus-encoded properties are mentioned to illustrate their diversity. i) Both CroV and Mimivirus, as well as other giruses, contain several CDSs involved in protein translation, including amino acyl tRNA synthases and translation initiation factors, as well as tRNAs ,1. ii) IEmiliania huxleyi, also has genes encoding this pathway and obviously horizontal gene transfer occurred between EhV and E. huxleyi [E. huxleyi/EhV interaction suggests that aquatic viruses may control their environment in ways virologists and ecologists are only just beginning to fathom. iii) Some chlorella viruses encode three enzymes, including hyaluronan synthase, involved in the synthesis of the extracellular matrix polysaccharide hyaluronan; hyaluronan consists of alternating β1,4-glucuronic acid and β1,3-N-acetylglucosamine residues. Hyaluronan  accumulates on the external surface of the infected chlorella cells [Examples of unusual putative pathways encoded by giruses include: i) The newly described CroV genome has a 38 kb genomic fragment that encodes an entire biosynthetic pathway for 3-deoxy-D-manno-octulosonate (referred to as KDO) . In Gram huxleyi . However huxleyi . This exla cells . Previoula cells . The funla cells .Examples of some other unexpected proteins and enzymes encoded by giruses include: i) the chlorella viruses encode several ion channel and transporter proteins . ii) TheIt should be emphasized that many of the virus encoded, unexpected CDSs mentioned above are still putative assignments because biochemical evidence is lacking. However, other girus encoded CDSs have been expressed and have the expected properties. Some chlorella virus encoded proteins are the smallest or among the smallest proteins of their family, e.g., a histone methyltransferase , an orni9.Xenopus oocytes on the 94 amino acid chlorella virus PBCV-1 encoded potassium ion channel protein Kcv . Kcv-li10.Arabidopsis [Although giruses are probably ancient, they are relatively new to virologists. Even with our limited knowledge, research efforts on large viruses are contributing scientific and economic benefits. For example, chlorella viruses, which encode as many as 400 CDSs, are sources of new and surprising proteins, including commercially important enzymes such as DNA restriction endonucleases . The chlbidopsis .The sequence of some girus host genomes have either recently been completed or are in the process. Annotation of these host sequences will contribute to studies on giruses. However, a major obstacle to studying these viruses is that currently none of them can be genetically modified by molecular techniques. The development of successful and reproducible host transformation procedures should lead to the molecular genetic analysis of these viruses, which would lead to major advances in the understanding of these fascinating viruses.


The respiratory disease caused by the Influenza A Virus is occurring worldwide. The transmission for new strain of the H1N1 Influenza A virus is studied by formulating a SEIQR  model to describe its spread. In the present model, we have assumed that a fraction of the infected population will die from the disease. This changes the mathematical equations governing the transmission. The effect of repetitive contact is also included in the model. Analysis of the model by using standard dynamical modeling method is given. Conditions for the stability of equilibrium state are given. Numerical solutions are presented for different values of parameters. It is found that increasing the amount of repetitive contacts leads to a decrease in the peak numbers of exposed and infectious humans. A stability analysis shows that the solutions are robust. Influenza virus can be divided into three types: influenza A, B, and C. The virus is spread when droplets from a cough or sneeze of an infected person is propelled through the air and deposited on the mouth, nose, or eye of persons nearby. The virus can also land on surfaces in the surrounding environment and survive on the surfaces for 24 hours if the surface is hard and around 20 minutes if the surface is soft. The subtype H1N1 (2009) influenza virus is one of the influenza viruses that can cause respiratory illnesses. It is related to the virus which caused the 1918 flu pandemic. It is now called the swine flu virus because the virus causing pigs in the United States to become sick shortly after the 1918 pandemic was identified as being the H1N1 virus . ABiological factors such as the duration of infectious period and social factors can influence the spread of this disease. Repetitiveness of contacts is also known to be the relevant factors effect to the transmission of droplet or contact transmitted diseases. In 2009, Smieszek et al. showed that random mixing models provide acceptable estimates of the total outbreak size if the number of contacts per day is high . KlinkenNishiura  showed tS denoted susceptible human; E, exposed; I, infectious; Q, quarantine, and R denotes a recovered human) [In this paper, we study the transmission of a new H1N1 influenza A virus in human with the effect of repetitive contact taken into consideration. A statistical model was developed to estimate the social contact network within a high school using friendship network data and a survey of contact behavior . We consd human)  to descrS(t) t;as being the number of susceptible persons at time  E(t) as the number of exposed persons at time t; I(t) as the number of infectious persons at time t; Q(t) as being the number of quarantine persons at time t; R(t) as being the number of recovered persons at time t.Our model is based on the standard SEIQR  model for contact transmission. A susceptible human becomes an exposed person if he comes in contact, direct or indirect . If C is the number of contacts a human makes with someone who could pass on the virus, then C(E(t) + I(t)) is the number of contacts which could result in a person being exposed to the virus. S(t)/N(t) (N(t) being the total number of humans at time t) is the probability that the person being contacted is a susceptible human. If a certain percentage of the contacts are repeated ones, then the increase in the number of exposed human is given byrc is the percentage of contacts which are repetitious. These contacts do not lead to a new exposed human. It should also be noted that just because a person comes in contact with an exposed person, he may not come in contact with the virus. We also do not know at what point a person exposed to a virus becomes infectious, we must assume that an exposed person could transmit the virus before he is placed in the infectious class.We consider the transmission of H1N1 influenza A virus' in human. We have included the effect of repetitive contact and taken into account that people having a prior illness have a higher mortality rate. We begin by definingλh is the birth rate of humans) minus the number of susceptibles being exposed to people  and theδdh is the additional death rate caused by the new virus for which the human population has no immunity. The change in the number of infectious humans is equal to the percentage of the exposed humans, who develop into infectious humans minus the number of infectious humans who are placed in quarantine, who die from any cause and who recover,t; that is,λh = dn, then we haveIh and Eh are not equal to zero, the human population will decline. The flow chart of this model is shown in x(t) = X(t)/N(t), thend/dt)N(t) = −δdh(E(t) + I(t)) in (Consider)=λhN(t)−−rcCS(t)Ethat is,N(t)=S(t)t), thenddtx(t)=d(i)E0 = ,the disease free steady state (ii)E1 = , wherethe endemic steady state where i* is the positive solution of the following equation:λ) are solutions of the characteristic equation:JE is the Jacobian matrix at the steady state and I is the identity matrix. If all eigenvalues produce the negative real parts, then the steady state is local stability.The steady state of our dynamical equations is given as follows:R0 < 1, whereThe characteristic equation is given bys*, e*, i*, q*, and r* are defined in (The characteristic equation is given byfined in -20)..(26)dn=165∗65,  rrc. We have set rc to 10%, 30%, 50%, 70%, and 90%. As rc increases, the effective contact rate to create newly exposed susceptible decreases. This is clearly seen in rc increases. The lower frame in rc causes the equilibrium value of the infectious population to be lower. The equilibrium values are reached at an earlier time as rc decreases  of infected humans placed into quarantine is increased. Exposed humans are not placed into quarantine since many of them will not become infectious and quarantine is a denial of a person's human rights. This denial cannot be justified on the basis of maybe he could become infectious. Looking at the time evolution of this population group, we see that the peak in the number and the number in the equilibrium state decrease as the fraction of infectious humans is increased.With a mathematical model to describe the progression of a disease, we can simulate the time course of an epidemic when the values of various parameters are different. This would allow us to gain insights into how the epidemic might respond to change in the practices which could change the values of the parameters. Recently, Aldila et al.,  in the cSensitivity Analysis. We apply the sensitivity analysis to show that the solutions are robust. We set all the parameters in the model to be the same but use different initial values of s*(0), e*(0), i*(0), q*(0), and r*(0). As we can see, all the trajectories (solutions) converge to the same epidemic equilibrium state  = . The results are as shown in rc) increases. Figures Figures


Measles virus induces an acute disease with rash and fever. Despite ongoing vaccination and elimination campaigns, the measles virus still sustains long-lasting transmission chains in Europe. Here we report the complete genome sequence of a wild-type measles virus isolated from a patient in Munich (MVi/Muenchen.DEU/19.13[D8]) during a German measles outbreak in 2013. Morbillivirus genus of the Paramyxoviridae family, remain a serious threat to human health worldwide. In 2011, approximately 160,000 deaths were caused by measles, although MV elimination is sought globally (1Infections caused by measles virus (MV), the prototype of the lobally 13. Infectlobally 1.MV wild-type (wt) infection chains have been recently observed all over Europe , 6. In 2de novo assembler Velvet  and for further refinement to that of a closely related MV isolate . Only reads mapping to these MV strains were used for genome assembly using the Genotyping according to the WHO protocol  using thKJ410048. The complete genome sequence of the MVi/Muenchen.DEU/19.13[D8] isolate is available at GenBank under the accession number


Mastomys natalensis mice carry arenaviruses. Analysis of 340 nt of the viral RNA polymerase gene showed sympatric occurrence of 3 distinct arenaviruses.To determine the specificity of Morogoro virus for its reservoir host, we studied its host range and genetic diversity in Tanzania. We found that 2 rodent species other than  Mastomys natalensis, is the reservoir host of Lassa virus in western Africa . This period corresponds to the end of the dry season, when the density M. natalensis mice and 44 individuals from 7 other small mammal species  showed 97.1%–100% amino acid homology with the Morogoro prototype L sequence (GenBank accession no. EU914104). In contrast, the 2 sequences derived from the blood samples of L. rosalia and M. minutoides mice showed only 69.3% and 65.2% aa homology with the Morogoro prototype L sequence. These sequences (320 bp) were compared with sequences of the Old World arenaviruses (M. minutoides mice clustered at 93.7% homology with that of the Kodoko virus, in the lymphocytic choriomeningitis clade (Figure; M. minutoides mice seems to be a strain of the Kodoko virus originally isolated from 2 M. minutoides mice in Guinea (M. minutoides mice as the true reservoir of Kodoko virus in Africa.Using RT-PCR selective for the arenavirus L gene, we obtained positive results for 43 mice: 41 aviruses . The vir Figure; : thus, tL. rosalia mice clusters with the Ippy virus sequence (Arvicanthis niloticus rodents (L. rosalia mice appears to be a new species of Old World arenavirus. The genus Lemniscomys is more closely related to the genus Arvicanthis than to the genera Mus and Mastomys.The amino acid sequence of the virus isolated from sequence . Ippy vi rodents . Thus, tM. natalensis mice, where Morogoro arenavirus transmission occurs, sympatric murine species do not seem to be secondary reservoirs for the virus. In contrast, 2 mouse species, L. rosalia and M.minutoides, seem to be reservoirs of 2 other Old World arenaviruses, 1 of which may be a new species. Our study emphasizes the complementary nature of serologic and genetic-based approaches for arenavirus detection. Because of the cross-reactivity of Morogoro antigens with immune serum from individuals infected with other arenaviruses, a serology-only approach might have led to the conclusion that an extended set of hosts exists for the Morogoro virus. Because of its high cost, a genetics-only approach might never have indicated the hot spot of arenavirus around Morogoro that was shown by IFA (In high-density habitats of L.rosalia mice and the strain of Kodoko virus, additional sampling and genotyping are being conducted. In particular, determining the sequence of the S segment will further clarify evolutionary relationships within the Old World group.Our study demonstrates the presence of 3 Old World arenaviruses in a single location. To date, only 5 Old World arenavirus species and 17 New World arenaviruses have been recognized by the International Committee for Taxonomy of Viruses (


Using a metagenomic approach and molecular cloning methods, we identified, cloned, and sequenced the complete genome of a novel circular DNA virus, porcine stool-associated virus (PoSCV4), from pig feces. Phylogenetic analysis of the deduced replication initiator protein showed that PoSCV4 is most related to a fur seal feces-associated circular DNA virus. In this study, fecal materials collected from swine exhibiting diarrhea between December 2009 and June 2010 were collected by the Indiana Animal Disease Diagnostic Laboratory, Purdue University, West Lafayette, Indiana, and subsequently submitted to the National Animal Disease Center for metagenomic analysis . Phylogenetically, PoSCV4 and FSfaCV form a unique clade of single-stranded circular viruses.KJ417134.The complete genome of PoSCV4 (isolate CP2) has been deposited at GenBank under the accession no.


Mastomys natalensis) among 263 captured. Full-genome sequences of the viruses suggested that they were new strains similar to Lassa virus–related arenaviruses. Analyzing samples from additional rodents and other species can elucidate epizootiologic aspects of arenaviruses.To investigate arenavirus in Zambia, we characterized virus from the kidneys of 5 arenavirus RNA–positive rodents ( Arenavirus, a bisegmented ambisense single-stranded RNA virus, is 1 of the viral pathogens responsible for hemorrhagic fever in Africa and South America. Until 2007, Lassa virus was the only known arenavirus to cause hemorrhagic fever in Africa. However, during September–October 2008, 5 hemorrhagic fever cases caused by a novel arenavirus named Lujo virus occurred in South Africa  were set up on dry, arable lands or scrublands surrounding the cities of Lusaka , Namwala , and Mfuwe . We captured 57, 48, and 158 rodents in each of these cities, respectively. Rodents were euthanized with diethyl ether, and kidney tissues were harvested and stored at −80°C.Mastomys spp. were identified by mitochondrial cytochrome b gene (cytb) (cytb. BLAST (www.ncbi.nlm.nih.gov/blast.cgi) search analysis against the amplified cytb fragments showed that 23 of 57 rodents from Lusaka, 24 of 48 from Namwala, and 143 of 158 from Mfuwe were M. natalensis.For species identification of the rodents, DNA was extracted by using the DNeasy Blood and Tissue Kit . The QIAGEN OneStep RT-PCR Kit (QIAGEN) was used to screen arenaviruses under the following conditions: 30 min at 50°C, 15 min at 95°C, 45 cycles of 20 s each at 95°C, 30 s at 50°C, 1 min at 72°C, and 10 at 72°C. The primer sequences used were 5′-CACATAGTTGGGCCCCACTTGCTGTGATC-3′ and 5′-AGGATAAGTGAAAGAGAGAGTAATTC-3′, which were designed on the basis of a consensus sequence of the large (L) gene among African arenavirus strains (Old World Arenaviruses [OWAs]), including Lujo virus. The region of the L gene has been reported as well conserved among OWAs . Reverse transcription (RT) PCR results indicated that 4 (17%) of the 23 rodents captured in Lusaka and 1 (4%) of the 24 captured in Namwala were positive for arenavirus, but none of the 143 rodents captured in Mfuwe were positive. Overall, 5 (3%) of the 190 We used RNA samples extracted from kidney tissues of representative Lusaka and Namwala strains to determine full-genome sequences. After several attempts to amplify the virus genome cDNA of OWAs by using deduced universal primers, we obtained some virus fragments and determined the full-genome sequence by the gap closing and rapid amplification of cDNA ends methods. GenBank accession numbers for the Lusaka strain are AB586644 and AB586645, and for the Namwala strain, AB586646 and AB586647. Sequence analysis indicated that the genome of the Lusaka and Namwala strains have a typical bisegmented structure containing 2 open reading frames in each segment, and the genes in the segments are separated by a stable stem-loop structure (data not shown). The small segments of the Lusaka and Namwala strains are both 3,377 bp, and the large segments are 7,230 and 7,236 bp, respectively. The shortage in the Lusaka strain genome, compared with the Namwala strain genome, was in a noncoding region between the stem-loop and the L gene.Lusaka-Namwala).We used MEGA5 software . The div,83TAP and P93PPY motifs, which are present in other OWAs, excluding Lujo, Dandenong, and lymphocytic choriomeningitis viruses  were well conserved in Luna virus. The Z protein has a critical role in arenavirus budding, and 2 conserved late-domain motifs, P(T/S)AP and PPPY, in the C-terminal have been reported .We attempted to isolate Luna virus from the 5 viral RNA–positive tissue samples. Each kidney homogenate was injected into Vero E6 cells in Dulbecco modified Eagle medium supplemented with 2% fetal bovine serum. The culture medium was changed every 6 days, and the supernatant was harvested after 28 days of cultivation. The harvested culture supernatant was injected into new Vero E6 cells. During the cultivation period, culture supernatant was sampled every 2 days and tested for the presence of Luna virus RNA by 1-step RT-PCR. Finally, the amount of viral RNA in the culture supernatant of 1 Lusaka sample was increased during days 6–12 . During M. natalensis rodents in Zambia. Comparison of the genetic backgrounds of Luna virus and Lujo virus, a novel pathogenic arenavirus also found in Zambia, showed that Luna virus is genetically different from Lujo virus. Luna virus was closely related to nonpathogenic arenaviruses that have been found from central to eastern Africa. To elucidate the epizootiologic aspects of arenaviruses in Zambia, the number of rodent and other species samples must be expanded. Such elucidation can lead to discovery of new arenaviruses, as demonstrated by isolation of a pathogenic New World arenavirus from bats during a study to increase knowledge of the geographic range and genetic diversity of arenaviruses naturally associated with the Mexican woodrat (Neotoma mexicana) in the western United States (We isolated a novel nonpathogenic arenavirus, which we propose be designated Luna virus, from


To determine seroprevalence of viruses in bats in Papua New Guinea, we sampled 66 bats at 3 locations. We found a seroprevalence of 55% for henipavirus (Hendra or Nipah virus) and 56% for rubulavirus (Tioman or Menangle virus). Notably, 36% of bats surveyed contained antibodies to both types of viruses, indicating concurrent or consecutive infection. Henipavirus in the family Paramyxoviridae contains 2 highly lethal viruses, Hendra virus (HeV) and Nipah virus (NiV), both of which use pteropid bats as their main natural reservoir  was isolated in Australia during a disease outbreak in pigs, with epidemiologic evidence suggesting the involvement of human patients as a result of pig-to-human transmission  in 2002 in Australia, but its disease-causing potential is unknown  isolated from a pteropid bat in 1968 in Australia ; for NiV, 55% (36/66); TioPV, 38% (25/66); MenPV, 56% (37/66); NBV-like viruses, 17% (13/66); and BroV, 6% (4/66). The seroprevalence of the 2 types of paramyxoviruses, HeV/NiV at 55% and TioPV/MenPV at 56%, is high. The most striking finding is the presence of antibodies to both groups of viruses in 36% (24/66) of the samples. Considering that VNT is a more specific and less sensitive assay, the actual positive rate could be >36%. Compared with results of the study conducted in Madgascar (P. conspicillatus bats (spectacled flying fox) from Madang than Dobsonia magna bats (bare-backed fruit bat) from Bensbach. We believe this difference has more to do with the bat species than the geographic location. For henipaviruses, seroprevalence is slightly higher for NiV than for HeV, which is also consistent with the Luminex inhibition assay showing a trend of slightly higher inhibition for NiV than for HeV (data not shown). This higher seroprevalence may suggest that henipaviruses circulating in Papua New Guinea are more NiV-like than HeV-like. However, in the absence of genetic sequence data, serologic findings are inconclusive.The positive rate for both groups of paramyxoviruses is much higher in In addition, for the 2 rubulaviruses, seroprevalence is higher for MenPV than for TioPV. This finding is noteworthy because our previous data indicated a one-way cross-neutralization, with MenPV antibodies failing to neutralize TioPV . The results from this study suggest either that the main strain of bat rubulavirus(es) circulating in bats in Papua New Guinea is more closely related to MenPV or that there are different strains circulating and the MenPV-like are more dominant.D. magna bats at Bensbach is not much different from the 20% (11/54) in P. conspicillatus bats at Madang, indicating the NBV group of orthoreoviruses is present in bats of different species in Papua New Guinea. Second, none of the 66 serum samples was positive for both NBV and BroV, which supports our previous conclusion that BroV is a new species in the genus Orthoreovirus and that no significant cross-reactivity occurs between BroV and orthoreoviruses of other species groups  of the NBV group viruses in Paramyxoviridae and 2 from the genus Orthoreovirus, family Reoviridae. The surprising finding of a high prevalence of antibodies to both henipaviruses and TioPV/MenPV in individual pteropid bats highlights the need for more structured studies to investigate the infection dynamics of zoonotic viruses in different bat populations across the world. It is not clear at this stage what factors are responsible for the vast difference in prevalence of antibodies to paramyxoviruses and orthoreoviruses. In the context of potential incursion of exotic bat viruses into Australia, it is noteworthy that the bat rubulavirus(es) circulating in Papua New Guinea are more MenPV-like , whereas the Papua New Guinea henipavirus(es) seem to be more NiV-like (Asia). These findings are consistent with those in Indonesia (In this study, a serologic survey was conducted for 4 groups of viruses, 2 from the family


To understand the progression of porcine epidemic diarrhea virus infection, we inoculated gnotobiotic pigs with a newly emerged US strain, PC21A, of the virus. At 24–48 hours postinoculation, the pigs exhibited severe diarrhea and vomiting, fecal shedding, viremia, and severe atrophic enteritis. These findings confirm that strain PC21A is highly enteropathogenic. Coronaviridae, genus Alphacoronavirus) was previously reported only in Europe and Asia. The first US outbreaks caused a high number of deaths among suckling pigs and, as a consequence, substantial economic losses (A highly contagious coronavirus that causes porcine epidemic diarrhea (PED) was first reported in the United States in May 2013 in Iowa. Since then, the virus—porcine epidemic diarrhea virus (PEDV)—has spread rapidly nationwide  selective for the nucleocapsid gene (229–557 nt). The partial nucleocapsid gene sequence of PC21A was identical to that of 2 US PEDV outbreak strains from Colorado, USA: USA/Colorado/2013 (GenBank accession no. KF272920) and 13-019349 (GenBank accession no. KF267450). Only coronavirus-like particles were observed in the fecal sample by electron microscopy . The sam10 genomic equivalents (GE) of PEDV strain PC21A; pig 4 was exposed to the virus by indirect contact with inoculated pig 3. For each sample, the quantity of PEDV RNA GE was ≈106 times higher than plaque assay results for a cell-adapted PEDV strain, PC22A. Clinical signs were monitored hourly. Pig 4 was monitored for longer-term clinical signs and virus shedding. Pigs were euthanized for pathologic examination at 3 stages of infection: acute, mid, and later stages . The Ohio State University Institutional Animal Care and Use Committee approved all animal-related experimental protocols.The sample was bacteriologically sterilized by using 0.22-μm syringe filters and then prepared as inoculum. Near-term gnotobiotic pigs were delivered aseptically by hysterectomy from a specific pathogen–free sow  as described  and accumulation of large amounts of yellowish fluid in the intestinal lumen , panel AImmunofluorescence-stained cells were observed mainly in the epithelium of atrophied villi of small (duodenum to ileum) and large intestines (10 GE/mL in serum samples (All infected pigs tested at acute or later stages of infection had viral RNA titers of 4.8–7.6 log samples . These t,In 2013, the first US outbreaks of the rapidly spreading porcine virus, PEDV, caused a high number of pig deaths and substantial economic losses (


Pneumonia remains a leading cause of child death in developing countries. The viruses in severe pneumonia remain poorly defined.The study was conducted at the Eastern Visayas Regional Medical Center in Tacloban City, Philippines from May 2008 to May 2009. Patients aged 8 days to 13 years old who were admitted to the Department of Pediatrics with severe pneumonia were enrolled for the study. Upon admission, polymerase chain reaction was performed using nasopharyngeal swabs and blood cultures to detect respiratory viruses and bacteria, respectively.Burkholderia cepacia group (n = 9), Streptococcus pneumoniae (n = 4), Staphylococcus aureus (n = 4), Haemophilus influenzae (n = 1), and Salmonella C1 (n = 1) were also isolated.Among the 819 patients enrolled, at least one virus was detected in 501 cases (61.2%). In addition, 423 cases were positive for a single virus while bacteria were detected in the blood culture sample of 31 cases. The most commonly detected viruses were human rhinoviruses (n = 189), including types A (n = 103), B (n = 17), and C (n = 69), and respiratory syncytial virus (RSV) (n = 165). Novel viruses such as human metapneumovirus, human coronavirus NL63, human bocavirus, and human polyomaviruses WU and KI were also detected. There were 70 deaths, and one or more viruses were detected in 35 (50%) of these cases. Positivity only for influenza A virus  was significantly associated with fatal outcome. From the blood culture, Viruses were commonly detected in children with severe pneumonia in the Philippines. Hence, viral etiologies should be considered while developing better effective strategies to reduce child pneumonia-related deaths in developing countries. Infant and under-five mortality rates are commonly considered as important health indicators. In 2000, the United Nations introduced the Millennium Development Goal 4 (MDG4) to reduce under-five mortality by two-thirds between 1990 and 2015. The under-five mortality rate has steadily decreased from 90 per 1000 live births in 1990 to 65 per 1000 live births in 2008,particulHistorically, the common viruses most closely associated with childhood pneumonia have included influenza virus (Flu), respiratory syncytial virus (RSV), parainfluenza virus (PIV), and adenovirus (AdV). The development of molecular techniques facilitated the detection of novel viruses in patients with respiratory infections. Human metapneumovirus (hMPV) was first documented in 2001. Since tIn this prospective study, we investigated the association between presence of a virus and severity of disease in hospitalized children with severe pneumonia to provide an indication of causality. With this aim, we detected viral as well as bacterial pathogens in children hospitalized with severe community-acquired pneumonia and assessed the clinical significance of these pathogens.The study was conducted at the Eastern Visayas Regional Medical Center (EVRMC) in Tacloban City, Philippines. Tacloban City is the capital of Leyte Province and the regional center for Region VIII, also known as “Eastern Visayas”. The climate of Leyte Province is classified as tropical monsoon, and it usually rains throughout the year with higher rainfall between November and January. Region VIII consists of two main islands, Leyte and Samar, with a population of approximately 4 million , stirred in 3 ml of viral transport medium (VTM), and stored at 4°C until sample transport. Swabs were shipped to the Research Institute for Tropical Medicine (RITM) for virus detection twice a week using the recommended biological transport containers with temperatures maintained at 4°C. The specimens were subjected to PCR within 10 days after sample collection. Common respiratory viruses were targeted. RNA and DNA present in 100 μl of VTM supernatant were purified using the QIAamp MinElute Virus Spin Kit . To synthesize complementary DNA, 11 μl of the final extract was used as a template with superscript III, random hexamers, and RNase inhibitor . PCR was performed to detect FluA, FluB, RSVs, hMPV, HCoVs, HBoV, WUV, and KIV as previously described. For PIVSerotypes or genotypes of RSVs, HRVs, and AdVs were determined by sequencing the hypervariable region of the G protein gene for RSVs, the VP4Burkholderia cepacia group, isolates were sent to Miroku Medical Laboratory in Nagano, Japan, and underwent sequencing for 16S rRNA.Blood culture for bacteria was performed by trained medical technologists in EVRMC as previously described. On admiAll data was encoded into the FileMaker database , and statistical analysis was performed using SPSS 18.0J . Univariate analysis was used to determine the viral pathogens associated with fatal outcomes. The risk of fatal outcome for each detected virus and virus combination was evaluated by estimating the odds ratio (OR) based on categorization in the “Discharged against medical advice-deteriorating” or “Died” category.The study protocol was approved by the Institutional Review Board of Tohoku University Graduate School of Medicine, RITM, and EVRMC. The parents or guardians gave written informed consent for their children to participate in the study.During the 13-month study period, a total of 1242 patients were screened and 1043 met with the inclusion criteria. Of the 1043 patients, 224 refused to participate. A total of 819 cases were subsequently enrolled in the study. Background information and physical findings of the 819 cases are summarized in TableWheezing was observed in half of the patients . One hundred seventy five patients (21.4%) had interstitial infiltrates on chest x-ray, and 157 patients (19.2%) had consolidation. The median time interval between the onset of illness and admission was 4 days. Furthermore, 32.2% patients (n = 264) had received antibiotics prior to hospital admission. Since 10.7% (n = 88) of the cases deteriorated during hospital admission, they were classified as “Died”  or “Discharged against medical advice-deteriorating” .Among the 819 respiratory samples, 501 samples 61.2%) were positive for at least one virus. These included 423 cases (51.6%); 72 cases (8.8%); and 6 cases (0.7%) who were positive for one, two viruses, and three viruses, respectively  was 10.7% [88/819], and one or more viruses were detected in 52.2% [46/88] of these fatal cases  were less likely to be associated with fatal outcomes compared to virus-negative cases (13.9% [42/321]), but there was insufficient evidence of an association between fatal outcome and detection of at least one respiratory virus . Second, the risk of fatal outcome according to specific viruses was evaluated (Table/498) werStreptcoccus pneumonia (n = 4), Staphylococcus aureus (n = 4), Staphylococcus lugdunesis (n = 3), Staphylococcus epidermidis (n = 2), and Staphylococcus haemolyticus (n = 2). Two isolates of S. aureus were methicillin resistant Staphylococcus aureus (MRSA). For gram-negative rods (n = 3), there was one case for Haemophilus influenza, Enterobacter cloacae, and Salmonella C1. For glucose non-fermentes gram-negative rods (n = 12), there were Burkholderia cepacia group (n = 9), Pseudomonas aeruginosa (n = 1), Pseudomonas stutzeri (n = 1), and Shewanella putrefaciens (n = 1). We confirmed isolation of Burkholderia cepacia group by 16S rRNA sequencing. For gram-positive rods, there was Bacillus species (n = 1). There were four fatal cases among bacterial culture-positive cases, including B. cepacia group (n = 3) and MRSA (n = 1), and one “Discharged against medical advice-deteriorating” case that was positive for MRSA (n = 1). There were concomitant viral infections in samples positive for S. pneumoniae , S. aureus , MRSA , H. influenzae , and P. aeruginosa .Blood culture samples were collected from 814 cases, 31 of which (3.8% [31/814]) were positive in blood culture. Among gram-positive cocci (n = 15), there were Our study highlighted the potential importance of viruses as pathogens of community-acquired pneumonia among hospitalized children in the Philippines. The CFR in our study setting was as high as 8%, which indicated that childhood pneumonia is still an important cause of death in the Philippines. We conducted our current study following the IMCI guideline, because IMCI has been implemented in various developing countries for nearly 20 years as well Although we have demonstrated an important potential role of viral pathogens, economical and effective options for the prevention and treatment of viral infections that are practical to implement in developing countries are unavailable. HRV- and RSV-positive cases accounted for approximately 55% of our study population and had significant effects including those on mortality. However, no effective and safe vaccines or antiviral drugs for these viruses are currently available. The monAlthough we previously demonstrated the presence of novel viruses in the Philippines, our curStudies on childhood pneumonia were conducted in the Philippines in the 1980s and 1990s,33,46. IS. pneumoniae (n = 4) and H. influenzae (n = 1), the most commonly isolated bacteria was B. cepacia group (n = 9). B. cepacia is a habitant of the natural environment, but can cause severe infection in the hospital settings especially among immunocompromised patients in ICU of the enrolled cases. However, a recent study indicated that the prevalence of bacteremia among children with community-acquired pneumonia in the emergency department was as low as 2.1%. Unlike S. pneumoniae, all three invasive isolates were susceptible to penicillin, as reported in a previous study in Bohol, Philippines[The treatment approach of IMCI recommends the empirical use of antibacterial drugs, but thelippines. Howeverlippines. As IMCIPolyomaviridae that frequently cause latent infection along with reactivation by immunosupression[There are some limitations of our study. Firstly, the detection of viruses by molecular techniques may not necessarily indicate that the detected virus is a causative agent of respiratory symptoms since it does not fulfill Koch’s postulate. We understand that the gold standard for the diagnosis of viral infection is virus isolation; however, many novel viruses cannot be isolated, and demonstrating the presence of the viral genome is the only available detection method. On the other hand, we identified multiple viruses in different combinations especially among patients positive for HBoV, WUV, and KIV Table. WUV andpression. Their ppression.Our study demonstrates a potentially important role for viral pathogens in the etiology of severe pneumonia among the children in the Philippines. Our findings suggest that viral etiologies should be always kept in mind while developing effective strategies to reduce child pneumonia-related deaths in developing countries.Flu: Influenza virus; RSV: Respiratory syncytial virus; hMPV: Human metapneumovirus; PIV: Parainfluenzavirus; HRV: Human rhinovirus; HCoV: Human coronavirus; AdVs: Adenoviruses; HBoV: Human bovavirus; WUV: Human WU polyomavirus; KVI: Human KI polyomavirus; IMCI: Integrated Management of Childhood Illness; MIC: Minimum inhibitory concentration; EQAS: External Quality Assurance System.The authors declare that they have no competing interests.AS, SL, RO, and HO conceived the study and designed it together with FY, NF, HG, LS, and RA. AS, YF, and NF established viral detection methods, and AS, YF, NF, and HG performed the test. LS and MM established and performed detection analysis for bacteria. RT performed statistical analysis. AS drafted the manuscript with assistance of SL and HO. All authors contributed to the final version of the manuscript, read and approved it.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2334/12/267/prepubMap of the Philippines and Region VIII. (a) Darker shaded area indicates Region VIII. (b) Darker shaded area indicates Tacloban city.Click here for fileAge distribution of children hospitalized with severe pneumonia in Eastern Visayas Regional Medical Center, May 2008 to May 2009.Click here for fileOutcome of the children hospitalized with severe pneumonia, Eastern Visayas Regional Medical Center, May 2008 to May 2009. Flu indicates influenzavirus; RSV, respiratory syncytial virus; hMPV, human metapneumovirus; PIVs, parainfluenzavirus; HRV, human rhinovirus; HCoV, human coronavirus; AdVs, adenoviruses; HBoV, human bovavirus; WUV, human WU polyomavirus; KVI; human KI polyomavirus.Click here for file