Induction of TNF-α in human macrophages by avian and human influenza viruses

The highly pathogenic avian influenza virus H5N1 is known to induce high level of tumor necrosis factor α (TNF-α) from primary macrophages. However, it is still unclear whether current H5N1 strains also induce high TNF-α production, as most of the data were derived from extinct clade 0 H5N1 strain. Here, we show that current clade 1 and 2 H5N1 strains induce variable levels of TNF-α that are not necessarily higher than those induced by seasonal influenza viruses. The result suggests that hyper-induction of TNF-α in human macrophages is not always associated with a highly pathogenic phenotype. We further tested the contribution of the NS gene segment from H5N1 isolates to TNF-α induction by using reverse genetics. While NS conferred some variation in TNF-α induction when incorporated into an H1N1 virus genetic background, it did not affect TNF-α induction in an H5N1 virus genetic background, suggesting that other viral genes are involved.     

Effects of oseltamivir phosphate (Tamiflu<Superscript>®</Superscript>) in human sera on results of microneutralization and hemagglutinin-inhibition tests for H5N2 avian influenza virus

To determine the influence of oseltamivir phosphate (Tamiflu) on the results of microneutralization and hemagglutinin-inhibition (HI) tests in human sera with H5N2 influenza virus, ten volunteers were administered Tamiflu and blood samples were collected. In the microneutralization test, no consistent effects were observed. However, in the HI test, specimens from all volunteers taken at 4 and 7 h after drug administration showed a higher titer as compared to 0 and 24 h after administration when mammalian cells (horse, guinea pig, and human) were used. These results suggest that the administration of Tamiflu may affect the results of HI tests for H5N2 virus.     

Telling “the truth” in dementia-Do attitude and approach of general practitioners and specialists differ?

OBJECTIVE: The prevailing opinion in the literature that disclosing the diagnosis of dementia to patients is important is not always put into practice. The purpose of this study was to investigate differences between GPs and specialists (neurologists and psychiatrists) in the German ambulatory care system concerning the disclosure of the diagnosis of dementia. METHODS: Thirty in depth interviews with randomly selected GPs were conducted. On this basis a standardised questionnaire was developed and sent to 389 GPs and 239 neurologists and psychiatrists. RESULTS: The postal survey revealed only minor differences between GPs and specialists, both groups being equally in favour of a timely disclosure. For example, 70% of the GPs and 77% of the specialists strongly agreed that "patients with dementia should be informed early because of the possibility to plan their lives". This positive attitude is pronounced among younger physicians, but is somewhat contradicted by difficulties in the communication with patients expressed in the interviews. In the interviews, what may be described as a "double taboo" emerges, in that GPs describe taboo topic areas related to dementia for them and for their patients. CONCLUSION: The postal survey shows the two professional groups to be very much in favour of a timely disclosure--an attitude that is pronounced among younger physicians. These findings can be interpreted as a recent change of attitudes regarding the disclosure of the diagnosis of dementia in the medical profession. PRACTICE IMPLICATIONS: Training opportunities are needed in order to overcome communication obstacles in the doctor-patient-communication about dementia.     

Phosphoinositides in Ca<Superscript>2+</Superscript> signaling and excitation–contraction coupling in skeletal muscle: an old player and newcomers

Since the postulate, 30 years ago, that phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P ₂) as the precursor of inositol 1,4,5-trisphosphate (Ins(1,4,5)P ₃) would be critical for skeletal muscle excitation–contraction (EC) coupling, the issue of whether phosphoinositides (PtdInsPs) may have something to do with Ca²⁺ signaling in muscle raised limited interest, if any. In recent years however, the PtdInsP world has expanded considerably with new functions for PtdIns(4,5)P ₂ but also with functions for the other members of the PtdInsP family. In this context, the discovery that genetic deficiency in a PtdInsP phosphatase has dramatic consequences on Ca²⁺ homeostasis in skeletal muscle came unanticipated and opened up new perspectives in regards to how PtdInsPs modulate muscle Ca²⁺ signaling under normal and disease conditions. This review intends to make an update of the established, the questioned, and the unknown regarding the role of PtdInsPs in skeletal muscle Ca²⁺ homeostasis and EC coupling, with very specific emphasis given to Ca²⁺ signals in differentiated skeletal muscle fibers.     

Genetic characterization of H5N1 avian influenza viruses isolated in southern China during the 2003–04 avian influenza outbreaks

Summary. The recent H5N1 avian influenza outbreaks in Asia spread over more than 8 countries. It has caused enormous economic loss and grand challenges for the public health. During these breakouts we isolated three strains of H5N1 Avian Influenza Virus (AIV) from chickens and one from duck in different farms of Southern China. We completely sequenced these four AIVs. Molecular characterization demonstrated that these strains retain the reported H5N1 AIV sequence properties relevant to virus virulence and host adaptation. Phylogeny results demonstrated that three of these isolates (except A/Chicken/Guangdong/174/04) were closely linked to other H5N1 AIVs isolated from the recent H5N1 outbreaks in Asia. Six of 8 segments (except PA and M) of A/Chicken/Guangdong/174/04 also shares a close linkage to other H5N1 AIVs isolated from the recent H5N1 outbreaks. However, the PA gene of A/Chicken/Guangdong/174/04 and another H5N1 strain forms a distinct subgroup along with an H6N1 AIV, and the M gene of A/Chicken/Guangdong/174/04 shows a close linkage to some H5N1 AIVs from aquatic species in China. Our findings suggest that a new genotype of AIV (in addition to previous reported ones) was present during the 2003–04 Asian bird flu outbreaks and that continuing virus surveillance of AIVs be conducted to monitor the evolutionary paths of the A/Chicken/Guangdong/174/04-like AIVs.     

Haemagglutinin and neuraminidase characterization of low pathogenic H5 and H7 avian influenza viruses isolated from Northern pintails (<Emphasis Type="Italic">Anas</Emphasis> <Emphasis Type="Italic">acuta</Emphasis>) in Japan,with special reference to genomic and biogeographical aspects

Pintails constitute an important host of avian influenza viruses (AIVs). Genetic, molecular, and antigenic characteristics of H5 and H7 AIVs, which we isolated from northern pintails (Anas acuta) wintering in Japan, were analyzed and found to be linked to various ecological features, chiefly in terms of gene geography, as shaped by various migratory aquatic host species. Although all the isolates were found to be of low pathogenicity (LP), we explored gene predispositions that may potentially underlie tentative transition to high pathogenicity (HP). Evolutionarily, the HA and NA genes of the isolates affiliated mostly with Eurasian lineage. The viruses closely related to ours were derived from China, Korea, Mongolia, Japan, and Australia. Comprehensive ecophylogenetic evaluations revealed that the pintail populations we sampled might have given rise to or been involved in the emergence of a LPAI H7N6 subtype that caused outbreaks in quail (Coturnix japonica) farms in Japan, as well as of the first H5N9 subtype ever isolated in Asia. The latter strain isolated by us showed, yet, notable affinity to certain North American and Australian strains, thereby signifying apparent intercontinental interfaces accounted for by extensive water-bird flyways. Noticeable conservation of certain antigenic sites within both Eurasian and North American H7 HAs is apparently an outcome of their advantageous survival value, in terms of restricted immunogenicity. Besides, the Japanese–Korean–Siberian regional axis seems to be particularly important for ongoing generation of novel viral strains due to conveyance of certain genes and genomes by migratory ducks, including such that circulate among pigs and human.     

The Genus Streptococcus – Part II: New Species and Pathogens in the “Miscellaneous” Streptococci and “Viridans” Streptococci Groups

In the second part of this two-part series on an update on the streptococci, new species and emergent human pathogens in the “viridans streptococci” and the “miscellaneous streptococci” groups are discussed. Among the “miscellaneous streptococci,” the most important organism in human infectious diseases is Streptococcus suis. This organism is primarily an agent of disease in swine and other animals and has now emerged as a significant human pathogen, causing bacteremia and meningitis, particularly among populations in Asia and the Far East. While S. suis capsular serotype 2 is the most frequently isolated agent, infections caused by other serotypes are increasingly being reported. In addition, disease caused by S. suis is now being described more frequently in North America in individuals with significant occupational exposure to swine. Among the viridans streptococci, several new species in the mitis-sanguinis group have been described as human pathogens, including Streptococcus pseudopneumoniae, Streptococcus oligofermentans, and Streptococcus tigurinus. Other newly reported viridans group species either comprise part of the human oral microbiome or have been isolated from animals. This article presents information on these bacterial agents, including characteristics for the recognition and identification of the more important clinical isolates in these streptococcal groups.     

Genetic characterization of avian influenza viruses isolated in Israel during 2000–2006

Our aim was to establish the phylogenetic and genetic relationships among avian influenza viruses (AIV) recently isolated from poultry in Israel. During this study we analyzed complete nucleotide sequences of two envelope (hemagglutinin and neuraminidase) and six internal genes (polymerase B1, polymerase B2, polymerase A, nucleoprotein, nonstructural, and matrix) of 29 selected H9N2 and six internal genes of five H5N1 viruses isolated in Israel during 2000–2006. Comparative genetic and phylogenetic analyses of these sequences revealed that the local H5N1 viruses are closely related to H5N1 viruses isolated in European, Asian, and Middle Eastern countries in 2005–2006. The H9N2 Israeli isolates, together with viruses isolated in Jordan and Saudi Arabia formed a single group. Our data support the claim that during recent years a new endemic focus of H9N2 has been formed in the Middle East. The introduction of H5N1 and co-circulation of these two subtypes of AIV in this region may augment the risk of potentially pandemic strains emergence.     

The avian influenza epidemic in Italy, 1999—2000: A review

During 1999, northern Italy has been affected by an epidemic of low pathogenicity avian influenza (LPAI) caused by a virus of the H7N1 subtype. Due to the characteristics of the poultry industry in the area and to the absence of specific legislative tools to eradicate infection, the virus continued to circulate for several months until a highly pathogenic virus of the same subtype emerged. The highly pathogenic virus had caused death, at the time of writing, of over 13 million birds in 3 months. The consequences of the highly pathogenic avian influenza (HPAI) epidemic appear to be devastating for the poultry industry and the social community. Several conditions generated the current situation, including the high density of susceptible animals and the structure of the poultry industry in the infected area. In addition, the circulation of LPAI virus for a number of months inevitably delayed the prompt identification of HPAI and complicated the interpretation of diagnostic results. A reconsideration of current European legislation and a reorganization of the poultry industry are suggested to prevent the occurrence of similar situations in countries of the European Union.     

Association between the σ protein of avian reovirus and virus-induced fusion of cells

Summary.  Monoclonal antibodies (MAbs) against a 39 kDa (σC) protein of the avian reovirus RAM-1 strain inhibited virus-induced fusion of cells and the protein was expressed on the surface of infected cells. The fusion-inhibiting activity of the three MAbs reacting with the σC protein suggest two putative epitopes were involved: one epitope recognised by antibody 6H1 and involved in fusion of both Vero and CK cells and a second epitope recognised by antibody 1G1 involved in fusion of Vero cells but not CK cells. The activity of the MAb 6E2 was intermediate, suggesting it may have been located in an intermediate position between the two putative epitopes and inhibited fusion by steric hindrance. Received May 6, 1998 Accepted July 30, 1998     

Human cytomegalovirus and Epstein–Barr virus in apical and marginal periodontitis: A role in pathology?

Periodontitis is presumably caused by bacterial infection, but it has been shown recently that affected tissue often contains human cytomegalovirus (HCMV) and Epstein-Barr virus (EBV). The present study was initiated to evaluate the role of these viruses in the pathogenesis of periodontitis. HCMV and EBV were quantified in 40 apical and 25 marginal periodontitis samples using real time PCR. In situ hybridization or immunohistochemistry was carried out on apical samples to detect viral presence within cells. A possible association with relevant bacteria was examined. Of the apical periodontitis samples, 50% contained EBV, while none contained HCMV. Of the marginal periodontitis samples, 40% were positive for EBV and 12% for HCMV. With one exception, however, the amount of virus was close to the detection limits. EBV was only detected in 1 out of 15 healthy periodontium samples. Immunohistochemistry and in situ hybridization were all negative. Significant associations were found between periodontal EBV and the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. Although there was an obvious association of the virus with clinical samples, it seems unlikely that these viruses play a major role in the pathogenesis of periodontitis of the average patient. Their presence may reflect that the clinical samples contain more blood or saliva compared to controls, or an accumulation of lymphoid cells harboring virus in the inflamed tissue.     

The genetic and antigenic diversity of avian influenza viruses isolated from domestic ducks, muscovy ducks, and chickens in northern and southern Vietnam, 2010–2012

To estimate the prevalence of avian influenza virus infection in Vietnam, surveillance was conducted in domestic and wild birds from households, live-bird markets, slaughtering sites, and bird sanctuaries in Vietnam between October 2010 and October 2012. Of the 4,550 samples collected, 226 influenza A virus isolates were obtained from domestic ducks, muscovy ducks, and chickens. Of these, 25 and 22 H5N1 highly pathogenic avian influenza viruses (HPAIVs) were isolated from apparently healthy domestic ducks in live-bird markets and slaughtering sites in northern and southern Vietnam, respectively. The HA genes of H5 viruses isolated from birds in northern Vietnam phylogenetically belonged to the genetic clade 2.3.2.1 and those in southern Vietnam belonged to the genetic clade 1.1. In addition, 39 H3, 12 H4, 1 H5, 93 H6, 2 H7, 18 H9, 3 H10, and 11 H11 viruses were isolated. Phylogenetic and antigenic analyses of the H6 and H9 viruses revealed that they were closely related to the isolates obtained from domestic poultry in China. Phylogenetic analyses of internal gene segments of these isolates revealed that these viruses were circulating in both domestic and wild birds in Asia and reassortment events had occurred frequently. Therefore, it will be important to continue the surveillance and strict controls over the movement and trade of poultry and poultry products in order to eradicate H5N1 HPAIV from Asia.     

“Speaking-for” and “speaking-as”: Pseudo-surrogacy in physician–patient–companion medical encounters about advanced cancer

Objective

To examine using audio-recorded encounters the extent and process of companion participation when discussing treatment choices and prognosis in the context of a life-limiting cancer diagnosis.

Methods

Qualitative analysis of transcribed outpatient visits between 17 oncologists, 49 patients with advanced cancer, and 34 companions.

Results

46 qualifying companion statements were collected from a total of 28 conversations about treatment choices or prognosis. We identified a range of companion positions, from “pseudo-surrogacy” (companion speaking as if the patient were not able to speak for himself), “hearsay”, “conflation of thoughts”, “co-experiencing”, “observation as an outsider”, and “facilitation”. Statements made by companions were infrequently directly validated by the patient.

Conclusion

Companions often spoke on behalf of patients during discussions of prognosis and treatment choices, even when the patient was present and capable of speaking on his or her own behalf.

Practice implications

The conversational role of companions as well as whether the physician checks with the patient can determine whether a companion facilitates or inhibits patient autonomy and involvement. Physicians can reduce ambiguity and encourage patient participation by being aware of when and how companions may speak on behalf of patients and by corroborating the companion's statement with the patient.     

Detection and quantitation of gallid herpesvirus 1 in avian samples by 5′ Taq nuclease assay utilizing Minor Groove Binder technology

A 5′ Taq nuclease assay utilizing Minor Groove Binder technology and targeting the thymidine kinase gene of gallid herpesvirus 1 (GaHV-1) was designed and optimized for use in diagnosing avian infectious laryngotracheitis. The assay was specific for GaHV-1 in that it did not react with other avian viral or bacterial pathogens. The detection limit was 1.0×10^?2 median tissue culture infectious dose per reaction or 90 target copies per reaction. Fifteen out of 41 diagnostic samples from sick birds reacted in the assay, five of which produced a typical alphaherpesvirus cytopathic effect (CPE) on chicken kidney (CK) cells. Sequencing, using amplicons generated by a polymerase chain reaction with primers flanking the 5′ Taq nuclease amplicon, confirmed the presence of GaHV-1 in six samples (two producing alphaherpesvirus CPE on CK cells, three not producing alphaherpesvirus CPE, and one that was not inoculated onto CK cells). Tracheal swabs taken from 18 healthy broilers did not react in the assay. The ability of the assay to determine viral load in samples was demonstrated. Overall the assay is suitable for the rapid diagnosis of infectious laryngotracheitis.     

Oxytocin and prostaglandins F<Subscript>2α</Subscript> and E<Subscript>2</Subscript> do not enhance HIV antigen production <Emphasis Type="Italic">in vitro</Emphasis>

Summary.  Oxytocin and prostaglandins (PGs) are hormones involved in labor and are used clinically for its induction. In this study the effect of oxytocin, PGF_2α, and PGE₂ on Humour immunodeficiency virus-1 production in acutely and persistently infected cells was measured. No significant effect on p24 antigen production was found with oxytocin or PGs, except for a transient decrease in persistently infected cells treated with 1 μM PGF_2α. These results showed that oxytocin and PGs could be used clinically for labor induction without any direct enhancement in viral production. Besides, the results with PGF_2α at the highest concentration studied may indicate a pharmacological effect. Received October 10, 2002; accepted October 28, 2002     

Detection and epidemiology of carbapenemase producing <Emphasis Type="Italic">Enterobacteriaceae</Emphasis> in the Netherlands in 2013–2014

Laboratory detection of carbapenemase-producing Enterobacteriaceae (CPE) is complicated. Screening with MIC values below clinical breakpoints followed by genotypic confirmation is recommended. We evaluated the application of recommended CPE screening and confirmation methods and provide an overview of CPE epidemiology in E. coli and K. pneumoniae in the Netherlands. Data on E. coli and K. pneumoniae isolates with elevated meropenem (>0.25 mg/L) and/or imipenem (>1 mg/L) MIC values in 2013–2014 were selected from the Infectious Disease Surveillance Information System for Antibiotic Resistance. Laboratories were requested to provide additional results of any confirmatory testing performed. Confirmation of elevated carbapenem MIC values using gradient testing was performed in 59.8 % of eligible isolates. Confirmatory testing showed elevated MIC values in 8 % of E. coli and 32 % of K. pneumoniae isolates. The overall proportion of confirmed non-susceptible E. coli and K. pneumoniae was 0.01 % and 0.16 %, respectively. Genotypic confirmation was performed in 61.0 % of isolates with confirmed elevated carbapenem MIC values. A carbapenemase gene was identified in 47 % of E. coli and 65 % of K. pneumoniae isolates. OXA-48, NDM and KPC were the most frequently found carbapenemase genes. The majority (62 %) of CPE isolates was detected through targeted screening. CPE are a rare finding in the Netherlands. Adherence to the national guideline is suboptimal and differs between laboratories, implying a risk of inadequate CPE detection. Since accurate identification of CPE is the first step in prevention of CPE spread, successful implementation of guidelines for testing and reporting of CPE is essential.