title: Biomarkers of Infection: Are They Useful in the ICU?

text: Biomarkers are increasingly used in patients with serious infections in the critical care setting to complement clinical judgment and interpretation of other diagnostic and prognostic tests. The main purposes of such blood markers are (1) to improve infection diagnosis (i.e., differentiation between bacterial vs. viral vs. fungal vs. noninfectious), (2) to help in the early risk stratification and thus provide prognostic information regarding the risk for mortality and other adverse outcomes, and (3) to optimize antibiotic tailoring to individual needs of patients (“antibiotic stewardship”). Especially in critically ill patients, in whom sepsis is a major cause of morbidity and mortality, rapid diagnosis is desirable to start timely and specific treatment. Besides some biomarkers, such as procalcitonin, which is well established and has shown positive effects in regard to utilization of antimicrobials and clinical outcomes, there is a growing number of novel markers from different pathophysiological pathways, where the final proof of an added value to clinical judgment and ultimately clinical benefit to patients is still lacking. Without a doubt, the addition of blood biomarkers to clinical medicine has had a strong impact on the way we care for patients today. Recent trials show that as an adjunct to other clinical and laboratory parameters these markers provide important information about risks for bacterial infection and resolution of infection. Moreover, biomarkers can help to optimize management of patients with serious illness in the intensive care unit, thereby offering more individualized treatment courses with overall improvements in clinical outcomes.


title: Optimal Ventilator Strategies in Acute Respiratory Distress Syndrome

text: Mechanical ventilation practices in patients with acute respiratory distress syndrome (ARDS) have progressed with a growing understanding of the disease pathophysiology. Paramount to the care of affected patients is the delivery of lung-protective mechanical ventilation which prioritizes tidal volume and plateau pressure limitation. Lung protection can probably be further enhanced by scaling target tidal volumes to the specific respiratory mechanics of individual patients. The best procedure for selecting optimal positive end-expiratory pressure (PEEP) in ARDS remains uncertain; several relevant issues must be considered when selecting PEEP, particularly lung recruitability. Noninvasive ventilation must be used with caution in ARDS as excessively high respiratory drive can further exacerbate lung injury; newer modes of delivery offer promising approaches in hypoxemic respiratory failure. Airway pressure release ventilation offers an alternative approach to maximize lung recruitment and oxygenation, but clinical trials have not demonstrated a survival benefit of this mode over conventional ventilation strategies. Rescue therapy with high-frequency oscillatory ventilation is an important option in refractory hypoxemia. Despite a disappointing lack of benefit (and possible harm) in patients with moderate or severe ARDS, possibly due to lung hyperdistention and right ventricular dysfunction, high-frequency oscillation may improve outcome in patients with very severe hypoxemia.


title: Efficiency of various cloned DNA probes for detection of bovine viral diarrhea viruses()

text: We have evaluated 24 cytopathic (CP) and 37 noncytopathic (NCP) strains of bovine viral diarrhea virus (BVDV) with a dot blot assay using four different genome segments of the NADL strain as hybridization probes (p80, p54, gp53, and gp62). The p80 and p54 probes hybridized to 23/24 (96%) and 2224 (92%), respectively, of CP strains examined. In contrast, these same two probes only detected 1636 (43%) and 536 (13%), respectively, of the NCP strains examined. The gp53 probe detected 1824 (75%) and the gp62 probe detected 1924 (79%) of the CP strains. In contrast, these latter two probes only detected 937 (24%) and 737 (20%), respectively, of NCP strains. This low detecteion rate of NCP strains suggests a need for developing a probe based on NCP sequences for identification of NCP strains.


Histopathological alterations of lymphatic tissues in cats without feline infectious peritonitis after long-term exposure to FIP virus


title: Growth of Mycoplasma bovis in organ cultures of bovine foetal trachea and comparison with Mycoplasma dispar

text: Inoculation of tracheal organ cultures from bovine foetuses with Mycoplasma bovis resulted in a loss of cellular structure of the lamina propria, followed 20–22 days later by lifting and detachment of overlying epithelium. The effect was associated with large numbers of M. bovis, identified by immunoperoxidase labelling and electromicroscopy, infiltrating between the epithelial cells and amassing in the lamina propria, especially in the region of the basement membrane of the epithelium. Ciliary activity was undiminished for up to 18 days following inoculation and little or no cytopathic effect on the ciliated epithelium was seen in spite of the close proximity of large numbers of organisms. In contrast, M. dispar was restricted to the margin of the ciliated epithelium where, as previously reported, it caused pyknosis, sloughing and flattening of the epithelium with consequent loss of ciliary activity. The cytopathology observed for each mycoplasma bore a close similarity to the behaviour of the two mycoplasmas in vivo and it is suggested that the organ culture system may be a useful and relevant system to elucidate the pathogenic mechanisms for each mycoplasma.


title: Detection of porcine rotavirus in stools by a latex agglutination test

text: We developed a simple agglutination test for the detection of porcine rotavirus in stools from pigs with diarrhea. The virus was detected with high sensitivity and specificity by a slide agglutination test using latex particles coated with antibody against the porcine rotavirus strain OSU (LA-antiOSU). The agglutination of LA-antiOSU with OSU on a glass slide evident macroscopically within 2 min. The sensitivity of this latex agglutination (LA) test was four times higher than that of the electron microscope method. The LA test is available for the rapid diagnosis of porcine rotavirus infections.


title: An ORF2 protein-based ELISA for porcine circovirus type 2 antibodies in post-weaning multisystemic wasting syndrome

text: Porcine circovirus type 2 (PCV2) plays a crucial role in the pathogenesis of post-weaning multisystemic wasting syndrome (PMWS) in swine. As PCV2 displays significant homology with PCV1 (a non-pathogenic virus) at the nucleotide and amino-acid level, a discriminative antigen is needed for specific serological diagnosis. The ORF2-encoded capsid protein from PCV2 was used to develop an indirect enzyme-linked immunosorbent assay (ELISA). GST-fused capsid protein from PCV2 and GST alone (both expressed in recombinant baculovirus-infected cells) were used as antigens for serodiagnosis. The specificity of the ELISA for detection of PCV2 antibodies was demonstrated in sera from pigs experimentally infected with PCV1, PCV2 and other swine viruses. The semi-quantitative nature of the test was evaluated versus an immunoperoxidase monolayer assay (IPMA). The ELISA was performed on 322 sera from pigs in eight Brittany herds and compared with IPMA. The sensitivity (98.2%) and specificity (94.5%) of this test were considered suitable for individual serological detection. High PCV2 seroprevalence was found in sows and pigs at the end of the growth phase (18–19 weeks) in all eight herds. The seroprevalence in piglets (11–17 weeks) was statistically correlated with clinical symptoms of PMWS (93% in affected versus 54%, in non-affected farms). A cohort study performed in PMWS-free farms showed that 57% of piglets exhibited active seroconversion after 13 weeks, indicating that PCV2 infection occurred earlier in PMWS-affected piglets.


title: Cross-reactivity between B. burgdorferi and other spirochetes affects specificity of serotests for detection of antibodies to the Lyme disease agent in dogs

text: Western immunoblots, the kinetics-based enzyme-linked immunosorbent assay (KELA), and the microagglutination test were used to evaluate cross-reactivity among antibodies to serovars of Leptospira interrogans (leptospiral serovars), and B. burgdorferi from naturally infected dogs, and to Serpulina (Treponema) hyodysenteriae from vaccinated rabbits. Whole-cell lysates from Borrelia spp., leptospiral serovars, and Serpulina spp. were used for SDS-PAGE, western blots, and KELA. Cross-reactivity occurred between the antibodies to B. burgdorferi and leptospiral serovars when tested on the heterologous antigens. Antibodies to leptospiral serovars tended to cross-react more strongly with antigens of B. burgdorferi spp. than did antibodies to B. burgdorferi when tested against antigens of leptospiral serovars. The antibodies against B. burgdorferi showed a lesser degree of cross-reactivity to the antigens of S. hyodysenteriae and S. innocens than they did to leptospiral serovars. We conclude that cross-reactivity occurs between B. burgdorferi and leptospiral serovars. Validation and interpretation of ELISA tests for detection of antibody activity to whole cell lysates of the Lyme agent must take this cross-reactivity into consideration. Conversely, dogs infected with the Lyme agent do not show significant cross-reactivity in the microagglutination test for antibody to the leptospiral serovars.


docid: ymciferk

title: Detection of feline immunodeficiency proviral DNA in peripheral blood lymphocytes by the polymerase chain reaction

text: Feline immunodeficiency virus (FIV) proviral DNA was detected by the polymerase chain reaction method (PCR). PCR products were detected by gel electrophoresis and ethidium bromide staining. The P-10, P-15 and P-24 regions of the gag gene of FIV were chosen as the target sequences for amplification, and three primer pairs were prepared. The PCR products subjected to amplification with each primer pair were found to possess sites of digestion by a restriction enzyme, as hypothesized. They did not react with feline leukemia virus (FeLV)-infected or feline syncytium-forming virus (FeSFV)-infected cell-derived DNA, and specifically amplified FIV-infected cell-derived DNA. FIV proviral DNA was detected by the PCR method with either primer pair (one-step amplification: single PCR) in DNA derived from peripheral blood lymphocytes (PBL) from 7 of 12 FIV antibody-positive cats. When PCR products in each of the 12 cats were subjected to a second amplification using the same primer pair (two-step amplification: double PCR), FIV proviral DNA was detected in all of the cats. When PBL samples collected from three cats that were negative and three that were positive in the single PCR were cultured for a few weeks in the presence of interleukin 2, FIV proviral DNA was detected in all six cats by the single PCR method. The results suggest that either the use of cultured PBL as the sample or the performance of the double PCR method enables simple and specific detection of FIV proviral DNA in PBL.


title: Transmissible gastroenteritis (TGE) of swine: In vitro virus attachment and effects of polyanions and polycations

text: Four transmissible gastroenteritis virus (TGEV) strains (Purdue-115, D-52, 188-SG and Gep-II) and two cell lines (swine testis-ST and pig kidney-RPD) were used to study virus attachment and cell susceptibility. Virus attachment was partially thermodependent and the rate varied, depending on the strain. Identical TGEV inocula produced a higher plaque number by plaque assay in the swine testis cell line (ST) than in the pig kidney cell line (RPD) but [(3)H]uridine-labèlled virus was found associated equally well with both cell lines. A field TGEV strain (Gep-II), which was unable to multiply in cell cultures, appeared able to inhibit the attachment of radiolabelled cell-passaged virus. Therefore, the susceptibility to TGEV infection was apparently not determined at the virus-to-cell attachment stage. The attachment sites on the cell surface were specific, however, differences in TGEV attachment determinant between strains were not observed. Attachment of all the virus strains tested was enhanced by DEAE-dextran and inhibited by dextran sulfate, poly-L-lysine (PLL), poly-L-α-ornithine (PLO) and protamine sulfate.


title: Inheritance of resistance to oedema disease in the pig: Experiments with an Escherichia coli strain expressing fimbriae 107

text: Inheritance of resistance to intestinal colonization with E. coli causing oedema disease is hypothetized to be under the control of one locus consisting of two alleles with susceptibility (S) dominating resistance (s). This mode of inheritance was investigated by matting pigs, resistant and susceptible to the disease, and examining the offspring. Weaned piglets were repeatedly inoculated orally with 5 × 10(5) CFU per pig per day of a streptomycin resistant strain of E. coli serotype O139:K12(B):H1:F(107) and susceptibility determined by daily semiquantitative cultural examination of rectal swabs. Using results obtained from offspring. 5 boars were retrospectively assigned the genotype ss, 1 was assigned Ss, and 2 were assigned SS. Nine sows were designated ss, 8 classified Ss and 4 SS. Ninety two pigs resulted from matings regarded as ss×ss; 89 (97%) of these were resistant to colonization and oedema disease. Of the 168 pigs from Ss×ss matings, 83 (49%) were resistant, while only 13 (9%) of 146 pigs from matings with at least one SS parent were classified resistant. The results are compatible with inheritance being controlled by one locus and with susceptibility dominating resistance to oedema disease.


title: Characterisation of rotavirus isolates from sub-clinically infected calves by genome profile analysis

text: Rotaviruses isolated from 43 sub-clinically infected calves from a single farm were analysed by genome profile analysis. The isolates showed genomic variation and eight different profiles were observed, including one which was atypical for Group A rotaviruses. The 3′ terminal labelling method for the analysis of genome profiles used in this study required only 1 ng of viral RNA, an increase of 1000-fold in sensitivity over ethidium bromide staining for detecting all rotavirus genome segments. However, dual infections involving two rotaviruses with distinct profiles could not be detected if the concentrations of the viruses differed by > 10-fold.


title: Effect of dual-subtype vaccine against feline immunodeficiency virus infection

text: Dual-subtype feline immunodeficiency virus (FIV) vaccine, consisting of inaactivated cells infected with subtypes A (Petaluma strain) and D (Shizuoka strain), was developed and tested for its vaccine efficacy against FIV infection in specific pathogen free (SPF) cats. Animals were monitored for proviral DNA by FIV-specific PCR and for FIV-specific antibody profiles by ELISA and virus-neutralization assays. In addition, blood from challenged cats was inoculated into naive SPF cats to confirm the viral status of the vaccinated cats. All cats immunized with Petaluma vaccine alone were protected against homologous Petaluma challenge, but only one of four cats was protected against heterologous Shizuoka challenge. More importantly, all cats immunized with the dual-subtype vaccine were protected against both Petaluma and Shizuoka challenges. These results suggest that a multi-subtype vaccine approach may provide the broad-spectrum immunity necessary for vaccine protection against strains from different subtypes.


docid: sprxbypw

title: The role of interferon in spontaneous cell-mediated cytotoxicity in pigs

text: Specific release of (51)Cr and the production of interferon (IFN) increased in parallel in a spontaneous cell-mediated cytotoxicity (SCMC) assay in which uninfected PK-15 cells or PK-15 cells persistently infected with transmissible gastroenteritis virus (PK-15-TGE cells) were used as targets, and peripheral blood lymphocytes (PBL) from a young adult pig were used as effector cells. Higher levels of both specific (51)Cr release and IFN were obtained in the assays containing PK-15-TGE cells. Co-cultivation of PBL from newborn piglets with PK-15-TGE cells yielded similar levels of IFN to those produced by co-cultivation of adult PBL and PK-15-TGE cells, but lower levels of IFN were produced by co-cultivation with uninfected PK-15 cells. Pretreatment of adult PBL with IFN augmented their SCMC effector activity for both PK-15 and PK-15-TGE cells in the (51)Cr release assay. Pretreatment of the PK-15-TGE target cells with IFN did not affect their release of (51)Cr in the SCMC reaction, while IFN pretreatment of PK-15 targets protected them against SCMC. In a single cell cytotoxicity assay the effects of IFN pretreatment on the effector adult PBL and on the PK-15 and PK-15-TGE target cells were confirmed, and SCMC incompetent PBL from neonatal piglets were rendered cytotoxic by pretreatment with IFN. PBL from newborn piglets bound to either target cell with the same frequency as PBL from SCMC competent adult pigs, and IFN pretreatment of either effector or target cells had no effect on target-binding frequency.


Detection of FCoV quasispecies using denaturing gradient gel electrophoresis


title: Lack of protection in vivo with neutralizing monoclonal antibodies to transmissible gastroenteritis virus

text: Monoclonal antibodies (Mabs) specific for the E1 and E2 surface glycoproteins of the transmissible gastroenteritis virus (TGEV) of swine were examined either alone or in combination to evaluate their potential value in protecting neonatal pigs against a lethal dose of TGEV. Cesarean-delivered colostrum-deprived (CDCD) piglets were given one pre-challenge dose of Mab and an equal dose of the same Mab at each successive feeding after challenge. In vivo challenge results demonstrated that neither Mabs given individually nor combinations of the Mabs were able to protect neonatal pigs against a lethal dose of TGEV. However, in parallel experiments, polyclonal antibodies from immune colostrum or serum were protective.


Why a European society for veterinary virology?


title: Antiviral activity of interferon against transmissible gastroenteritis virus in cell culture and ligated intestinal segments in neonatal pigs

text: Segments of jejunum in 5 to 6 days old piglets were surgically ligated, inoculated with transmissible gastroenteritis virus (TGEV) and 18 hours later the segments were fixed for histology or suspensions were prepared for plaque assay in swine testis (ST) cell cultures to determine the yield of virus. When the virulent Purdue strain of TGEV was used, villous atrophy was seen and TGEV antigen was demonstrated immunohistochemically in the villous enterocytes. The Miller M6 strain of virus produced less extensive lesions in the segments, but since it was titratable by plaque assay it was used in the subsequent yield reduction assays to determine the antiviral activity of interferon. When intestinal segments were inoculated simultaneously with either 3200 units of natural porcine interferon-α or up to 100000 units of recombinant human interferon-α(2a), and TGEV, there were no reductions in virus yield, although the same cytokines exerted an antiviral effect in ST cells treated in a similar way. However, virus yields were significantly reduced in intestinal segments in piglets treated parenterally with teh synthetic interferon inducer polyinosinic: polycytidylic acid 6 hours before challenge of the segments with TGEV. There was also a trend for the antiviral effects of interferon induction before challenge to be augmented by the inclusion of interferon with the virus inoculum. It was concluded that interferon would be ineffective as a therapeutic for TGEV, although it might be useful prophylactically.


title: Inhibitory effects of ribavirin alone or combined with human alpha interferon on feline infectious peritonitis virus replication in vitro()

text: The antiviral activities of ribavirin (1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxamide; virazole), either alone or in combination with recombinant human leukocyte (alpha) interferon (rHuIFN-α), were evaluated against feline infectious peritonitis virus (FIPV) in feline kidney-cell cultures. The 50% inhibitory dose (ID(50)) of ribavirin for uninfected, rapidly dividing cells was ∼17 μg ml(−1) whereas the ID(50) for FIPV was 2.5 μg ml(−1). The therapeutic index (TI) of ribavirin (i.e. the ratio of the minimum cell-toxic dose to minimum virus-inhibitory dose) was 6.8. Although a dose-dependent inhibition of viral infectivity occurred at non-toxic doses, maximum antiviral effects (⩾4 log(10) reduction in FIPV) occurred at cytotoxic doses. When low or moderate doses of ribavirin were combined with either 10 or 100 U of rHuIFN-α ml(−1), the resulting antiviral effects were significantly greater than the sum of the observed effects from either ribavirin or rHuIFN-α alone. Significant synergistic interactions with rHuIFN-α occurred at ribavirin doses of 1, 5, 12.5 and 25 μg ml(−1). Synergistic combinations of rHuIFN-α and ribavirin produced up to an 80-fold or a 200-fold relative increase in FIPV antiviral activities compared with that produced by equivalent doses, respectively, or ribavirin or rHuIFN-α alone. In cell growth studies, the addition of either 10 or 100 U of rHuIFN-α ml(−1) to test doses of ribavirin did not increase the anticellular effect observed with ribavirin alone; seemingly, the potentiation of ribavirin antiviral activity by rHuIFN-α was independent of any additive cytotoxic effects. Potentially, synergistic combinations of the two antiviral agents in vivo may decrease the therapeutic dose of ribavirin required for inhibition of FIPV and thus reduce drug toxicity.


title: Endocrinology and paracrinology: Cell surface peptidases in human endometrium

text: Three cell surface peptidases have been shown to be present in the human endometrium. Aminopeptidase N and neutral endopeptidase are detected on the endometrial stromal cells and decidual cells, while dipeptidyl peptidase IV is detected on the endometrial glandular cells and surface epithelium. As these cell surface peptidases can degrade a variety of biologically-active peptides including cytokines and growth factors, they are considered to be involved in the local metabolism of these molecules. In addition, recent studies have indicated that they are involved in local immune responses, cell attachment, and cellular maturation/differentiation of endometrial cells, and suggest an important role of these endometrial cell surface peptidases in implantation processes.


docid: ydrnanug

title: Detection of bovine viral diarrhea virus in serum from cattle by dot blot hybridization assay

text: A dot blot hybridization assay was developed for use as a rapid screening test to detect bovine viral diarrhea virus (BVDV) in serum from infected cattle. A 1.1. kilobase cDNA, prepared from the BVDV genome, was molecularly cloned and used in this study. Insert cDNA was removed from the pUC9 plasmid vector by Pst-I restriction endonuclease digestion and purified from plasmid DNA by agarose electrophoresis and electroelution. The hybridization probe was prepared by nick translation in the presence of gamma dCT(32)P and labelled to a specific activity of 2 × 10(8) cpm/μg of DNA. Specificity was determined by dot blot hybridization of infected cell culture supernate from nine different BVDV strains. The probe hybridized equally with all strains of BVDV tested, which included four cytopathic and five noncytopathic strains of BVDV. Serum was collected from veal calves with respiratory tract disease, unthriftiness, anorexia, and/or poor conditions. Serum samples were treated with nonidet P40 detergent and denatured with formaldehyde and heat prior to application on 1.2 micron nylon membrane filters using a vacuum dot blot apparatus. Hybridization was done under relatively stringent conditions (50% formamide at 42°C). A total of 141 serum samples from different calves were tested and of these samples, 55 (39%) were positive by dot blot hybridization for BVDV RNA. Eight calves (33%) out of 24, tested 3 to 4 weeks later, remained positive for BVDV RNA.


title: Animal immunodeficiency viruses

text: Feline immunodeficiency virus (FIV) has morphological, physical and biochemical characteristics similar to human immunodeficiency virus (HIV), the cause of AIDS in man. However, it is antigenically and genetically distinct from HIV; an antigenic relatedness with equine infectious anaemia virus has been demonstrated. FIV has been molecularly cloned and sequenced. Diagnostic tests are commercially available and attempts at preparing inactivated, subunit and molecularly engineered vaccines are being made in different laboratories. During FIV infection a transient primary illness can be recognized, with fever, neutropenia and lymphadenopathy. After a long period of clinical normalcy a secondary stage is distinguished with signs of an immunodeficiency-like syndrome. The incubation period for this stage can be as long as 5 years, during which gradual impairment of immune function develops. Many FIV-infected cats are presented for the first time showing vague signs of illness: recurrent fevers, emaciation, lack of appetite, lymphadenopathy, anaemia, leucopenia and behavioural changes. Later, the predominant clinical signs observed are chronic stomatitis/gingivitis, enteritis, upper respiratory tract infections, and infections of the skin. Neoplasias, neurological, immunological and haematological disorder are seen in a smaller proportion. The immunodeficiency-like syndrome is progressive over a period of months to years. Concomitant infection with feline leukaemia virus has been shown to accelarate the progression of disease. In vitro, phenotypic mixing between FIV and an endogenous feline oncovirus (RD114) has been demonstrated which leads to a broadening of the cell spectrum of the lentivirus. Bovine immunodeficiency virus (BIV) has been isolated only once, and all attempts to obtain additional isolates have failed; it has been recovered from the leucocytes of cattle with persistent lymphocytosis, lymphadenopathy, lesions in the central nervous system, progressive weakness and emaciation. As with the feline representative, BIV also was found to possess a lentivirus morphology and to encode a reverse transcriptase with Mg(++) preference; it replicates and induces syncytia in a variety of embryonic bovine tissues in vitro. Antigenic analyses have demonstrated a conservation of epitopes between the major core protein of BIV and HIV. The original isolate has been molecularly cloned and sequenced. Besides the three large open reading frames (ORFs) comprising the gag, pol, and env genes common to all replication-competent retroviruses, five additional small ORFs were found. Numerous point mutations and deletions were found, mostly in the env-encoding ORF. These data suggest that, within a single virus isolate, BIV displays extensive genomic variation. When random cattle sera from the Netherlands were tested by indirect immunofluorescence, Western blot and/or radio-immunoprecipitation, 0.7% appeared seropositive; thus BIV infection is not uncommon in one European cattle population.


title: Development of specific nucleic acid probes for the differentiation of porcine rotavirus serotypes

text: A dot blot hybridization assay is described for the detection and differentiation of porcine rotavirus serotypes. Recombinant complementary DNA (cDNA) representing gene 9 (the gene encoding the neutralization antigens in VP7 glycoprotein) from OSU (porcine rotavirus serotype 1) and Gottfried (porcine rotavirus serotype 2) strains were used to determine the optimal hybridization conditions which allow specific detection of group A porcine rotaviruses. Probes were prepared by excision of the inserts from the recombinant plasmids and radiolabeling of cDNA with (32)P by the random primer extension method. Probes were hybridized at various stringencies with viral RNA from different rotavirus serotypes bound to nylon membranes. Hybridization at low stringency (26% base pair mismatch for stable hybrid formation) had high sensitivity but low specificity. Hybridization at high stringency (16% base pair mismatch for stable hybrid formation) produced high specificity but decreased the sensitivity observed at low stringency. Probes were specific for rotavirus at both stringencies and did not hybridize with nucleic acids from other porcine viruses. Subgenomic gene 9 fragments were then tested to provide more specific probes. A 322 bp fragment from OSU gene 9 between nucleotides 382 and 704 and a 266 bp fragment from Gottfried gene 9 between nucleotides 230 and 496 were found to be specific as hybridization probes. These studies demonstrated the feasibility of the dot blot hybridization assay using subgenomic fragments of gene 9 to detect and differentiate serotypes of porcine rotavirus. Additional studies are warranted to further evaluate the sensitivity and the capability of these probes to detect porcine field isolates of the same serotype.


title: Different stabilities to bile among feline calicivirus strains of respiratory and enteric origin

text: Feline calicivirus (FCV) strains isolated from feces (E-FCV) were compared with FCV strains of respiratory origin (R-FCV). All strains were shown to be labile at pH 3.0. All strains except one strain of E-FCV were found to be sensitive to the action of trypsin. When exposed to bile salt (deoxycholic acid sodium salt), all R-FCV strains were markedly inactivated, but none of the E-FCV strains was inactivated. It was possible to select bile-resistant substrains from a bile-sensitive strain.


title: Antigenic differences between European and American isolates of porcine reproductive and respiratory syndrome virus (PRRSV) are encoded by the carboxyterminal portion of viral open reading frame 3

text: Antigenic differences between European and American isolates of porcine reproductive and respiratory syndrome virus (PRRSV) were revealed by serologic analysis of a recombinant protein derived from PRRSV open reading frame 3 (ORF 3). The hydrophilic carboxyterminal 199 amino acids encoded by the ORF 3 of a European (Lelystad) isolate of PRRSV were expressed as a recombinant fusion protein (BP03-P) in a baculovirus gene expression system. Sera from gnotobiotic swine exposed to prototypic reference European and American isolates of PRRSV and sera from conventionally reared European and American swine convalescing from naturally acquired PRRSV infections were used to characterize the BP03-P protein. Sera from gnotobiotic and conventionally reared swine exposed to European isolates of PRRSV were significantly more reactive (P < 0.01) with BP03-P than were the corresponding American PRRSV antisera using the indirect immunoperoxidase monolayer assay (IPMA). Prototypic European, but not American, PRRSV antisera also recognized BP03-P using western immunoblotting and radioimmunoprecipitation assay (RIPA) procedures. However, gnotobiotically derived antiserum to an atypical American-origin PRRSV was reactive with BP03-P by both IPMA and western immunoblot. Despite a predicted potential for N-linked glycosylation, studies with tunicamycin and peptide-N-glycosidase F (PNGase F) indicated that BP03-P was not N-glycosylated in either insect cell cultures or Trichoplusia ni larvae infected with the recombinant baculovirus. Sera from rabbits inoculated with BP03-P failed to neutralize both the European (Lelystad) and American (ATCC VR-2332) reference isolates of PRRSV and did not react by IPMA with PRRSV-infected cell cultures. Taken together, the data suggest that the carboxyterminal portion of PRRSV ORF 3 encodes a non-neutralizing viral peptide that is partially responsible for the serologic differences noted between European and most American isolates of PRRSV.


Feline leukemia virus vaccine: New developments


title: Contribution of molecular biology to the study of the porcine interferon system()

text: We have performed molecular studies on the pig interferon (IFN) system (i) to analyse the role played by endogenous IFN in neonatal viral enteritis such as transmissible gastroenteritis and possibly to obtain, via recombinant DNA technology, a new anti-infectious and immunomodulatory agent in this species, (ii) to characterize the structure and biological functions of the IFN-like antiviral activity produced by the porcine embryo at the time implantation in the uterus. By probing porcine genomic libraries with human and porcine IFN-α probes to isolate related genes, we have shown that the porcine IFN-α multigene family included, like several other mammalian species, two subfamilies of related but distinct genes. Class I subfamily contains at least 11 loci, located on chromosome no. 1, among which nine have been cloned and two (potentially functional) sequenced. Class II subfamily, which is specifically expressed by the embryo of ruminants before implantation, contains at least seven loci among which six have been cloned. One of the sequenced class I loci: PoIFN-α1 encodes a 189 amino acids (AA) preprotein. After removal of the sequence encoding the putative signal peptide (23 N-terminal AA) this gene was inserted into an Escherichia coli bicistronic expression vector allowing intracellular synthesis of mature porcine IFN-α1 (methionyl IFN-α1). Expression of the recombinant protein was optimized by insertion of a seven base pairs long random synthetic sequence in the intercistronic region, followed by cloning in E. coli and immunodetection of clones expressing high amounts of recombinant protein. The E. coli strain obtained produced high levels of a 18 000 Da protein exhibiting the same in vitro overall biological properties as leucocyte derived porcine IFN (LeuIFN). However, it had a stronger antiviral effect on porcine cells than LeuIFN. After immunoaffinity purification to a specific activity of 5−10 × 10(7) International Units (IU)/mg of protein, pharmacokinetic and pharmacological studies were realized to determine the in vivo half life of this rIFN-α in the pig. These experiments revealed no major toxic effects in newborn (given 5 × 10(6)IU/kg) or adult (1 × 10(6)IU/kg) pigs. A significant pyrogenic effect (+ 1.5°C) was noted only in the adults.


title: Serological and genotypic characterization of group a rotavirus reassortants from diarrheic calves born to dams vaccinated against rotavirus()

text: Two strains of bovine rotavirus (BRV), designated strain Nebraska Scottsbluff-1 (NS-1) and NS-2, were isolated from 2 neighboring cow-calf beef cattle ranches where dams had been vaccinated with a commercial vaccine containing group A BRV strain Neonatal Calf Diarrhea Virus (NCDV)-Lincoln (P1:G6). Nothern blot by hybridizations using whole genomic RNA probes indicated that strains NS-1 and NS-2 had identical group A RNA electrophoretic patterns and were homologous at all gene segments. Strain NS-1 was compared with reference group A BRV strains using serological and genotypic methods. In vitro virus neutralization assays indicated that strain NS-1 was neutralized by a G6-specific neutralizing monoclonal antibody (mAb) and guinea pig hyperimmune serum (GPHS) raised against BRV strain B641 (P5:G6), but not by G10-specific neutralizing mAb or GPHS raised against BRV strain BRV strain B223 (P11:G10). Nucleic acid hybridization experiments using whole-genomic RNA probes revealed that gene segment 4 of strain NS-1 differed from BRV strains NCDV-Lincoln and B223, but hybridized with strain B641. Conversely, gene segment 5 of strain NS-1 hybridized with BRV strain B223, but not with BRV strains NCDV-Lincoln and B641. A G-specific cDNA probe produced by reverse transcription polymerase chain reaction (RT-PCR) amplification of strain NS-1 hybridized specifically only with G6 strains NCDV-Lincoln and B641, but not with G10 strain B223. Co-electrophoresis experiments using strains NS-1, B641, and B223 further confirmed these results, suggesting that strain NS-1 was a naturally-occurring reassortant BRV between strains B641 and B223. Taken together these results indicated that a naturally-occurring group A BRV reassortant with a P gene different from the vaccine virus was responsible for the diarrheal syndrome observed on both ranches. Results from this study also indicate the existence of at least 2 different gene segments 5 among group A BRV infecting cattle.


title: Seroepidemiology of Breda virus in cattle using ELISA

text: Two direct blocking enzyme linked immunosorbent assays (ELISA) for the detection of antibodies to Breda virus in sera of cattle were compared. An ELISA with consecutive addition of antigen and test serum to an antibody-coated plate gave higher positive: negative absorbence ratios than an ELISA in which antigen and test serum were added simultaneously. Sera collected from breeding and fattening herds in The Netherlands (n = 1313) and the F.R.G. (n = 716) were tested, and antibodies to Breda virus were demonstrated in 94% of adult cattle. Ninety percent of newborn calves had high levels of maternal antibodies, which waned until the age of 3 months. Active seroconversion occurred between 7 and 24 months in most animals.


title: Dysentery in calves caused by an atypical strain of Escherichia coli (S102-9)

text: Dysentery lasting 4–8 days was produced in five 4-day-old colostrum-fed calves, after inoculation with an atypical strain of Escherichia coli S102-9; peak excretion of S102-9 occurred during the period of dysentery. Two calves were killed when clinical signs were most severe and bacteria were seen attached to the surfaces of enterocytes in the large intestine; microscopic lesions were seen in these areas. The lesions were identical to those previously reported in a natural outbreak of dysentery in calves, from which E. coli S102-9 was isolated, and to those seen in gnotobiotic calves experimentally infected with S102-9. Reinfection of the three surviving calves 16–20 days later with S102-9 and primary infection of two calves aged 24 and 51 days did not cause dysentery. Four of 659 coliforms isolated from field outbreaks of calf diarrhoea resembled the atypical strain S102-9. These four isolates and S102-9 did not produce heat-stable enterotoxin, but all produced a toxin cytopathic for Vero and HeLa cells. Two of the four isolates were inoculated alone into 4-day-old gnotobiotic calves deprived of colostrum; neither calf developed dysentery but microscopic lesions identical to those produced by S102-9 were detected in the large intestines of both animals.


title: Detection of avian infectious bronchitis viral infection using in situ hybridization and recombinant DNA

text: A recombinant DNA probe with specificity for the 3′ end of genomic RNA from the Ark 99 strain of infectious bronchitis virus (IBV) was found to hybridize with extracted RNA of three strains with the Ark serotype, as well as the Mass41, Holl52, Gray, JMK, Conn, Fla and SE17 strains of IBV. Viral infection was detected in the cytoplasm of chicken embryo kidney cells inoculated with Mass41, Ark99, SE17 or two recent field isolates of IBV using in situ cytohybridization and a biotinylated probe. In vivo infections were detected in individual cells of tracheas and lungs 2, 4, and 6 days after inoculation of chicks with Mass41 and Ark99. In situ hybridization of Ark99 infected tissue sections using (32)P-dATP labelled probe indicated that more viral replication was present in the trachea on day 4 than either days 2 or 6; whereas more viral RNA was found in the lungs on day 6 than days 2 or 4 after inoculation.


title: Molecular biology of transmissible gastroenteritis virus

text: The causative agent (TGEV) of porcine transmissible gastroenteritis belongs to the Coronaviridae, a family of enveloped viruses with a positive, single-stranded RNA genome. Important progress has recently been made concerning the molecular biology of TGEV. The research work of our group has been focused on two main aspects: genome structure and functional domains of the envelope proteins. TGEV genomic RNA is organised into seven regions. The sequence of six of them, i.e. the 3′ most 8300 nucleotides, has been established from cDNA clones. Three genes encoding the structural proteins, the peplomer protein E2, the transmembrane protein E1 and the nucleoprotein, have been identified. Additional open reading frames allowed for the prediction of four non-structural polypeptides, the role of which remains to be discovered. The remaining part of the genome (estimated length 20 kb) is thought to encode the polymerase. Expression of TGEV genes involves the production of six subgenomic mRNAs, which together with the virion RNA, form a 3′ terminal nested set. The peplomer glycoprotein E2 (220 kDa) is 1431 residues long and highly glycosylated. Several domains were identified, including a C-terminal anchoring region and at least four major antigenic sites, which cluster in the amino half part of the molecule. Two sites containing most of the critical neutralisation determinants are highly conserved among TGEV strains. The glycoprotein E1 (29kDa) is mostly embedded in the membrane and plays a crucial role in the virion architecture. However, a short N-terminal domain protruding out of the particle mediates complement-dependent neutralisation, and induces alpha interferon synthesis, likely through a direct interaction with the lymphocyte membrane.


title: Characteristics of vero cytotoxin producing Escherichia coli associated with intestinal colonization and diarrhea in calves

text: Isolates of Escherichia coli which produce Vero cytotoxin (VTEC) were obtained during 1983–1989 from calves raised in 5 north-central states of the USA. All of the calves experienced intestinal epithelial colonization by VTEC, diarrhea or both; twelve of the calves had bloody diarrhea. Twenty one isolates were serogroup O111 and the others were O103, O69, O45, O26, O5, or non-typable (4 isolates). All but one of the isolates hybridized with the CVD419 probe which identifies most VTEC strains. Thirty two isolates hybridized with the VT1 probe, 3 with both the VT1 and VT2 probes, and one with neither probe. The culture filtrate of the VT probe negative isolate was partially neutralized by SLT I monoclonal antibody. For the other isolates, the results of toxin neutralization by anti-SLT I and anti-SLT II monoclonal antibodies corresponded exactly with the VT1 and VT2 probe hybridization results. Three of the strains adhered in a localized manner to HEp-2 cells and Intestine 407 cells.


title: The sites of early viral replication in feline infectious peritonitis

text: The sites of early replication of feline infectious peritonitis virus were tudied following oral inoculation of specific-pathogen-free (SPF) cats with virus grown in cell cultures. Viral antigen was first detected by immunofluorescence in the tonsils and small intestine within 24 h of inoculation, and was later found in caecum, colon, mesenteric lymph nodes and liver. However, histological changes in the gut did not appear until relatively late in the course of infection. Virus was recovered from the oropharynx and the faeces from as early as the second or third day after inoculation, and shedding continued until euthanasia.


title: In vitro properties and experimental pathogenic effect of three strains of feline immunodeficiency viruses (FIV) isolated from cats with terminal disease

text: Three strains of virus isolated from peripheral blood mononuclear cells (PBMC) of sick cats were identified as feline immunodeficiency virus (FIV) on the basis of in vitro cytopathic effect, T-lymphotropism, ultrastructural morphology and magnesium-dependent reverse-transcriptase activity. The pathogenic properties of two isolates were studied in 13 experimentally infected cats. The primary phase of infection was characterised by a range of haematological (neutropenia, lymphopenia, presence of atrypical lymphocytes) and clinical alterations (fever, various signs lasting several weeks, generalised lymphadenopathy persisting for several months) and specific seroconversion. A correlation between the inoculated dose of virus and the intensity and duration of clinical signs was observed. The primary phase was followed in the 10 surviving cats by a stage of asymptomatic seropositivity of undetermined duration but which has persisted for over 35 months for the earliest infections. Viruses reisolated several weeks or months after experimental infection retained the same in vitro properties as the initial isolates.


title: The neurobiology of canine distemper virus infection

text: Canine distemper virus (CDV) invades the nervous system and replicates in neurons and glial cell of the white matter during a period of severe viral induced immunosuppression. Demyelination occurs in infected white matter areas in the absence of inflammation. The mechanism of demyelination is not apparent because there is no ultrastructural evidence of viral replication in the oligodendrocytes, the myelin producing cells. However, brain tissue culture studies have shown that oligodendrocytes support transcription of all CDV genes and later on degenerate, although no viral proteins can be found in these cells. It remains to be shown how such a restricted infection leads to demyelination. Concomitant with immunologic recovery during the further course of the disease, inflammation occurs in the demyelinating lesions with progression of the lesions in some animals. A series of experiments in vitro suggested that chronic demyelination is due to a bystander mechanism associated with the virus-induced immune response in which antibody dependent cell-mediated reactions play an important role. The progressive, or even relapsing, course of the disease is associated with viral persistence in the nervous system. Persistence of CDV in the brain appears to be due to non-cytolytic selective spread of the virus with very limited budding. In this way CDV escapes immune surveillance.


title: Antigenic and biological diversity among transmissible gastroenteritis virus isolates of swine

text: Twenty-four field isolates of transmissible gastroenteritis virus (TGEV) were isolated and examined for antigenic and biological characteristics. Most TGEV isolates produced a typical cytopathic effect (CPE) in swine testis (ST) cell culture, which included a ballooning or lifting away of the infected cells from the cell monolayer with heavy granulation evident. Minor variations in CPE were observed with one isolate, IA-145. Protein profiles of the TGEV isolates as determined by SDS-PAGE were essentially identical, with the exception of the isolate IA-101. The TGEV isolate IA-101 presented a higher molecular mass M protein and lacked an N protein doublet that was present in all other TGEV isolates. The TGEV isolates were shown to be closely related antigenically by using hyperimmune sera in a virus neutralization (VN) test. Some antigenic diversity was detected by utilizing monoclonal antibodies (mAbs) in a VN test. Titers of the mAbs were highest with the homologous Miller TGEV, and one virus isolate, IA-156, was very poorly neutralized with the mAbs used in this study. Indirect immunofluorescence assay (IFA) results were similar to those obtained by the VN test. These studies show that some biologic and antigenic diversity exists among TGEV isolates.


title: Intestinal changes associated with rotavirus and enterotoxigenic Escherichia coli infection in calves

text: Newborn calves inoculated with rotavirus, enterotoxigenic Escherichia coli (ETEC) serotype 020:K′ × 106′:K99:HNM, either alone or in combination, became depressed, anorectic, diarrhoeic and dehydrated. ETEC did not adhere to the intestine although there was extensive proliferation in the lumen. Only slight mucosal changes were induced by ETEC and the activity of membrane bound lactase remained normal. More severe mucosal damage and a decrease in lactase activity were found in newborn calves inoculated with either rotavirus or rotavirus and ETEC in combination. The most severe clinical illness was found in calves inoculated with both rotavirus and ETEC. Calves inoculated at 1 week of age with either rotavirus or ETEC remained clinically normal. Rotavirus infection produced slight mucosal changes and a reduction of lactase activity. In contrast, colostrum-fed or suckling calves up to 2 weeks old inoculated with both rotavirus and ETEC became clinically affected, showed severe mucosal damage and decreased lactase activity. There was no bacterial adhesion to the intestinal mucosa as observed by immunofluorescent labelling and light microscopy.


title: Characterization of rotaviruses isolated from pigs with diarrhoea in Venezuela

text: The prevalence of porcine rotavirus infection was studied in 15 different herds located in the north-western region of Venezuela. The presence of rotavirus was studied by direct electron microscopy (EM) and by an enzyme-linked immunosorbent assay (ELISA). From 136 samples analyzed during the six months of the study (September 1983–February 1984), 38 (27.9%) were found to be positive for rotaviruses, with infection more common in animals that were 4–6 weeks old. Atypical rotaviruses were not detected in any of the samples examined. Most rotavirus positive specimens were subgrouped using specific monoclonal antibodies in an ELISA test. The majority of the samples (26 out of 38) were found to exhibit Subgroup I antigenicity. Only two specimens, collected from the same herd in two consecutive months, were found to belong to Subgroup II. To characterize further the circulating rotaviruses, electrophoretic analysis of the RNA genome was performed on samples selected from nine different herds. Great variability in the RNA electropherotypes was observed. No correlation was found between subgroup specificity and the migration of the two smaller segments (Genes 10 and 11), as has been described for human rotaviruses.


title: Binding of porcine transmissible gastroenteritis virus by enterocytes from newborn and weaned piglets

text: Enterocytes were harvested by chelation in a series of seven fractions from the tips of the villi to the crypts of the jejunum of newborn or weaned piglets. Binding of the low cell culture passaged Miller-6 strain of transmissible gastroenteritis virus (TGEV) to villous enterocytes from newborn piglets was at a high level, similar to that observed to culturedswine test is (ST) cells. Binding of the virus to cryptal enterocytes from newborn piglets or to villous or cryptal enterocytes from weaned piglets was significantly lower. In a competitive virus binding assay with radiolabelled virus, the binding of TGEV to ST cells was found to be saturable, while binding to MDBK cells, in which the virus fails to replicate, was at a lower level and was non-saturable. In the same assay, virus binding to the villous enterocytes from the jejunum of a newborn piglet was saturable, while binding to cryptal enterocytes from a newborn piglet, and to villous and cryptal enterocytes from a weaned piglet, was non-saturable. It was concluded that the high susceptibility of newborn piglets to TGEV infection, and the tropism of the virus for villous enterocytes, may relate to the presence of specific, saturable binding sites on the plasma membrane of villous enterocytes.


title: Reduction in morbidity due to diarrhea in nursing beef calves by use of an inactivated oil-adjuvanted rotavirus — Escherichia coli vaccine in the dam

text: An outbreak of neonatal diarrhea occurred among beef calves (2000 animals) from one large Argentinian farm in 1985. Rotavirus was detected in 78% (106136) and enterotoxigenic Escherichia coli in 1.5% of the samples (2136) obtained from sick calves. In comparison rotavirus was identified in only 1.6% (163) of the samples from clinically healthy calves. The rotavirus strain responsible for the outbreak was characterized as serotype 6 belonging to group A. In the following three years the protective capacity of a combined rotavirus—E. coli inactivated vaccines administered to the dams during the last third of the gestation period was evaluated on this farm by comparison of morbidity due to diarrhea in calves from vaccinated vs. placebo cows within the same year. The morbidity due to diarrhea among calves from dams in the vaccinated and placebo groups was 34% and 77%, respectively in 1986; 23% and 47% in 1987, and 15% and 34%, in 1988. In 1987 morbidity of diarrhea in calves born from vaccinated heifers was 54% and 74% in calves from placebo heifers. In 1988 morbidity from diarrhea was 41% and 54%, respectively among calves in these two groups. In all experiments, calves from heifers showed significantly greater morbidity than calves from cows. Differences in diarrhea morbidity between the vaccinated and placebo groups were statistically significant (P<0.05). Additional studies showed that the diarrhea had a significant influence (P<0.05) on the average live weight of the calves at weaning (5 to 7 months) with an average weight loss of 7.8 kg per calf among the calves affected with diarrhea.


docid: zgeorzsf

title: Virus shedding and immune responses in cats inoculated with cell culture-adapted feline infectious peritonitis virus

text: Eight specific pathogen-free cats were inoculated orally or parenterelly with a cell culture-adapted strain of feline infectious peritonitis virus (FIPV). Faeces and oropharyngeal swabs were monitored daily for infectious virus by inoculation of feline embryo long cells. Virus was recovered from both sites for ∼ 2 weeks after inoculation, before clinical signs of disease developed. Peripheral blood lymphocytes collected from these cats were tested in an in-vitro blastogenic assay using concanavalin A (con A) and FIPV antigen. All cats showed a profound suppression of the response to con A which only recovered to pre-inoculation levels in 2 cats, one of which survived. These 2 cats also responded to FIPV antigen on the 21st day after infection, the greater response being in the survivor. The other cats, surviving 16–18 days, developed no response to FIPV antigen. Antibody titres, measured by immunofluorescence and by virus neutralization, rose rapidly to very high levels in all cats, regardless of the route of inoculation.


title: Interferon induction in porcine leukocytes with transmissible gastroenteritis virus

text: Leukocytes were harvested from the peripheral blood, mesenteric lymph node and small intestinal lamina propria from groups of three piglets before, and 1,2 and 3 weeks after infection with virulent transmissible gastroenteritis virus (TGEV) at 2 weeks of age. The donor piglets developed clinical signs of transmissible gastroenteritis which persisted for up to 3 days, and they developed peak serum titres of TGEV-neutralizing antibodies 2 weeks post-infection. The leukocytes were cultured in the presence of pokeweed mitogen (PWM), various dilutions of purified TGEV, or control media for 3 or 5 days, and the culture supernatants were tested for antiviral activity in MDBK cells challenged with vesicular stomatitis virus. The antiviral activity was characterized as porcine interferon (IFN)-α or porcine IFN-τ on the basis of its stability at pH 2.0 and neutralization by anti-human IFN-α antibodies. Viability of the leukocytes in culture, determined by trypan blue exclusion, was highest for the peripheral blood leukocytes and lowest for the mesenteric lymph node leukocytes. There were no consistent differences in antiviral activity between cultures incubated for 3 or 5 days. Porcine IFN-α was found in the supernatants of the leukocyte cultures stimulated with TGEV antigen, harvested before or after infection of the donor piglets with TGEV. Porcine IFN-τ was demonstrated in the supernatants of the leukocyte cultures stimulated with PWM, more frequently when the leukocytes were harvested post-infection. This was the first demonstration of IFN induction in vitro in leukocytes from porcine gut-associated lymphoid tissue.


title: Antibodies to berne virus in horses and other animals

text: After inoculation into 2 foals, Berne virus induced neutralizing antibody, but did not cause clinical symptoms. In a horizontal study of seropositive mares and their offspring, a decline of maternal antibodies and a sudden synchronous seroconversion in all foals were observed, again without clinical symptoms. The virus is widespread in the Swiss horse population and has been so during the last decade; rises in antibody titers were noted in 9% of paired sera sampled at random. Positive reactions were also obtained in serum neutralization tests and ELISA using small numbers of horse sera from Germany, France and the U.S.A. The results of neutralization tests and ELISA were correlated in 83% of random samples tested; 13% were neutralization-positive and ELISA-negative and in 4% the inverse was observed. Neutralizing activity was found in the sera of other ungulates (cattle, goat, sheep and pig), laboratory rabbits and 2 species of wild mice (Clethrionomys glareolus and Apodemus sylvaticus). Inconclusive results were obtained with feline and human sera; those from dogs and foxes (Vulpes vulpes) were consistently negative. The probable occurrence of antigenic variants in Berne-type viruses is discussed.


title: Resistance of Berne virus to physical and chemical treatment

text: Thermal inactivation of Berne virus proceeded at a linear rate between 31°C and 43°C. Storage at temperatures lower than −20°C preserved the infectivity, while at 4°C appreciable loss occurred between 92 and 185 days. Freeze-drying or desiccation at 22°C caused only insignificant loss of infectivity. Virus preparations were not affected by pH values between 2.5 and 10.3. Inactivation by UV occurred more rapidly than with herpes, toga and rhabdoviruses. Berne virus infectivity was sensitive to pronase and B. subtilis proteinase. It was not inactivated by trypsin and chymotrypsin treatment, which resulted in enhancement of infectivity; low concentrations of pronase (< 10 μg ml(−1)) had a similar effect on Berne virus. Neither phospholipase C or RNase, alone or in combination, nor sodium deoxycholate (0.1%) inactivated the virus; in contrast, Triton X-100 (0.1%, 1.0%) caused rapid inactivation with a constant level of residual infectivity.


title: Survey of rotavirus infection in a dairy herd: comparison between polycrylamide gel electrophoresis and two commercial tests

text: A survey of rotavirus infection in a dairy herd with a history of neonatal diarrhoea was carried out. Faecal samples taken from 15 cows before and after calving as well as faeces taken from their calves daily from birth to two weeks of life were tested for rotavirus by polyacrylamide gel electrophoresis (PAGE) and compared with an ELISA and a latex agglutination commercial test. Rotavirus excretion was not detected in faeces from cows around parturition by any of the three tests. However, all of their calves shed rotaviruses during the observation period. The onset of rotavirus excretion determined by PAGE ranged from day 2 to day 8 of life (day 4.8± 1.8 on average) and lasted for 4 to 7 days (5.3±1.1 days on average). Chi-square test showed a significant association (P=0.0001) between the presence of rotavirus and the altered consistency of calves faeces. All the three tests showed similar results (overall agreement 92.5%) but discrepancies were detected mainly at the beginning or at the end of the rotavirus excretion period. Results obtained with both commercial kits closely paralleled each other and parameters other than sensitivity, specificity, diagnostic accuracy or predictive values have to be considered as selection criteria.


title: A case-control questionnaire survey of risk factors for porcine reproductive and respiratory syndrome (PRRS) seropositivity in Danish swine herds

text: Sixty-eight case herds seropositive to porcine reproductive and respiratory syndrome (PRRS) were compared to 128 seronegative controls in a double-blinded questionnaire survey. The study indicated no increased risk of PRRS seropositivity for herds using artificial insemination with semen from PRRS seropositive AI-stations. Also the herd-size was non-related to the risk of PRRS seropositivity, indicating that air-borne spread of PRRS may not have been a predominant feature in Denmark. Introduction of replacement breeding animals from seropositive breeding- and multiplying herds significantly increased the risk of a herd being PRRS seropositive, as did introduction of 25 kg pigs for feeding. PRRS seropositivity was in the farmers' opinions associated with abortions in sows, early farrowing, high postweaning mortality and low weight gain in fattening pigs. However, the reported frequencies of problems were relatively low.


title: Effect of experimentally-induced villus atrophy on adhesion of K88ac-positive Escherichia coli in just-weaned piglets

text: Three- to four-week-old, just-weaned piglets were infected with transmissible gastroenteritis (TGE) virus and the next day with K88ac(+) enterotoxigenic Escherichia coli (ETEC). Histological examination of caudal jejunum and ileum of piglets killed 2–3 days after virus challenge (1–2 days after ETEC infection) revealed severe villus atrophy especially in the jejunum compared with controls (P<0.05). Four-5 days after TGE virus infection villus length increased and after 7 days it was near normal. Villi scraped from jejunal and ileal mucosa of the piglets were incubated in vitro with K88ac(+)E. coli and the number of bacteria adhering to 250 μm villus brush border was counted. Attachment of bacteria to villi of piglets killed 2–3 days after TGE virus infection was significantly decreased in comparison with adhesion to villi of non-infected piglets or of piglets killed 7 days after the virus infection. Correlation between in vitro adhesion and villus height was 0.6649 (P<0.001). The results suggest that the experimentally-induced villus atrophy was attended with a temporarily diminished susceptibility of villus enterocytes to adhesion of K88ac(+)E. coli.


title: Evaluation of an enzyme-linked immunosorbent assay for the detection of transmissible gastroenteritis virus antibodies

text: An enzyme-linked immunosorbent assay (ELISA) using a detergent-solubilized antigen of purified virus was developed for detection of antibody against porcine transmissible gastroenteritis (TGE) virus in swine serum. The ELISA demonstrated antibody responses in pigs immunized intramuscularly with the attenuated TO-163 strain of TGE virus and in pigs orally infected with the virulent Shizuoka strain of the virus. The results of the ELISA were well correlated with those of the neutralization test. These results indicate the usefulness of the ELISA as a serological tool for TGE virus antibody.


title: Immunoblot studies in the differential diagnosis of porcine brucellosis: an immunodominant 62 kDa protein is related to the mycobacterial 65 kDa heat shock protein (HSP-65)

text: Smooth Brucella spp. share certain lipopolysaccharide antigens with other bacteria, resulting in serological cross-reactions which can prevent the definitive diagnosis of brucellosis. To identify other antigens with serodiagnostic potential, immunoblot studies following sodium dodecyl sulphate - polyacrylamide gel electrophoresis were carried out. Sera from pigs experimentally infected with Brucella suis and naturally infected feral pigs, sera from pigs from a farm with a known history of Yersinia enterocolitica 0:9 infection, Brucella Complement Fixation Test (CFT) reactor pigs (aetiology unknown) and pigs from consistently Brucella CFT negative farms were examined. Although B. suis infected pigs recognized a total of nine B. melitensis antigens, individual pigs rarely recognized more than three antigens in the range. A 62 kDa antigen was recognized by the majority (73%) of the Brucella infected pigs, but only by 10 to 23% of pigs from the other groups. This antigen was shown to be the Brucella homologue of the ubiquitous 65 kDa heat shock protein (HSP-65) family by immunoblot studies with 14 monoclonal antibodies to the Mycobacterium leprae HSP-65. Only four of these monoclones (Y1.2, ML-30, D7C and IIIC8) identified the B. melitensis 62 kDa protein suggesting that unshared, potentially Brucella specific, regions exist. Sera from Y. enterocolitica 0:9 infected pigs, CFT reactor pigs (aetiology unknown), CFT negative pigs and hyperimmune pig serum raised to Y. enterocolitica 0:9 also recognized B. melitensis antigens, most notably a 17 kDa protein. This antigen appears to be a common cross-reactive protein.


title: Homotypic and heterotypic serum and milk antibody to rotavirus in normal, infected and vaccinated horses

text: The homotypic and heterotypic antibody response to rotavirus was determined in three pony mares and their foals. The normal concentrations of anti-rotavirus antibodies in mares ‘milk and mares’ and foals′ serum over the first 10 weeks post-partum were measured using IgA, IgG and rotavirus sero-type-specific enzyme linked immunosorbent assays. Experimental infection of the foals with serotype 3 equine rotavirus produced a rapid, serotype-specific response which peaked 10 days after infection and a slower heterotypic response which peaked 32 days later. In contrast, vaccination of the mares with an inactivated, adjuvanted serotype 6 bovine rotavirus produced a heterotypic response similar to that of the homotypic response in both serum and milk, although the predominant response in serum was IgG, while in milk it was IgA. These results suggest that non serotype-restricted passive protection of foals against rotavirus may be achieved by parenteral vaccination of mares.


title: An ELISA for detection of antibodies against porcine epidemic diarrhoea virus (PEDV) based on the specific solubility of the viral surface glycoprotein

text: Viral proteins of porcine epidemic diarrhoea virus (PEDV) were extracted from the cytoplasm of infected Vero cells using hypotonic conditions and a non-ionic detergent. Both the pH and the NaCl concentration of the extraction buffer were varied in attempts to increase the solubility of the virion spike glycoproteins (S-protein) and of the nucleocapsid proteins (N-protein). Monoclonal antibodies, hyperimmune sera and convalescent pig sera were used to identify and monitor these proteins by immunoprecipitation and Western blots. The solubility of the S-protein was optimal at pH 4, whereas that of the N-protein was optimal at pH 9. Consequently, it was possible to enrich for either S-protein or N-protein; increases in the NaCl concentration of the buffer were of no advantage in this respect. Enriched preparations of the S-protein and N-protein were used as ELISA antigen for the S-ELISA and N-ELISA, respectively. The S-ELISA proved to be the more effective of the two immunoassays. Antibodies against S-protein remained detectable for longer periods of time than anti-N-protein antibodies in the sera of PEDV-infected pigs. Using this ELISA of increased sensitivity, it was observed that only a small number of farms in Switzerland had been infected with PEDV.


docid: bpeondjd

title: Studies on bovine breda virus

text: Diarrheic feces from 21 calves were examined by electron microscopy and 16 contained particles morphologically similar to those of Breda virus. The particles were spherical or elongated, 60–270 nm in greatest dimension and had surface spikes 9–13 nm long. Convalescent serum from a human patient with Breda virus-associated diarrhea reacted with one of the bovine viruses by immune electron microscopy, suggesting a serological resemblance between human and bovine Breda-like viruses. Immune electron-microscopy and immunofluorescence demonstrated that isolates of bovine Breda virus from the U.S.A. were related to the French virus. One of the viruses had a density in sucrose solution of 1.16, similar to the value for Berne virus.


title: Antigenic variation of porcine transmissible gastroenteritis virus detected by monoclonal antibodies

text: Mouse myeloma cells (SP2/O) were fused with spleen cells from BALB/c mice immunized with detergent-solubilized antigen of purified virus, and 21 monoclonal (MC) antibodies reactive in enzyme-linked immunosorbent assay with the TO-163 strain of porcine transmissible gastroenteritis (TGE) virus were obtained. Of these MC antibodies, 14, 6 and 1 were IgG1, IgG2a and IgM, respectively. All of the MC antibodies contained light chains of the kappa type. Of these MC antibodies, 8 were found to have neutralization (NT) activity against the TO-163 strain. Comparison of 7 strains of TGE virus by NT tests using our panel of MC antibodies confirmed their close antigenic relationships, but also revealed the occurence of distinct antigenic differences. These results suggest that there may be at least 6 different epitopes involved in NT reaction on the virion of the TO-163 strain. This notion was confirmed by the competitive binding assay.


title: Applications of nucleic acid probes in veterinary infectious diseases

text: Nucleic acid probe technology is increasingly being used in basic research in veterinary microbiology and in diagnosis of infectious diseases of veterinary importance. This review presents an overview of nucleic acid probe methodology and its applications in veterinary infectious diseases. The major applications of nucleic acid probes include detection of pathogens in clinical samples, especially those organisms which are fastidious and difficult to cultivate, differentiation of virulent from a virulent organisms and vaccine strains from wild type isolates, typing of microorganisms mapping genes, screening libraries of cloned DNA for specific genes, detection of latently infected or carrier animals, study of mechanisms of pathogenesis, epidemiological studies and food safety.


docid: zkjnzqax

title: Inducement of cytopathic changes and plaque formation by porcine haemagglutinating encephalomyelitis virus

text: ESK cells were shown to be a good medium for propagating the 67N strain of porcine haemagglutinating encephalomyelitis virus, although no cytopathic effect was observed. The virus induced a readily recognizable cytopathic effect in ESK cells, when a non-cytotoxic amount of diethylaminoethyl-dextran (DEAE-dextran) was incorporated in the culture medium. Based on this finding, a sensitive, practical assay method for the virus was developed. When DEAE-dextran was incorporated in the agar overlay medium, 67N virus formed plaques in ESK cell monolayers. The cytopathic effect as well as the plaque formation were specifically inhibited by antisera against the virus. Neutralization tests were developed on the basis of these findings. Neutralization and haemagglutination-inhibition tests on swine serum samples indicated a wide dissemation of haemagglutinating encephalomyelitis virus or antigenically-related viruses in Japanese pigs.


title: Interaction of rotavirus and enterotoxigenic Escherichia coli in conventionally-reared dairy calves

text: A study was made of the effects of rotavirus and/or enterotoxigenic Escherichia coli (ETEC) on dairy calves born and suckled on the farm and subsequently reared in isolation. Calves were orally inoculated at 6 days old with either rotavirus (5), ETEC (7), rotavirus and ETEC (5) or remained uninoculated controls (4), and their reactions were recorded by clinical, microbiological, and pathological observations. Rotavirus infection consistently produced diarrhoea, while ETEC inoculated alone did not colonise the intestine. In dual infections, both rotavirus and ETEC multiplied, although the severity of diarrhoea was not greater than that caused by rotavirus alone. Some ETEC-inoculated calves developed subsequent naturally-acquired rotavirus infections, but in these no ETEC multiplication occurred. The results suggest that prior or simultaneous rotavirus infection is necessary to enable ETEC colonisation of the intestine in convenstional calves of this age.


title: More than one component of the Newcastle disease virus particle is capable of interferon induction

text: The interferon (IFN)-inducing capacities of intact NDV virions, β-propiolactone-inactivated particles and several structural components were compared, using human PBML as the IFN producing cells. Intact and inactivated virions as well as the nucleocapsid fraction did not differ significantly in their IFN-inducing capacity. In contrast, genomic RNA as well as M protein fraction and envelopes induced IFN titres to a level of about 10% of those achieved with virions. NDV-induced IFN production could be blocked specifically by incubation with polychonal anti-NDV-monoclonal antibodies (mAbs) and with two of three anti-HN-mAbs, but not with anti-NDV-mAbs directed against the F, M or NP protein. In addition, IFN induction by fixed MDBK cells, expressing NDV surface proteins after infection with NDV Ulster, was inhibited by one of two anti-F-mAbs. The results suggest that the induction of IFN synthesis in human PBML is a complex process involving not only the HN protein but also the uncleaved F protein precursor, a component of the M protein fraction and — once having entered the cell — the genomic RNA.


title: Prospects for the treatment of drug-resistant malaria parasites

text: Widespread parasitic resistance has led to an urgent need for the development and implementation of new drugs for the treatment of Plasmodium falciparum malaria. Artemisinin and its derivatives are becoming increasingly important, used preferably in combination with a second antimalarial agent to increase the efficacy and slow the development of resistance. However, cost, production and pharmacological issues associated with artemisinin derivatives and potential partner drugs are hindering the implementation of combination therapies. This article reviews the molecular basis of the action of, and resistance to, different antimalarials and examines the prospects for the next generation of drugs to combat this potentially lethal human pathogen.


title: CYP2C8 and antimalaria drug efficacy

text: Malaria is a major infectious disease. In the last 10 years it has killed more than 20 million people, mainly small children in Africa. The highly efficacious artemisinine combination therapy is being launched globally, constituting the main hope for fighting the disease. Amodiaquine is a main partner in these combinations. Amodiaquine is almost entirely metabolized by the polymorphic cytochrome P450 (CYP) isoform 2C8 to the pharmacologically active desethylamodiaquine. The question remains whether the efficacy of amodiaquine is affected by the gene polymorphism. Genotype-inferred low metabolizers are found in 1–4% of African populations, which corresponds to millions of expected exposures to the drug. In vivo pharmacokinetic data on amodiaquine is limited. By combining it with published in vitro pharmacodynamic and drug metabolism information, we review and predict the possible relevance, or lack of, of CYP2C8 polymorphisms in the present and future efficacy of amodiaquine. Chloroquine and dapsone, both substrates of CYP2C8, are also discussed in the same context.


title: Tracking progress towards malaria elimination in China: Individual-level estimates of transmission and its spatiotemporal variation using a diffusion network approach

text: In order to monitor progress towards malaria elimination, it is crucial to be able to measure changes in spatio-temporal transmission. However, common metrics of malaria transmission such as parasite prevalence are under powered in elimination contexts. China has achieved major reductions in malaria incidence and is on track to eliminate, having reporting zero locally-acquired malaria cases in 2017 and 2018. Understanding the spatio-temporal pattern underlying this decline, especially the relationship between locally-acquired and imported cases, can inform efforts to maintain elimination and prevent re-emergence. This is particularly pertinent in Yunnan province, where the potential for local transmission is highest. Using a geo-located individual-level dataset of cases recorded in Yunnan province between 2011 and 2016, we introduce a novel Bayesian framework to model a latent diffusion process and estimate the joint likelihood of transmission between cases and the number of cases with unobserved sources of infection. This is used to estimate the case reproduction number, Rc. We use these estimates within spatio-temporal geostatistical models to map how transmission varied over time and space, estimate the timeline to elimination and the risk of resurgence. We estimate the mean Rc between 2011 and 2016 to be 0.171 (95% CI = 0.165, 0.178) for P. vivax cases and 0.089 (95% CI = 0.076, 0.103) for P. falciparum cases. From 2014 onwards, no cases were estimated to have a Rc value above one. An unobserved source of infection was estimated to be moderately likely (p>0.5) for 19/ 611 cases and high (p>0.8) for 2 cases, suggesting very high levels of case ascertainment. Our estimates suggest that, maintaining current intervention efforts, Yunnan is unlikely to experience sustained local transmission up to 2020. However, even with a mean of 0.005 projected up to 2020, locally-acquired cases are possible due to high levels of importation.


Session orale (1)


Infections inhabituelles


Infection non bactérienne


title: Investigation d’une épidémie hospitalière et règles de prise en charge des cas suspects de rougeole ou d’infection invasive à méningocoque en établissement de soins

text: Outbreaks of infectious diseases within healthcare institutions must be detected early and controlled. Hospitals should develop a plan for coordinating all hospital components to respond to these critical situations. The knowledge of the different steps in an outbreak investigation can help identify the source of ongoing outbreaks and prevent additional cases. Outbreak investigation is based on a multidisciplinary approach and is an opportunity for research, training and program considerations.


title: Early respiratory manifestations of severe burn patient

text: Respiratory dysfunction with hypoxemia is common in early phase of severe burn, with or without smoke inhalation injury. In this article, we discuss the mechanisms associated with the occurrence of pulmonary injury in burn intensive care patients, diagnostics approaches and therapeutics options to be implemented based on identified causes.


title: Prise en charge multidisciplinaire des pneumopathies nécrosantes ventilées

text: Pneumonia may be complicated by necrosis and destruction of lung tissue due to factors related to both pathogen and host as well as to their interactions. Lung necrosis may lead to two main entities sharing common features, but also several clinical and pathological differences: lung abscesses and necrotizing pneumonia. Necrotizing pneumonia is characterized by diffuse, possibly bilateral, lung parenchyma inflammation with multiple cavitations and necrosis. Necrotizing pneumonia is usually associated with severe sepsis and acute respiratory failure. Adequate antibiotics, mechanical ventilation, pleural drainage, and prolonged supportive care are mandatory. Adult patients with necrotizing pneumonia may require surgery. In our practice, indications for surgery are: (1) uncontrolled sepsis in spite of medical therapy and chest drainage; (2) major air leaks responsible for ventilation difficulties with serious hypoxemia/hypercapnia; and (3) hemodynamic disturbances by compression of vena cava and/or right heart cavities by tumor-like forms. Surgical treatment should be adapted to each case. Despite serious morbidity, massive parenchyma damage and prolonged hospitalization, long-term outcome following necrotizing pneumonia seems good when multidisciplinary care management is used in these patients with unusual but severe respiratory infectious disease.


title: Formes graves de la grippe H1N1 2009 chez l’adulte: l’expérience française

text: The REVA-Flu-SRLF register allowed collection of data from 562 patients infected with H1N1 influenza virus 2009 and hospitalized in the intensive care unit (ICU). The overall mortality of these patients was 20%. The use of invasive ventilation, heart failure, and immunosuppression were associated with mortality. Three hundred forty-one (82%) among the 417 mechanically ventilated patients had an acute respiratory distress syndrome (ARDS). One hundred sixty-nine (30%) had a bacterial co-infection. Corticosteroid therapy was associated with an increased mortality in patients with ARDS. The occupancy rate associated with influenza patients crossed the threshold of 15% in many ICUs.


title: Faut-il contrôler la fièvre dans les infections sévères ?

text: Temperature control during severe sepsis is currently used in intensive care and involves 66% and 70% of severe sepsis and septic shock, respectively. Nevertheless, the conclusive evidence of the benefit of such a strategy is still lacking.We might wonder, with regards to experimental works and recent noninterventional studies, about the risk of a control strategy on an ongoing infectious process, the patient’s outcome, and the safety of the means implemented to obtain temperature control. On the other hand, it is also demonstrated that fever increases oxygen consumption, which may lead in some clinical situations to tissular ischemia and that fever may be associated with a deleterious focal inflammatory process. Methods to control the temperature include external and/or internal cooling and/or antipyretic medications such as paracetamol and nonsteroidal antiinflammatory drugs. In septic patients, external cooling and paracetamol are the mains means used to control temperature. Despite the uncertainties about the benefit to control or not the temperature, it could be stated that extreme temperature (hypo- or hyperthermia) should be avoided and that the benefit/risk of temperature control must be individually weighted.


title: Dysfonction immunitaire induite par la ventilation mécanique

text: Although mechanical ventilation is an essential support in patients admitted to the intensive care unit, clinical and experimental studies have shown that it could be harmful and could induce lung injury. Pulmonary and immune cells can convert mechanical stimuli into biological signals that will lead to inflammation. This sterile inflammation both locally and systemically will cause immunosuppression.


docid: hipvvezt

title: Glucocorticoids in the treatment of acute respiratory distress syndrome

text: Acute respiratory distress syndrome (ARDS) is characterized by local inflammation and an intense systemic inflammatory reaction. Glucocorticoid administration has been suggested due to their anti-inflammatory properties. However, results from the initial studies of glucocorticoids in ARDS, which evaluated high-dose and short-term treatments, were negative. More recent studies have evaluated the effect of lower doses of glucocorticoids administered over longer periods, but the results thus far have been inconclusive.


title: Organization of extracorporeal membrane oxygenation for acute respiratory distress syndrome in the United Kingdom

text: This article reviews the evolution of extracorporeal membrane oxygenation (ECMO) in the United Kingdom to treat patients with refractory acute respiratory distress. The UK centralized commissioning of public health care has delivered a coherent high-quality national adult ECMO service and defined the key factors in the designation as adult ECMO centre. This strategy seems adequate to provide for the needs of the population and avoid the danger of occasional practice by teams who do not undertake ECMO regularly.


title: Nouveautés thérapeutiques sur le SDRA

text: Treatment of acute respiratory distress syndrome (ARDS) has been subject to many researches, sometimes leading to intense controversy. New findings in this field are varied. Effects on prognosis of commonly used treatments for ARDS have recently been investigated. Consistently, prone position, previously known to improve oxygenation without effect on mortality, has been shown to improve survival of the most severely hypoxemic patients. Administration of neuromuscular blocking agents in the acute phase of ARDS has been also shown to be beneficial on survival. In contrast, the exact place of extracorporeal membrane oxygenation (ECMO) in ARDS management remains to be defined despite data suggesting its possible efficiency. In addition, a new era of research has emerged with the advent of cell therapy. Mesenchymal stem cells are able to both promote alveolar epithelium repair and prevent infections. Their efficacy in animal models of ARDS still needs to be confirmed by clinical trials. Finally, other promising therapies including beta-2 adrenergic agonists and omega-3 fatty acids have shown significant limitations in large clinical studies on ARDS.


title: Conduite et complications de l’oxygénation extracorporelle veinoveineuse

text: This article reviews the management of patients treated with venovenous extracorporeal membrane oxygenation (ECMO) for acute respiratory failure refractory to the conventional therapies. The period of extracorporeal respiratory support can be divided in three successive periods: the period of ECMO initiation, the period of treatment with ECMO, and the period of ECMO weaning. We will describe the main technical aspects of ECMO as well as the monitoring of the extracorporeal circuit and the ECMO-treated patient. The most frequent complications in each period of the management of ECMO-treated patients will be described and the possible adequate solutions will be considered.


title: Particularités de l’ECMO pour le syndrome de détresse respiratoire aiguë en pédiatrie

text: Since the 80’s, extracorporeal techniques have been used in pediatrics for the most severe acute respiratory distress syndromes. In 2012, data from the International Register of the Extracorporeal Life Support Organization revealed that more than 5,000 children had undergone extracorporeal membrane oxygenation (ECMO) with an increase in the number of annual cases since the epidemics of 2009. The survival rate (56%) is stable while the number of children with comorbidities increases due to the improvements in the equipment. Although there are no randomized studies, results encourage considering ECMO in pediatric ARDS. If veno-venous ECMO should be preferred in respiratory diseases, veno-arterial ECMO may be necessary and is still used frequently in children (50% of cases). In pediatrics, the technical features are first related to the physiological characteristics of the child and secondly to the technical limitations of the available equipment in the different ages. ECMO is a technique requiring specific expertise and specialized pediatric teams due to children’s particularities.


title: Assistance par circulation extracorporelle veinoveineuse dans le traitement du syndrome de détresse respiratoire aiguë : rationnel et objectifs cliniques

text: Extracorporeal circulation techniques can be used for the management of severe respiratory failure complicating the acute respiratory distress syndrome with three objectives: 1) to ensure satisfying oxygenation by bypassing the sick lung using a veno-venous circulation with high blood flows; this technique easily extracorporeal permit CO(2) elimination; 2) to ensure partial elimination of CO(2) with the aim to protect the lungs from risky mechanical ventilation. Four to five times lower blood flows are sufficient with a veno-venous circulation or pumpless arterio-venous techniques; 3) associated cardiac failure may exceptionally require veno-arterial extracorporeal circulation. Physiological studies and clinical trials are absolutely needed to better delineate the indications of these techniques.


title: Preterm infant feeding performance at term equivalent age differs from that of full-term infants

text: OBJECTIVE: To identify differences in feeding skill performance among preterm infants at term equivalent age compared with full-term infants. STUDY DESIGN: Ninety-two infants (44 preterm infants born ≤32 weeks gestation at term equivalent age and 48 full-term infants within 4 days of birth) had a standardized oral feeding assessment. RESULT: Preterm infants at term equivalent age had lower Neonatal Eating Outcome Assessment scores (67.8 ± 13.6 compared with 82.2 ± 8.1; p < 0.001) and were more likely to have poor arousal (p = 0.04), poor tongue positioning (p = 0.04), suck–swallow–breathe discoordination (p < 0.001), inadequate sucking bursts (p = 0.01), tonal abnormalities (p < 0.001), discoordination of the jaw and tongue during sucking (p < 0.001), lack of positive engagement with the feeder and/or discomfort (p < 0.001), signs of aspiration (p < 0.001), difficulty regulating breathing (p < 0.001), and have an inability to maintain an appropriate state (p < 0.001), and complete the feeding (<0.001). CONCLUSION: A broad range of feeding-related difficulties appear to remain evident in preterm infants at term equivalent age.


Updated rapid risk assessment from ECDC on coronavirus disease 2019 (COVID-19) pandemic: increased transmission in the EU/EEA and the UK


title: Electron-microscopic appearance of the DA virus, a demyelinating murine virus()

text: The DA virus is a neurotropic murine virus which can induce acute encephalomyelitis in suckling mice and a chronic myelopathy in weanlings. The agent has been attenuated by serial passage in baby hamster kidney (BHK-21) cells. When attenuated virus is inoculated in 8-week-old C(3)HeJ mice a myelopathy of delayed onset with prominent demyelination of lateral and anterior columns occurs. The DA virus is believed to be related to the Theiler murine encephalomyelitis (TME) viruses because of the similar clinical and pathological conditions which it causes, and because neutralization tests indicate shared antigens between it and GD7, a TME virus. This paper reports electron-microscopic studies of BHK-21 cells infected with DA virus. The cells were prepared 24 and 48 hr after inoculation. Cytopathic effects were observed and infected cells contained plaques consisting of numerous 25 nm virus particles in crystalline array. The virions were exclusively intracytoplasmic and were morphologically indistinguishable from human poliomyelitis virus. These observations appear to establish DA as a picorna virus, related to the TME virus group. The chronic myelopathy caused by DA may prove relevant to chronic demyelinative myelopathies in man, such as multiple sclerosis, and also to amyotrophic lateral sclerosis.


title: Ultrastructural study of myelinating cells and sub-pial astrocytes in developing rat spinal cord()

text: The anterior funiculus of the spinal cervical cord of post-natal rats was examined ultrastructurally. The myelinating cells found one day after birth contained a large amount of evenly distributed ribosomes up to the outer tongue of mesaxons, representing the cytoplasmic density. These cells were separated by astrocytic processes from the pial basement membrane, even when they were located on the pial surface. Astrocytes contained glial fibrils from one day onwards and often attached their processes to the pial basement membrane. Although the cytoplasmic processes of astrocytes occasionally wrapped axons, they were never shown to form the initial layer of myelin sheaths. However, the tenuous processes of the sub-pial astrocytes were occasionally rolled in myelin lamellae, as if a part of the myelin sheaths was constructed by astrocytic processes. The interpretation for this finding is discussed in relation to function and potency of the astrocytes, and variations and anomalies of nervous ontogeny.


Expanding nursing's role in responding to global pandemics 5/14/2018


title: Myelination by oligodendrocytes isolated from 4–6-week-old rat central nervous system and transplanted into newborn shiverer brain

text: Oligodendrocytes isolated from 4–6-week-old rat brains were transplanted into newborn shiverer brains. Cells were identified as mature oligodendrocytes both by immunocytological and ultrastructural criteria. Normal myelin was detected using immunolocalisation (with an anti-MBP antiserum) and electron microscopy (presence of the major dense line). Patches of normal myelin (made by transplanted oligodendrocytes), widely spread throughout the host brains, were detected between 20 and 130 days after grafting. No sign of acute rejection was observed, but the graft became progressively delimited by astrocytic processes forming a continuous basal lamina.


docid: mfglfgdw

title: Management of Refugees and International Adoptees

text: Refugee children and international adoptees have special medical considerations that must be addressed. Providers must be aware of the immigration history, where, and under what circumstances the child lived and migrated to the United States. Federal guidelines exist regarding which infections should be screened for, and how and when and which vaccines should be administered.


docid: ahiekwch

title: Special assorted cardiovascular topics


Tackling two pandemics: a plea on World Tuberculosis Day


Are We Ready for Anything?


One more paper towel, longer protection


COVID-19: global consequences for oncology


title: Haploidentical bone marrow transplant with posttransplant cyclophosphamide for sickle cell disease: An update

text: Hematopoietic cell transplant (HCT) can cure both children and adults with sickle cell disease. Outcomes have historically been poor for the vast majority of patients who lack a matched sibling donor. However, the development of haploidentical HCT (haplo-HCT) with high doses of posttransplant cyclophosphamide (PTCy) has allowed for curative long-term potential with favorable transplant-related outcomes, though this has not obviated the potential for graft rejection from human leukocyte antigen mismatch and repeated red blood cell transfusions. Accordingly, multiple strategies have been developed to improve outcomes, the majority of which are based on the Johns Hopkins platform from 2012. Presently, we aim to discuss results from pertinent studies and compare outcomes with the two most recent approaches involving either thiotepa plus 200-cGy total body irradiation or 400-cGy total body irradiation. Direct comparisons are required to determine the optimized curative potential. Transplant-eligible patients must be referred to tertiary medical centers for consideration of haplo-HCT.


COVID-19 in Italy: Ageism and Decision-Making in a Pandemic


docid: odupmsen

title: True epidemic growth construction through harmonic analysis

text: In this paper, we have proposed a two-phase procedure (combining discrete graphs and wavelets) for constructing true epidemic growth. In the first phase, a graph-theory-based approach was developed to update partial data available and in the second phase, we used this partial data to generate plausible complete data through wavelets. We have provided two numerical examples. This procedure is novel and implementable and adaptable to machine learning modeling framework.


Porcine reproductive and respiratory syndrome virus (PRRSV) inhibition with engineered Cas13d


COVID-19 et personnes suivies pour un cancer du sein : recommandations françaises pour la pratique clinique de Nice-St Paul de Vence, en collaboration avec le Collège Nationale des Gynécologues et Obstétriciens Français (CNGOF), la Société d’Imagerie de la FEMme (SIFEM), la Société Française de Chirurgie Oncologique (SFCO), la Société Française de Sénologie et Pathologie Mammaire (SFSPM) et le French Breast Cancer Intergroup-UNICANCER (UCBG)


I think I got “That Disease”: An interview with a medical student who had recovered from SARS in Guangzhou, China


Rewards, risks, and responsibilities of globalization for the cardiothoracic surgeon


Rabbit Gastroenterology


Pathologies virales émergentes en pneumologie


The effectiveness of training and taste testing when using respirator masks


Asthme du nourrisson : monopole du virus respiratoire syncytial ?


Counting the cost of cancelled surgery: a system wide approach is needed


High justification for universal stringent precautions in lung function testing


title: Solidarity, vulnerability and mistrust: how context, information and government affect the lives of women in times of Zika

text: BACKGROUND: The public health response to Zika outbreak has mostly focused on epidemiological surveillance, vector control, and individual level preventative measures. This qualitative study employs a social-ecological framework to examine how macro (historical, legislative, political, socio-economic factors), meso (sources of information, social support, social mobilization) and micro level factors (individual actions, behavioral changes) interacted to influence the response and behavior of women with respect to Zika in different contexts. METHODS: A qualitative study was carried out. Women were recruited through the snowball sampling technique from various locations in Brazil, Puerto Rico, and the United States. They were of different nationalities and ethnicities. Data were collected through semi-structured interviews. The data transcripts were analyzed using thematic analysis. RESULTS: Women in this study deemed the information provided as insufficient, which led them to actively reach out and access a variety of media sources. Social networks played a vital role in sharing information but also resulted in the spread of hoaxes or rumors. Participants in our research perceived socio-economic inequities but focused on how to remedy their microenvironments. They did not engage in major social activities. Lack of trust in governments placed women in vulnerable situations by preventing them to follow the guidance of health authorities. These impacts were also a result of the response tactics of health and government administrations in their failed attempts to ensure the well-being of their countries’ populations. CONCLUSIONS: Our findings call for public health interventions that go beyond individual level behavioral change campaigns, to more comprehensively address the broader meso and macro level factors that influence women’ willingness and possibility to protect themselves.


Feasibility and safety of watershed detection by contrast-enhanced ultrasound in patients receiving peripheral venoarterial extracorporeal membrane oxygenation: a prospective observational study


SARS, emerging diseases, healthcare workers and respirators


SARS in a hospital visitor and her intensivist


SARS and masks


Laboratory evaluation of selective mannitol broth for MRSA screening


Coming of age in travel medicine and tropical diseases: a need for continued advocacy and mentorship


Decrease in the incidence of meticillin-resistant Staphylococcus aureus nosocomial bloodstream infections in Taiwan


Radiological Findings in Severe Acute Respiratory Syndrome


Can we increase public awareness without creating anxiety about Corona viruses?


Vitamins C and D