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Indiana chronic disease management program risk stratification analysis.

OBJECTIVE: The objective of this study was to compare the ability of risk stratification models derived from administrative data to classify groups of patients for enrollment in a tailored chronic disease management program. SUBJECTS: This study included 19,548 Medicaid patients with chronic heart failure or diabetes in the Indiana Medicaid data warehouse during 2001 and 2002. MEASURES: To predict costs (total claims paid) in FY 2002, we considered candidate predictor variables available in FY 2001, including patient characteristics, the number and type of prescription medications, laboratory tests, pharmacy charges, and utilization of primary, specialty, inpatient, emergency department, nursing home, and home health care. METHODS: We built prospective models to identify patients with different levels of expenditure. Model fit was assessed using R statistics, whereas discrimination was assessed using the weighted kappa statistic, predictive ratios, and the area under the receiver operating characteristic curve. RESULTS: We found a simple least-squares regression model in which logged total charges in FY 2002 were regressed on the log of total charges in FY 2001, the number of prescriptions filled in FY 2001, and the FY 2001 eligibility category, performed as well as more complex models. This simple 3-parameter model had an R of 0.30 and, in terms in classification efficiency, had a sensitivity of 0.57, a specificity of 0.90, an area under the receiver operator curve of 0.80, and a weighted kappa statistic of 0.51. CONCLUSION: This simple model based on readily available administrative data stratified Medicaid members according to predicted future utilization as well as more complicated models.

Adult↗

Heterotypic exclusion between vesicular stomatitis viruses of the New Jersey and Indiana serotypes.

Co-infection of cells with vesicular stomatitis viruses of the Indiana and New Jersey serotypes results in interference. Using specifically-labelled immunofluorescent antibodies, it was demonstrated that within any one co-infected cell, one virus serotype replicated to the relative exclusion of the other serotype. This result was further substantiated by an examination of the virus serotypes released by infectious centres co-infected with both viruses. Dominance of one serotype over the other was shown to be a function of the relative multiplicity of the two viruses. Superinfection by the second serotype at a higher multiplicity resulted in dominance by the second virus during the early period (up to 1-5 h) post-infection. After this time, the minority virus was able to overcome this dominance. Dominance of the majority virus was also abolished by u.v; inactivation. Cell protein synthesis appeared to be less affected in cells infected with both serotypes than when infection was with a single serotype.

Fluorescent Antibody Technique↗

Conservation of potential phosphorylation sites in the NS proteins of the New Jersey and Indiana serotypes of vesicular stomatitis virus.

A full length cDNA copy of the NS mRNA of the Missouri strain (Hazelhurst subtype, New Jersey serotype) of vesicular stomatitis virus (VSV) has been cloned and sequenced. The mRNA is 856 nucleotides long (excluding polyadenylic acid) and encodes a protein of 274 amino acids (mol. wt. 31 000). Comparison with the NS gene of the Ogden strain (Concan subtype, New Jersey serotype) showed 15% difference at the nucleotide level and 10% difference at the amino acid level; the majority of the changes were located in the 3' half of the mRNA. Comparison with the NS genes of two strains representing the Indiana serotype showed about 50% nucleotide and 33% amino acid sequence homology between the serotypes. In a four-way comparison of the proteins, two regions of higher homology were noted which may be of functional importance. Eighteen potential phosphorylation sites (Ser or Thr) were conserved between the four proteins; five of these sites correspond to the residues which have been suggested to be constitutively phosphorylated and may be essential for NS activity.

Base Sequence↗

Full-length genome analysis of natural isolates of vesicular stomatitis virus (Indiana 1 serotype) from North, Central and South America.

Most studies on the molecular biology and functional analysis of vesicular stomatitis virus Indiana 1 serotype (VSV-IN1) are based on the only full-length genomic sequence currently deposited in GenBank. This sequence is a composite of several VSV-IN1 laboratory strains passaged extensively in tissue culture over the years and it is not certain that this sequence is representative of strains circulating in nature. We describe here the complete genomic sequence of three natural isolates, each representing a distinct genetic lineage and geographical origin: 98COE (North America), 94GUB (Central America) and 85CLB (South America). Genome structure and organization were conserved, with a 47 nucleotide 3' leader, five viral genes -- N, P, M, G and L -- and a 59 nucleotide 5' trailer. The most conserved gene was N, followed by M, L and G, with the most variable being P. Sequences containing the polyadenylation and transcription stop and start signals were completely conserved among all the viruses studied, but changes were found in the non-transcribed intergenic nucleotides, including the presence of a trinucleotide at the M-G junction of the South American lineage isolate. A 102-189 nucleotide insertion was present in the 5' non-coding region of the G gene only in the viruses within a genetic lineage from northern Central America. These full-length genomic sequences should be useful in designing diagnostic probes and in the interpretation of functional genomic analyses using reverse genetics.

Base Sequence↗

Crystallization and preliminary X-ray analysis of a proteinase-K-resistant domain within the phosphoprotein of vesicular stomatitis virus (Indiana).

Several stable domains of the phosphoprotein (P) of vesicular stomatitis virus (Indiana) were identified by limited proteolysis of purified recombinant P protein expressed in Escherichia coli. The proteinase-K-resistant domain could be crystallized using ammonium sulfate as a precipitant and ethylene glycol as an additive. The crystals belong to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 74.50, c = 156.84 A. X-ray diffraction data were collected to 2.75 A resolution at a synchrotron-radiation source.

Crystallization↗

Fetal mortality in oral cleft families: data from Indiana and Montreal.

Analysis of 418 sibships of oral cleft probands from Indiana, and 288 sibships from Montreal indicate that the incidence of fetal mortality is significantly greater in sibships of probands with cleft lip and cleft palate (CLP) as compared to that in sibships of probands with cleft lip (CL) alone. These findings support a multifactorial-two-threshold concept, according to which a lower level of liability results in clefting whereas a higher level of liability causes fetal deaths. They add confirmatory evidence in support of the authors' earlier work, utilizing data from two different sources.

Cleft Lip↗

Atrazine retention and degradation in the vadose zone at a till plain site in central Indiana.

The vadose zone was examined as an environmental compartment where significant quantities of atrazine and its degradation compounds may be stored and transformed. The vadose zone was targeted because regional studies in the White River Basin indicated a large discrepancy between the mass of atrazine applied to fields and the amount of the pesticide and its degradation compounds that are measured in ground and surface water. A study site was established in a rotationally cropped field in the till plain of central Indiana. Data were gathered during the 1994 growing season to characterize the site hydrogeology and the distribution of atrazine, desethylatrazine, deisopropylatrazine, didealkylatrazine and hydroxyatrazine in runoff, pore water, and ground water. The data indicated that atrazine and its degradation compounds were transported from land surface to a depth of 1.5 m within 60 days of application, but were undetected in the saturated zone at nearby monitoring wells. A numerical model was developed, based on the field data, to provide information about processes that could retain and degrade atrazine in the vadose zone. Simulations indicated that evapotranspiration is responsible for surface directed soil-moisture flow during much of the growing season. This process causes retention and degradation of atrazine in the vadose zone. Increased residence time in the vadose zone leads to nearly complete transformation of atrazine and its degradation products to unquantified degradation compounds. As a result of macropore flow, small quantities of atrazine and its degradation compounds may reach the saturated zone.

Atrazine↗

The influence of preservice instruction in health education methods on the health content taught by elementary teachers in Indiana.

This study examined whether receiving preservice instruction in health education methods affects the health instruction provided by elementary classroom teachers. Using a researcher-constructed self-report questionnaire, 800 third-, fourth-, and fifth-grade teachers in Indiana were randomly selected to complete the questionnaire. Returns included 366 usable responses from an effective sample of 772, for an overall response rate of 48.06%. Analysis of variance indicated a statistically significant difference among respondents in the mean number of 10 health content areas taught, the depth of coverage of all content areas except Community and Environmental Health, the mean number of health lessons taught per week, and the minutes spent teaching health per week. Pairwise comparisons indicated respondents who completed both a health education methods course and a physical education methods course taught a greater number of health content areas, and taught more topics in eight of the 10 health content areas, than did respondents who did not complete preservice instruction in health education methods.

Adult↗

Laboratory vector competence of black flies (Diptera:Simuliidae) for the Indiana serotype of vesicular stomatitis virus.

In previous experiments we have demonstrated that colonized and wild black flies are competent laboratory vectors of different Mexican and Western USA isolates of vesicular stomatitis virus, serotype New Jersey (VSV-NJ). We have recently demonstrated biological VSV-NJ transmission by black flies using animal models. In the study described here, we tested the vector competence of colonized and wild black flies for the vesicular stomatitis virus, serotype Indiana (VSV-IN). A 1998 equine isolate was used. After a 10 day incubation period, saliva from experimentally infected Simulium vittatum and S. notatum was individually collected and tested for the presence of infectious virus. Virus was detected in the saliva of both species following oral infection, indicating that they are competent laboratory vectors of VSV-IN. In addition, the results suggest that the black fly gut may exert evolutionary pressures on the virus.

Animals↗

Quantitative multiplex assay for simultaneous detection and identification of Indiana and New Jersey serotypes of vesicular stomatitis virus.

In order to establish a rapid and reliable system for the detection of vesicular stomatitis virus (VSV), we developed a quantitative reverse transcription-PCR assay for the detection, quantification, and differentiation of the major serotypes, VSV Indiana and VSV New Jersey, using a closed-tube multiplex format. The detection system is based on the recently invented primer-probe energy transfer (PriProET) system. A region of the gene encoding the RNA-dependent RNA polymerase was amplified by using VSV-specific primers in the presence of two serotype-specific fluorescent probes. By incorporating nucleotide analogues in the primers, both serotypes were amplified with similar efficiencies. The generation of specific amplicons resulted in fluorescent signals for either of the two serotypes, and the specificities of the reactions were confirmed from the melting temperature profiles of the fluorescent probes. The limits of detection were found to be less than 10 50% tissue culture infective doses/ml for both serotypes. The diagnostic value of the new method was tested with clinical materials from experimentally infected pigs, and it is concluded that the method is a powerful tool for the rapid identification of VSV.

Animals↗

Comparisons of nucleotide sequences in the genomes of the New Jersey and Indiana serotypes of vesicular stomatitis virus.

Nucleotide sequences of around 200 residues were determined adjacent to the 3' terminus of the genome RNA of vesicular stomatitis virus, New Jersey serotype, and adjacent to the 3'-terminal polyadenylic acid tract of the N protein mRNA of the same virus. These sequences were compared with the corresponding sequences previously determined for the Indiana serotype of vesicular stomatitis virus. The sequences obtained for the two strains were readily aligned, showing 70.8% homology overall. Examination of the sequences allowed identification of the translation initiation and termination codons for the N mRNA of each serotype. The deduced N-terminal and C-terminal amino acid sequences of the two N polypeptides were each similar, and most of the differences between them consisted of substitution by a clearly homologous amino acid. It was proposed that these nucleotide sequences, within limits imposed by their functions, comprise reasonably representative measures of the extent of sequence homology between the genomes of the two serotypes, and that this is higher than previously estimated, but with little exact homology over extended regions.

Amino Acid Sequence↗

Monoclonal antibodies to the M protein of vesicular stomatitis virus (Indiana serotype) and to a cDNA M gene expression product.

Twenty-nine independent hybridomas producing monoclonal antibodies to the matrix (M) protein of vesicular stomatitis virus (Indiana serotype) were prepared by fusion of SP2/0 myeloma cells with spleen lymphocytes obtained from BALB/c mice which had been immunized with the purified M protein. The specific reactivity of each monoclonal antibody was determined by an enzyme-linked immunosorbent assay and a competitive binding assay. Most of the antibodies were of the immunoglobulin G2a and G2b isotypes, although some were immunoglobulin M. By measuring the competitive binding of 125I-antibody, we identified four antigenic determinants in the M protein of the virus; two of these determinants, however, exhibited a large degree of overlap. Western blot analysis revealed little or no cross-reactivity of the antibodies with other viral proteins or with the M protein of the New Jersey serotype. Prolonged trypsin proteolysis removed the first 43 amino acids from the amino-terminal region of the M protein, but it retained its reactivity with monoclonal antibodies to each epitope, except for diminished reactivity with one. To aid in future mapping of these epitopes, we inserted a cDNA clone of the mRNA encoding the M protein of vesicular stomatitis virus into an inducible lac expression vector; the M protein produced in the JM103 strain of Escherichia coli under induced conditions was found to be approximately the same size as native M protein and was recognized by the monoclonal antibodies. These monoclonal antibodies and the cDNA clone should be useful for studying the role of M protein in virus maturation and the regulation of viral transcription.

Animals↗

Polymerase errors accumulating during natural evolution of the glycoprotein gene of vesicular stomatitis virus Indiana serotype isolates.

We report the entire glycoprotein (G) gene nucleotide sequences of 26 vesicular stomatitis virus Indiana serotype (VSV IND) type 1 isolates from North and Central America. These sequences are also compared with partial G gene sequences of VSV IND type 2 (Cocal) and type 3 (Alagoas) viruses and the complete G gene sequences of the more distantly related VSV New Jersey (NJ) and Chandipura viruses. Phylogenetic analysis of the G gene sequences by maximum parsimony revealed four major lineages or subtypes within the classical VSV IND (type 1) viruses, each with a distinct geographic distribution. A high degree of VSV genetic diversity was found in Central America, with several virus subtypes of both VSV IND and NJ serotypes existing in this mainly enzootic disease region. Nineteen percent sequence variation but no deletions or insertions were evident within the 5' noncoding and the coding regions of the VSV IND type 1 G genes. In addition to numerous base substitutions, the 3' noncoding regions of these viruses also contained numerous base insertions and deletions. This resulted in striking variation in G gene sizes, with gene lengths ranging from 1,652 to 1,868 nucleotides. As the VSV IND type 1 subtypes have diverged from the common ancestor with the NJ subtypes, their G mRNAs have accumulated more 3' noncoding sequence inserts, ranging up to 303 nucleotides in length. These primarily consist of an imprecise reiteration of the sequence UUUUUAA, apparently generated by a unique polymerase stuttering error. Analysis of the deduced amino acid sequence differences among VSV IND type 1 viruses revealed numerous substitutions within defined antigenic epitopes, suggesting that immune selection may play a role in the evolution of these viruses.

Amino Acid Sequence↗

Vesicular stomatitis virus Indiana glycoprotein as a T-cell-dependent and -independent antigen.

The neutralizing immunoglobulin M (IgM) response to vesicular stomatitis virus (VSV) has been shown to be largely T-cell independent in several T-cell-deficient models of mice. By using different antigen froms of VSV, VSV antigen doses could be graded in vivo (infectious > > UV inactivated > formalin inactivated). The present study reveals a T-cell-dependent component of the neutralizing IgM response in nude mice given intravenous injections of low doses of noninfectious UV-inactivated VSV serotype Indiana (VSV-IND) only if the mice are transfused with VSV-IND-specific helper T cells. Instead, nude mice immunized with infectious VSV, which leads to greater antigen doses in vivo, were able to mount an IgM response in the absence of T cells. These results indicate that the IgM response to low doses of VSV-IND glycoprotein (G) is T-cell dependent. Nude mice immunized with infectious VSV also made a variable but low VSV-IND-neutralizing IgG response. A VSV-IND matrix (M)-specific helper T-cell line rendered this response more consistent, much higher, and longer lasting. Thus (i) VSV-G induces a mostly T-cell-independent but partially T-cell-dependent IgM (the latter can be visualized best at low doses of antigen) and (ii) the antibody response to VSV in nude mice proceeds through steps, i.e., IgM and IgG, that are dose dependent. The results suggest that the predominant role of helper T cells may be to expand and maintain the individual steps of differentiating B cells.

Animals↗

Expression of L protein of vesicular stomatitis virus Indiana serotype from recombinant baculovirus in insect cells: requirement of a host factor(s) for its biological activity in vitro.

The 241-kDa large (L) protein of vesicular stomatitis virus (VSV) Indiana serotype, a multifunctional catalytic subunit of the viral RNA polymerase, has been expressed in Spodoptera frugiperda cells infected with recombinant baculovirus BacPAK6-L containing the L gene under the control of a polyhedrin promoter. The recombinant L protein was biologically active and supported viral mRNA synthesis in vitro. When the expressed L protein was purified by phosphocellulose column chromatography, it eluted in two peaks, one at 0.4 M NaCl (peak I) and the second at 0.75 M NaCl (peak II). The L protein in peak I showed significant transcriptional activity in an in vitro transcription reconstitution experiment, whereas the L protein in peak II was inactive. Interestingly, the addition of cytoplasmic extract from uninfected Sf21 cells to peak II completely restored transcription in vitro, indicating the requirement of a host factor(s) for the activity of the L protein. This factor is relatively heat stable and is dissociable from the recombinant L protein. It is also present in BHK, COS, and HeLa cells in detectable levels. The role of the putative host protein(s) in the activation of the L protein is discussed.

Animals↗

Comparison of 3'-end encoding regions of turkey coronavirus isolates from Indiana, North Carolina, and Minnesota with chicken infectious bronchitis coronavirus strains.

OBJECTIVE: To analyze the 3'-end structural protein-encoding region of turkey coronavirus (TCoV) isolates associated with outbreaks of acute enteritis in Indiana, North Carolina, or Minnesota. METHODS: Four isolates of TCoV were sequenced over the entire 3'-end structural protein-encoding region and compared phylogenetically along with the corresponding sequences of infectious bronchitis virus (IBV) strains. RESULTS: The sequence similarity between TCoV and IBV was lower than that among TCoV isolates or that among IBV strains. The variation of sequences between TCoV and IBV was mainly contributed by the S protein gene. The sequence similarity of S gene between TCoV and IBV was lower than that among TCoV isolates or that among IBV strains. The phylogenetic tree based on the S protein region was similar to that based on the entire 3'-end structural protein-encoding region with TCoV isolates and IBV strains grouped in two separate clusters. The phylogenetic tree based on other genes had a very different topology with TCoV isolates randomly forming groups with different IBV strains. CONCLUSIONS: These results suggested that TCoV probably shared the same origin with that of IBV and acquired sequences of S gene for turkey intestine tropism during the process of evolution in a separate environment.

Animals↗

Quality improvement in the diagnosis and treatment of heart failure by participating Indiana and Kentucky hospitals.

Acute-care hospitals in Kentucky and Indiana collaborated with Health Care Excel, the Medicare peer review organization, to improve the care of patients admitted with heart failure. Current guidelines for the treatment of heart failure support a focus on quality indicators including confirmation of diagnosis, choice of therapeutic agents, education of patients and their caregivers, and discharge planning. Significant improvement occurred in use of diagnostic tests to confirm diagnosis and in patient education and discharge planing. Improvement in use of therapeutic agents was minimal or lacking in most hospitals. This project demonstrated an opportunity and need for continued improvement efforts in the care of heart failure patients.

Chi-Square Distribution↗

Circovirus-like viral associated disease in weaned pigs in Indiana.

Inclusion bodies with staining affinity and ultrastructural characteristics typical of circoviruses that stained positive for porcine circovirus (PCV)-like virus were demonstrated in association with granulomatous lesions in multiple tissues of three clinically ill 10- to 12-week-old pigs. A syndrome of poor growth and wasting in 5-15% of weaned pigs was an intermittent problem on a 450-sow one-site farrow-to-finish swine farm in Indiana. Routine diagnostic testing did not demonstrate a cause. Gross examination of three representative weaned pigs from two farrowing groups over a 1-month period revealed generalized lymphadenopathy and interstitial pneumonia. A unique microscopic finding for all three pigs was granulomatous inflammation of lymphoid tissues associated with large numbers of multinucleate giant cells and characteristic viral inclusions in the cytoplasm of macrophages. These inclusions were round, homogeneous, and magenta to basophilic, varied in size (5-25 microm), and either were single or formed botryoid clusters. Ultrastructurally, these inclusions were composed of electron-dense paracrystalline arrays of small nonenveloped icosahedral viral particles that were approximately 17 nm in diameter. The sizes and shapes of the virus particles, the unique microscopic appearance of the inclusions, and the positive staining of the intracytoplasmic viral inclusions by the Feulgen technique are consistent with circoviruses. Immunohistochemistry for PCV-like virus demonstrated viral antigen in the cytoplasm of macrophages that were within inflammatory infiltrates in a variety of organs. The described inclusion bodies stained positively.

Animals↗