Evaluation of the Cepheid Xpert Flu Assay for rapid identification and differentiation of influenza A, influenza A 2009 H1N1, and influenza B viruses.

The Xpert Flu Assay cartridge is a next-generation nucleic acid amplification system that provides multiplexed PCR detection of the influenza A, influenza A 2009 H1N1, and influenza B viruses in approximately 70 min with minimal hands-on time. Six laboratories participated in a clinical trial comparing the results of the new Cepheid Xpert Flu Assay to those of culture or real-time PCR with archived and prospectively collected nasal aspirate-wash (NA-W) specimens and nasopharyngeal (NP) swabs from children and adults. Discrepant results were resolved by DNA sequence analysis. After discrepant-result analysis, the sensitivities of the Xpert Flu Assay for prospective NA-W specimens containing the influenza A, influenza A 2009 H1N1, and influenza B viruses compared to those of culture were 90.0%, 100%, and 100%, respectively, while the sensitivities of the assay for prospective NP swabs compared to those of culture were 100%, 100%, and 100%, respectively. The sensitivities of the Xpert Flu Assay for archived NA-W specimens compared to those of Gen-Probe ProFlu+ PCR for the influenza A, influenza A 2009 H1N1, and influenza B viruses were 99.4%, 98.4%, and 100%, respectively, while the sensitivities of the Xpert Flu Assay for archived NP swabs compared to those of ProFlu+ were 98.1%, 100%, and 93.8%, respectively. The sensitivities of the Xpert Flu Assay with archived NP specimens compared to those of culture for the three targets were 97.5%, 100%, and 93.8%, respectively. We conclude that the Cepheid Xpert Flu Assay is an accurate and rapid method that is suitable for on-demand testing for influenza viral infection.

Overt and relational aggression in Russian nursery-school-age children: parenting style and marital linkages.

Maternal and paternal parenting styles and marital interactions linked to childhood aggressive behavior as described in Western psychological literature were measured in an ethnic Russian sample of 207 families of nursery-school-age children. Results corroborated and extended findings from Western samples. Maternal and paternal coercion, lack of responsiveness, and psychological control (for mothers only) were significantly correlated with children's overt aggression with peers. Less responsiveness (for mothers and fathers) and maternal coercion positively correlated with relational aggression. Some of these associations differed for boys versus girls. Marital conflict was also linked to more overt and relational aggression for boys. When entered into the same statistical model, more marital conflict (for boys only), more maternal coercion, and less paternal responsiveness were found to be the most important contributors to overt and relational aggression in younger Russian children.

Comparative study of maternal and paternal disciplinary strategies.

Parents (109 mothers, 109 fathers) of 109 middle-class preschool-age children were interviewed separately in individual taperecorded home interviews to assess whether either parent was prone to use assertion of power or inductive reasoning as disciplinary strategies. Fathers reported using more power-assertive disciplining strategies with their preschool-age children than mothers.

Acute infectious mononucleosis stimulates the selective expression/expansion of V beta 6.1-3 and V beta 7 T cells.

Acute infectious mononucleosis (AIM) is caused by the Epstein-Barr virus (EBV) and is characterized by a proliferation of atypical lymphocytes, predominantly CD8+ T cells. Various diseases associated with T-cell activation have been shown to stimulate the selective expansion of certain V beta (variable region of the T-cell receptor beta chain) expressing T-cell populations. The purpose of this investigation was to determine if the proliferation of T cells accompanying AIM is associated with selective expression/expansion of distinct populations of V beta T cells. We determined V beta expression in eight patients with clinical and laboratory evidence of AIM, including an atypical lymphocytosis. Gel electrophoresis and quantitative analysis were performed on cDNA amplified by the polymerase chain reaction (PCR) using different V beta region primers. Gel electrophoresis analysis showed prominent V beta 6.1-3 and V beta 7 bands in all eight patients with AIM but not in the controls. Quantitative PCR analysis showed that the V beta 6.1-3 and V beta 7 mean PCR ratios increased, respectively, from 163.0 +/- 22.5 and 142.3 +/- 5.5 in controls to 339.9 +/- 38.8 (P < .03) and 396.1 +/- 45.6 (P < .01) in the eight patients with AIM. Two of the eight patients who had increased V beta 6.1-3 and V beta 7 expression were retested after clinical resolution of AIM and no longer had evidence of increased V beta 6.1-3 and V beta 7 T-cell expression. AIM is associated with a selective increased expression of V beta 6.1-3 and V beta 7 T cells present at the time of initial clinical symptoms and atypical lymphocytosis. This increased expression resolves following recovery from AIM. This V beta-specific selective expression resembles the super-antigen response seen after staphylococcal toxin stimulation and may be caused by EBV triggering of selective expansion of V beta 6.1-3 and V beta 7 T-cell subsets.

Human interleukin-1 receptor antagonist. High yield expression in E. coli and examination of cysteine residues.

The human IL-1 receptor antagonist (IL-1ra) was produced in a high yield E. coli expression system, and was purified in a rapid two-step purification. This recombinant IL-1ra molecule possessed full binding activity to the IL-1 receptor (type I) and totally inhibited IL-1-induced PGE2 production by human dermal fibroblasts. Radioalkylation and analysis of V8-derived IL-1ra peptides indicate that the four cysteines present in the IL-1ra are not disulphide-linked.

Multiple delivery methods for an interdisciplinary audience: assessing effectiveness.

Recent federal legislation has provided guidelines for intervention services for infants and toddlers with handicaps, birth to age three, and their families. Many disciplines, including nursing, will be involved in providing these services. Serving on an interdisciplinary team or acting as a consultant requires appropriate preparation. Many professionals are not comfortable with their roles in serving this new population, due in part to the limited emphasis on the handicapped infant and toddler in most basic preservice professional education programs. The project described here was useful in helping an interdisciplinary audience develop knowledge and skills needed for intervention services for infants and toddlers.

An epidemiologic study of sudden death at work in an industrial county, 1979-1982.

The descriptive epidemiology of sudden death at work was studied in an urban, industrial county. County coroner's records were used to identify the 212 deaths that occurred at work among employed white males in Allegheny County, Pennsylvania in 1979-1982. Occupations and industries with increased risks for either sudden natural deaths or fatal injuries at work were identified by comparison with the white male county employed population. Men employed in service occupations had the highest age-adjusted sudden natural death rate at work (27.0 per 100,000). This was 2.5 times as high as the overall county rate. Men employed in the construction industry had the highest age-adjusted rate of fatal injuries at work (24.3 per 100,000). This was 4.4 times as high as the overall county rate. Twenty-five per cent (17/68) of occupational fatalities involved multiple fatalities or injuries. Only 1 per cent (2/144) of natural deaths at work and 7 per cent (5/68) of fatal injuries had blood alcohol levels exceeding 0.1 mg/100 ml, the level of intoxication. Improvements in the prevention and surveillance of sudden deaths that occur at work are suggested. Coroner's records are suggested for use in future surveillance on sudden deaths at work because they identified more sudden deaths at work than death certificates did.

An E1A mutant of adenovirus type 3: Ad3hr15 has reiterated DNA sequences 5' to its E1A gene.

Defective variants of adenovirus type 3 (Ad3) have been isolated from a heterogeneous, high multiplicity passage stock of the virus. A strikingly defective variant, Ad3hr15, fails to propagate on normally permissive A549 cells, yet has greater infectivity than wild type Ad3 in the adenovirus type 5 (Ad5) DNA-transformed 293 cell line. Investigation of its genomic alterations revealed that Ad3hr15 bears two short tandem duplications of viral DNA sequences near its left end, 5' to the E1A gene. The variant also bears a large tandem triplication at its right end. Marker rescue experiments with plasmid-cloned left end DNA sequences of Ad3 implicate that the duplications 5' to E1A are responsible for the Ad3hr15 defect and the E1A structural gene of the variant is functional. Northern analysis revealed no detectable E1A transcripts early after Ad3hr15 infection of A549 cells. The 293 cell line, however, supports high levels of transcription of the Ad3 E1A gene by the mutant Ad3hr15 E1A promoter.

Polar encapsidation of adenovirus DNA: evolutionary variants reveal dispensable sequences near the left ends of Ad3 genomes.

Repeated passage of adenovirus type 3 in HeLa cells has led to a novel stock of variant genomes, characterized by deletions and substitutions of DNA sequences within the left-end 750 base pairs. This heterogeneous stock retains few if any parental genomes--the majority of variants appear viable. Analysis of viable variants with deleted sequences reveals the 182 nucleotides proximal to the left-end inverted terminal repeat (136-318 bp) are not required for Ad3 infectivity in cultured human cell lines nor for maintenance of viral DNA encapsidation polarity.

Simian virus 40 A gene function: further characterization and growth of tsA transformed chinese hamster cells.

Chinese hamster embryo cells transformed with the tsA 58 mutant of Simian virus 40 express the transformed phenotype at the permissive temperature (33 degrees C or 37 degrees C) and a "normal" phenotype at the nonpermissive temperature (40.5 degrees C). Immunofluorescence and immunoprecipitation of T antigens demonstrated that the "T" antigen (100 K) has an increase rate of synthesis and degradation at 40.5 degrees C. However, the cells continue to replicate at the nonpermissive temperature when assayed by flow cytometry and autoradiography. This DNA synthesis was cellular, not viral, and not owing to an increase in DNA repair. When the cell cycle distributions of G1, S, and G2 + M were assayed by the fraction labeled mitoses method, no differences were evident at the permissive and nonpermissive temperature; however, the doubling time was lengthened at 40.5 degrees C (13 hours vs. 100 hours). These results suggest that at 40.5 degrees C, the tsA transformed cells are cycling and dying. However, if the transformed cells are seeded onto monolayers of normal Chinese hamster cells at 40.5 degrees C, the cells are growth arrested when measured by growth assays, flow cytometry, autoradiography, and immunofluorescence for T antigen. Therefore, growth arrest can be obtained in tsA 58 transformed Chinese hamster cells when cocultured with normal Chinese hamster cells.

Replication of Chinese hamster embryo cells transformed by temperature-sensitive T-antigen mutants of simian virus 40.

Chinese hamster embryo cells transformed by simian virus 40 temperature-sensitive T-antigen mutants replicated when confluent at 40.5 degrees C, regardless of the selection method, selection temperature, or virus strain used.

Cytogenetic studies of a simian virus 40 temperature-sensitive mutant-transformed Chinese hamster cell clone at the permissive and nonpermissive temperature.

A clone of Chinese hamster embryo cells transformed by tsA58, the temperature-sensitive mutant of simian virus 40, was analyzed for chromosome abnormalities at the permissive temperature (37 degrees C) and nonpermissive temperature (40.5 degrees C). Trypsin-Giemsa-banded metaphases were analyzed 1 week after the temperature shift. The metaphases from cells at both temperatures were pseudodiploid, with numerous chromosome changes primarily affecting chromosomes no. 1 and 2. Other chromosomes (no. 6, 11, and the X) were also frequently involved. A marker chromosome, LM, was present in 35% of the cells at 40.5 degrees C.

Simian virus 40 A gene function: DNA content analysis of Chinese hamster cells transformed by an early temperature-sensitive virus mutant.

Replication of two Chinese hamster embryo cell lines transformed by an early temperature-sensitive mutant of simian virus 40, tsA58, was examined by flow microfluorometry and autoradiography of [3H]thymidine-labeled cells in order to determine whether transformed cell DNA synthesis is initiated by the virus A gene. At the permissive temperature (37 degrees), cells transformed by the mutant were like the wild-type virus transformants in appearance, colony-forming ability, high saturation density, and rapid replication. At the nonpermissive temperature (40.5 degrees), the tsA58 transformed cells resembled normal embryo fibroblasts and seem to return to normal growth patterns. Although both mutant transformed cell lines at 40.5 degrees appeared to cease growth at low saturation density, the cells did not enter a resting state, but continued to replicate. The cultures were maintained at low densities by a balance among cell replication, cell death, and sloughing of dead cells into the supernatant. These results suggest that the simian virus 40 A gene function effected by the tsA58 mutation does not prevent Chinese hamster embryo transformed cells from entering a resting state, although the gene may control other phenotypic characteristics of transformation.