Surveillance of penicillin-resistant Streptococcus pneumoniae in one American metropolitan area, 1989-1998.

The emergence of invasive penicillin-resistant (PRSP) and multidrug-resistant (MDRP) Streptococcus pneumoniae was tracked from 1989 to 1998 in one medium-sized metropolitan area in the USA, encompassing western West Virginia, including Huntington, the only major city, and neighboring sections of Kentucky and Ohio. Capsular serotyping and antibiotic sensitivity tests were performed on 350 community-acquired isolates comprising 93.1% of all pneumococcal isolates identified. The incidence of PRSP increased from 3 to 10% during the 10 years of the study. Twenty-nine (22.1%) of 131 isolates of serotypes 6, 9, 14, 19, and 23 were PRSP (one-fourth were MDRP) and 1 (0.5%) of 219 other serotypes was PRSP (serotype 35). Invasive PRSP occurred most frequently in young children and in adults aged 80 years and older, 8.9 and 10.9 cases per 100,000 persons, respectively.

A 20-year epidemiological study of pneumococcal meningitis.

We conducted a retrospective analysis of 55 community-acquired Streptococcus pneumoniae meningitis illnesses in Huntington, West Virginia, from 1978 to 1997. Fourteen (36.8%) of 38 adults and 2 (11.8%) of 17 children died. Serotypes 6, 23, 3, and 18 accounted for 20 (41.7%) of 48 strains available for serotyping. Of 40 strains available for antimicrobial susceptibility testing, 1 serotype 19 and 1 serotype 23 strain showed intermediate resistance and a second serotype 23 strain showed high resistance to penicillin; all three patients survived. The case-fatality rates among adults who received penicillin alone, gentamicin in combination, or vancomycin and cephalosporin together were 57.1%, 55.5%, and 60%, respectively, and among those who received chloramphenicol or a third-generation cephalosporin, they were 11.1% or nil, respectively. No child died who received chloramphenicol or vancomycin. Two (33%) of 6 children died who received a third-generation cephalosporin; both were critically ill when initially treated. No child and one adult had received pneumococcal vaccine prior to becoming ill.

Bacteremic pneumococcal pneumonia in one American City: a 20-year longitudinal study, 1978-1997.

A surveillance of bacteremic pneumococcal pneumonia was conducted in Huntington, West Virginia, from 1978 to 1997 to investigate case-fatality rates, incidence of disease, capsular types, and antibiotic usage. Our study population comprised consecutive inpatients admitted to the hospitals in Huntington, West Virginia, and included 45 children younger than 15 years and 328 adults. All blood isolates were serotyped by capsular swelling procedures; clinical characteristics, treatment, and outcome for all patients were abstracted from hospital charts. The overall case-fatality rate was 20.3%, with most deaths occurring among adults older than 50 years. Case-fatality rates peaked at 37.7% among patients 80 years of age and older. Only 1 of 45 (2.2%) children died. Case-fatality rates declined in each successive 5-year period, from 30.2% in 1978-1982 to 15.6% in 1993-1997. In that same period, incidence rates increased severalfold among children younger than 4 years to 44.5 cases per 100,000 population and among adults 70 years and 80 years of age and older to 38.5 and 76.2 cases per 100,000, respectively. Of the 34 serotypes isolated, 10 accounted for two thirds of the cases of pneumonia: 1, 4, 9, 14, 3, 6, 12, 5, 23, and 19 (in rank order). Chronic renal disease and arteriosclerotic heart disease increased the risk of death. Treatment regimens that included a macrolide and a penicillin or cephalosporin resulted in the lowest case-fatality rate in adults older than 50 years: 6% in 1993-1997. In conclusion, as bacteremic pneumococcal pneumonia evolved over time, the case-fatality rate decreased, its incidence increased, predominant capsular types changed, and treatment regimens that included a macrolide resulted in the lowest fatality rates.

Identification of a variant subgroup A strain of respiratory syncytial virus.

During epidemiologic surveillance of children with respiratory syncytial virus (RSV) disease in Huntington, W.Va., we identified seven strains of a new variant subgroup A RSV (subgroup A-Var) by their reactions in an enzyme immunoassay with two anti-F monoclonal antibodies (MAbs) specific for two epitopes, F1 and F4, generated against the subgroup B RSV. The prototype strain of subgroup A and all other subgroup A field strains from that epidemiologic year failed to react with these two subgroup B MAbs. Additional enzyme immunoassays with 18 subgroup B anti-F MAbs specific for 14 epitopes showed that subgroup A-Var strains also reacted with a MAb specific for the subgroup B F2 epitope. In a radioimmune precipitation assay, the molecular size of the subgroup A-Var F2 subunit of the fusion (F) protein clearly differed from those of both prototype strains of subgroup A and subgroup B RSV. The molecular size of the F2 subunit of subgroup A-Var (24 kDa) was intermediate between the size of the F2 subunit of subgroup A (25 kDa) and that of subgroup B (23 kDa). However, the molecular sizes of the F1 subunits of both subgroup A and subgroup A-Var were identical (54 kDa) and slightly larger than those of the F1 subunits of both subgroups B1 and B2 (53 kDa). These data suggest that subgroup A-Var may represent a distinct RSV A subgroup, analogous to subgroup B1 and B2 RSV, and it is the first-identified naturally occurring subgroup A RSV with an F protein different from that of the prototype A RSV.

Emergence of penicillin-resistant invasive pneumococci in a single American community.

Three decades ago, penicillin-resistant strains of Streptococcus pneumoniae were first reported. Since then, myriad penicillin resistant strains of S. pneumoniae have been identified worldwide and in the United States. No resistant or intermediate resistant strains have been reported in West Virginia because testing has not been done. Between 1983 and 1994, the authors' surveillance of invasive pneumococcal disease in metropolitan Huntington, West Virginia, identified 356 pneumococcal strains from blood and other usually sterile sites, including 110 strains belonging to serotypes 6, 9, 14, 19, and 23, the main serotypes exhibiting penicillin resistance. The authors tested these serotypes for penicillin susceptibility by the E-test. Sixteen (14.5%) strains of types 6, 9, 14, 19, and 23 exhibited intermediate resistance to penicillin. No highly resistant strains were identified. Most of the intermediate resistant strains of types 9, 14, and 23 were detected in epidemiologic years 1992-1994. The increasing number of intermediate resistant penicillin strains signals the need for routine testing of invasive pneumococcal strains for penicillin susceptibility and necessitates appropriate antibiotic usage.

Reactivities of a monoclonal antibody directed against an immunosuppressive molecule in boar seminal plasma.

Using a monoclonal antibody (LIS-4) to an immunosuppressive factor isolated from boar vesicular gland secretion it was determined that this gland secretes a tissue-specific immunosuppressive molecule that is absorbed onto the acrosome of spermatozoa during ejaculation. Absorption of the immunosuppressive molecule onto murine embryos at the 2-, 4-, 8-cell, morula and blastocyst stages in vitro was evaluated by indirect immunofluorescence. In vivo absorption was detected on the zona pellucida of murine embryos obtained from oviducts injected with the immunosuppressive molecule. Immunofluorescence revealed that the immunosuppressive molecule was not absorbed onto murine embryos after solubilization of the zona pellucida. There was no effect of the antibody to the immunosuppressive molecule on the ability of boar spermatozoa to penetrate the porcine zona pellucida.

Characterization of a new porcine differentiation antigen involved in the proliferative response to mitogens.

The cytotoxic monoclonal antibody (Mab) LIL 13 reacts with a widely distributed antigen that is expressed on 80-95% of swine peripheral blood mononuclear cells (PBMC) and a variety of lymphoid cell types. Using indirect immunofluorescence (IIF) the positive cells (75-100%) were divided between the bright, intermediate and dull populations. The remaining negative cell population contained B-cells, T-cells and probably null cells. Mab LIL 13 did not react with swine major histocompatibility complex (MHC) class I antigens (SLA) and did not inhibit E-rosette formation. Reactivity of LIL 13 with leukocyte function antigen 1 (LFA-1) was excluded by competitive IIF and cytotoxicity tests with cross-reacting anti-human CD 18 or anti-swine LFA-1-specific antibodies. Mab LIL 13 and complement treatment severely reduced mitogen-induced proliferative response to phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM) and lipopolysaccharide (LPS) (90-100%). In the absence of complement, LIL 13 partially reduced proliferation of cells by interfering with the capability of mitogens to bind to the corresponding surface receptor (LIL 13 followed by mitogens), and partially inhibited mitogenic proliferative response following post-treatment (mitogens followed by LIL 13). Biochemical analysis of the antigen using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed bands of approximately 180-190 kDa and 46-50 kDa under reducing conditions and 200 kDa and 46-50 kDa under nonreducing conditions.

Effect of boar seminal plasma immunosuppressive factor on NK cell activity and skin graft survival.

The B 10 strain of mice was used to test the effect of the boar seminal vesicle immunosuppressive factor on the female mouse response to the male-specific transplantation antigen. Influence of this factor on human natural killer (NK) cell activity was also studied. No inhibitory effect of the immunosuppressive factor on graft survival was apparent during a time of more than 200 days, nor did the factor suppress NK cell activity.

High performance ion exclusion chromatographic characterization of the vaginal organic acids in women with bacterial vaginosis.

Vaginal organic acids have previously been detected by gas-liquid chromatography, but we have applied an ion exclusion high performance liquid chromatographic procedure to the analysis of vaginal discharge samples. This procedure has the advantage of not requiring derivitization of non-volatile acids and provides the convenience of a technique which does not require the use of flammable gasses, while allowing the identification of at least 18 different acids from the same chromatographic analysis. Vaginal discharge from women with symptoms of bacterial vaginosis was collected on weighed swabs and analysed for the presence of organic acids. The results were compared to the organic acid content of samples obtained from the same cohort of women after treatment with metronidazole. In addition, samples were obtained from asymptomatic women and these samples were analysed in the same manner. The number of organic acids present in samples from women with bacterial vaginosis was greater than the number found after treatment or among asymptomatic women. Succinic acid appeared to be inversely related to lactate concentration and succinate:lactate ratios were greater among women with bacterial vaginosis before treatment than after treatment. Liquid chromatography has proven useful as a means of evaluating the metabolic end-products of vaginal microorganisms in situ.

Identification of subgroups of bovine respiratory syncytial virus.

The occurrence of antigenic variation among nine isolates of bovine respiratory syncytial virus (BRSV) was determined by examining their reaction patterns to human respiratory syncytial virus (HRSV) subgroup A and B monoclonal antibodies (MAbs) by enzyme immunoassay and radioimmunoprecipitation with fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis by using MAbs and polyclonal antisera to HRSV and BRSV. Shared epitopes were demonstrated on four of five structural proteins between BRSV and both subgroups A and B of HRSV. The nine isolates of BRSV showed different patterns of reactivity in enzyme immunoassays with panels of MAbs to HRSV subgroups A and B. Major variations in the molecular weights of the P (phosphoprotein) and F (fusion protein) proteins were demonstrated among the BRSV isolates tested. These results suggest that BRSV belongs to a different antigenic grouping than HRSV and that BRSV is composed of two distinct subgroups.

Distinguishing between respiratory syncytial virus subgroups by protein profile analysis.

We subgrouped 75 strains of respiratory syncytial virus by a protein profile method (PPM) which relies on different mobilities of the phosphoprotein in one-dimensional polyacrylamide gel electrophoresis and does not require monoclonal antibodies. When compared with enzyme immunoassay, PPM correctly subgrouped 54 of 56 subgroup A and all 19 subgroup B strains.

Effects of chlorhexidine gluconate douche on normal vaginal flora.

The effects of a 0.5% aqueous chlorhexidine gluconate (CHG) douche on the normal vaginal flora of twenty healthy nonpregnant volunteers were investigated. The douche was applied in a premeasured 180-ml quantity daily for 7 consecutive days. Specimens for aerobic and anaerobic culture of the vaginal flora were obtained on 3 occasions from each volunteer, once before and twice after CHG use. The culture results were analyzed qualitatively and quantitatively. A blood sample was also collected within 24 h of the last CHG application to determine serum CHG concentration. Lactobacillus spp., Gardnerella vaginalis and Staphylococcus epidermidis were the most prevalent aerobic bacteria at all phases of the study and Bacteroides spp. were the most prevalent anaerobes. The composition of the normal flora was essentially the same 30 days after the last dose as the pretreatment flora. Small amounts of CHG were detected in the serum of all but one subject. No untoward effects on the participants were observed. Candida albicans counts were slightly higher, but prevalence was not significantly increased at the end of the study.

Immunosuppressive factor in cow follicular fluid.

The activity of the immunosuppressive fraction and proteinase inhibitor isolated from cow follicular fluid was investigated. The immunosuppressive factor was separated from the accompanying proteinase inhibitor by Sepharose 6B and Sephacryl S-200 column chromatography. In vitro, the fraction significantly reduced mitogen-induced lymphocyte proliferation. In vivo, the fraction inhibited mouse plaque formation. However, the factor had no effect on the development of intact or zona free embryos. The active components have a molecular mass of about 110,000.

Lack of effect of boar seminal vesicle fluid immunosuppressor factor on embryo development.

The effects of an immunosuppressive factor (ISF) isolated from boar seminal vesicle fluid on in vitro and in vivo mouse development were investigated. It was found that the zona pellucida of porcine and mouse oocytes pre-incubated with ISF reacted in indirect immunofluorescence with antibodies against ISF. Further results indicated that the boar ISF had no effect on embryo development.

In vivo and in vitro immunosuppression by boar seminal vesicle fluid fraction.

Boar seminal vesicle fluid inhibits blast transformation of porcine lymphocytes. Boar seminal vesicle fluid was precipitated in 8% ethanol and the dissolved precipitate separated into two peaks upon chromatography on a Sephacryl S-200 column. The inhibitory activity was found predominantly in the second peak. This peak was rechromatographed on the Sephacryl S-200 column, and the fractions with inhibitory activity were pooled and passed down a Sephadex G-75 column. This chromatography run separated the accompanying protease inhibitor from the immunosuppressive fraction. The molecular mass, estimated by chromatography on the Sephadex G-75 column, was 25,000-27,000 for the immunosuppressive factor and 7,500 for the protease inhibitor. Activity for the immunosuppressive fraction was also demonstrated in vivo.