Prevalence of abnormal oral cytology and impact of sexual behavior in women with abnormal cervical cytology.

UNLABELLED: PURPOSE OFINVESTIGATION: To assess the frequency of oral cytological abnormalities in women who have cervical intraepithelial lesions, and transmission of infection depending on their sexual behavior. The authors also aimed to investigate the oral cytological changes in male partners. MATERIAL AND METHODS: Thirty patients with abnormal cervical cytological results via punch biopsy formed the case group, and 68 patients constituted the control group with normal cervical smear results. The Bethesda system was used for classification of the cytological alterations. RESULTS: Oral dysplasia was significantly higher in the squamous intraepithelial lesion (SIL) group. Oral sex percentage was 43.3% in SIL group, whereas it was 19.1% in the control group. History of genital warts in women with SIL was also significantly higher in the case group. Three patients were diagnosed with abnormal oral cytology in the SIL group (10%), however abnormal oral cytology was not detected in the control group. No oral dysplastic changes was identified in the male partners of women with oral lesions. CONCLUSION: The authors detected oral dysplastic changes in the SIL group, especially in the (low grade squamous intraepithelial lesion (LGSIL) patients. Interestingly they could not find any oral dysplastic changes in the male partners of the study population.

Usefulness of elevated red cell distribution width for predicting systemic inflammatory response syndrome after extracorporeal circulation.

OBJECTIVES: Cardiac surgical operations performed by using extracorporeal circulation (ECC) lead to a systemic inflammatory response (SIR). Sometimes SIR may turn into a severe state, the systemic inflammatory response syndrome (SIRS) that usually has a poor outcome with no specific clinical tools described for its prediction. Red cell distribution width (RDW) is a routine hematological parameter. It has been proposed as a marker of morbidity and mortality in various clinical conditions. We aimed to investigate the relationship between high RDW and SIRS which is triggered by ECC. METHODS: Eleven hundred consecutive patients who underwent elective heart surgery with the use of ECC were retrospectively analyzed. A total of 19 patients fulfilled the described SIRS criteria and 20 consecutive patients were selected as the control group. RDW and other laboratory parameters, preoperative clinical status, operative data and postoperative data were compared between the SIRS and the control groups. RESULTS: Baseline characteristics of the patient groups were similar. Significant mortality was found in the SIRS group; 18 (94.73%) patients and 2 (10%) patients in the control group (p < 0.002). RDW was found to be significantly higher in the SIRS group vs the control group (15.02 ± 2.03 vs 13.01 ± 1.93, respectively, p < 0.003). Multiple logistic regression analyses showed an association between high RDW levels and SIRS development (OR for RDW levels exceeding 13.5%; 95% confidence limits of 1.0-1.3; p < 0.04). Total operation time and the need for inotropic support were also found to be significant against the SIRS group (p = 0.049). CONCLUSION: Increased RDW was significantly associated with increased risk of SIRS after ECC. The results of this study suggest that paying attention to RDW might provide valuable clinical information for predicting SIRS development among patients who are candidates for open heart surgery, without incurring additional costs.

Efficacy of a live attenuated Escherichia coli O78:K80 vaccine in chickens and turkeys.

A candidate live vaccine for avian pathogenic Escherichia coli (APEC) was constructed from a virulent field APEC 078 strain by mutation of the aroA gene. The mutant was highly similar to the parent wild-type strain in respect of colony morphology, motility, growth in suspension, hemagglutination, Congo Red binding, HEp-2 cell adhesion, and the elaboration of surface antigens type 1 fimbriae and flagella, although production of curli fimbriae was reduced marginally. The mutant proved avirulent when inoculated into 1-day-old chicks by spray application and when presented again in the drinking water at 7 days of age. Chickens and turkeys vaccinated with an 078 aroA mutant were protected against a challenge at 6 wk of age by virulent APEC strains.

Efficacy and safety of an attenuated live QX-like infectious bronchitis virus strain as a vaccine for chickens.

The attenuation of infectious bronchitis (IB) QX-like virus strain L1148 is described. The virus was passaged multiple times in embryonated specific pathogen free (SPF) chicken eggs, and at different passage levels samples were tested for safety for the respiratory tract and kidneys in 1-day-old SPF chickens. There was a clear decrease in pathogenicity for the respiratory tract and kidneys when the virus had undergone a large number of passages. Passage level 80 was investigated for safety for the reproductive tract in 1-day-old and 7-day-old SPF chickens. In 1-day-old chickens, 12.5% of the vaccinated birds had macroscopic lesions. No lesions were observed if the chickens had been vaccinated at 7 days of age. Passage level 80 was investigated for its ability to spread from vaccinated to non-vaccinated chickens and for dissemination in the body. The virus was able to spread from vaccinated chickens to groups of non-vaccinated chickens, and in the vaccinated birds the virus was found frequently in oro-pharyngeal and cloacal swabs. A fragment of the hypervariable region of the S1 protein of passage level 80 was sequenced and revealed nucleotide changes resulting in two amino acid substitutions. Passage level 80 was given additional passages to levels 82 and 85. Both passage levels were tested for efficacy in SPF chickens and passage level 85 was tested for efficacy in commercial chickens with maternally derived antibodies (MDA) against a challenge with QX-like strain IB D388. In both SPF chickens and chickens with MDA, the vaccines based on strain IB L1148 were efficacious against challenge.

In vivo and In vitro interferon induction in chickens by S -28828, an imidazoquinolinamine immunoenhancer.

Imiquimod and its analogs belonging to a class of imidazoquinolinamines, activate immune system via cytokine induction, and have antitumor and antiviral effects in mammals. In this study, we showed that a related analog, designated S-28828, induced interferon (IFN) and macrophage activating cytokine(s) (macrophage activating factor, MAF) in chickens in vivo, ex vivo, and in vitro. IFN and MAF were detectable in the serum of chickens following oral administration. Serum IFN levels were the highest at 2 h after treatment. Although there was no detectable IFN in sera of chickens at 8, 24, and 48 h after treatment, high levels of interferon inducible enzyme, 2'-5' oligoadenylate synthase (2'5'OAS) were present at these time points. In vitro and ex vivo studies showed that spleen cells, bone marrow (BM) cells, and peripheral blood leukocytes (PBL) were capable of producing IFN and MAF, although spleen cells produced the highest levels. Our results suggest that S-28828 administered orally may be a useful immunoenhancing and antiviral agent for chickens.

Nitric oxide inducing factor as a measure of antigen and mitogen-specific T cell responses in chickens.

We describe here an assay to measure responses of T cells to in vitro stimulation with antigens and a T cell mitogen (ConA). Spleen cells from chickens immunized with live viruses and an inactivated antigen produced macrophage activating factors (MAF) in response to in vitro stimulation with homologous antigens. The production of MAF, quantitated by the induction of NO in a retrovirus transformed macrophage cell line, HD11 (Beug et al., 1979, Cell 18, 375) was antigen-specific and correlated well with T cell proliferation. Further studies showed that production of MAF was abrogated by cyclosporin A, anti-CD4 and anti-CD8 monoclonal antibodies. These data suggested that production of MAF required T cell activation and can be used as measure of antigen and mitogen-specific T cell responses in chickens.

Recombinant canarypoxvirus vaccine carrying the prM/E genes of West Nile virus protects horses against a West Nile virus-mosquito challenge.

An ALVAC (canarypoxvirus)-based recombinant (vCP2017) expressing the prM and E genes derived from a 1999 New York isolate of West Nile virus (WNV) was constructed and assessed for its protective efficacy in horses in two different experiments. In the first trial, a dose titration study was conducted to evaluate both serum neutralising antibody responses to WNV and duration of immunity. In the second trial the onset of protection was determined. Twenty-eight adult horses received two doses of vCP2017 administered intramuscularly at 5-week intervals and sixteen horses comprised age-matched non-vaccinated controls. Individual sera were taken periodically and tested for neutralising antibodies against WNV. Horses were challenged by allowing WNV-infected Aedes albopictus mosquitoes to feed on them two weeks (second trial) or one year (first trial) after the second vaccination. After challenge, horses were monitored for clinical signs of disease, and blood samples were collected for detection of WNV viremia and antibody. In both trials, all vaccinated horses developed neutralising antibodies against WNV. None of the vaccinated or control horses developed clinical signs of WNV disease upon challenge. None of the nine horses challenged 2 weeks after primary vaccination and only one of the ten vaccinated horses challenged 1 year after vaccination developed detectable viremia after challenge, whereas more than 80% of the controls became infected. Results from these studies demonstrated that a primary course of two doses of vCP2017 provides both antibody response and an early immunity in horses against WNV viremia.

Oligonucleotide probes in infectious bronchitis virus diagnosis and strain identification.

Genomic RNA fingerprints of infectious bronchitis virus (IBV) strains M41 and Conn46 were prepared to identify T1 RNase-resistant oligonucleotides 'unique' to each of the two IBV strains. Such oligonucleotides were subsequently eluted from the gels and their nucleotide sequences determined. When oligonucleotide probes of those sequences were synthesized and used in a dot-blot hybridization assay, the probes lacked IBV strain-specificity and reacted with the RNAs of homologous as well as heterologous IBV strains. Based on these results, the methods used in this study need to be applied to a large number of oligonucleotide probes, to find one or a few that might be suitable as IBV strain- or serotype-specific oligonucleotide probes.

A monoclonal antibody blocking ELISA to detect serotype-specific infectious bronchitis virus antibodies.

A monoclonal antibody (Mab) blocking enzyme-linked immunosorbent assay (B-ELISA) was developed and compared to a conventional indirect ELISA (I-ELISA) and a virus-neutralization (VN) test for detection of specific antibodies to avian infectious bronchitis virus (IBV) serotypes. Sera used in this study were derived from chickens experimentally inoculated with the three most prevalent IBV serotypes, Arkansas (Ark), Connecticut (Conn), and Massachusetts (Mass). Overall, there was good correlation between the results of B-ELISA and the VN test. Both detected serotype-specific antibodies in chicken sera during primary and secondary phases of the immune response. Results of both tests indicated that the antibodies produced during the primary response to IBV serotypes are strongly serotype-specific. Those produced during the secondary response react more strongly with the homologous virus, but do exhibit some level of cross-reactivity with heterologous antigens. I-ELISA detected IBV group-specific and not serotype-specific antibodies. The B-ELISA which both offers the convenience of the conventional I-ELISA and the serotype specificity of the VN test, hold excellent promise for field application in IBV diagnosis and evaluation of response to IBV vaccines.

Suppressor macrophages mediate depressed lymphoproliferation in chickens infected with avian reovirus.

A previous study indicated that spleens from reovirus-infected chickens contained macrophages that were primed to produce nitric oxide (NO). The presence of these primed macrophages correlated with depressed in vitro T cell mitogenesis. The current studies indicated that splenic adherent macrophages from virus-exposed chickens inhibited concanavalin A (ConA) induced proliferation of normal spleen cells. ConA-stimulated spleen cells from uninfected chickens, but not virus-exposed chickens, produced large quantities of interleukin-2 (IL-2) and a factor that induced NO production. This factor was tentatively named NO inducing factor (NOIF). The removal of macrophages from the spleens of virus-exposed chickens by plastic adherence resulted in partial recovery of ConA-induced proliferation and the production of normal levels of IL-2 and increased levels of NOIF, although these remained below normal. However, nonadherent spleen cells produced substantial quantities of NO, which indicated an incomplete removal of macrophages. Because removal by plastic adherence did not result in the depletion of all macrophages, spleen cells were panned with anti-CD3 antibody to obtain an almost pure population of T cells. Fractionated T cells from virus-exposed chickens proliferated vigorously to ConA and produced normal levels of IL-2 and NOIF. When splenic adherent cells from virus-exposed chickens were added to purified T cells, the T cells failed to respond to ConA. Addition of splenic adherent cells from virus-free chickens did not induce mitogenic inhibition. Further, the addition of purified T cells from the spleens of reovirus-infected chickens to T cells from virus-free birds did not adversely affect T cell mitogenesis. These data indicated that reovirus infection in chickens does not compromise the functional capabilities of T cells but induces suppressor macrophages that inhibit T cell functions.

Enhanced expression of cytokine genes in spleen macrophages during acute infection with infectious bursal disease virus in chickens.

We examined the effects of infectious bursal disease virus (IBDV) on splenic T cells and macrophages. In acute IBDV infection, splenocytes responded poorly to Con A stimulation. However, when T cells were isolated from whole spleen cells, purified T cells responded normally to Con A. This result indicated that functional T cells were present in the spleen but mitogen-induced proliferation of T cells was being suppressed by other cells. Previous studies indicated that soluble factors from suppressor cells may mediate this inhibition of T cell mitogenesis. We thus examined the effects of IBDV on spleen adherent cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to quantitate the expression of several cytokine genes in splenic macrophages. In acute IBDV infection, splenic macrophages exhibited enhanced gene expression of type I interferon (IFN), chicken myelomonocytic growth factor (cMGF), an avian homolog of mammalian IL-6, and 9E3/CEF4, an avian homolog of mammalian IL-8. Mitogen-stimulated spleen cell cultures also produced elevated levels of nitric oxide. The elevation of cytokine gene expression by macrophages occurred transiently during the acute phase of viral infection and coincided with in vitro inhibition of T cell mitogenic response of spleen cells.

Effect of temperature-sensitive Mycoplasma gallisepticum vaccine preparations and routes of inoculation on resistance of white leghorns to challenge.

One-week-old chickens were vaccinated with live or formalin-killed temperature-sensitive (TS) Mycoplasma gallisepticum (MG) either intranasally (IN) or subcutaneously (SQ). Live TS MG protected chickens against S6 strain challenge directly into the air sacs, regardless of route of vaccination. Killed MG, however, protected chickens only when administered SQ. Antibody to MG was detected in sera and in the tracheal and air-sac washings of only the chickens given live vaccine IN. The antibody present in tracheal and air-sac washings may be one of the mechanisms that play a role in resistance to MG challenge.

A radioimmunoassay for the detection of antibodies to Mycoplasma gallisepticum.

A radioimmunoassay technique was developed to determine the antibodies to Mycoplasma gallisepticum (MG) in sera, yolk fluids, and tracheal washings.

Efficacy of commercial Mycoplasma gallisepticum bacterin (MG-Bac) in preventing air-sac lesions in chickens.

One-week-old chickens were vaccinated with commercial Mycoplasma gallisepticum bacterin subcutaneously and challenged with the S6 strain by the intra-air-sac route 3 weeks later. Significantly fewer vaccinated chickens had air-sac lesions than controls.

Production and characterization of monoclonal antibodies to three infectious bronchitis virus serotypes.

Three panels of monoclonal antibodies (MAbs) were prepared against the spike (S) proteins of infectious bronchitis virus (IBV) strains Arkansas 99, Connecticut 46, and Massachusetts 41. Based on enzyme-linked immunosorbent assay (ELISA), the MAbs were grouped into three categories: 1) group-specific, which reacted with a broad spectrum of homologous and heterologous IBV serotypes; 2) serotype-specific, which reacted only with strains of the homologous serotype; and 3) strain-specific, which reacted "selectively" with only certain strains of homologous and heterologous serotypes. MAbs that displayed serotype specificity were all specific to S1 fractions of the homologous serotype, confirming that epitopes that determine virus serotype are associated with the S1 protein. An excellent correlation was found when the results of IBV serotyping by MAb-based indirect ELISA were compared with those from the conventional virus-neutralization test. This confirms that the MAbs described here will serve as valuable tools in epizootiological studies and serotype-specific diagnosis of IBV infection.

Response of chickens to inoculation with a temperature-sensitive mutant of Mycoplasma gallisepticum.

Newly hatched chickens were inoculated intranasally with either the S6 or TS 100 strain of Mycoplasma gallisepticum (MG) or they were left uninoculated. The three groups of chickens did not differ discernibly in body, spleen, or bursa weight during the 27-day sampling period. However, the S6-inoculated chickens showed a more pronounced cellular response in the nasal passages and had nearly complete lymphoid depletion in the spleens. The TS 100-inoculated birds expressed only a mild cellular reaction, which was localized in the nasal passages. Uninoculated chickens appeared normal histologically. Serologic tests such as rapid serum plate agglutination, hemagglutination-inhibition, and radioimmunoassay were able to detect antibody responses of chickens to MG inoculations yet could not differentiate the response to TS 100 from the response to S6. Tracheal secretions in intact TS 100-inoculated chickens contained antibodies to MG, yet only one-half of the bursectomized inoculated chickens contained detectable antibody, which appeared to be IgG. This led to the conclusion that bursectomy suppresses the appearance of locally synthesized IgG antibodies to MG in tracheal washings. The locally produced antibody was considered important in the development of resistance induced by intranasal inoculation of TS mutants.

An antiviral effect of nitric oxide: inhibition of reovirus replication.

We have previously shown that macrophages from chickens infected with avian reovirus are primed to produce nitric oxide (NO) in response to T cell cytokines and bacterial lipopolysaccharide (LPS). We now show that NO exerts potent antireovirus effects. Reovirus replication was substantially reduced in a chicken macrophage cell line, HD11, induced to make NO by stimulation with LPS or conditioned medium from concanavalin A-stimulated spleen cells. The use of a competitive inhibitor of nitric oxide synthase, NG-monomethyl-L-arginine, reduced the antiviral effect of LPS-stimulated HD11 cells. Cytostatic effects were concurrent with the observed antiviral effects of NO. Among these cytostatic effects were reduction in DNA synthesis, protein synthesis, and mitochondrial metabolism. These results indicated that a potential consequence of macrophage priming following virus infection is the protection of cells against virus-induced replication and cytopathic effects, and this protection may be mediated by the cytostatic effects of NO on the host cell.

Serological and molecular characterization of three enteric isolates of infectious bronchitis virus of chickens.

Three coronaviruses isolated from the intestines of laying chickens were partially characterized. Serological and molecular assays indicated that the enteric coronaviruses are infectious bronchitis virus (IBV) isolates. Although the three isolates were recovered from three unrelated chicken flocks, their RNase T1-resistant oligonucleotide fingerprints were almost identical. The three isolates were not neutralized by antisera specific to IBV serotype Connecticut, but their RNase T1-resistant oligonucleotide fingerprints closely matched the fingerprints of strain Conn-46, a Connecticut serotype. This and the co-fingerprinting data suggested that the three field isolates may have emerged from the Connecticut virus through mutation(s). The mutation(s) apparently involved the S1 protein gene that determines the virus serotype.

Role of harderian glands in resistance against Mycoplasma gallisepticum challenge.

Harderian glands of one-day-old chickens were surgically removed. At one week old, these chickens and controls from which these tissues were not removed, were vaccinated intranasally with a temperature-sensitive mutant of Mycoplasma gallisepticum. Humoral and local immunity were measured by means of antibody in sera and tracheal washings, respectively. Protection was measured by resistance to intra-air-sac challenge with the S6 strain of M gallisepticum. There was no discernible difference in either humoral or local antibody response between vaccinated chickens from which the glands had been removed and control birds. In addition, both groups were significantly protected against air-sac challenge compared with unvaccinated controls. These results indicate that removal of the Harderian glands neither affects the production of antibody to M gallisepticum, nor alters the effectiveness of temperature-sensitive M gallisepticum vaccination. The role that the Harderian glands play in resistance to M gallisepticum is therefore questioned.

Characterization of a novel monoclonal antibody which identifies chicken secretory component.

A mouse monoclonal antibody (MAb) produced against chicken biliary IgA (bIgA) was characterized. This MAb, designated A63, reacted with purified chicken biliary IgA (bIgA) but not with serum IgA in ELISA. In Western blot analysis, it recognized an 80-kDa protein associated with bIgA. MAb A63, but not a chicken IgA-specific MAb, reacted strongly with chicken embryo allantoic fluid, a rich source of SC, in immunoblots. In immunohistostained sections of chicken intestines A63 stained intracytoplasmic vacuoles in the enterocytes consistent with goblet cells, whereas the IgA-specific MAb predominantly stained plasma cells in the lamina propria. Whereas the IgA-specific MAb only reacted with the homologous IgA, MAb A63 reacted with bile samples of chicken, turkey, and quail but not of duck or pheasant. These data collectively indicated that MAb A63 recognized an avian secretory component (sc).