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Human papillomavirus (HPV) hybrid virus-like particles (VLPs) were prepared using complementary regions of the major capsid L1 proteins of HPV-11 and -16. These hybrid L1 proteins were tested for assembly into VLPs, for presentation and mapping of conformational neutralizing epitopes, and as immunogens in rabbits and mice. Two small noncontiguous hypervariable regions of HPV-16 L1, when replaced into the HPV-11 L1 backbone, produced an assembly-positive hybrid L1 which was recognized by the type-specific, conformationally dependent HPV-16 neutralizing monoclonal antibody (N-MAb) H16.V5. Several new N-MAbs that were generated following immunization of mice with wild-type HPV-16 L1 VLPs also recognized this reconstructed VLP, demonstrating that these two hypervariable regions collectively constituted an immunodominant epitope. When a set of hybrid VLPs was tested as immunogens in rabbits, antibodies to both HPV-11 and -16 wild-type L1 VLPs were obtained. One of the hybrid VLPs containing hypervariable FG and HI loops of HPV-16 L1 replaced into an HPV-11 L1 background provoked neutralizing activity against both HPV-11 and HPV-16. In addition, conformationally dependent and type-specific MAbs to both HPV-11 and HPV-16 L1 VLP were obtained from mice immunized with hybrid L1 VLPs. These data indicated that hybrid L1 proteins can be constructed that retain VLP-assembly properties, retain type-specific conformational neutralizing epitopes, can map noncontiguous regions of L1 which constitute type-specific conformational neutralizing epitopes recognized by N-MAbs, and trigger polyclonal antibodies which can neutralize antigenically unrelated HPV types., ((C)2001 Elsevier Science.)

The high-molecular-weight sulfated or sulfonated polysaccharides or polymers cellulose sulfate, dextran sulfate, and polystyrene sulfonate were tested for microbicidal activity against bovine papillomavirus type 1 (BPV-1) and human papillomavirus type 11 (HPV-11) and type 40 (HPV-40). In vitro assays included the BPV-1-induced focus-forming assay and transient infection of human A431 cells with HPVs. The compounds were tested for microbicidal activity directly by preincubation with virus prior to addition to cell cultures and indirectly by addition of virus to compound-treated cells and to virus-coated cells to test inactivation of the virus after virus-cell binding. The data indicated that all three compounds showed direct microbicidal activity with 50% effective concentrations between 10 to 100 microg/ml. These concentrations were nontoxic to cell cultures for both assays. When a clone of C127 cells was tested for microbicidal activity, approximately 10-fold-less compound was required to achieve a 50% reduction in BPV-1-induced foci than for the uncloned parental C127 cells. Pretreatment of cells with compound prior to addition of virus also demonstrated strong microbicidal activity with dextran sulfate and polystyrene sulfonate, but cellulose sulfate required several orders of magnitude more compound for virus inactivation. Polystyrene sulfonate prevented subsequent infection of HPV-11 after virus-cell binding, and this inactivation was observed up to 4 h after addition of virus. These data indicate that the polysulfated and polysulfonated compounds may be useful nontoxic microbicidal compounds that are active against a variety of sexually transmitted disease agents including papillomaviruses.

Malignant progression is a life-threatening consequence of human papillomavirus-associated lesions. In this study, we tested the efficacy of papillomavirus early-gene-based vaccines for prevention of carcinoma development of papillomavirus-induced skin papillomas on rabbits. Rabbit skin papillomas were initiated by infection with cottontail rabbit papillomavirus (CRPV). The papillomas were allowed to grow for 3 months without any treatment intervention. Rabbits were then immunized by gene gun-mediated intracutaneous administration of four DNA plasmids encoding CRPV E1, E2, E6, and E7 genes, respectively. All eight control rabbits receiving vector alone developed invasive carcinoma within 8 to 13 months. In contrast, only two of eight vaccinated rabbits developed carcinoma at 12 and 15 months, respectively. Papilloma growth was suppressed in the majority of vaccinated rabbits but not completely eradicated. These results indicate that gene gun-mediated immunization with papillomavirus early genes may be a promising strategy for prevention of malignant progression of human papillomavirus-associated lesions in humans.

We previously demonstrated that gene gun-based intracutaneous vaccination of rabbits with a combination of, but not with individual papillomavirus E1, E2, E6 and E7 genes provided complete protection against cottontail rabbit papillomavirus (CRPV) infection. In the present study, we tested whether vaccination of inbred and outbred rabbits with a combination of CRPV E1 and E2 genes could provide complete protection against virus infection. In the first experiment, gene gun-based intracutaneous vaccination with E1 and E2 genes prevented papilloma formation in the majority of inbred rabbits and promoted systemic papilloma regression in one non-protected rabbit. In contrast, needle-mediated intramuscular injection of E1 and E2 genes did not prevent papilloma formation nor promoted systemic papilloma regression, indicating an absence of strong protective immunity. In the second experiment, six outbred rabbits were immunized by gene gun-based intracutaneous administration of the E1 and E2 genes. Prevention of papilloma formation or systemic papilloma regression was observed in three vaccinated rabbits. Papillomas persisted on the remaining three rabbits, but were significantly smaller than that on control rabbits. These results suggested that gene gun-based intracutaneous vaccination with the combination of papillomavirus E1 and E2 genes induced strong protective antivirus immunity but may be insufficient for complete protection in an outbred population.

Rabbit oral papillomavirus (ROPV) infects mucosal tissues of domestic rabbits. The viral genomic sequence has been determined and the most related papillomavirus type was the cutaneous cottontail rabbit papillomavirus (CRPV). Homologies between the open reading frames (ORFs) of ROPV and CRPV, however, ranged from 68% amino acid identity for L1 to only 23% identity for E4. Shared features unique to the two rabbit viruses included a large E6 ORF and a small E8 ORF that overlapped the E6 ORF. Serological responses to ROPV L1 viruslike particles (VLPs) were detected in rabbits infected at either the genital or oral mucosa with ROPV. The antibody response was specific to intact ROPV L1 VLP antigen, was first detected at the time of late regression, and persisted at high levels for several months after complete regression. Both oral and genital lesions regressed spontaneously, accompanied by a heavy infiltrate of lymphocytes. ROPV infection of rabbit genital mucosa is a useful model to study host immunological responses to genital papillomavirus infections., (Copyright 2000 Academic Press.)

In this study, cottontail rabbit papillomavirus infection of domestic rabbits was used as an animal model to develop papillomavirus early gene-based vaccines. Groups of rabbits were intracutaneously vaccinated with single papillomavirus early genes E1, E2, E6, and E7 or with a combination of these four genes. Only a fraction of rabbits were protected from subsequent viral challenge when vaccinated with the E1 or E6 gene. Viral tumor growth in those rabbits vaccinated with the E1 or E2 gene was suppressed compared to that in controls. In contrast, seven of nine rabbits vaccinated with the combination of the E1, E2, E6, and E7 genes were completely protected against viral challenge. These data indicated that intracutaneous genetic vaccination with the combination of the E1, E2, E6, and E7 genes can be an effective strategy for immunoprophylaxis of papillomavirus infection.

To test the efficacy of genetic vaccination against papillomavirus infection, plasmid DNA encoding cottontail rabbit papillomavirus (CRPV) E1, E2, E6, E7 or without insert were intramuscularly injected into five groups of rabbits. Peripheral blood mononuclear cells (PBMCs) showed specific proliferation upon in vitro stimulation with E1, E2, E6 or E7 proteins in a majority of vaccinated rabbits but Western blot analysis did not detect antibodies specific for these viral proteins in rabbit serum. All rabbits grew papillomas after virus challenge and none of the rabbits showed systemic papilloma regression. These observations showed that intramuscular injection of plasmid DNA encoding CRPV E1, E2, E6 or E7 induced CD4+ T cell-mediated but not humoral immune responses, and did not result in the protection of rabbits from virus infection.

Sodium dodecyl sulfate (SDS), an alkyl sulfate surfactant derived from an organic alcohol, possesses surfactant properties but also denatures and unfolds both monomeric and subunit proteins. In preliminary experiments, we demonstrated that SDS is a potent inactivator of herpes simplex virus type 2 and human immunodeficiency virus type 1 at concentrations comparable to those used for the surfactant nonoxynol-9. We hypothesized that SDS might be capable of denaturing the capsid proteins of nonenveloped viruses. In this report, we demonstrate inactivation of rabbit, bovine, and human papillomaviruses after brief treatment with dilute solutions of SDS. Effective concentrations were nontoxic to rabbit skin and to split-thickness grafts of human foreskin epithelium. This is the first report of a microbicidal surfactant that will inactivate papillomaviruses. We propose that SDS is now a candidate microbicide for formulation and testing with humans.

The economic impact of disability on employment, earnings, and education appears to be more devastating for women than for men. Women with disabilities who are making the transition either back into the workforce or into the workforce for the first time often face barriers that are unique to this population. Many researchers have described women with disabilities as having a "double disadvantage" that results in social and psychological barriers to their transition back to work. The purpose of this article is to help vocational and career development programs better address the psychosocial needs of women with disabilities by (a) describing key psychosocial barriers faced by women with disabilities in their transition back to work and (b) providing career development strategies designed to ease this transition process for women with disabilities and enhance their employment outcomes.

Cottontail rabbit papillomavirus (CRPV) induces rabbit skin papillomas, which progress to invasive carcinoma in some animals. Two early genes, E7 and E6, have been demonstrated previously to be oncogenes. In this study, we identified two additional transforming genes, E8 and E5. Both E8 and E5 stimulated C127 and BALB/c A31 (A31) cell proliferation and affected cell cycle transition. The E8 and E5 transfectants lost cell contact inhibition, reaching a high saturation density when cultured up to 2 weeks. E8-C127 transfectants formed colonies in soft agar in the presence of platelet-derived growth factor (PDGF) while E5-C127 transfectants formed colonies without the requirement for PDGF. E8-C127 transfectants were highly tumorigenic whereas E5-C127 transfectants showed a weak tumorigenicity in nude mice. Both E8 and E5 A31 transfectants failed to form colonies in soft agar even in the presence of platelet-derived growth factor (PDGF) and did not develop tumors in nude mice. These results clearly showed that CRPVE8 and E5 are oncogenes and that the PDGF beta-receptor signaling pathway may be involved in E8-mediated C127 cell transformation. The difference in colony formation in soft agar and tumorigenicity in nude mice between C127 and A31 cell lines indicates that additional alterations in cellular gene expression are needed for E5- and E8-transfected cells to acquire a malignant phenotype., (Copyright 1998 Academic Press.)

Individuals diagnosed with Crohn's disease are at an increased risk for developing low bone density. The exact cause of low bone mineral density in Crohn's disease patients has not been determined. The purpose of this study was to assess the incidence of low bone mineral density in premenopausal women with Crohn's disease and to determine the role diet plays in bone mineral density for this population. Bone mineral density of the lumbar spine (L2-L4), proximal femur, and forearm was measured in 51 female controls and 50 females with Crohn's disease using dual energy X-ray absorptiometry (Lunar DPXPlus, Madison, WI). Dietary intake for all Crohn's disease participants was analyzed using both 3-d dietary records and a food frequency questionnaire. When compared to healthy controls, bone mineral density values of Crohn's disease participants were decreased for all sites, particularly the spine (1.169 +/- 0.114, p = 0.054), Ward's area (0.831 +/- 0.128, p = 0.052), and the femoral neck (0.927 +/- 0.100, p = 0.01). Factors associated with lower bone density in Crohn's participants were weight, corticosteroid usage, length and age of diagnosis, history and length of resection, and dietary intakes of magnesium, copper, magnesium, vitamin K, and zinc. The results of this study indicate for the first time that diet plays a role in the development of low bone density in premenopausal women with Crohn's disease.

The purpose of this study was to determine if differences exist in premenopausal women between z-scores for lumbar spine and proximal femoral bone mineral densities (BMD). Participants were 237 women ranging in age from 20 to 45 years. BMDs of the lumbar spine and proximal femur (femoral neck, Ward's area, and trochanter) were assessed using dual-energy X-ray absorptiometry (Lunar DPX). Mean (+/-SD) age, height, and weight of the participants were 29.4 +/- 6.9 years, 164.4 +/- 6.1 cm, and 64.9 +/- 12.1 kg, respectively. Lumbar spine BMD and BMD at the femoral neck, Ward's area, and trochanter were significantly correlated with large SEEs (r = 0.59-0. 65; SEE = 0.09-0.11). No positive correlation with age and BMD at any site was seen in this population but a significant negative correlation with age was seen in the proximal femur beginning at age 30. Twenty to 24% of the 20-29-year-olds exhibited a difference in z-scores of greater than 1 between the spine and sites in the proximal femur. This percentage increased to 32-46% in the 30-45-year-olds but the nature of the observed differences changed. The differences in spine and proximal femoral z-scores that are seen in the older age group appear to be the result of the earlier onset of bone loss in the proximal femur rather than an initial difference in peak bone mass which has been maintained.

The athymic mouse xenograft system was used to prepare infectious stocks of two additional anogenital tissue-targeting human papillomaviruses (HPVs) in a manner similar to that for the development of infectious stocks of HPV-11. An anal condyloma from a transplant patient was used as material for extraction of infectious virus, and human foreskin fragments were incubated with the virus suspension and transplanted subrenally into athymic mice. Partial viral sequencing indicated that two rare HPV types (HPV-40 and HPVLVX82/MM7) were concurrently present in both the patient condyloma and the foreskin xenografts, and passage of both types was achieved as a mixed infection with HPV-40 predominating. Xenografts that developed from simultaneous infection of human foreskin fragments with HPV-11, -40, and -LVX82/MM7 virions produced regionally separate areas of HPV-11 and -40 infection as determined by in situ hybridization. In addition, in situ hybridization with HPV-40 and HPVLVX82/MM7 DNA probes demonstrated that both of these HPV types were present as adjacent but separate infections within the same anal condyloma of the transplant patient. These studies indicate that multiple HPV types can simultaneously infect genital tissue and that each HPV type predominantly maintains regional separation within the same papilloma.

This article provides an overview of the philosophical and practical aspects of supported employment. The authors describe: (1) the primary supported employment clientele; (2) the disciplines and professions involved in delivering community-based employment services for the population; and (3) selected intervention strategies that maximize supported employees' job tenure and satisfaction. Guidelines for interdisciplinary collaboration in identifying and implementing supported employment services are offered.

A set of 13 monoclonal antibodies (MAbs) was generated against HPV-6 L1 virus-like particles (VLPs), screened for reactivity to HPV-6 and HPV-11 L1 VLPs by ELISA, and tested for neutralization of HPV-11 infection. Both cross-reactive and type-specific epitopes were detected such that 4 of 13 MAbs reacted to surface conformational sites on HPV-6 L1 VLPs and the remaining 9 MAbs were cross-reactive to both HPV-6 and HPV-11 L1 VLPs. four of the 9 cross-reactive MAbs were able to neutralize HPV-11 infectivity, and 3 of 4 of these neutralizing MAbs (N-MAbs) identified shared surface conformational sites. One N-MAb therefore recognized a surface linear epitope as judged by positive binding to L1 in a Western blot assay. The neutralization status of these cross-reactive MAbs with regard to HPV-6 could not be assayed. These results demonstrated that the closely related HPV types 6 and 11 contain both type-specific and shared neutralizing epitopes.

A panel of 24 monoclonal antibodies (MAbs) was generated against human papillomavirus (HPV) types 16 and 18 L1 virus-like particles (VLPs). The MAbs were screened for reactivity to a variety of VLPs prepared from HPV-6, -11, -16, -18, -31, -33, -35, and -45, cottontail rabbit papillomavirus, bovine papillomavirus type 1, and a set of 35 overlapping 20-amino-acid peptides spanning the entire HPV-16 L1 gene. Type-specific linear and conformational surface epitopes were detected as well as several cross-reactive linear epitopes that showed various levels of cross-reactivity between different genital HPV and animal papillomavirus L1s. Most of the linear epitopes were mapped using synthetic peptides, and the epitopes were identified as being either surface or buried within the VLP as defined by the pattern of reactivity in ELISA using intact and disrupted VLP antigen. These MAbs may be useful reagents to help define neutralizing epitopes of HPV-16 and -18 when infectivity assays become available, and to define the regions of L1 that are exposed on the surface or buried within the assembled capsid.

Several small, raised lesions from the underside of the tongue of domestic rabbits were isolated, and an extract prepared and tested for the presence of rabbit oral papillomavirus (ROPV). Two weeks after inoculation of this extract into the underside of rabbit tongues, multiple small discrete, grey-white nodules were observed that reached a maximum size of 2 mm in diameter by 5 weeks. These lesions showed typical ROPV pathology, and nuclei stained positive for papillomavirus (PV) group-specific antigen (GSA) by immunocytochemistry. Tissue fragments from rabbit tongues were incubated with a suspension of ROPV and placed subrenally into athymic mice. After 60 days, cysts were removed, sections cut for histology, and a virus stock prepared. GSA staining and in situ hybridization demonstrated that the xenografts were morphologically transformed with areas showing strong nuclear staining for viral capsid antigen and ROPV DNA. Extracts prepared from the pooled xenografts contained infectious ROPV as demonstrated by inoculation into the undersurface of tongues of nonimmune New Zealand White rabbits. The results demonstrated that stocks of infectious ROPV can be prepared in the athymic mouse xenograft system for use in studies on the experimental transmission of a mucosal-targeting animal papillomavirus.

Rabbits were immunized with recombinant baculovirus-produced virus-like particles (VLPs) of cottontail rabbit papillomavirus (CRPV) to determine whether these antigens could induce long-term protection against experimental challenge with CRPV. Infectious CRPV and human papillomavirus type 11 L1 VLPs were used as positive and negative control immunogens, respectively. Three groups of immunized animals were challenged with 10-fold serial dilutions of infectious CRPV at 2 weeks, 6 months, and 12 months after immunizations. Antibody titers in serum reached 1:10,000 immediately after the final booster immunization and then decayed to 1:150 at 6 months and 1:100 at 12 months in unchallenged rabbits. Serum neutralization titers followed similar kinetics. Papillomas grew on control-immunized rabbits at sites challenged with 10(-1) (100% of sites), 10(-2) (96% of sites), 10(-3) (63% of sites), and 10(-4) (13% of sites) dilutions of virus. At 2 weeks after CRPV L1 VLP immunizations, the rabbits were completely protected against virus challenge. At both 6 and 12 months after CRPV L1 VLP immunizations, strong protection was also observed. In the last two groups, three of seven rabbits were completely protected and only 4 of 14 or 29% of sites challenged with 10(-1 dilution of virus grew papillomas. Papillomas growing at these four sites were also reduced in size (3.5 +/- 0.7 mm) at 50 days postchallenge compared with sites challenged with 10(-1) dilution on control-immunized rabbits (13.2 +/- 4.2 mm). The results demonstrate that strong and long-lasting protection against experimental challenge with papillomaviruses can be achieved with VLP immunogens.

Data indicate that midlevel providers are more likely to provide health promotion and disease prevention counseling, health education, and preventive health and screening services as well as use more community resources. Although the literature is sparse regarding cancer screening activities by midlevel providers, such activities are routinely taught in midlevel educational programs, and midlevel providers see these activities as consistent with their roles. Therefore, it is only logical to assume that use of midlevel providers, who have already been shown to focus on health promotion and disease prevention, would be an effective way to provide quality, cost-effective cancer screening. In fact, models of this sort exist around the United States at the present time, such as the one at Moffitt Cancer Center's Lifetime Cancer Screening Program in Tampa, Florida. In such programs, midlevel providers are successfully conducting comprehensive cancer screening activities. Outcome data from these programs have yet to be published, however. In a collaborative practice environment, the delivery of preventive care, including cancer screening activities, is best accomplished when both the physician and the midlevel provider agree on the importance of these screening activities and work together as a team to integrate these preventive health activities into their office practice. Office systems that are sensitive to patient preferences and that promote preventive care, such as protocols and checklists or health maintenance flowsheets, along with appropriate education tailored to patients' ages and to their social, cultural, and educational backgrounds also help to improve compliance with cancer screening guidelines. More than ever before, health care reform has been the focus of national debate. Most of the health care reform proposals that have been advanced recognize the importance of primary and preventive health care and the role that midlevel providers could and should be playing in such a system. As competent providers, NPs, CNMs, and PAs have the ability to enhance medicine's effectiveness in preventive care, through improved outreach and more thorough screening.

Anti-idiotypic antibodies (anti-Ids) were generated against three mouse monoclonal antibodies (MAbs) which neutralized three different papillomaviruses. The neutralizing MAbs (N-MAbs) were generated against infectious human papillomavirus type 11 (HPV-11), cottontail rabbit papillomavirus (CRPV), and bovine papillomavirus type 1 (BPV-1), and all recognized surface conformational epitopes that were lost upon denaturation of the virions. The polyclonal anti-Ids were screened in an ELISA using a panel of 10 N-MAbs (4 HPV-11 neutralizing, 5 CRPV neutralizing, and 1 BPV-1 neutralizing), and cross-reactive idiotypic reactivity was observed. Affinity-purified anti-H11.B2 (HPV-11 N-MAb) cross-reacted with the 3 other HPV-11 N-MAbs, was unreactive to a nonneutralizing HPV-11 MAb and all 5 CRPV N-MAbs, but surprisingly was reactive to the BPV-1 N-MAb. In contrast, affinity-purified anti-CRPV-1A (CRPV N-MAb) reacted with all 5 CRPV N-MAbs, but not with any other MAb. Affinity-purified anti-B1.A1 (BPV-1 N-MAb) also showed unexpected cross-reactivity with the 4 HPV-11 N-MAbs, but not with any of the CRPV N-MAbs. Two of the three polyclonal rabbit anti-Ids induced strong anti-viral antibody responses (including virus-neutralizing antibodies) in BALB/c mice following immunization. The results indicate that within a mouse strain, papillomavirus-neutralizing antibodies share cross-reactive idiotopes that group the N-MAbs into several subsets. One group includes the CRPV N-MAbs, and another group includes N-MAbs directed to apparently unrelated papillomaviruses, including HPV-11, BPV-1, and possibly HPV-6.

Postattachment neutralization of papillomaviruses (PVs) was analyzed in three PV-infectivity models: (i) the BPV-1-induced focus-forming assay using C127 cells; (ii) in vitro abortive infection of rabbit RK-13 and Sf1Ep cells with CRPV; and (iii) HPV-11-induced morphological transformation of human foreskin chips in the athymic mouse xenograft system. In each assay system, aliquots of infectious virus were added to the appropriate target cells and incubated at 37 degrees, followed at various postinfection time intervals with neutralizing monoclonal antibodies (N-MAbs) that target surface conformational epitopes. In all three model systems, the N-MAbs were able to neutralize PV infection when added as late as 8 hr after addition of infectious PV to host cells. These results imply that papillomaviruses attach to but do not penetrate inside host cells for a significant period of time and that the bound virus is thus still susceptible to neutralization by neutralizing antibodies.

Baculovirus-expressed human papillomavirus type 11 (HPV-11) major capsid protein (L1) virus-like particles (VLPs) were produced in insect cells and purified on CsCl density gradients. The VLPs retained conformational neutralizing epitopes that were detected by a series of HPV-11-neutralizing monoclonal antibodies. Electron microscopy determined that the HPV-11 L1 VLPs were variable in size with a surface topography similar to that of infectious HPV-11. The VLPs were very antigenic, and induced high titres of neutralizing antibodies in rabbits and mice when used as an immunogen without commercial preparations of adjuvant. These VLP reagents may be effective vaccines for protection against HPV infections.

By July 26, 1994, employers with 15 or more employees must comply with the Americans with Disabilities Act. By understanding the ADA regulations, you can recruit qualified, dedicated employees from a virtually untapped segment of the population.

The crystal structure of the tri-isopropyl silyl species, i-Pr(3)Si(Br(6)-CB(11)H(6)), where the brominated carborane Br(6)-CB(11) H(6)(-) is perhaps the least nucleophilic anion presently known, has revealed the highest degree of silylium cation character (R(3)Si(+)) yet observed. The average C-Si-C angle is 117 degrees , only 3 degrees short of the planarity expected of a pure silylium ion(120 degrees ). This value compares to 114 degrees recently reported for a toluene-solvated silyl cation, [Et(3)Si(toluene)](+) by Lambert and co-workers. The greater silylium ion character of i-Pr(3)Si(Br(6)-CB(11)H(6)) versus [i-Pr(3)Si(toluene)](+) is also reflected in the larger downfield shift of the silicon-29 nuclear magnetic resonance, 109.8 versus

We have investigated three factors that may be related to the recovery of cytomegalovirus (CMV) using the shell vial culture procedure. First, we compared fluorescent-antibody staining of shell vial cultures using a monoclonal antibody to a CMV immediate early antigen at 16 vs 40 hr after inoculation. Of 332 routinely submitted specimens cultured in duplicate and stained at the different times, 25 (7.5%) were positive at 16 hr and 32 (9.6%) were positive at 40 hr. The increased yield was 28%. Second, we analyzed the effect of using duplicate shell vials (both stained at 40 hr) for all routinely submitted CMV cultures. During a 6-month period, 272 (12.5%) of the 2157 cultures processed with duplicate shell vials were positive, including 222 positive in both vials and 50 positive in only one. Assuming that a single-vial setup would have detected 50% of those positive in only one of the two vials, the increased yield attributable to the duplicate vial was estimated at 10% (25/(222 + 25)). Third, we investigated the effects of seeding density and culture age on the shell vial assay. Cell age of greater than 1 day was associated with a decrease in sensitivity both in cultures that were confluent and in those that were subconfluent at the time of inoculation. Incorporating these findings in the routine shell vial culture procedure used in our Clinical Virology Laboratory has resulted in a greater overall detection of CMV in shell vial cultures than in conventional 6-week tube cultures.

Povidone-iodine is an effective broad-spectrum disinfectant with no reported toxicity to the cornea and conjunctiva when applied topically in single dose to the ocular surface. We challenged four strains of Neisseria gonorrhoeae, a clinical isolate of Chlamydia trachomatis, and one strain of herpes simplex virus type II with three different concentrations of povidone-iodine (5%, 1%, and 0.1%) for one minute. The challenge inoculum of Neisseria gonorrhoeae and herpes simplex virus type II were completely sterilized by all three solutions. The chlamydia titer was reduced by two log units at the 5% and 1% concentrations, but not the 0.1% concentration. Povidone-iodine may be of potential use in the prophylaxis of newborns against ophthalmia neonatorum.

We compared two commercially available tests, a direct immunofluorescence assay and an enzyme-linked immunosorbent assay (ELISA), to viral isolation in tissue culture for the laboratory diagnosis of untreated and partially treated experimental herpes simplex virus keratitis. New Zealand albino rabbits were inoculated bilaterally with herpes simplex virus-1 McKrae strain after corneal scarification. One eye of each rabbit was treated with a 1% trifluorothymidine solution daily, starting on the third day after inoculation. The direct immunofluorescence assay showed lower sensitivity for herpes simplex virus detection than viral isolation in tissue culture for both untreated and partially treated eyes. The Herpchek ELISA demonstrated similar sensitivity to tissue culture in detecting herpes simplex virus in untreated eyes. In the treated group, however, the Herpchek ELISA showed a higher percentage of eyes positive for herpes simplex virus than did viral isolation in tissue culture. After the initiation of antiviral therapy, eyes that no longer harbor infectious virus that can be isolated in tissue culture may remain herpes simplex virus antigen-positive and thus be more amenable to laboratory diagnosis using the rapid ELISA method.

The synthesis and structural characterization of a six-coordinate (carbonmonoxy)(porphyrin)iron(II), carbonyl(deuteroporphinato)(tetrahydrofuran)iron(II), is described. The choice of tetrahydrofuran as the sixth ligand represents one of the weakest field and weakest binding ligands possible and leads to stronger than usual bonding of CO to the heme. The Fe--C(CO) bond distance is 1.706 A, unusually short. The Fe--O(THF) bond distance in the low-spin complex is 2.127 A. The Fe--C--O and O--Fe--C groups are essentially linear with angles of 178.3 degrees and 177.4 degrees, respectively. The iron atom is displaced out of plane 0.10 A toward the tightly bound CO. The results suggest that the different CO stretching frequencies observed in myoglobin might result from modulating the bonding of the histidine trans to CO. Crystal data are as follows: a = 11.524 (3) A, b = 14.915 (5) A, c = 10.583 (2) A; alpha = 91.51 (2) degrees, beta = 108.99 (2) degrees, gamma = 102.86 (3) degrees; triclinic, space group P1, Z = 2; rho calcd = 1.363 g/cm3, rho obsd = 1.370 g/cm3.