Early prenatal androgen exposure reduces testes size and sperm concentration in sheep without altering neuroendocrine differentiation and masculine sexual behavior.

Prenatal androgens are largely responsible for growth and differentiation of the genital tract and testis and for organization of the control mechanisms regulating male reproductive physiology and behavior. The aim of the present study was to evaluate the impact of inappropriate exposure to excess testosterone (T) during the first trimester of fetal development on the reproductive function, sexual behavior, and fertility potential of rams. We found that biweekly maternal T propionate (100 mg) treatment administered from Day 30-58 of gestation significantly decreased (P < 0.05) postpubertal scrotal circumference and sperm concentration. Prenatal T exposure did not alter ejaculate volume, sperm motility and morphology or testis morphology. There was, however, a trend for more T-exposed rams than controls to be classified as unsatisfactory potential breeders during breeding soundness examinations. Postnatal serum T concentrations were not affected by prenatal T exposure, nor was the expression of key testicular genes essential for spermatogenesis and steroidogenesis. Basal serum LH did not differ between treatment groups, nor did pituitary responsiveness to GnRH. T-exposed rams, like control males, exhibited vigorous libido and were sexually attracted to estrous females. In summary, these results suggest that exposure to exogenous T during the first trimester of gestation can negatively impact spermatogenesis and compromise the reproductive fitness of rams.

Salivary human herpesvirus 8 shedding in renal allograft recipients with Kaposi's sarcoma.

Renal allograft recipients in the Middle East are at high risk of developing Kaposi's sarcoma. This report describes the extent of oral human herpesvirus 8 shedding and the genomic diversity of the virus in five Saudi Arabian kidney transplantation patients in whom Kaposi's sarcoma had developed. PCR protocols were applied to amplify three fragments of the viral genome from whole-mouth saliva, parotid saliva, buccal and palatal exfoliates, plasma, peripheral blood leukocytes and biopsy of the Kaposi's sarcoma lesion, and to quantify the viral load in whole-mouth saliva. Viral DNA was detected in all plasma and biopsy samples, 80% of whole-mouth saliva, 20% of each of the other oral samples, and none of the leukocyte samples. The viral load in the cell-free fraction of whole-mouth saliva ranged between approximately 1.2 x 10(3) and 2.2 x 10(6) genome-copies/ml. Genotypically distinct viral strains were evident: intra-lesionally in 1 patient; intra-orally in one patient; between an oral sample and biopsy in two patients; and in four patients, between an oral sample and plasma, and between plasma and biopsy. Thus, in the patients studied, salivary shedding of human herpesvirus 8 was frequent and could be extensive, and they were prone to multiple infections. Measures to curtail salivary viral transmission to pre- and post-transplantation patients might reduce the incidence of post-transplantation Kaposi's sarcoma.

Genotypic profile of human herpesvirus 8 (Kaposi's sarcoma-associated herpesvirus) in urine.

Human herpesvirus 8 (HHV-8) open reading frame K1 sequences amplified from the urine of 5 of 78 (6.4%) infected people in Malawi were monotypic. In two people, urinary and oral sequences were genotypically different. Comprehensive evaluation of HHV-8 transmission may require characterization of HHV-8 shed both in urine and orally.

Multiple human herpesvirus-8 infection.

In Malawian patients with Kaposi sarcoma (KS) and their relatives, we investigated nucleotide-sequence variation in human herpesvirus-8 (HHV-8) subgenomic DNA, amplified from oral and blood samples by use of polymerase chain reaction. Twenty-four people had amplifiable HHV-8 DNA in >1 sample; 9 (38%) were seropositive for human immunodeficiency virus type 1, 21 (88%) were anti-HHV-8-seropositive, and 7 (29%) had KS. Sequence variation was sought in 3 loci of the HHV-8 genome: the internal repeat domain of open-reading frame (ORF) 73, the KS330 segment of ORF 26, and variable region 1 of ORF K1. Significant intraperson/intersample and intrasample sequence polymorphisms were observed in 14 people (60%). For 3 patients with KS, intraperson genotypic differences, arising from nucleotide sequence variations in ORFs 26 and K1, were found in blood and oral samples. For 2 other patients with KS and for 9 people without KS, intraperson genotypic and subgenotypic differences, originating predominantly from ORF K1, were found in oral samples; for the 2 patients with KS and for 4 individuals without KS, intrasample carriage of distinct ORF K1 sequences also were discernible. Our findings imply HHV-8 superinfection.

Effect of human immunodeficiency virus-1 protease inhibitors on the clearance of human herpesvirus 8 from blood of human immunodeficiency virus-1-infected patients.

The effect of human immunodeficiency virus-1 protease inhibitors on the frequency of human herpesvirus 8 DNA detection from peripheral blood of human immunodeficiency virus-positive persons was evaluated. Thirty-three human immunodeficiency virus-seropositive male patients were studied longitudinally. DNA from open reading frame 26 of the human herpesvirus 8 genome was amplified by the polymerase chain reaction from the CD45+ fraction of peripheral blood before and after the introduction of protease inhibitor therapy. Human herpesvirus 8 IgG status, CD4+ cell counts, and human immunodeficiency virus-1 plasma viral load were also assessed before and after therapy. When both reverse transcriptase inhibitor and protease inhibitor treatment were introduced at the same time, there was an increase in CD4+ T cell counts (P=0.0041), a decrease in human immunodeficiency virus plasma load (P=0.0584), and a decrease in the detection rate of human herpesvirus 8 DNA (P=0.0077). Introducing protease inhibitor to patients already receiving reverse transcriptase inhibitor treatment was associated with an increase in CD4+ T cell counts (P=0.0003), a decrease in human immunodeficiency virus plasma viral load (P=0.0911), and a decrease in the human herpesvirus 8 detection rate (P=0.0412). No significant changes in the titters of anti-human herpesvirus 8 IgG were observed. Treatment with human immunodeficiency virus-1 protease inhibitors is therefore associated with the clearance of human herpesvirus 8 DNA from peripheral blood of human immunodeficiency virus-infected patients. The concomitant decrease in the human immunodeficiency virus plasma load and increase in the peripheral CD4+ cell count suggest that an amelioration in the immune defect following reduction in the burden of human immunodeficiency virus-1 infection is responsible for the clearance of human herpesvirus 8 by protease inhibitors.

Tissue distribution of Epstein-Barr virus genotypes in hosts coinfected by HIV.

BACKGROUND: In healthy people, oral and pharyngeal epithelium preferentially carries Epstein-Barr virus (EBV) belonging to a genotype that possesses three copies of a 29 base-pair repeat in the first intron of the BZLF-1 gene, while peripheral blood mostly carries a genotype that bears two copies. Whether EBV shows differential tropism in HIV-1-coinfected hosts, who are prone to develop oral hairy leukoplakia, has not been studied. METHODS: Tongue scrapings and CD45+-enriched peripheral blood cells of 20 HIV1-infected patients and 40 healthy controls were examined. EBV-specific DNA was amplified from segments in the first intron of the BZLF-1 gene, in exon C of the LMP-1 gene, and the type A/B-specifying domain of the EBNA-3C gene. Size polymorphisms of these amplicons were assessed by agarose gel electrophoresis, and DNA sequence differences among BZLF-1 gene amplicons by single-strand conformation polymorphism analysis. RESULTS: The predominant EBV genotype in peripheral blood as well as tongue carried two copies of the BZLF-1 repeat. In controls, although the BZLF-1 genotype with two copies was exclusively detected in the blood, the genotype with three copies predominated in the tongue. The findings could not be correlated with EBV genotyped according size polymorphisms in the EBNA-3C or LMP-1 genes. DNA sequences of a proportion or all of the clones derived from the BZLF-1 amplicons in the tongues of HIV-1-infected patients were identical to those in the blood. CONCLUSIONS: These findings are consistent with EBV haematogenous superinfection of the tongue of HIV-positive individuals. Such superinfection may precede or lead to the development of oral hairy leukoplakia.

Diversity of naturally occurring Epstein-Barr virus revealed by nucleotide sequence polymorphism in hypervariable domains in the BamHI K and N subgenomic regions.

The extent of nucleotide sequence microheterogeneity varies among subgenomic regions of Epstein-Barr virus (EBV). We examined, in EBV-carrying lymphoid cell lines, the extent of polymorphism in EBV DNA fragments amplified from the BamHI E, K, N and Z regions, and then investigated the diversity of the more hypervariable regions in tissues and body fluids. In cell lines, sequence dissimilarities in a genotype-specifying fragment of the EBNA-3C gene varied from < 1-4% within each genotype; dissimilarities in the first intron of the BZLF- 1 gene were < 2% within each genotype. By contrast, dissimilarities in a C-terminal unique domain of the EBNA-1 gene, and in a fragment that encompasses and is upstream of the LMP-1 start codon, varied between 2 and 7% and were not genotype-specific. The sequence diversity in BamHI K and N regions was then examined in tissues and body fluids by single-strand conformation polymorphism (SSCP) analysis and cycle sequencing. Extensive inter-host diversity was observed, whether the host was co-infected by human immunodeficiency virus (HIV) or not. In the oral cavity of HIV-infected patients, inter-compartmental EBV diversity could be demonstrated, even between sites that were anatomically proximate. Studies of BamHI K clones derived from EBV in oral lesions revealed infection by multiple variants. Identification of hypermutable loci within the EBV genome such as those located in the BamHI K and N regions should permit fine discrimination of individual EBV variants.

Human herpesvirus 8 variants in sarcoid tissues.

BACKGROUND: The cause of sarcoidosis is unknown, although mycobacteria have been implicated. We examined sarcoid tissues for human herpesvirus 8 (HHV-8) in addition to mycobacterial genomic sequences. METHODS: Biopsy samples from 17 patients with sarcoidosis were studied (eight transbronchial, 27 lymph node, two skin, and two oral mucosa). We used tissues (n = 137) from 96 patients without sarcoidosis as negative controls. A nested PCR was applied to amplify a segment of open reading frame (ORF) 26 of the HHV-8 genome, and a heminested PCR was to amplify a segment of ORF 25 of HHV-8 and of the 16 S rRNA gene of mycobacteria. Differences in base sequences of the amplified fragments were resolved with single-strand conformation polymorphism and dideoxy sequencing. FINDINGS: HHV-8 ORF 26 DNA was detected in significantly higher proportions of sarcoid than of non-sarcoid tissue samples from lung (8/8 vs 0/54; p < 0.0001), lymph nodes (26/27 vs 6/29; p < 0.0001), skin (2/2 vs 0/17; p = 0.006), and oral tissues (2/2 vs 1/13; p = 0.029). 31 (82%) of the 38 ORF 26 DNA-positive sarcoid specimens were also positive for ORF 25 DNA. For mycobacteria-like 16 S rRNA DNA, the proportion positive was significantly higher in sarcoid than non-sarcoid tissues for lymph node samples (11/27 vs 2/29; p = 0.003) but not for other tissues (lung 3/8 vs 22/54; skin 2/2 vs 15/17; and oral tissues 1/2 vs 0/13). Overall, the prevalence of HHV-8 ORF 26 sequences was higher in sarcoid tissues than in non-sarcoid tissues (p < 0.0001). When patients whose tissues were included in a masked phase of the study were treated as units of analysis, eight of eight sarcoidosis patients were positive for HHV-8 ORF 26 DNA, compared with three of 56 control patients (p < 0.0001); for mycobacteria-like sequences, three of eight sarcoidosis patients were positive, compared with four of 56 controls (p = 0.0464). The HHV-8 ORF 26 sequences, ten of which were unique, could be segregated into four groups according to peptide motifs. In seven of nine patients from whom biopsy samples were taken from various sites, different sequences were recovered. The mycobacterial sequences amplified from sarcoid tissues were also varied, but none was homologous to those of known species. INTERPRETATION: Variant HHV-8 DNA sequences are found in a wide range of sarcoid but not non-sarcoid tissues. Mycobacteria-like 16 S rRNA sequences are more frequently present in sarcoid lymph nodes and not in other tissue types, but do not indicate infection by a particular mycobacterial species.

Detection of human herpesvirus-8 DNA in oral ulcer tissues of HIV-infected individuals.

OBJECTIVE: To determine the frequency of detection of human herpesvirus-8 (HHV-8) in HIV-related oral ulcers. DESIGN: Analysis of archived biopsy material. METHODS: Nested polymerase chain reaction of DNA extracts. RESULTS: HHV-8 DNA was detected in six of 10 oral ulcers of HIV-positive patients without oral Kaposi's sarcoma (KS) lesions and five of 11 oral KS lesions. The positive non-KS samples were derived from various oral sites. CONCLUSIONS: In HIV-positive people, HHV-8 can infect oral tissues that are not affected by KS.

Presence of human herpesvirus 8 variants in the oral tissues of human immunodeficiency virus-infected persons.

A 210-bp DNA segment specific to the human herpesvirus 8 (HHV-8) genome was amplified by nested polymerase chain reaction from 10 of 14 archived oral biopsy samples of HIV-positive patients in London who had no evidence of oral Kaposi's sarcoma (KS). Various oral sites were represented. Oral tissues from 20 general dental patients not known to be HIV-infected were negative. When DNA sequences of these products were compared with sequences derived from 5 oral KS tissues of AIDS patients in London and 10 skin biopsies of Italian patients with Mediterranean KS (total number of positive tissues = 25), 11 were found to be unique. DNA and predicted peptide motifs of these sequences were also different from those in 28 of 36 HHV-8-positive lesions previously reported from American and African patients. HHV-8 is tropic for the oral mucosa of HIV-infected persons, and HHV-8 variants, though diverse, may be geographically restricted.

Erythema multiforme involving gingiva.

Erythema multiforme is a vesiculobullous condition that may affect skin and/or mucosa. Oral lesions are characterized by hemorrhagic crusting of the lips and ulceration mainly of the non-keratinized mucosa. This paper describes a patient who presented with gingival lesions as well as the more typical oral signs of erythema multiforme.

Viral infections affecting periodontal health.

A wide spectrum of viruses can give rise to oral and periodontal manifestations. This article reviews viral infections of clinical importance to the periodontologist and confirms the important role of the periodontologist in the recognition and treatment of common viral infections of the mouth.

Linear IgA disease manifesting as recalcitrant desquamative gingivitis.

A case of desquamative gingivitis caused by adult linear IgA disease is presented. Management initially proved to be difficult, however, the introduction of sulfapyridine caused rapid resolution of the gingival problem. This is one of the first reports of desquamative gingivitis caused by linear IgA disease successfully treated with sulfapyridine.

Antimicrobial prophylaxis of infective endocarditis: effect of BSAC recommendations on compliance in general practice.

General dental practitioners in the South West Region of the U.K. were surveyed to assess compliance with the recently published recommendations of the British Society for Antimicrobial Chemotherapy on the antimicrobial prophylaxis of infective endocarditis. For the majority of dental patients, the level of prophylaxis had improved compared with previous reports: 75% of practitioners organized the prophylaxis themselves, 80% gave oral amoxycillin to penicillin non-allergic patients, and 86% gave erythromycin to penicillin-allergic patients. Some 51% complied to an acceptable level with recommended amoxycillin schedules but only 2% complied with erythromycin schedules. The BSAC recommended prophylactic regimens have certainly improved compliance although they are not yet universally accepted.

Clinical evaluation of benzydamine, chlorhexidine, and placebo mouthwashes in the management of recurrent aphthous stomatitis.

Eighteen patients with minor recurrent aphthous stomatitis were given alcoholic benzydamine hydrochloride, aqueous chlorhexidine, or a benzydamine-free placebo mouthwash in random order. Each patient used each preparation for a 3-month period. Ulcer diaries were marked at the same time each week during the 9-month test period. Records were made of the number, size, sites, and pain severity of any ulcers present. Statistical analysis of the results showed no significant differences between any of the treatments tested. Stinging of the oral mucosa was the only consistent side effects noted by nine patients using benzydamine, nine patients using placebo, and three patients using chlorhexidine. Eight patients stated a personal preference for benzydamine because of the transient local anesthetic effect of benzydamine, which gave pain relief.

In situ characterisation of the oral mucosal inflammatory cell response of rats induced by 4-nitroquinoline-N-oxide.

The inflammatory infiltrate induced in palatal and lingual mucosae of Sprague-Dawley rats after treatment with the water-soluble carcinogen 4-nitroquinoline-N-oxide (4NQO) was investigated using immunohistochemical methods on acetone-fixed frozen sections. Tissues from untreated and solvent-painted control rats were similar to each other and contained small numbers of OX-19+ T cells and larger numbers of W3/25+, LCA+, OX-19- cells within the lamina propria. This latter population could be divided into Ia+ and Ia- subpopulations. Although mucosal specimens from carcinogen-treated rats showed significantly increased numbers of T lymphocytes cells expressing the Ia+ and Ia-, W3/25+, LCA+, OX-19- antigenic phenotypes formed the two predominant cell populations beneath treated epithelium and surrounding tumour islands. Ia+ cells were often detected as focal collections adjacent to or within overlying epithelium which itself appeared to express Ia. None of the tumours showed this focal Ia+ cell infiltrate or expressed Ia. The infiltrates did not contain significant numbers of cytotoxic T cells (OX-8+ and OX-19+) or NK cells (OX-8+, OX-19[-], large granular lymphocytes) and, except for increased cell numbers, the populations present appeared similar to those found in normal mucosa. These results indicate that although immune reactions are stimulated by 4NQO treatment the effector cells necessary for controlling tumour development and growth are absent, perhaps reflecting a passive or stimulatory role in this experimental carcinogenesis model.

Oral lichen planus: an immunoperoxidase study using monoclonal antibodies to lymphocyte subsets.

Sixteen biopsies from patients with oral lichen planus (10), simple keratosis (3) and lichenoid reactions (3) were studied using monoclonal antibodies directed against lymphocyte markers. T lymphocytes predominated in the cellular infiltrates of the epithelium and lamina propria of all biopsies, and cells positive with Leu 3a and Leu 2a antibodies were also present at both sites in all specimens. However, many of the Leu 3a positive intraepithelial cells had a dendritic appearance and distribution consistent with their being Langerhans cells. Limited results from cell counts on three lichen planus specimens suggested that the majority of intraepithelial T cells were of the cytotoxic/suppressor phenotype (OKT3 and Leu 2a positive).