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Pseudomonas aeruginosa is the leading cause of morbidity and mortality in cystic fibrosis (CF). This study examines the role of organism-specific factors in the pathogenesis of very early P. aeruginosa infection in the CF airway. A total of 168 longitudinally collected P. aeruginosa isolates from children diagnosed with CF following newborn screening were genotyped by pulsed-field gel electrophoresis (PFGE) and phenotyped for 13 virulence factors. Ninety-two strains were identified. Associations between virulence factors and gender, exacerbation, persistence, timing of infection and infection site were assessed using multivariate regression analysis. Persistent strains showed significantly lower pyoverdine, rhamnolipid, haemolysin, total protease, and swimming and twitching motility than strains eradicated by aggressive antibiotic treatments. Initial strains had higher levels of virulence factors, and significantly higher phospholipase C, than subsequent genotypically different strains at initial isolation. Strains from males had significantly lower pyoverdine and swimming motility than females. Colony size was significantly smaller in strains isolated during exacerbation than those isolated during non-exacerbation periods. All virulence factors were higher and swimming motility significantly higher in strains from bronchoalveolar lavage (BAL) and oropharyngeal sites than BAL alone. Using unadjusted regression modelling, age at initial infection and age at isolation of a strain showed U-shaped profiles for most virulence factors. Among subsequent strains, longer time since initial infection meant lower levels of most virulence factors. This study provides new insight into virulence factors underpinning impaired airway clearance seen in CF infants, despite aggressive antibiotic therapy. This information will be important in the development of new strategies to reduce the impact of P. aeruginosa in CF. [ABSTRACT FROM AUTHOR]

The persistence of Langerhans cells, melanocytes and ABH red cell antigens in human epidermal cell cultures derived from neonatal foreskin explants has been investigated. Langerhans cells were recognised via the detection of ATPase activity and by immunohistochemical staining for T6 and HLA-DR antigens. Melanocytes were detected by the dihydroxyphenylalanine (DOPA) technique; whilst the presence of red cell antigens was confirmed by immunohistochemical tests using monoclonal anti-A, anti-B, and anti-H antibodies. Langerhans cells were not detected in the cultured epidermis, and few melanocytes were found more than 0-2 mm from the periphery of the explants. However, both Langerhans cells and melanocytes were found to persist for up to 4 weeks in the explants themselves. Red cell antigens, consistent with the blood group of the donor infant, were consistently detected on the surface of the differentiating keratinocytes throughout their in vitro lifespan. The significance of these findings is discussed. [ABSTRACT FROM AUTHOR]

Aims: Pleomorphic adenomas of the salivary gland have gender and age distributions suggesting that oestrogen has a causal role. However, oestrogen receptor (ER)alpha is expressed at low levels in normal salivary gland tissues and data from salivary gland tumours are conflicting. There is preliminary evidence that the recently described ERbeta may be the major ER in salivary gland tissue. The aim of this study was to determine the nature and extent of ERbeta expression in pleomorphic adenomas of the salivary gland., Methods: Pleomorphic adenomas and normal tissues of the parotid gland from 49 patients were tested for ERalpha and ERbeta expression by semiquantitative immunohistochemistry. Associations were sought with patient age and gender., Results: ERalpha and ERbeta expression was localised mainly to the nuclei of ductal cells in normal tissues and the epithelial components in pleomorphic adenomas. Within each tissue and receptor type there were no associations between ER positivity and patient age or gender. ERbeta was expressed in almost twice as many normal tissues and pleomorphic adenomas as ERalpha. Expression of ERbeta was also significantly higher in tumour compared with normal tissues., Conclusions: This is thought to be the first study of ERbeta in pleomorphic adenomas of the salivary gland. Findings support ERbeta as the major ER in salivary glands, and provide evidence that ERbeta may have a role in the development of pleomorphic adenomas of the salivary gland.

Background: Burkholderia pseudomallei is an important cause of acute fulminant pneumonia and septicaemia in tropical regions of northern Australia and south east Asia. Subacute and chronic forms of the disease also occur. There have been three recent reports of adults with cystic fibrosis (CF) who presumably acquired B pseudomallei infection during extended vacations or residence in either Thailand or northern Australia., Methods: The clinical course, molecular characteristics, serology and response to treatment are described in four adult CF patients infected with B pseudomallei. Polymerase chain reaction (PCR) based methods were used to confirm B pseudomallei and exclude B cepacia complex. Genotyping was performed using randomly amplified polymorphic DNA (RAPD) PCR and pulsed field gel electrophoresis (PFGE)., Results: Four patients are described with a mean duration of infection of 32 months. All but one patient lived in tropical Queensland. Two patients (with the longest duration of infection) deteriorated clinically and one subsequently died of respiratory failure. Both responded to intravenous treatment specifically targeting B pseudomallei. Another patient suffered two severe episodes of acute bronchopneumonia following acquisition of B pseudomallei. Eradication of the organism was not possible in any of the cases. PFGE of a sample isolate from each patient revealed the strains to be unique and RAPD analysis showed retention of the same strain within an individual over time., Conclusions: These findings support a potential pathogenic role for B pseudomallei in CF lung disease, producing both chronic infection and possibly acute bronchopneumonia. Identical isolates are retained over time and are unique, consistent with likely environmental acquisition and not person to person spread. B pseudomallei is emerging as a significant pathogen for patients with CF residing and holidaying in the tropics.

The functional significance of sequence variation within the upstream regulatory region (URR) of six human papillomavirus type 16 (HPV16) cervical cancer isolates from Australia was investigated. Specific changes in transcription factor binding sites leading to increased promoter activity may explain the transforming ability of some episomal HPV16 isolates.

We present an immunocompetent man with extensive warts on the hands, refractory to a number of conventional treatment modalities and causing substantial morbidity and impairment of normal function. Isolated limb infusion (regional intra-arterial chemotherapy) with melphalan and actinomycin D was performed, with substantial clearing of the warts within 2 months. Treatment-induced morbidity was limited to mild local erythema and oedema which resolved within 3 weeks. After 9 months' follow up, the patient had only a few residual warts and was able to resume normal activities.

Squamous cell carcinoma (SCC) of the head and neck remains a major health problem worldwide. Recent advances in cell biology suggest that cancer results from the accumulation of specific genetic mutations, many of which have now been identified. These mutations can cause the activation of genes that promote cellular proliferation or inhibit cell death (oncogenes), or they may inactivate genes that inhibit proliferation or promote cell death (tumour suppressor genes). Although there is no known set sequence of events leading to the formation of SCC of the head and neck, there is evidence that many of the genomic mutations implicated in other forms of cancer have an aetiological role in these tumours. Certain viruses, notably Epstein-Barr virus and some types of human papillomaviruses, are causally related to some head and neck cancers. There is now the prospect of using molecular markers to achieve earlier diagnosis and to aid in the prediction of both tumour behaviour and likely responses to particular treatment modalities.

High rates of cervical cancer have been reported from parts of China and this may reflect a predominance of cervical infection with particularly aggressive human papillomavirus (HPV) variants. This PCR-based investigation of cervical tumours from Sichuan province in central China demonstrated an HPV positivity rate of 88%. HPV 16 was most common (21/34, 61%), followed by HPV 18 (3/34, 9%), while types 33, 45, 58 and 59 were each identified in one specimen. Sequencing of up to 1349 bases of the 21 HPV 16-positive isolates, encompassing the enhancer/promoter of the upstream regulatory region (URR) and the E6 and E7 genes, revealed distinct patterns of genomic stability and variability. An overall mutation rate of 5% was seen in the URR. One isolate had a large deletion of 436 bases in the enhancer; while varying combinations of 21 point mutations were identified in the remainder, impacting several YY1, NF1, TEF-1 and Oct-1 sites. More sequence variations were found in E6 compared to E7 (81% vs. 52% of isolates showing at least one mutation), some of which resulted in changes to the translated amino acids. Since the E6/E7 genes encode the oncogenic proteins essential for malignant transformation, and as their expression is controlled by the URR, it is possible that some of the identified mutations altered the oncogenicity of the virus: either directly by changing amino acid sequences of the E6 or E7 oncoproteins, or indirectly through alterations to transcription factor binding sites in the URR., (Copyright 2000 Wiley-Liss, Inc.)

An optimal method for the processing of archival cervical Papanicolaou (pap)-stained smears for the amplification of human papillomavirus (HPV) DNA by polymerase chain reaction (PCR) was developed. This methodology was then applied to a series of 44 pap smears designated as HPV positive or negative (on the basis of both major and minor cytological criteria) or cervical intraepithelial neoplasia (CIN)-cancer. For the detection of HPV DNA, each sample was tested with the consensus GP5/6 primers, and when negative, with CPI-IIG primers. The HPV DNA was detected in 100% (8 of 8) of CIN-cancer smears using the GP5/6 primers. In smears with cytological evidence of HPV without CIN. the use of both sets of primers yielded positive results in 100% (19 of 19) of the samples. Direct sequence analysis of PCR products showed that 16 of the 27 HPV-positive samples contained more recently described HPV types. When tested with both primer combinations, all 17 cytologically negative smears were positive for beta-globin but negative for HPV DNA. The findings show the value of using archival pap smears for further investigations to address issues such as latency, but they indicate that cytological criteria and DNA technology will be critical factors in the reliability of the results.

This study describes sequence variation in both the enhancer and promoter segments of the upstream regulatory region (URR) of 28 human papillomavirus (HPV) type 18 isolates from cervical cancers, 25 from women treated at an Australian center and three from overseas included for comparison. No large-scale changes were found in either region. Fourteen substitutions were identified in the enhancer region with the number in individual isolates ranging from one to eight. Four substitutions impacted cellular transcription factor binding sites but there were no obvious associations with clinicopathological variables. The promoter segment was found to be more highly conserved than the enhancer, but four of the five point mutations identified involved cellular transcription factor binding motifs including a substitution of C for T at nt 104 which affected 21 samples. This change was found to impact upon a previously unrecognized Yin Yang (YY1) binding site. Electrophoretic mobility shift assays (EMSA) showed that this substitution significantly reduced protein-DNA binding and evidence was sought for its possible clinical implications. Of the 24 women with less than Stage III disease and known clinical outcome, tumor recurrence was observed in all of the 6 women whose isolates had the "prototype" T at nt 104, whereas only 8 of the 18 cancers with the mutation at this YY1 site recurred. This is the first report on URR variation in HPV 18 isolates from the South Pacific region. The study also provides initial data on diversity in the promoter region and preliminary evidence suggesting that a specific point mutation in this region may be clinically significant.

Objective: Our purpose was to determine the prognostic significance of human papillomavirus deoxyribonucleic acid in cervical cancers., Study Design: The polymerase chain reaction was used to detect human papillomavirus deoxyribonucleic acid types 6, 11, 16, 18, 31, 33, 52, or 58 in tumors from 148 patients (equal numbers of whom were disease free or had relapses) surgically treated for stage IB or IIA cancers in a major Australian hospital. Cox regression modeling was used to assess the effect of human papillomavirus status on tumor recurrence, taking into account patient age, clinical stage, histologic node status, and type of tumor., Results: Seventy of 74 (95%) of the recurring tumors and 62 of 74 (84%) of the nonrecurring tumors were human papillomavirus deoxyribonucleic acid positive. The rates of positivity of types 16 and 18 were 64% versus 31% in the recurrers and 65% versus 14% in the nonrecurrers. Human papillomavirus type 18 positivity was associated with a greater risk of recurrence than was type 16 positivity (hazard ratio 1.8; p = 0.03). Clinical stage, nodal metastasis, and young age (< or = 35 years) also had adverse effects on relapse (hazard ratio for each approximately 2)., Conclusion: Human papillomavirus type 18 positivity is a risk factor for tumor recurrence in surgically treated cervical cancer.

The polymerase chain reaction (PCR) was used to detect human papillomavirus (HPV) DNA in Formalin-acetic acid alcohol (FAA)-fixed paraffin-embedded tissue from 40 patients whose primary cervical cancers showed a tentacular pattern of invasion at their advancing edges, and 40 patients (matched by age, International Federation of Obstetrics and Gynecology (FIGO) stage and histology type) whose tumors showed broad front invasion. The rate of HPV DNA positivity was the same in both the tentacular and the broad front tumors (83%), but the ratios of HPV 16 to HPV 18 in the two groups were markedly different (20:10 versus 27:4, respectively). HPV type 18 was detected more frequently in tentacular than broad front tumors (P = 0.03). The overall rates of recurrence and mortality were 15 and 9%, respectively (18 and 10% in the tentacular group compared with 13 and 8% in the broad front group). Univariate analysis showed statistically significant associations between HPV 18 and tumor recurrence (P = 0.04), but not between a tentacular pattern of invasion and tumor recurrence (P < 0.05). The findings to emerge from this survey indicate that the presence of particular HPV types may, in part, mediate the histological and clinical behavior of cervical cancers.

Studies indicate the presence of four different forms of human papillomavirus (HPV) E6/E7 mRNA resulting from differences in transcription patterns and post-transcriptional nuclear splicing. Through a retrospective analysis of 28 cervical cancer patients, correlations were sought between E6/E7 transcription patterns and histologic type, FIGO stage, and tumor aggression. A combined reverse transcription/polymerase chain reaction was used to study E6/E7 transcription patterns in fresh and/or fixed paraffin-embedded tissues of HPV 16-positive cervical cancers. Random cervical biopsies from nine women with no history of cervical disease were included as controls. Two sets of primers used in the investigations detected the full-length (FL) E6, E6*I, and E6*II mRNAs and the FL E6 and E6*I mRNAs, respectively; eight of the tumors were also analyzed using a third primer combination designed to identify the E6*III mRNA. At least two of the transcripts were detected in all of the tumors, whereas E6/E7 mRNAs were not identified in any of the control cervical biopsies. Overall, the transcription patterns were consistent, but the major E6*I mRNA was not detected in two of the tumors. No relationship was found between the E6/E7 transcription patterns and the histological type and differentiation of the tumors, nor with the FIGO stage and the clinical behavior of the disease. The study confirms that E6/E7 transcription is a constant feature of HPV 16-related carcinoma, but the findings indicate that there may be no clinical advantage in adding E6/E7 mRNA detection to the routine assessment of cervical tumors.

To investigate the possibility of human cytomegalovirus (HCMV) involvement in the aetiology of cervical carcinoma or in the development of a more clinically aggressive cancer cell phenotype, biopsies of cervical cancer from 103 women undergoing primary therapy for invasive (Stage Ia to IV) cervical cancer were investigated using a polymerase chain reaction (PCR) designed to detect sequences from the IE mtrII region of HCMV. PCR assays were also used on the same specimens to identify the presence of common human papillomavirus (HPV) types associated with cervical cancer (HPV 16, 18, 31, 33, 52, and 58). Of the 103 cancers examined, only 4 contained detectable HCMV DNAs, a proportion lower than that found by another Australian group investigating the cervical carriage of HCMV in women with normal cervices. In contrast, 89 of these cancers were positive for HPV DNA sequences, with HPV 16 (65/103) and HPV 18 (17/103) being most commonly detected. Three of the 4 HCMV-positive tumors were also positive for HPV 16 DNA. An examination of the relevant histopathological and clinical data revealed no evidence to support a contention that cancers positive for HCMV are associated with any unusual histologic cell types or with aggressive clinical behavior.

Human papillomavirus (HPV) 16 E6/E7 mRNAs have been detected in paraffin-embedded sections of histologically cancer-free pelvic lymph nodes from four of six patients with HPV 16-associated cervical cancer. The cDNA obtained from the viral mRNA by reverse transcription was amplified by the polymerase chain reaction (PCR). Transcripts were present in a small but significant proportion (6/42, 14%) of the histologically negative/HPV 16 DNA-positive lymph node blocks; but neither HPV 16 DNA nor transcripts were found in 12 lymph node blocks from two patients whose cervical cancers were not HPV-related. Both of the HPV 16 mRNAs detectable by the PCR primers used in the assay (the E6*I and the full-length E6 transcript) were found in the primary tumors, but the transcription patterns in the lymph nodes were variable. The presence of HPV E6/E7 mRNAs in lymph nodes of patients with HPV-related cancer may be a more sensitive indicator of metastasis than conventional histology. In addition, their detections is likely to be more significant than that of HPV DNA sequences alone. The practical significance of the findings, however, awaits correlation with the ultimate clinical outcome.

The polymerase chain reaction (PCR) was used to investigate the prevalence and distribution of human papillomavirus (HPV)-16 DNA in paraffin sections of all pelvic lymph nodes removed from 14 patients with Stage Ib-cervical cancer at the time of resection of their primary tumours. The results were compared with those obtained from 8 women with no known history of cervical abnormality. In all, 22 cervical biopsies and 40 I lymph nodes (296 paraffin blocks) were examined. Nine of the 14 cervical cancer patients had primary tumours that were positive for HPV-16 DNA: only 3 of these had lymph nodes with histological evidence of metastasis, and HPV 16 DNA was detected in each of the corresponding paraffin blocks. HPV 16 DNA was also detected in varying proportions (8%-92%) of the histologically-negative lymph nodes from these women. There was no correlation between the HPV DNA-positive lymph nodes and their proximity to the primary tumour. HPV-16 DNA was not identified in any of the lymph nodes from the 5 women whose cancers were not HPV-16-related, or in those of women with no evidence of cervical abnormality. This preliminary survey suggests that HPV DNA is frequently transported from HPV-16-related cervical tumours to regional lymph nodes. However, its practical significance will not be clear until sufficient time has elapsed for correlation of the results with the clinical outcome.

An archival survey of 98 cervical cancer specimens dating from the 1920s to the 1980s was undertaken to determine whether changes had occurred in the prevalence of human papilloma-virus (HPV) DNA. HPV DNA was detected in paraffin sections of cancers fixed in 10% formalin by in situ hybridisation (ISH) using HPV 6, 11, 16, and 18 32P-labelled DNA probes under conditions of high stringency; and by the polymerase chain reaction (PCR) using 20-mer oligonucleotide primers to amplify 109 bases of the E6 region of HPV 16. In 30 instances results obtained from Southern blot hybridisations which had been carried out on specimens of fresh tissue from the same cancers collected during the 1980s were available for comparison. The rates of HPV DNA detection in cervical cancers ranged from 83% (by Southern or PCR) and 70% (by ISH) on specimens from the 1980s, to 50% and 63% (by ISH and PCR, respectively) on specimens from the 1920s. HPV 16 was by far the most common type, being identified by Southern or ISH in approximately 92% of HPV DNA-positive specimens. No significant change in the prevalence of HPV DNA, or of HPV types, in cervical cancers was found over the 65 year period examined.

A simple, and sensitive method for the detection of human papillomavirus (HPV) 16 E6/E7 transcripts in RNA purified from formalin acetic alcohol (FAA)-fixed, paraffin embedded cervical cancer (CaCx) tissue is described. The entire procedure, including polymerase chain reaction (PCR) amplification of cDNA obtained from mRNA by reverse transcription, can be completed within a day. The results obtained compare favourably with those using total nucleic acids extracted from the same fixed material, and are consistent with results using RNA purified from corresponding fresh tumour tissue. Although developed for investigating the expression of HPV in tumours, this method should find general application in studies of gene expression and viral detection.

As part of a study of archival cervical cancer specimens (1920s to 1980s) to determine whether changes have occurred in the prevalence of human papillomavirus (HPV) DNAs, investigations were performed on tissues which had been fixed in either 10% buffered formalin (NBF) or formalin-acetic acid-alcohol (FAA). HPV DNA was detected by in situ hybridization (ISH) using HPV 6, 11, 16 and 18 32P-labelled DNA probes under conditions of high stringency; and by the polymerase chain reaction (PCR) using 20-mer oligonucleotide primers to amplify 109 bases of the E6 region of HPV 16. In some instances results obtained from Southern blot hybridizations, which had been carried out on specimens of fresh cancer tissue, were available for comparison. When tissues had been fixed in NBF, HPV DNA sequences were detected in 53% of specimens by ISH and in 72% by PCR. In comparison, the rates of detection of HPV by ISH and PCR in tissues fixed in FAA were 17% and 21% respectively. These results indicate that FAA is clearly inferior to NBF for the preservation of detectable HPV DNA sequences in tissue sections.

The aim of the present study was to compare the recently developed polymerase chain reaction (PCR) technique with conventional dot-blot DNA hybridization for human papillomavirus (HPV) detection. Cells were collected by cervicovaginal lavage from a study group of 109 women attending a sexually transmitted diseases clinic. Using a machine that we developed for alternation of temperature cycles, HPV was detected in 51% of patients by PCR. By dot-blot hybridization, 44% of the patients were positive. Concordance of combined positive and negative results between PCR and dot blot was 69%. The greater sensitivity of PCR may have accounted for 19% of specimens that were PCR positive but dot-blot negative. Unexpectedly, however, 12% of specimens were dot-blot positive but negative by PCR, and several specimens were discordant for type of HPV. Both HPV DNA tests agreed with cytology in 41% of women, and in 33% cytology was negative in the face of positive PCR and dot blot. Concordance of cytology with just PCR was 59%, and only with dot blot was 56%. Cervicography agreed with both HPV DNA tests in 41% of patients, with PCR alone in 55%, and with dot blot alone in 58%. Biopsy results did not reveal a strong correlation between histopathological criteria of HPV infection and detection of HPV DNA by either PCR or dot-blot hybridization. Thus the present study has shown that PCR is a slightly more sensitive indicator of HPV infection than dot-blot hybridization. Agreement of HPV DNA results with conventional screening tests was not strong, an observation consistent with many comparative studies by others. In conclusion, PCR is slightly more sensitive than DNA hybridization for detection of HPV, it can be used in conjunction with specimen collection by gentle lavage of the cervicovaginal epithelium, and the possibility remains that it may prove suitable as a screening test.

A culture method has been developed which enables a reasonably large quantity of human epidermal cells to be grown in vitro without the use of irradiated 3T3 feeder layers or collagen substrates. These cultures become confluent within 2 or 3 weeks of initiation, and may be sub-cultured several times over their 16-week average lifespan provided that the calcium levels in the cultured medium are controlled. Some of the potential uses of these cultures are briefly discussed.

Explant-derived cultured autologous epidermis was used as a graft in a 41-year-old female patient with burns, the first subject in a clinical trial of the technique. A small full-thickness biopsy specimen which was taken on Day 2 of the hospital admission was used to initiate epidermal cultures, four of which were grafted onto the patient's back and right leg 29 and 35 days later. Three of these epidermal cultures engrafted successfully, in spite of infection with Pseudomonas aeruginosa, which resulted in the loss of some of the conventional, split-thickness meshed autografts that were applied concurrently. The fourth graft, which may have been oriented incorrectly onto the graft bed, was largely unsuccessful, and only small islets of epithelial cells remained after 10 days. The successful grafts produced full-thickness, epidermal coverage with a good cosmetic result and little evidence of contraction during a six-months' follow-up.

The results of a clinical and virological survey of anal warts from 85 predominantly homosexual or bisexual men is presented, together with an improved technique for the surgical treatment of these lesions. Two types of anal warts, which were classified as either "acute" or "chronic" on the basis of their appearance and clinical behaviour, were recognized commonly. However, our laboratory investigations--which consisted of the routine histopathological examination of all specimens, together with immunohistochemical testing for the common wart antigen of specimens from 30 patients and papillomavirus typing by human papillomavirus DNA probing of 27 specimens--failed to reveal any significant differences between the two classes of anal warts. By means of a dot hybridization technique with mixed human papillomavirus DNA probes for types 6 and 11 and types 16 and 18, all wart biopsy specimens that were tested were shown to contain human papillomavirus types 6/11; two specimens also contained human papillomavirus types 16/18. Southern blot hybridization studies of eight specimens revealed that five warts contained human papillomavirus type 6 only and three warts contained human papillomavirus type 11 only. Although the surgical technique that is described was successful in terms of patient acceptance and the eventual eradication of anal warts, there was a high rate of recurrence of the lesions, which necessitated repeat operations in two-thirds of the patients. The need for counselling before surgery and for regular follow-up examinations after surgery is discussed.

A simple, reliable culture system for keratinocytes from anogenital warts is described. Using this technique we found that it was possible to produce multiple confluent keratinocyte cultures from two-thirds of the surgically-excised anogenital wart specimens received in our laboratory. Some morphological and cultural differences between these cells and normal keratinocytes derived from neonatal foreskins were observed, although there was no evidence that wart-derived keratinocytes were 'transformed'. The cultures were tested for evidence of HPV-DNA replication using 32P-labelled HPV-DNA probes, for the production of viral capsid proteins using peroxidase-antiperoxidase staining and for whole virus particles using electron microscopy. Fifty-seven per cent (8 of 14) of the wart cultures tested showed persistence of HPV-DNA (5-100 copies HPV-DNA/cell genome equivalent). However, no viral proteins or particles were detected in any culture. This system may prove to be a useful in vitro model for the study of virus-cell interaction and the role of HPV in the malignant conversion of epithelial cells.

Human papillomavirus infection currently is accepted as a major factor in the etiology of carcinoma of the cervix, vagina and vulva. While the nature of genital human papillomavirus infection in women is well documented, detailed knowledge of the disease in the male partners is lacking. Therefore, a prospective study was done to define the disease in the genitals of heterosexual men and to formulate an appropriate plan of management. We studied 52 men during an 8-month period for evidence of genital human papillomavirus infection. The majority of the lesions occurred on the shaft of the penis and on the foreskin of uncircumcised men. Deoxyribonucleic acid dot hybridization of biopsies of macroscopic warts and suspected warty lesions with mixed human papillomavirus types 6 and/or 11 and 16 and/or 18 probes revealed that 87 and 55 per cent, respectively, were positive for 1 or more of these human papillomavirus types. Of the macroscopic warts and subclinical lesions 52 and 29 per cent, respectively, contained the more potentially oncogenic types 16 and/or 18, either alone or in combination with types 6 and/or 11. There was no evidence of human papillomavirus in any semen or urine sample but human papillomavirus deoxyribonucleic acid sequences were detected in 31 per cent of the biopsies from apparently normal penile skin and in 18 per cent of the urethral mucosal biopsies. We suggest that management of human papillomavirus infection be directed toward prevention and contact screening, together with ablation of localized lesions.

The persistence of Langerhans cells, melanocytes and ABH red cell antigens in human epidermal cell cultures derived from neonatal foreskin explants has been investigated. Langerhans cells were recognised via the detection of ATPase activity and by immunohistochemical staining for T6 and HLA-DR antigens. Melanocytes were detected by the dihydroxyphenylalanine (DOPA) technique; whilst the presence of red cell antigens was confirmed by immunohistochemical tests using monoclonal anti-A, anti-B, and anti-H antibodies. Langerhans cells were not detected in the cultured epidermis, and few melanocytes were found more than 0.2 mm from the periphery of the explants. However, both Langerhans cells and melanocytes were found to persist for up to 4 weeks in the explants themselves. Red cell antigens, consistent with the blood group of the donor infant, were consistently detected on the surface of the differentiating keratinocytes throughout their in vitro lifespan. The significance of these findings is discussed.

Specific varieties of human papillomavirus (HPV) infecting the anogenital region were detected in clinical samples by use of a filter hybridization technique suitable for rapid screening of cervical and anal scrapes. In this way possibly benign types (HPV6 and HPV11) could be differentiated from types thought to be capable of malignant transformation (HPV 16 and HPV 18). Cervical or anal canal cells were applied directly to nylon filters and fixed by u.v. irradiation before hybridization with mixed viral DNA probes under both low- and high-stringency conditions. In addition, probe for the human Alu-repeated DNA sequence was used to assess the relative amount of total nucleic acids in each sample applied to the filter. HPV DNA was detected in 3 of 19 cervical scrapes from patients with no past or present history of wart virus infection or cervical dysplasia. Within a positive study group totalling 71 patients, HPV (6/11 or 16/18) was detected in cervical scrapes from 24% of 41 patients who did not have visible genital dysplasia, 30% of 27 patients with visible genital dysplasia or cervical intraepithelial neoplasia (CIN) I, and in 1 of 3 patients with past CIN II/III. In addition, HPV6/11 or 16/18 DNA was detected in anal scrapes from 3 of 6 male patients and in 85% of genital biopsies. A notably high proportion (4/6) of vaginal condylomata were positive with both the HPV6/11 and the HPV16/18 mixed viral DNA probes. Of the biopsies prepared for histopathology and positive for HPV DNA, the HPV group-specific antigen could be detected in only 60%.