Prevalence of human papillomavirus in cervical swabs in the Okinawa Islands, Japan.

Although human papillomavirus (HPV) is the main risk factor for invasive cervical cancer, the mechanisms of developing carcinoma are not entirely understood. In particular, the biology and natural history of HPV infection are still unknown. Therefore, we have investigated the incidence of human papillomavirus infection by polymerase chain reaction (PCR) in cervical swabs obtained from women who had cervical cancer screening in three separate areas of the islands of Okinawa. The prevalence of HPV in women aged between 30 and 85 years in the three local areas, Yonashiro-town, Naha-city, and Hirara-city, was consistent (9 to 10%). HPV of various oncogenic types, including HPV16, 18, 31, 33, 35, 35, and 58, were identified in HPV-positive swabs and the prevalence of these types varied in the three areas.

Detection of human papillomavirus DNA in primary and metastatic lesions of carcinoma of the cervix in women from Okinawa, Japan.

Of 351 patients with invasive cervical cancer treated at Ryukyu University Hospital, Okinawa, 293 who were tested for human papilloma virus (HPV) DNA of the primary cervical lesion before the initiation of treatment were considered for the study. The polymerase chain reaction (PCR) was performed using fresh specimens, immediately after sampling. In 250 of 293 patients (85.3%), HPV DNA was detected in cervical tumor by PCR using L1 consensus primer. The positive rate by histology was 89.9% in squamous cell carcinoma, 93.8% in adenosquamous carcinoma, and 51.4% in adenocarcinoma. The former two figures were significantly higher than the latter (p < 0.001 and p = 0.002). Concerning identification of HPV types, HPV 16 was most predominant in squamous cell carcinoma, whereas type 18 was relatively high in adenocarcinoma. However, the type distribution of HPV was different to some extent from those in other countries. During treatment, 489 nodal and other tissue samples were obtained from 113 of 250 HPV DNA-positive patients, and were submitted to an assay of HPV DNA. HPV DNA was amplified in all 55 metastatic samples and also in 12 of 434 nonmetastatic tissues (2.8%). HPV types specified in these samples were always identical with the HPV types determined in their primary tumors. In 154 samples from 29 of 43 HPV DNA-negative patients, HPV DNA was not detected, either in 14 metastatic samples or in 140 histologically benign samples. Cancer-free, but HPV DNA-positive nodal, liver, and pulmonary tissues could be interpreted to be already involved at the time of examination, by observing the clinical course of the disease over time.

Persistence of human papillomavirus infection after therapeutic conization for CIN 3: is it an alarm for disease recurrence?

OBJECTIVE: The aims of this study were (1) to examine whether HPV DNA is persistently detected in the cervix after therapeutic conization for CIN 3 and (2) to explore whether a patient with persistence of HPV infection is at risk of developing recurrent disease. METHODS: Of 74 patients referred with CIN 3, 58 who were tested for HPV DNA in the pretreatment cervical lesions were enrolled in the study. After standard therapeutic conization, patients were followed prospectively at the outpatient clinic. Our follow-up protocol was to follow patients without therapeutic intervention as long as they developed no recurrence or recurrence of CIN 1 or 2, while patients who experienced recurrence of CIN 3 were recommended for reconization or hysterectomy. The polymerase chain reaction for detecting HPV DNA was performed using fresh cell samples from the cervix. RESULTS: In 56 of 58 patients (96.6%), HPV DNAs were detected in their primary cervical lesions prior to conization. With regard to the distribution of HPV types, HPV type 16 family (types 16, 31, and 35) was identified in 28 cases (50.0%), type 18 family (types 18, 33 and 58) in 15 (26.8%), and type X in 18 (32.1%). Up to August 1999, all of the 58 patients have been followed with a mean follow-up period of 31.8 months (range: 12 to 73 months). After treatment, HPV DNA was persistently detected in 11 (19.6%) but negative in 45 (80.4%) of 56 HPV DNA-positive patients. HPV DNA was not detected in both HPV DNA-negative patients. Five of 11 persistently HPV DNA-positive patients (45.5%) developed CIN recurrence, while none of 45 persistently HPV DNA-negative patients did. Thus, there was a significant difference between the recurrence rates of these two groups (P < 0.0001). Both patients who were initially HPV DNA-negative developed no recurrence. Accordingly, the overall recurrence following conservative treatment for CIN 3 was 5 of 58 patients (8.6%). CONCLUSIONS: Patients with persistent HPV infection after conization for CIN 3 should be especially closely followed because they are at increased risk of developing disease recurrence.

A possible interaction of thioredoxin with VDUP1 in HeLa cells detected in a yeast two-hybrid system.

Human thioredoxin (hTrx), a small ubiquitous protein with strong reducing potential, has multiple biological functions, including signal transduction and regulation of the activity of transcription factors. hTrx expression is enhanced in HPV-transformed cancer cells; however, the role of hTrx in the malignant cells is not fully understood. We employed a yeast two-hybrid system to search for proteins that bind to hTrx in HeLa cells, a type of HPV-transformed human cervical cancer cell. In a screen of 1.62 x 10(6) yeast cotransformed with a HeLa cDNA library and an hTrx vector, 13 clones were identified as candidates for hTrx-binding proteins. Among them, 3 clones were found to code in frame for the carboxyl-terminal portion of VDUP1 protein, lacking at most the first 155 residues from the start codon. A reconstructed clone carrying the full-length VDUP1 coding sequence also showed the ability to bind to an hTrx fusion protein. Loss of interaction between VDUP1 and hTrx was observed either when two cysteines (Cys 32 and 35) in hTrx were substituted by serines or when the deletion in VDUP1 was extended from amino acid position 155 to 225 or beyond. The 71-mer peptide fragment (position 155-225) of VDUP-1 alone did not bind to hTrx.

Alterations of protein kinase C, 8-hydroxydeoxyguanosine, and K-ras oncogene in rat lungs exposed to passive smoking.

To investigate the effects of exposure to sidestream cigarette smoke (CS) on the initiation and promotion of lung cancer, two groups of 8 or 10 rats were exposed to CS for a 1 h period twice a day for 8, 12, or 20 weeks. The protein kinase C (PKC) activity of the lung exhibited significant changes of 120, 86 and 81% in the CS groups, compared with the respective control group values in the three exposure periods. The in vitro activation of PKC by the active oxygens was efficiently eliminated by hydroxyl radical scavengers, indicating that hydroxyl radicals are responsible for the PKC activation. For the alterations in the lung nucleus caused by passive smoking, the 12- and 20-week exposure CS groups showed significant increases in the accumulation of 8-hydroxydeoxyguanosine. One rat with K-ras activation by G:C transversion (GGT-->GCT) at codon 12 was found among 26 rats of the CS groups in the three exposure periods. These results show that active oxygens introduced by passive smoking may contribute to K-ras activation as an initiator of a tumor model, possibly through the oxygen-induced DNA damage, and may also contribute to an initial activation and the subsequent down-regulation of PKC as a promoter.

Detection of human papilloma virus type 56 in extragenital Bowen's disease.

A case of Bowen's disease arising on the medial part of the first metatarsal bone of an 81-year-old Japanese woman is described. Histopathologically, proliferation of atypical cells was found throughout the epidermis. Electronmicroscopy revealed virus particles 40-50 nm in diameter in the nuclei of tumour cells at the granular cells just on or below the horny layer. Positive bands were obtained by polymerase chain reaction using a consensus primer of human papilloma virus L1 portion. Sequencing analysis of the amplified DNA revealed the same base sequences and homology as human papilloma virus 56. To the best of our knowledge, this case is the first report in which human papilloma virus 56 was found in a case of extragenital Bowen's disease. We consider it important to understand that human papilloma virus 56, often found in cervical lesions, can be detected in extragenital Bowen's diseases.

Detection of human papilloma virus type 58 in a case of a perianal Bowen's disease coexistent with adult T-cell leukemia.

A case of Bowen's disease (BD) that appeared in the perianal region of a 65-year-old Japanese woman coexistent with chronic adult T cell leukemia (ATL) is described. Histopathological findings revealed that irregularly arranged tumor cells with atypical nuclei throughout the epidermis, which itself disclosed hyperkeratosis, dyskeratotic cells, and clumping cells. Positive staining for HPV antigens was immunohistochemically seen in several nuclei of the tumor cells. Electron microscopic study of the tumor tissue disclosed virus particles of about 50 nm in diameter form the squamous cells. A positive band at 256 bp was obtained by PCR using HPV-L1 primer. The amplified DNA by L1 primer completely corresponded to that of HPV-58.

Association of human papillomavirus type 16 with malignant melanoma.

We report a case of malignant melanoma associated with human papillomavirus (HPV) in a 37-year-old woman. The patient has had numerous brown papular and nodular tumors, 5 to 30 mm in diameter, on her left leg for > 15 years, some of them coalescing rapidly in the last 12 months to a multilobulated black nodule diagnosed as malignant melanoma by histology and immunohistochemistry. HPV type 16 DNA was detected in the melanoma specimen by reverse transcriptase polymerase chain reaction (rt-PCR) and in situ hybridization (ISH) of the tumor tissues. This is the first report of melanoma associated with HPV 16.

[Clinical assessment of rotational digital angiography for the diagnosis of hepatocellular carcinoma].

PURPOSE: The purpose of the present study was to investigate whether rotational digital angiography add new diagnostic informations concerning the localization and feeding artery of hepatocellular carcinoma. MATERIALS AND METHODS: Rotational digital angiography was performed in 100 patients with hepatocellular carcinoma. There are 70 males and 30 females, ranging in age from 40 to 81 years. At first, abdominal anterior-posterior digital subtraction angiography was performed in C-arm system using 12/9/7 inch I.I.-TV camera. After that, rotational digital angiography was carried out using SF-VA 100 system in all patients. In the processing of images, the pulse modes were taken during 360 degrees transverse rotation of the X-ray system at a speed of 4.8 sec. The pictures were displayed in real time on a CRT screen with high resolution, and 3 D images were observed. RESULTS: New diagnostic informations concerning the localization and the feeding artery of the tumor were added in 20 patients using this rotational digital system. The tumors in segment 7/8 (n = 10) were clearly visualized by -90.0 degrees and 90.0 degrees to 135.0 degrees, while tumors in segment 4 (n = 5) were revealed by -45.0 degrees, 49.0 degrees and 60.0 degrees. On the basis of this information, segmental transcatheter arterial embolization could be performed. CONCLUSION: The device has, therefore, been found to be of great value in diagnosing hepatocellular carcinoma, particularly for the feeding artery and the localization of the tumor.

Changes of the hepatic CT absorption value in hemodialysis patients.

The effect of hemodialysis on the CT absorption value of the liver was examined in 14 hemodialysis patients. In the high ultrafiltration group [10 patients with a weight loss of -1.7 to -3.5 (2.9 +/- 0.6) kg], the hepatic CT absorption value increased from 49.6 +/- 7.0 to 58.7 +/- 8.7 Hounsfield units (p less than 0.001) during hemodialysis. The increase in hepatic CT absorption value during dialysis correlated with the weight loss (p less than 0.001, r = 0.798). A 5.3% body weight loss by ultrafiltration resulted in an 18.5% increase in the CT absorption value of the liver. This study suggests that one should be careful in making an early diagnosis of hepatic disease, especially hemosiderosis, in hemodialysis patients based on the hepatic CT absorption value because it is affected by hemodialysis.

[Backward flow signal in the left atrium studied by Doppler echocardiography. Differentiation from mitral regurgitation].

A backward flow signal in the left atrium masquerading as mitral regurgitation was studied by a pulsed Doppler method. The subjects consisted of 20 normal volunteers, 12 cases with mitral valve prolapse syndrome, five cases with rheumatic mitral regurgitation, five cases with lone atrial fibrillation, four cases with asymmetric septal hypertrophy and three cases with the Björk-Shiley tilting disc valve in the mitral position. In two-dimensional echocardiography combined with pulsed Doppler method, a Doppler signal was recorded by locating a sample volume in the left atrium. In all of the cases with mitral valve prolapse syndrome and the cases with the prosthetic valve as well as in all of the normal subjects, the backward flow signal was observed in the left atrium. In three cases with mitral valve prolapse syndrome, it was differentiated from a transvalvular regurgitant flow signal. In all cases with rheumatic mitral regurgitation, the backward flow signal was masked by a turbulent flow signal representing regurgitation. In cases with mitral stenosis, the backward flow signal was scarcely recognized. The duration of the backward flow signal had no relationship with heart rate. The histogram of incidence on the scale of R-R interval revealed normal distribution with a mean value of 0.24 sec (+/- 0.09 sec). Therefore, in cases with tachycardia, the backward flow signal was seen throughout systole. The peak backward flow velocity of Doppler signals was correlated (r = 0.71, p less than 0.01) with the peak forward flow velocity in diastole. The faint backward flow signal seen in cases with mitral stenosis and post-extrasystolic potentiation of the backward flow signal were suggestive of the foregoing relationship. The mechanism producing the backward flow was postulated as a water hammer phenomenon caused by closure of the mitral valve.

Psychological correlates of stress-induced cortisol and growth hormone releases in neurotic patients.

The mirror drawing test was performed in 23 neurotic patients and plasma cortisol (Fk) and growth hormone (GH) were measured serially throughout the test. The patient's personality was assessed by the Minnesota Multiphastic Personality Inventory and related to the endocrine findings. The results indicated that 1) Fk and GH have different personality correlates; 2) the Fk responders tend to be more "defensive" trying to maintain good "self-control" over inner mental processes; 3) high GH level seems to be associated with personality features such as exaggeration, hostility, and a distorted thinking process; 4) psychoendocrine control mechanisms for Fk and GH tend to offset, rather than enhance, each other.

Prolactin and growth hormone responses to psychological stress in normal and neurotic subjects.

In order to study the response of plasma prolactin (PRL) to acute psychological stress and to compare it with that of growth hormone (GH), the mirror drawing test (MDT) was performed in 20 normal controls (11 male, 9 female) and 22 neurotic patients (12 male, 10 female). Plasma PRL and GH were measured serially before, during and after the test. In controls, the test caused no significant change in plasma levels of either hormone. In neurotic males, the response of PRL to the test was not consistent, whereas, in neurotic females, plasma PRL level rose significantly following the test. Increase of GH, on the other hand, was apparent in the neurotics of both sexes. The correlation between the responses of the two hormones in the neurotics was low and non-significant. The results indicate that although the psychoendocrine coping mechanism in the neurotics works less effectively for both PRL and GH, the two hormones may have different psychological correlates.

[Metabolism of synthetic corticosteroid esters in man (author's transl)].

Metabolism of water-soluble synthetic corticosteroid esters, hydrocortisone hemisuccinate (H-H); hydrocortisone phosphate (HP), prednisolone hemisuccinate (P-H), prednisolone phosphate (P-P), dexamethasone phosphate (D-P) and dexamethasone sulfate (D-S) was studied by i.v. administration of steroids in a dose of 100 mg to 6 healthy volunteers and consequtive urine collections over 24 hrs. Uniary metabolites were separated into free, glucuronide, sulfate and "unhydrolyzed" fractions and measured by means of Porter-Silber (P-S) reaction and isonicotinic acid (INH) reaction. In H-H, H-P and D-S, glucuronide conjugates constituted the largest fraction of P-S positive metabolites in 24 hr-urine. In contrast, P-H, P-P and D-P were excreted mainly as free metabolites. Small but variable portions were found as sulfate and "unhydrolized" conjugates. Time course study revealed that, in H-H, H-P and D-S, the free/glucuronide ratio of P-S positive metabolites decreased progressively or remained low throughout the collection periods, whereas in P-H, P-P and D-P the ratio increased markedly in the 4--12 hr period. For all steroids, the INH/P-S ratio was found to be high in the free fraction and low in the glucuronide fraction; the tendency was especially marked in P-H, P-P and d-p. INH-positive metabolites were also predominant in the sulfate and "unhydrolyzed" fractions. The results indicate that the metabolism of synthetic corticosteroids, in general, is characterized by diminised rate of ring A reduction followed by glucuronide conjugation and compensatory increase in free metabolites, the bulk of which consists of ring A intact, C-20 reduced metabolites. 6-Hydroxylated metabolites may also be increased. Conjugation with sulfuric acid occurs but to a small extent. The consistent presence of "unhydrolyzed" metabolites suggests that portions of esters can be excreted unsplitted. It is also noted that the structure of steroid moieties and type of esters were important factors determining the quantitative and qualitative difference in their metabolic fates.