Using two direct introduction methods, DNA synthesis or cell proliferation activities of three purified proteins from E. coli, namely, human papillomavirus (HPV) E7 proteins of type 16, a mutant type 16 (24 C-G) (transformation defective) and type 6b, were measured in mouse fibroblast, C127 cells. By a microinjection method, the order of the cell mitotic indexes for the three E 7 proteins as determined by 5-bromo-2'-deoxy-uridine (BrdU) staining was type 16, 6b and 16 (24 C-G). By the osmotic shock method, the ³H-TdR incorporation and coloration by (3-carboxymethoxy phenyl)-2-(4-sulfophenyl)-2H-tetolazorium (MTS) for the three proteins correlated with the pRb binding and focus forming activities previously reported (hunger et al. 1991). These results indicate that the simple osmotic shock method for direct protein introduction may be generally useful for transformation assays of oncoproteins.

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It has been previously reported that infection by a bovine enterovirus (MZ-468), which does not infect normal human cells, was remarkably effective for the therapy of adult T cell leukemia (ATL) in experimental rabbit models (Shingu et al. 1991). Therapeutic treatment studies were performed twice on 30 and 20 autochthonous rat ovarian carcinomas induced by DMBA. The average tumor size of the virus treatment group was significantly smaller than the control group when analyzed by a nonparametric Mann-Whitney method (p<0.06 and 0.002 for the 1st and 2nd experiments, respectively). These results indicate that an enterovirus infection may be clinically useful for the therapy of human ovarian cancers with bad prognoses.

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A 68-year-old woman presented a one-month history of lower abdominal pain and weight loss, and was admitted to our hospital. On physical examination, a large hard mass was palpated in her right lower abdomen. An ultrasonograph and computed tomographic (CT) scan revealed a right ovarian tumor that measured 6.9.4.9 cm in size. A total hysterectomy and bilateral salpingo-oophorectomy were performed. The postoperative diagnosis of the tumor was squamous cell carcinoma (SCC) of the ovary. She died of infection and disseminated intravascular coagulation 5 months after surgery. The clinical and autopsy examinations did not show the primary lesions of SCC except in the right ovary. Mature cystic teratoma, Brenner tumor and endometriosis, which are ordinary regarded as the histogenesis of ovarian SCC, were not found, but a few surface epithelial inclusion cysts with squamous metaplasia were observed in non-cancerous area of the right ovary, and the contiguous transition from the metaplastic cyst wall to SCC was confirmed by stepwise serial sections. The present case suggests that the surface epithelium of ovary could be the fourth possibility in the histogenesis of the ovarian SCC.

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A patient with a Sertoli-stromal cell tumor (SSCT) that contained metastatic lung cancer tissue is described, the first such report. The left ovarian tumor, which developed in a patient known to have lung cancer, mainly exhibited the histology of SSCT with a retiform pattern, and included a component of well-differentiated adenocarcinoma. lmmunohistochemical specific stain for surfactant distinguished the adenocarcinoma from the primary ovarian tumor.

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The clinical usefulness of tissue reflection spectrophotometry in the prognosis of ischemic limbs was evaluated. The diseased group (n=12) consisted of 7 cases of femoral artery thrombosis, 4 cases of chronic obstructive arteriosclerosis and 1 case of femoral vein thrombosis. The normal control group consisted of 14 limbs in 7 normal subjects (n=14). Index of tissue hemoglobin concentration (IHb) was 37.3±4.5 (n=14) in the normal control group and 38.5±16.9 (n=12) in the diseased group, which was not significantly different. In a subgroup of patients who required amputation or developed necrosis (severe subgroup), the value was 54.2±1.7 (n=6). This was significantly higher than the value of 22.8±8.5 (n=6) obtained in a subgroup of patients who received the conservative treatment of thrombectomy (mild subgroup) (p<0.05). Index of tissue oxygen saturation (ISO₂) was 28.0±4.9 (n=14) in the normal control group, and 9.1±2.7 (n=12) in the diseased group (p<0.05). 1502 was 7.0±1.6 (n=6) in the severe subgroup which was significantly lower than 11.2±1.9 (n=6) in the mild subgroup (p<0.05). Tissue reflection spectrophotometry therefore appears to provide clinically useful information for the prognosis of ischemic limbs.

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A 55-year-old man was admitted because of weakness in the legs of a few days duration. Neurological examination revealed paraparesis, ataxia of the left limbs, superficial hypoesthesia below the T-10 dermatome level and urinary retention. The antibody titers to human T-lymphotropic virus type I (HTLV-I) were ×640 in the serum and ×16 in the cerebrospinal fluid (CSF). The HTLV-I gag proteins, p19, p24, p46 and p53 were identified by Western blotting analysis of the serum and CSF. The CSF contained 691 white cells/c.mm; protein, 260 mg/dl; IgG, 84 mg/dl; and IgG index, 2.49 . Although the paraparesis and urinary retention disappeared within 10 days, he developed right uveitis which responded well to corticosteroid treatment . With improvement of the sensory impairment, cerebellar signs and CSF pleocytosis as well as uveitis, he was discharged eight weeks after admission. HTLV-I uveitis is a recently established disease entity. The case emphasizes the need to test blood and CSF for HTLV-I antibodies if patients developed myelopathy and cerebellar signs of acute onset, especially when associated with uveitis.

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A case of benign intracerebellar cyst is reported in a 61-yearold woman who presented progressive neurological deficits. MRI and operative findings could not reveal any evidence of neoplastic growth. The biopsied cyst wall revealed normal cerebellar tissue. The simple cyst of the cerebellum is rare, thus we discuss the clinical features and ethiology of this rare entity.

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From 1966 to 1992, a total of 3, 118 gastric cancer (GC) patients were referred to the First Department of Surgery, Kurume University Hospital. Fifty-six (0.02%) of these patients also had liver cirrhosis (LC). Forty-nine (87.5%) of these patients with LC underwent GC resection. Postoperative complications occurred in 32.7%, but the operative mortality rate was zero using our preoperative assessment of liver function to decide the operation. The 5-year survival rate following curative resection was 69% in early cancer and 17% in advanced cancer. Deaths were mainly cirrhosisrelated in the early cancer cases, while cancer recurrence and hepatic failure were the main causes of mortality in the advanced cancer cases.

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In situ hybridization has been used for the detection of viral and bacterial mRNA and DNA which has infected tissue sections, but the sensitivity of this method is very limited. In this paper, a new, sensitive and simple non-isotopic in situ hybridization method in combination with a polymerise chain reaction (PCR) technique is described, in which the genomic or viral DNA can be amplified directly on formalin-fixed and paraffin-embedded tissue sections. As low as two copies of target DNA per cell can be detected by this in situ hybridization after in situ PCR. The essential modification concerns a two step cycling PCR procedure that was applied using single primer pairs. The first step, annealing, was achieved at a low temperature for a relatively long time. The second step was performed at a high temperature for a short period of time. This procedure greatly decreased the non-specific amplification of DNA and increased the sensitivity of detection.

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The pathological assessment of the pulmonary vascular bed is essential for the diagnosis and management of congenital heart disease with pulmonary hypertension. To investigate the relationship between intravascular ultrasound (IVUS) findings and the Heath-Edwards (HE.) grade of human pulmonary arteries, 40 pulmonary arterial segments from 17 autopsies and 5 patients with pulmonary hypertension were evaluated using a high-frequency IVUS imaging system. Histological observations were carried out at the same sites that were examined with IVUS and at the distal pulmonary arterioles of the same branches. The subjects were subdivided into 3 groups -group I: 7 patients with pulmonary hypertension (20 segments); group II: 10 patients without pulmonary hypertension (20 segments); and group III: 5 patients with pulmonary hypertension who were studied in vivo. All 20 segments from group I had a three-layered appearance, and 3 patients with H.E.grade <3 had an increased thickness of the echolucent zone due to media hypertrophy without an intimal reaction. In contrast, IVUS findings from patients with severe pulmonary hypertension (H.E. grade ≥3) showed an increased thickness with a bright inner layer due to intima hyperplasia. Evidence for a plaque-like lesion was defined in one patient with severe pulmonary hypertension. The percentage of wall thickness derived from IVUS was significantly correlated with that determined by histological examination in group I. The pulmonary artery walls in group II had a one-layer appearance. Similar IVUS findings were obtained from the in vivo study regardless of whether pulmonary hypertension was present. These findings indicate that the IVUS may be an unique technique for evaluating the morphology of the pulmonary artery or arteriole in pulmonary hypertension and may provide objective and quantitative evaluation without a lung biopsy.

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We reported on three unrelated Japanese families with carbonic anhydrase II (CA II) deficiency syndrome. In the present study, the CA II gene was sequenced in the family of a patient with hybrid type renal tubular acidosis whose parents were nonconsanguineous, and a T to G transition at exon 2 was identified. The change results in the substitution of the stop codon (TAG) at position 40 for Tyr (TAT). The maternal and paternal mutations were the same suggesting that they were obligate heterozygotes. This is a novel mutation in the CA II deficiency syndrome, which has not been described before.

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