Microfluorometric study of nuclear DNA in normal human epidermis.

Quantitative and qualitative changes of nuclear DNA were analyzed by microfluorometry in normal human epidermis during terminal differentiation. Normal human epidermis from five healthy volunteers was separated by an EDTA-trypsin method and fractionated by Percoll density gradient. Staining was carried out with either Feulgen reaction, Acridine Orange or 4'-6-diamidino-2-phenylindole (DAPI). When the cells with lower density (granular layer cells) were stained with Acridine Orange or DAPI, the greatest population was found at lower value of DNA than that in the cells stained with Feulgen reaction. After thermal denaturation procedure, a ratio of double-stranded DNA to single-stranded DNA was highest in the cells with lower density. These results seem to suggest the qualitative and quantitative changes of nuclear DNA during terminal differentiation of human epidermal keratinocytes.

Effect of a helium-neon laser on cutaneous inflammation.

A Helium-Neon (He-Ne) laser with a wavelength of 632.8 nm is known to have photobiological effects and is widely used for reducing the pain of herpes zoster and accelerating wound healing, however the cellular mechanism and effect of the He-Ne laser are poorly understood. The present study was designed to examine the influence of He-Ne laser irradiation on irritant and allergic contact dermatitis of the mouse ear and on histamine release from rat peritoneal mast cells. Irradiation was applied with a He-Ne laser (12.2 J/cm2) at 1 h, 10 min, 5 min and 0 min before, and 5 min, 6 hs and 24 hs after a challenge of an irritated contact dermatitis (ICD) or allergic contact dermatitis (ACD) was made on the right ears of ICR-mice. Twenty-four hours after the challenge, the swelling of the ear was measured with a dial thickness gauge, and the anti-inflammatory effect of He-Ne laser irradiation was expressed as an ear thickness ratio (ETR). Although the laser did not decelerate the ETR from ICD, the allergic response was decelerated. Irradiation at 5 min after the challenge of contact dermatitis increased the thickness ratio. Next, the influence of the He-Ne laser on histamine release from Wistar-rat peritoneal mast cells was observed. The spontaneous histamine release was inhibited by laser irradiation, while substance P and compound 48/80-induced histamine release were not inhibited. From these results, it can be suggested that He-Ne laser irradiation has an anti-inflammatory effect on cutaneous inflammation.

Proteoglycans in albo-papuloid lesions of the Pasini form of dominant dystrophic epidermolysis bullosa.

Epidermolysis bullosa is a group of inherited blistering diseases classified into three main sub-groups on the basis of the level of cleavage within the skin. In dominant dystrophic epidermolysis bullosa, characterized by cleavage below the basal lamina, two variants can be distinguished by the presence (Pasini form) or absence (Cockayne-Touraine form) of albo-papuloid lesions. The present study was undertaken to investigate the glycosaminoglycan chains of proteoglycans in the albo-papuloid lesions of a patient with the Pasini form, using histochemical and immunohistochemical methods. Histological examination revealed no dermo-epidermal separation. In the dermis, the papillary and subpapillary layers were slightly homogeneous, and exhibited a strong affinity towards alcian blue, which was abolished by treatment with chondroitinase ABC or in the presence of MgCl2 0.3M, but was resistant to digestion with streptomyces hyaluronidase. The papillary and subpapillary layers were intensely stained with a monoclonal antibody against small size proteoglycan with dermatan sulfate. These results suggest the presence of degraded dermatan sulfate proteoglycan in the papillary and subpapillary dermis of albo-papuloid lesions in the Pasini form of dystrophic epidermolysis bullosa.

Chemotherapeutic approach to uterine papillary serous carcinoma.

Uterine papillary serous carcinoma (UPSC) is a recently recognized histologic variant of endometrial carcinoma with an aggressive clinical course. This study reviews 7 patients with the diagnosis of UPSC treated at the National Kokura Hospital between 1987 and 1989. The mean age was 60 years. Clinically, 4 patients presented as Stage I, one as Stage II, one as Stage III, and one as Stage IV. Surgery upstaged 28.7% (2/7) of these patients. All 7 patients underwent a staging laparotomy followed by chemotherapy. Deep myometrial invasion was found in 5/7 (72%) of the specimens. Four of these 5 specimens demonstrated up to the uterine serosa. Lymph nodal involvement was observed in 1/4 (25%) of the specimens. Furthermore, positive peritoneal washings were obtained in 4/7 (57%) of the specimens. All patients received an adjuvant combination chemotherapy with cisplatin 50 mg/m2 on day 1, adriamycin 50 mg/m2 on day 1, and ifosfamide 1.0 g/m2 on day 1 to day 5 (PAI). Recurrence and relapse of the disease developed in the 2 patients within 6 months. Two patients had a negative second look operation with no evidence of disease (NED) at 18 months after the initial treatment. Serial determination of the serum CA125 was seen to have great value in monitoring treatment in 4 patients. However, additional accrual and follow-up are needed to determine whether or not PAI therapy has an impact on this disease.

Cisplatin, adriamycin and ifosfamide-based combination chemotherapy for gynecological cancers derived from the extended müllerian system.

Their histogenetic similarity suggests that a group of malignant tumors may have a common sensitivity to a cytotoxic chemotherapy. Seventy patients with a variety of gynecological cancers arising from the uterine cervix, endometrium, ovary and pelvic peritoneum were treated with a combination of cisplatin, adriamycin and ifosfamide (PAI). As schedule modifications, PAI plus bleomycin for cancers containing squamous components and PAI plus etoposide for nonepithelial malignancies were recommended. In twenty-five evaluable cases, including 12 recurrent tumors after previous radiation therapy or PAC (cisplatin, adriamycin and cyclophosphamide) chemotherapy, the total response rate was 95% for epithelial cancers (vaginal cancer: 1/1, cervical: 9/10, endometrial: 2/2, ovarian: 6/6 and peritoneal: 2/2), and 100% for nonepithelial malignancies including one uterine leiomyosarcoma, one uterine mixed Müllerian tumor and one extragonadal mixed Müllerian tumor. The survival rates of patients with non measurable lesion were 100% for cervical cancer (the observation period: 65-879 days), 92.9% for endometrial cancer (96-975 days) and 88.9% for ovarian cancer (148-976 days). The hematological toxicity of this treatment was severe but acceptable. The results obtained indicate that a wide range of gynecological cancers originating in the primary and secondary Müllerian tissues (extended Müllerian system) must have a similar sensitivity to cytotoxic treatment with a PAI-based combination chemotherapy.

Effects of 2,4-dinitrophenol and other metabolic inhibitors on the thermograms of Ehrlich ascites carcinoma cells registered with a microcalorimeter.

The heat produced by Ehrlich ascites carcinoma cells was measured microcalorimetrically to investigate the characteristics of the energetics of these cells. 2,4-Dinitrophenol and other inhibitors were used to obtain the standard thermograms. The heat level of starved Ehrlich ascites carcinoma cells (endogenous) was almost constant and linear and depended on the cell number used in the experiments (about 1 cal/1 X 10(8) cells/hr). The endogenous heat generation was suppressed strongly by potassium cyanide but not by iodoacetic acid or 2,4-dinitrophenol. On the other hand, heat evolution was directly related to the amount of glucose added to the medium but not to the number of cells (19 cal/mmol of glucose). The addition of iodoacetic acid markedly suppressed heat generation, but potassium cyanide did not. In contrast, glucose with dinitrophenol increased heat production markedly to about 10% higher than that of the control, possibly due to the uncoupled energy from oxidative phosphorylation. Malonic acid (1 mM) suppressed slightly the endogenous heat but caused a 15% reduction in exogenous heat. The amount of heat produced by the addition of glucose (19 cal/mmol) was equivalent to 40% of the theoretical value of energy released through glycolysis (47 cal/mmol). Thus, the heat produced by exogenous glucose appears to be mainly dependent on glycolysis.

Computer experiment for selection of optical isomer during prebiotic polymerization.

Computer experiments simulating prebiotic polymerization were carried out using the Monte Carlo method. A high degree of single optical isomers from racemic monomers occurred by assuming a small asymmetry in their chemical natures, which was included to investigate an amplification of the asymmetry due to parity non-conservation. The asymmetry developed much more slowly than the growth of polymers, and the degree of asymmetry in the polymers increased with the degree of polymerization. Competition between the preferential selection and the disturbance by the statistical fluctuation are discussed.