Studies on eleven kidney transplants from non-heart-beating donors.

This study was performed to analyze postoperative courses and complications, retrospectively, following transplants from non-heart-beating donors and to examine the correlation between early graft function and clinical parameters. We experienced 11 cases of kidney transplants from non-heart-beating donors during the period from April 1995 to May 2003. Warm ischemic time was less than 30 min in all cases, and total ischemic time ranged from 8.4 hours to 27.9 hours. Rejection reactions occurred in seven cases, two of which were vascular rejections. Infectious disease complications included CMV in two cases, interstitial pneumonia in one case and fungal infection in one case. One patient died from interstitial pneumonia, and three patients had to be restarted on dialysis due to loss of function of the grafted kidney. The remaining seven patients all made full recoveries. All of the 16 patients who underwent living related kidney transplantations during the same period made full recoveries. Both the donor's gender and the latest creatinine level of the donor influenced the posttransplant dialysis period. The posttransplant dialysis period significantly influenced the creatinine level one month after transplant. These results suggest that patients who undergo kidney transplants from non-heart-beating donors have higher rates of complications than patients who undergo living related kidney transplantation. It is important that, in cases where the donor's creatinine level is high, especially when the donor is male, the kidney is carefully retrieved and transported to the recipent hospital to shorten the ischemic period as much as possible.

Clinical and biological studies of 26 cases of xeroderma pigmentosum in northeast district of Japan.

Twenty-six patients with xeroderma pigmentosum (XP), who live in the Northeast (Tohoku) District of Japan, were examined for the clinical characteristics of UV-induced DNA synthesis (unscheduled DNA synthesis, UDS) and UV sensitivity of skin fibroblasts or lymphoblastoid cells, or both. A history of consanguineous marriage within two generations was found in 19 of 26 cases (73%). Two pairs of siblings showed similar manifestations and almost the same levels of UDS and of UV sensitivity. Squamous cell carcinoma, basal cell carcinoma, or both were observed on the exposed skin in 14 patients, but no malignant melanoma was found. Cancer had developed in approximately 71% (10/14) of the cancer-bearing patients by the age of 20, and 8 of them belonged to the UDS-deficient group. Neurological manifestations were associated with nine patients, including 3 with typical de Sanctis-Cacchione syndrome (DSC), and most of the cells derived from these patients had a UDS level less than 10% of that of the normal cells. A clear correlation between the levels of UDS and UV sensitivity, on the one hand, and the severity of clinical manifestations on the other could not be detected, but it seems that the UDS-deficient group is generally much more sensitive to UV in terms of cell killing and the induction of sister chromatid exchange (SCE) than the UDS-proficient group. After a photosensitivity test, one patient with mild skin manifestations showed distinct skin tanning without preceding erythema.

Radical production during tyrosinase reaction, dopa-melanin formation, and photoirradiation of dopa-melanin.

It has been suggested that superoxide anion (O2-) may be produced during eumelanin formation and during the photoirradiation of eumelanin , but no direct evidence for this has yet been reported (although O2- production during photoirradiation of pheomelanin has been shown). In this report, the production of O2- was investigated during the formation and photoirradiation of dopa-melanin, a synthetic eumelanin . It was found that cytochrome c was reduced during the tyrosinase reaction and dopa-melanin formation in vitro; this reduction could not be inhibited by superoxide dismutase (SOD). When dopa-melanin was irradiated by UV radiation or by visible light, high nitroblue tetrazolium (NBT) reduction was observed; this reduction was proportional to the light energy and the amount of dopa-melanin. NBT reduction by visible light could be slightly inhibited by SOD, but a 12% decrease of NBT reduction by UV radiation could be shown with the addition of SOD. These observations indicate that some radicals were produced during the tyrosinase reaction and dopa-melanin formation. Further, when dopa-melanin was irradiated, radicals were also produced, some of which were thought to consist of O2-, but others were unknown.

Inhibition of aminoacyl-transfer RNA formation by low-molecular substances from melanoma extract.

Two partially purified fractions of the ethanol precipitate (70-95%) of the water extract of Harding-Passey mouse melanoma, which inhibit protein and DNA syntheses of B-16 melanoma cells in culture, also inhibit protein synthesis in various cell-free systems. By examining their inhibitory effects on limited reactions of protein synthesis, it was found that one of them (ME II) inhibits protein synthesis by blocking aminoacyl-tRNA formation, while the other (ME IV) does not. This inhibition of aminoacyl-tRNA formation was not limited to specific amino acids. Since the amino acid-dependent pyrophosphate (PPi)-ATP exchange reaction catalyzed by aminoacyl-tRNA synthetases was not inhibited, it was concluded that some factor(s) in ME II inhibits amino acid transfer from aminoacyl-AMP to tRNA. ME II contains more than 20 proteins from 10,000 to 90,000 daltons. EDTA treatment of this fraction caused the release of low-molecular substances with inhibitory activity from the proteins. The molecular weights of the active substances are less than 5,000 daltons. The active low-molecular substances are apparently not peptides or nucleotides.

Effect of macrophages on elimination of dermal melanin from the dermis.

To facilitate the mobilization of dermal melanin from the skin, we studied the role of macrophages in the elimination of dermal melanin in incontinentia pigmenti. The protein and melanin moieties of mouse-melanoma melanosomes were labeled with 14C and 3H, respectively. Labeled melanosomes were injected intracutaneously into the backs of guinea pigs. Carrageenan, a macrophage toxic agent, showed an inhibitory effect on the degradation and elimination of protein moiety of melanosomes from the dermis, when carrageenan was injected i.p. at the time of inoculation of melanosomes. Delayed-type inflammatory infiltrates in contact dermatitis showed some increase in the elimination of melanin moiety of melanosomes from the dermis.

Transfer of tyrosinase to melanosomes in Harding-Passey mouse melanoma.

The transfer of tyrosinase from microsomes into melanosomes, without passing through the cytosol in the Harding-Passey mouse melanoma cell, was confirmed by experiments carried out using a combination of radioisotope tracer techniques and immunoprecipitation. 3H-Labeled amino acid incorporation into tyrosinase present in the microsome, melanosome, and soluble fractions confirmed the precursor-product relationship of the enzyme in the microsome fraction and in the melanosome fraction. However, two forms of the enzyme, Ts1- and Ts2-tyrosinase, separated from the soluble fraction by polyacrylamide gel electrophoresis, were shown to play no role in the transfer since little or no incorporation of radioactivity into tyrosinase in this fraction was found. It is suggested that most tyrosinase observed in the soluble fraction does not leak from the melanosomes or the microsomes during homogenization, but comes from necrotic tumor cells. It appears that melanosomal and microsomal tyrosinase might be released from the membrane of necrotic cells modified by various degradation enzymes, considering the data on the recovery of tyrosinase from the soluble fraction, where one-third of total enzyme activity in the postnuclear fraction could not be increased, even when the postnuclear fraction of the tumor was further homogenized radically.

Immunoelectron microscopic examination of IgA deposition in dermatitis herpetiformis.

Ultrastructural localization of IgA in the skin of three Japanese patients with dermatitis herpetiformis (DH) was studied with the immunoelectron microscopic technique using periodate-lysine-paraformaldehyde fixation. In direct immunofluorescence studies, two of the three cases showed fine fibrillar deposition of IgA and the other case fine granules in the dermal papillae. In the former, the reaction products of IgA were present in the upper dermis forming various-sized aggregates which were occasionally arranged perpendicularly to the epidermis and appeared to be associated with microfibrillar bundles of the elastic tissue. Reaction products were also deposited to a lesser extent on the microfibrillar component of the elastic fibers at the lower part of the dermal papillae. However, in the latter, the reaction products were found to form smaller aggregates on and around the collagen fibrils rather than on the elastic tissue. Such a localization of IgA reaction products has not yet been reported in DH. The difference of the distribution patterns of IgA and the possible singularity of Japanese DH cases are discussed.

Acral melanoma in Japan.

Clinical records and histologic materials from 81 patients with malignant melanoma at the Department of Dermatology at Tohoku University School of Medicine were reviewed. In addition, a statistical study on 1597 cases of malignant melanoma collected from the Japanese literature from 1961 to 1982 was performed. The annual mortality rate has been increasing almost linearly over the past 20 years. The mortality rate per year for 1980 was 0.21 per 100,000. Five-year survival rate at Tohoku University was 35 percent. The most common site of melanoma was acral, especially the plantar surfaces. The clinical and histologic study of acral melanomas showed that clinicopathologic features are the same as those reported for acral lentiginous melanoma in the United States.

Incontinentia pigmenti achromians (Ito).

Three children had incontinentia pigmenti achromians (IPA). A study of these cases, together with a review of 70 previously documented cases, suggests that this dermatosis is closely related to systematized depigmented nevus and should be categorized as one of the congenital neurocutaneous diseases. There is also a relationship between IPA and incontinentia pigmenti.

Pigmentary incontinence in fixed drug eruptions. Histologic and electron microscopic findings.

Pigmentary incontinence is a phenomenon observed in some inflammatory skin disorders. Clinically it may be seen as a slate-colored pigmentation. Histologically it is seen as an accumulation of melanin in the upper dermis. The possible mechanism for development of pigmentary incontinence is discussed based on a review of the literature and electron microscopic studies of fixed drug eruption.

Histidine-rich protein as a possible origin of free amino acids of stratum corneum.

The origin of free amino acids and/or their metabolites of the stratum corneum was investigated by pulse-chase experiments using hairless mice. Two, 24, 48, and 72 hrs after the animals were pulse-chased with 3H-histidine or 3H-arginine, radioactivity was determined in the following three fractions of the epidermis: the 0.1 N HClO4 soluble, ethanol soluble fraction (Fr. I); the 0.1N NClO4 soluble, ethanol insoluble fraction (Fr. II); and the 0.1 N HClO4 insoluble, 8M urea soluble fraction (Fr. III). Radioactivity of the epidermal proteins was also determined in SDS-PAGE gels by the gel slicing method. At first, 3H-histidine and/or 3H-arginine were predominantly incorporated into Fr. III, especially into a certain epidermal protein which showed little mobility on SDS-PAGE. Subsequently, 3H, once incorporated into Fr. III, appeared to shift from Fr. III to Fr. II and, simultaneously, from the top band(s) to a band of 32,000 daltons. As a result of amino acid analysis, a protein of Fr. II was considered a histidine-rich protein corresponding to HRP-II of the new born rats of Ball et al. After 48 hrs, as the radioactivities of Fr. III and Fr. II decreased remarkably, Fr. I gradually increased in radioactivity. The major radioactive substances in Fr. I at 72 hrs were identified as 3H-histidine and 3H-urocanic acid when 3H-histidine was used as a tracer, and as 3H-arginine, 3H-ornithine, and 3H-citrulline in the case of 3H-arginine. The amino acid composition of the histidine-rich protein was very similar to that of the free amino acids of the stratum corneum in the hairless mouse, in which some amino acid metabolites were considered as their precursor amino acids. These results strongly suggest that the free amino acids and/or their metabolites of the stratum corneum might be the final products of a degradation of the histidine-rich protein.

Effect of beta-carotene on ultraweak chemiluminescence of UVB-irradiated squalene.

An emission spectral analysis was carried out on chemiluminescence emitted from UVB-irradiated squalene. The main emission species produced by the transition of (1 delta g) (1 delta g) and (1 delta g) (1 sigma g +) to (3 sigma g -) (3 sigma g -) were found by spectroscopic analysis of the chemiluminescence. When beta-carotene was added to the irradiated squalene, its spectral pattern changed drastically and many peaks disappeared.

Chemiluminescence of UV-irradiated linolenic acid and squalene.

An emission spectral analysis was carried out on ultraweak chemiluminescence emitted from UVB-irradiated linolenic acid and squalene. The main emission species produced by the transition of (1 delta g) (1 delta g) dimer and an additional weak band near 477.5 nm (0, 0) by the transition of (1 delta g (1 epsilon g+) to (3 epsilon g-) (3 epsilon g-) were found by spectroscopic analysis of chemiluminescence in both cases of irradiated linolenic acid and of squalene. A distinct peak around 410-420 nm was observed in irradiated squalene and the emitter seems to be due to the excited carbonyl compound.