The polymorphism of transforming growth factor-beta1 gene in Japanese patients with systemic sclerosis.

BACKGROUND: Transforming growth factor (TGF)-beta has been shown to be a potent stimulator of collagen production by fibroblasts, and could play a role in the pathogenesis of systemic sclerosis (SSc). OBJECTIVE: To study the possible involvement of TGF-beta1 gene polymorphism in Japanese patients with SSc. METHODS: Fifty-nine patients with SSc and 110 normal subjects were studied. Genomic DNA was extracted from skin tissues, and was amplified in a thermal cycler, generating a TGF-beta1 gene fragment with a size of 294 bp. The T to C transition at T869C (Leu10Pro) and the G to C transition at G915C (Arg25Pro) were identified by digestion with MspA1I and BglI, respectively. RESULTS: At T869C (Leu10Pro), the frequency of the C allele in SSc (65.3%) was significantly higher than in normal controls (50.5%) (P < 0.01). SSc showed C/C allele 42.4%, C/T 45.8% and T/T 11.2%. Normal controls showed C/C allele 26.4%, C/T 48.2% and T/T 25.5%. The frequency of the C/C allele in SSc was significantly higher than in normal controls, in comparison with the T/T allele (P < 0.02), but no significant difference was found between the frequency of the C/C allele vs. the C/T allele. The frequency of the C/C allele showed no significant difference between diffuse and limited SSc. At G915C (Arg25Pro), all the normal controls and SSc patients showed only the G/G allele. These results are different from a previous study in which the frequency of the T/T allele was high in SSc at T869C (Leu10Pro). CONCLUSIONS: This discrepancy may indicate that Japanese patients with SSc show a different genetic predisposition to TGF-beta1.

The polymorphism of telomerase RNA component gene in patients with systemic sclerosis.

BACKGROUND: The molecular basis of senescence and immortalization is not still understood, but one hypothesis for which there has recently been much evidence involves the shortening of telomeres. It can be hypothesized that abnormalities of telomerase contribute to the emergence of abnormal fibroblast clones in systemic sclerosis (SSc). OBJECTIVES: To study possible telomere abnormalities with respect to polymorphism of the telomerase RNA component gene. METHODS: Fifty-three patients with SSc and 98 normal controls were studied. Polymerase chain reaction was used to amplify 598 bp of the telomerase RNA component gene. Amplified fragments were digested with restriction enzyme BsrDI. RESULTS: The frequency of the A allele in SSc (41.5%) showed no significant difference from that in the normal controls (32.1%). The frequency of the A/A alleles in SSc (18.9%) was significantly higher than in normal controls (5.1%), compared with G/G (35.8% and 40.8%, respectively; P < 0.02), G/A (45.3% and 54.1%, respectively; P < 0.01) and G/G plus G/A (81.1% and 94.9%, respectively; P < 0.01). CONCLUSIONS: These results showed the possible involvement of a telomerase abnormality in the emergence of abnormal fibroblast clones in SSc skin-derived fibroblasts.

Quantitative analysis of microchimerism in systemic sclerosis skin tissue.

It has been reported that more male DNA of presumed fetal origin is present in the blood and skin of women with systemic sclerosis (SSc) as compared with healthy controls after delivery, but these findings are controversial. We sought to determine whether male cell DNA is present in SSc using a quantitative polymerase chain reaction for Y chromosome DNA. The study groups comprised 57 healthy women, 49 patients with SSc and 30 patients with connective tissue diseases other than SSc who had given birth to at least one son and/or had experienced fetal loss. The intensity of the PCR bands on negatives of gel photographs was quantified with a video densitometer linked to a computer analysis system. Positive Y chromosome DNA was found in 20 healthy women, 14 SSc patients and 6 patients with connective tissue diseases other than SSc. The occurrence rate of DNA equivalents of male cells among the three groups showed no significant differences. The number of male cell DNA equivalents per 80 ng tissue DNA in SSc patients (4.59+/-9.63), however, was significantly higher than in healthy women (1.83+/-4.96; P < 0.05) and in patients with connective tissue diseases other than SSc (0.27+/-0.64; P < 0.01). The occurrence rate of fetal loss in male cell DNA-positive SSc (eight) was significantly higher than in male cell DNA-negative SSc patients (four; P < 0.01). No correlation was found between the number of male cell DNA equivalents and birth of sons or clinicolaboratory findings. These results indicate that the elevated amount of male cell DNA in SSc skin tissue may contribute to the pathogenesis of SSc.

Sarcoidosis with giant parotomegaly.

We report the case of a 63-year-old man with bilateral parotid gland sarcoidosis. Giant, elastic, hard, subcutaneous tumors had been present on the right parotic and submaxillary regions for 11 years and on the left for 1 year. The patient had had diabetes mellitus for 8 years. Noncaseating epithelioid cell granulomata were revealed histopathologically in the periductal area of the parotid gland. Bilateral hilar lymphadenopathy was noted on chest x-ray studies. Serum levels of lysozyme were increased. Levels of serum angiotensin-converting enzyme were within normal limits. Tuberculin skin reaction was positive. The tumors gradually improved after treatment with oral minocycline. Giant parotomegaly, as it occurred in this case, is very rare.

Upregulation of histidine decarboxylase mRNA expression in scleroderma skin.

The involvement of histamine in the pathogenesis of systemic sclerosis (SSc) has been suggested. Possible involvement of histamine and histidine decarboxylase (HDC), the synthesizing enzyme for histamine, in the formation of the skin abnormalities in SSc was studied. Skin histamine concentrations in SSc were significantly lower than in normal controls (P < 0.02). In situ hybridization with an HDC probe revealed that the expression of the HDC gene in SSc was greater than in normal controls. The number of cells and the mean grain number per cell expressing HDC mRNA were both significantly greater in SSc than in normal controls (both P < 0.01). These results show a reduction in histamine concentration and an elevated HDC gene expression in SSc skin, indicating an increase in both histamine release and HDC gene expression. The upregulation of histamine turnover appears to be involved in the formation of the skin abnormalities of SSc.

[A case of acute urticaria from hydroxyzine hydrochloride used for preanesthetic medication].

We report a case of acute urticaria probably caused by hydroxyzine hydrochloride. A 66-year-old male was scheduled for cardiac cystectomy by thoracic endoscopy. The patient was premedicated with atropine and hydroxyzine intramuscularly 30 min before arrival in the operating room. Abrupt general exanthema was observed when intravenous infusion was started. Although his general condition was stable, the operation was postponed. Three days later, the skin test was performed using several drugs. Only hydroxyzine hydrochloride gave positive result. It is assumed that hydroxyzine hydrochloride may have caused his urticaria.

Simultaneous analysis of [3H]-thymidine uptake and alpha 1(I) procollagen mRNA expression in systemic sclerosis skin fibroblasts--an in situ hybridization study.

Heterogeneity of DNA synthesis and collagen synthesis has been reported in skin fibroblasts from systemic sclerosis (SSc) patients. The uptake of [3H]-thymidine and expression of alpha 1(I) procollagen mRNA by cultured skin fibroblasts from four normal controls and four SSc patients was analyzed simultaneously. The grains overlying the cytoplasm representing alpha 1(I) procollagen mRNA and overlying the nucleus representing [3H]-thymidine uptake were counted using computer-aided image analysis. The results were analyzed statistically. Procollagen mRNA expression by SSc fibroblasts was significantly greater than by control fibroblasts (P < 0.01). The distribution curve of [3H]-thymidine uptake showed two peaks representing low- and high-uptake cells. Significantly more SSc fibroblasts than control fibroblasts showed high [3H]-thymidine uptake (P < 0.05). The number of SSc fibroblasts expressing low amounts of alpha 1(I) procollagen mRNA was significantly lower than the number of control fibroblasts (P < 0.05). [3H]-thymidine uptake by SSc fibroblasts expressing high amounts of alpha 1(I) procollagen mRNA was significantly lower than by those expressing low amounts (P < 0.05). These results indicate that elevated DNA synthesis and elevated collagen mRNA synthesis in SSc skin fibroblasts are due to different clones with high DNA-synthesizing ability and high collagen-producing ability.

Digital ulcers and necroses: novel manifestations of angiocentric lymphoma.

We describe a patient with angiocentric lymphoma whose presenting features were multiple areas of digital ulceration and necrosis, including deep ulcers on both great toes. He lacked the lateral halves of both earlobes because of multiple ulcers. Skin biopsy revealed a patchy and diffuse infiltrate of lymphoid cells with nuclear atypia in the dermis and subcutaneous tissue. Angiocentric and angiodestructive features of the lymphoid cells, a prominent histiocytic infiltrate and some epithelioid cell granulomas were found. The results of immunohistochemical staining showed a T-cell phenotype, and showed positive staining for apoptosis. He died in July 1999. Peripheral vascular disturbances including Raynaud's phenomenon, digital skin ulcers and necroses are novel clinical symptoms in patients with angiocentric lymphoma, which should be added to the differential diagnosis in patients with peripheral vascular disturbances.

Correlation between diffuse pigmentation and keratinocyte-derived endothelin-1 in systemic sclerosis.

BACKGROUND: The precise mechanism of diffuse pigmentation in systemic sclerosis (SSc) is still unknown. We suspected the participation of endothelin-1 (ET-1), which is produced by keratinocytes, in the hyperpigmentation in SSc. The aims of this study are to demonstrate the hyperproductivity of ET-1 from epidermal cells in SSc by in situ hybridization histochemistry, and to show a correlation between the hyperproductivity of ET-1 in keratinocytes and skin hyperpigmentation. METHODS: In situ hybridization histochemistry was performed on nine SSc specimens (five cases of diffuse scleroderma (dSSc), four cases of acrosclerosis (lSSc)), and compared with four normal control specimens. We counted the grains on 10 x 10 microm(2) of epidermis and microvessels in each histology and examined the degree of skin pigmentation using the skin reflectance factor (Y). RESULTS: In the specimens of the SSc patients, the number of grains on the epidermis was remarkably higher than those of the control specimens (P < 0.01). We found a close correlation between the number of grains and the skin reflectance factor in dSSc patients (P = 0.02). Correlations were not identified between serum ET-1 and skin pigmentation and between serum ET-1 and the frequency of grains on the epidermis. As for grains on microvessels, lSSc patients showed a greater frequency than dSSc patients. CONCLUSIONS: The findings of this study suggest that an increase in the ET-1 productivity of keratinocytes is experienced in SSc patients, especially in dSSc patients. The results suggest a strong correlation between the ET-1 productivity of keratinocytes and skin pigmentation in severe cases of SSc. We conclude from these results that keratinocyte-derived ET-1 plays an important role in the pathogenesis of the hyperpigmentation of the skin in SSc patients.

Lipodystrophia centrifugalis abdominalis infantilis-- immunohistochemical demonstration of an apoptotic process in the degenerating fatty tissue.

We report a case of lipodystrophia centrifugalis abdominalis infantilis (LCAI) showing apoptosis during the lipoatrophic process by immunohistochemical staining. A 3-year-old boy was seen with a 3-month history of a centrifugally spreading depressed eruption without pain and pruritus on his abdomen. He had fever, diarrhea and vomiting for a week at the beginning of his eruption. Physical examination revealed a palm-sized well-demarcated atrophic plaque on his lower abdomen and left inguinal fold. His plaque showed fine erythema peripherally. A skin biopsy specimen showed remarkably decreased subcutaneous fatty tissue. Inflammatory cell infiltration including lymphocytes and histiocytes was noted in the degenerating fatty tissue. In the degenerating subcutaneous fatty tissue, positive staining for HLA-DR, Fas, bcl-2, p53 and transferase-mediated uridine nick end labeling in mononuclear cells were observed. He was diagnosed as having LCAI. The present case showed the possible involvement of apoptosis in the fatty tissue degeneration in LCAI.

A case of prurigo and lichenified plaques successfully treated with proton pump inhibitor.

A case of prurigo and lichenified plaques successfully treated with proton pump inhibitor is presented. She presented with pruritic eruptions, which showed marked lichenification and prurigo nodules, on her trunk and extremities. She had been treated with steroid ointment and H1-histamine receptor antagonist without success. Laboratory examinations revealed increased eosiophils and elevated lactate dehydrogenase. The skin biopsy specimen showed moderate acanthosis with spongiosis and lymphocytic and eosinophilic infiltration into the upper dermis. Because of vomiting and epigastralgia, endoscopical examination was performed, and an ulcer was found at the angle of her ventricle. A biopsy specimen disclosed a benign gastric mucosa with moderate inflammation within the lamina propria, and organisms consistent with Helicobacter pylori. Treatment for gastric ulcer with proton pump inhibitor (omeprazole) and aluminium hydroxide gel improved her eruptions and her pruritus resolved. She was discharged with complete cure of her eruption and ventricular ulcer. Our case indicates that gastric lesions induced by Helicobacter pylori infection may play an important role in dermatological diseases. Proton pump inhibitors including omeprazole are one of the choices for the treatment of some dermatological diseases including prurigo and lichenified plaques.

Dermal mucinosis and musculoskeletal symptoms simulating polymyositis as a presenting sign of hypothyroidism.

A case of dermal mucinosis and musculoskeletal symptoms simulating polymyositis as a presenting sign of hypothyroidism is presented. The patient presented muscle weakness and edema of the face including the eyelids. Laboratory examination revealed elevated creatinine phosphokinase, decreased free-T4, decreased free-T3, elevated TSH, positive anti-microsome antibody and positive anti-TSH receptor antibody. The skin biopsy specimen revealed swelling of the collagen bundles with the bundles splitting up into individual fibers, with some blue threads and granules of mucin interspersed. Alucian blue stain demonstrated vast amounts of mucin throughout the whole dermis, which was completely removed on incubation with Streptomyces hyaluronidase. The patient was diagnosed as having hypothyroidism due to Hashimoto's disease with possible polymyositis complications. After two months of thyroid hormone replacement therapy, she was euthyroidic and discharged. These results indicate that our case was a rare case of severe generalized myxoedema due to hypothyroidism of Hashimoto's disease simulating the musculoskeletal symptoms of polymyositis.

Quantitative analysis of alpha 1(I) and alpha 1(III) procollagen mRNA expression in systemic sclerosis skin tissue--an in situ hybridization study.

Human alpha 1(I) and alpha 1(III) procollagen mRNA expression in skin tissue from 15 systemic sclerosis (SSc) patients and from 7 normal control subjects was quantitatively analyzed using in situ hybridization. The grains accumulating in each area, representing procollagen mRNA expression per cell, were counted. To normalize the results from each subject, the number of cells and the number of grains per cell were divided by the area of the skin specimen (in square millimeters). The number of cells per square millimeter expressing alpha 1(I) and alpha 1(III) procollagen mRNA in SSc skin was significantly elevated compared with normal control skin (both P < 0.01). The number of grains per cell per square millimeter expressing alpha 1(III) procollagen mRNA in SSc skin was also significantly elevated compared with normal control skin (P < 0.01). The relationship between procollagen mRNA expression and the histological findings in SSc was also studied. The numbers of cells and grains per cell per square millimeter expressing alpha 1(I) procollagen mRNA in fibrotic zone SSc skin were significantly elevated compared with normal control skin (both P < 0.01). The numbers of cells and grains per cell per square millimeter expressing alpha 1(III) procollagen mRNA in SSc skin were significantly elevated compared with normal control skin (both P < 0.01) and with border zone SSc skin (number of cells P < 0.01, number of grains P < 0.05). These results indicate an increase in the number of cells showing elevated expression of alpha 1(I) and alpha 1(III) procollagen mRNA, and a close relationship between alpha 1(I) and alpha 1(III) procollagen mRNA expression and the histological findings in SSc.

Bandlike melasma mimicking linear morphea ("en coup de sabre" type).

A case of bandlike melasma on the median line of the forehead of a middle-aged woman is described. It was an atypical case for its clinical shape and distribution. At initial examination, it was diagnosed as linear morphea. Epidermal nevus was another differential diagnosis. Based on the presence of minor protuberance and the absence of atrophy, clinically, and the presence of sebaceous glands and the absence of swelling of collagen fibers, documented histologically, we revised our diagnosis to that of an atypical type of melasma.

Elevated expression of epidermal ornithine decarboxylase mRNA in scleroderma.

Using in situ hybridization techniques, we examined the expression of ornithine decarboxylase (ODC) mRNA in the skin of five patients with systemic sclerosis (SSc) and five normal controls. Sections treated with an anti-sense probe showed concentrated grains exclusively in the epidermis of SSc patients, but not in that of normal controls. Because our subcloned anti-sense probe specifically hybridizes with ODC mRNA, these findings indicate that the expression of ODC mRNA is elevated in SSc epidermis. Possibly polyamines have an important part to play in the skin changes of SSc.