Small invasive colonic cancer occurring in a hyperplastic polyp.

We report a case of advanced colon cancer which was supposed to have arisen from a hyperplastic polyp in a 68-year-old man. Colonoscopy revealed a depressed reddish area with a surrounding elevated lesion that was of a faded color compared with the normal mucosa. After the mucosal surface had been sprayed with crystal violet dye, magnifying colonoscopy showed an amorphous area in the central depression and the surrounding, slightly elevated lesion had an asteroid pattern. The depressed area was therefore considered to be a colonic cancer surrounded by a hyperplastic polyp. Endoscopic ultrasonography showed that the lesion was infiltrating further than the deep submucosal layer and it was therefore decided to treat the patient by laparoscopically assisted right hemicolectomy. The depressed lesion was found to be a well-differentiated adenocarcinoma invading the muscularis propria (diagnosed as IIc + IIa-like advanced adenocarcinoma). The surrounding flat elevated lesion was found to be hyperplastic mucosa. No adenomatous lesions were found. There have been few reported cases in which a preoperative diagnosis of carcinoma in a hyperplastic polyp has been made, but the possibility of carcinogenesis from hyperplastic polyps has come under consideration recently. This case was considered to be important because it raises the possibility that nonpolypoid cancer can develop from a hyperplastic polyp.

Synthesis of calcitonin gene-related peptide (CGRP) by rat arterial endothelial cells.

We investigated the protein and mRNA expression of calcitonin gene-related peptide (CGRP) in endothelial cells of the rat thoracic aorta and femoral artery. Light microscopic immunocytochemistry revealed that immunoreactivity for CGRP was preferentially located in the endothelium of both vessels. Immunoelectron microscopy showed that CGRP-immunoreactive gold particles were preferentially localized on cisterns of the rough endoplasmic reticulum and on the Weibel-Palade (WP) bodies in the endothelial cells. Prepro CGRP mRNA signals were also detected on the endothelium. Our results are the first to demonstrate that endothelial cells of both elastic and large muscular arteries synthesize CGRP and store it, in part, in WP bodies, implying that CGRP may act as an endothelium-derived relaxing factor in these vessels.

Synthesis and receptor sites of endothelin-1 in the rat liver vasculature.

Immunocytochemical localization of big endothelin-1 (big ET-1), ET-1, and ET receptor A and B (ET(A) and ET(B)), and gene expression of prepro ET-1 mRNA were examined on the rat liver vasculature. Immunoreactivities for big ET-1 and ET-1 were preferentially seen along the endothelium of interlobular veins (IV) and artery (IA), although the staining intensity was more pronounced in IV. Expression of preproET-1 mRNA was detected in both vascular endothelia and the signal intensity was more prevalent in IV. Immunoelectron microscopy showed that rough endoplasmic cisterns were immunoreactive for big ET-1, while Weibel-Palade (WP) bodies, a storage site for ET-1, were immunoreactive for ET-1 in endothelial cells of IV. These results indicate that endothelial cells of IV are the major site of synthesis of ET-1, which is extracellularly secreted by degranulation and/or exocytosis of WP bodies. Hepatic stellate cells (HSCs), especially of the plasma membrane of perisinusoidal and interhepatocellular processes, were immunoreactive for both ET(A) and ET(B) receptor antibodies. These findings suggest that ET-1 receptor-mediated HSC contraction is involved in the regulation of hepatic sinusoidal blood flow as previously cited in mammalian liver cirrhosis. We also showed that sarcolemma and caveoles in the smooth muscle cells of the media of IV, and its branches before reaching the hepatic sinusoids, were immunoreactive for ET(A) receptor antibody. The results suggest that such vessels, which contains a large amount of hepatic blood inflow, participate in pump mechanism toward hepatic sinusoidal circulation in a receptor-mediated paracrine fashion.

Regressing Merkel cell carcinoma-a case showing replacement of tumour cells by foamy cells.

Merkel cell carcinoma (MCC) is a malignant neuroendocrine tumour with a high rate of recurrence and metastasis. However, some cases of spontaneous regression have recently been reported. We describe the clinical course of an 80-year-old Japanese woman with regressing MCC. We also report histological findings of the regressing tumour for the first time. After the patient's first visit to our hospital, the lesion was a rapidly progressive tumour, but suddenly began decreasing in size, and rapidly regressed. The surface of the tumour flattened, the colour changed from red to dark red, and finally the lesion appeared as a small yellowish plaque. Histopathological analysis of the completely regressed tumour revealed that the tumour cells were completely replaced by numerous foamy cells. This is the first report demonstrating the histopathological features of regressing MCC.

Pilomatricoma associated with several hair follicles.

We report two cases of pilomatricoma in which the neoplasm was connected with several hair follicles. A 17-year-old boy developed an erythematous nodule, 1 cm in diameter, in the right temporal region, histologically showing basophilic cells connected with at least two hair follicles. In the other case, a 20-year-old man had a centrally ulcerated tumor, 2 cm in diameter, in the left lower back. Basophilic cells proliferated in connection with at least six hair follicle-like structures, accompanied by the development of shadow cells underneath. The histopathologic findings in these two cases suggest the existence of a subset of pilomatricomas that involve several pre-existing hair follicles.

Expression of neuronal nitric oxide synthase in dermal infiltrated eosinophils in eosinophilic pustular folliculitis.

Nitric oxide (NO) is produced by a wide variety of human cells and affects physiological and pathophysiological processes. In this study, we demonstrated for the first time that neuronal NO synthase (NOS) is expressed in eosinophils infiltrating into the dermis and follicular epidermis in eosinophilic pustular folliculitis (EPF). EPF is a cutaneous inflammatory follicular disorder first described in Japan. The clinical and histological features of EPF are characterized by erythematous papules, infiltration of numerous eosinophils, and a spongiotic appearance of the follicular epidermis, but the pathophysiology of EPF remains unclear. Our results suggest that NO produced from eosinophils plays an important part in the pathogenesis of EPF. Furthermore, we speculate that NOS inhibitors may be useful in the management of EPF.

Effects of endothelin-1 on vasoactivity and its synthesis, storage, and acting sites in the rat superior mesenteric vasculature: an ultrastructural and immunocytochemical study.

Vasoactivity after treatment with endothelin (ET)-1 and immunoreactivity for ET-1 and its receptors were investigated in the rat superior mesenteric vasculature (SMV). By measurements of corrosion cast images of the SMVs, it was seen that ET-1 induces remarkable vasocontraction of the distal arterial branches, consisting of small arteries and arterioles, and localized vasoconstriction throughout the venous branches which possess localized medial thickenings. Immunoreactivity for ET-1 was preferentially seen along the endothelia of the proximal arterial branches. Cisterns of the rough endoplasmic reticulum and Weibel-Palade (WP) bodies of the endothelial cells were immunoreacted. Immunoreactivity for the ETa receptor was preferentially seen on the media of the distal arterial branches. These findings indicate that endothelial cells of the proximal arterial branches synthesize ET-1 and store it in the WP bodies. Because WP bodies are involved in the release of ET-1, this suggests that this endogenous ET-1, which is released from the proximal arterial branches, may be involved in the regulation of blood flow through the distal arterial branches by mediation of the ETa receptor. In addition, it seems likely that ET-1-induced vasoconstriction of the venous branches may act to impel the portal blood flow.

Two cases of pilomatrixoma in Turner's syndrome.

Multiple and familial cases of pilomatrixomas have been reported in myotonic dystrophy. We report 2 cases of pilomatrixoma in Turner's syndrome. Between 5 and 19 years of age, 1 patient developed 6 pilomatrixomas including atypical occurrences on the chest and the posterior aspect of the leg. This is the first report of multiple pilomatrixomas in Turner's syndrome.

A case of superficial leiomyosarcoma, and a review of Japanese cases.

We report a case of leiomyosarcoma in a 55-year-old Japanese female. She developed an erythematous nodule, 1.5 cm in diameter, on her right thigh. The totally excised specimen contained tumor cells with variously sized, chromatin-rich nuclei. Gitter staining revealed the so-called encased picture, immunohistopathological staining was positive for smooth muscle actin, and electron microscopy revealed filaments with focal densities. A diagnosis of leiomyosarcoma was made based on these findings, which suggested a muscle cell origin. Normal arrector pili muscles were continuous from the tumor, further suggesting the tumor's origin. The presence of Epstein-Barr virus-encoded RNA (EBER) was examined by in situ hybridization with negative results.

Larva currens: the usefulness of the agar plate method.

Creeping eruption caused by Strongyloides stercoralis is called larva currens, while those caused by other parasites are called larva migrans. To detect the parasite, the direct smear method was used. However, this method is not very sensitive in detecting S. stercoralis. A 62-year-old male with polyarteritis nodosa cutanea had an erythematous creeping eruption in various regions, such as his lumbar region, abdomen and thigh. The diagnosis of larva currens was made by the clinical appearance and the detection of S. stercoralis in his feces using the direct smear method and the agar plate method. This is the first reported case of larva currens in Japan. The agar plate method was useful and more sensitive than the usual direct smear method for detecting the S. stercoralis in feces and for elucidating the effects of treatment with antiparasitic drugs.

Activation of bullous pemphigoid antigen gene in mouse ear epidermis by ultraviolet radiation.

Bullous pemphigoid (BP) is an autoimmune blistering disease and is a photoaggravated dermatosis, but the mechanism of the aggravation is still unknown. Since damage to DNA initiates transcription of some genes, we investigated in epidermis of mouse ears the relationship between DNA damage by ultraviolet (UV) radiation and BP antigen (BP-Ag) gene activation. For this, albino male mice were irradiated with 254 nm wavelength UV for a total dose of 500 J m-2. At fixed times (0.5, 2, 24, 48 and 72 h) post-UV irradiation, mouse ears were cut off, frozen and sectioned. In the sections, it was found that immunohistochemically detectable pyrimidine dimers were observed in nuclei of all epidermal cells at 0.5 h that were almost repaired by 72 h; a frequency of single strand breaks in DNA detected by in situ nick translation started to increase in nuclei of all epidermal cell layers at 0.5 h and the increase continued up to 24 h; mRNA for BP-Ag localized by non-radioactive in situ hybridization appeared in nuclei of basal cells at 0.5 h and in both nuclei and cytoplasm at 2 h; and immunoreactive BP-Ag started to increase in the basal cell cytoplasm and in the basement membrane zone at 2 h. BP-Ag started to accumulate in the basement membrane zone at 2 h. It is suggested that UV radiation increased BP-Ag synthesis through BP-Ag synthesis through BP-Ag gene activation and that this reaction is a factor which aggravates BP following UV irradiation in BP patients.

The role of endothelin-1 in regulation of rat mesenteric microcirculation.

Immunocytochemistry of endothelin-1 (ET-1) and the ETA receptor, and scanning electron microscopy using cast resin after treatment with ET-1, were carried out in the rat mesenteric microvasculature. Immunoreaction of ET-1 was preferentially seen along the endothelium of the proximal portion of the anterior mesenteric artery, the endothelial cells of which contain abundant Weibel-Palade (WP) bodies. However, the arterioles and small veins distal to the artery showed little immunoreactivity but showed heavy immunoreaction of ETA receptors in the media. By scanning electron microscopy after treatment with ET-1, the mesenteric microvasculature became slightly narrower compared to the control exhibited of localized constricted areas, especially in the region of the small veins. Light microscopy of such areas revealed localized thickening of the medial muscle cells. Because the release of ET-1 from endothelial cells depends in part on extracellular discharge of the WP bodies, the results indicate that ET-1, discharged from the proximal portion of the anterior mesenteric artery, induces vasoconstriction of the arterioles and small veins, mediated by ETA receptors. The localized thickening in areas in the media of the small veins may participate in the maintenance of blood flow through the portal circulation.

Hereditary perioral pigmented follicular atrophoderma associated with milia and epidermoid cysts.

Eight members of a single family all presented the characteristic changes of facial, especially perioral, pigmented follicular atrophoderma, with numerous milia and epidermoid cysts. For this condition. diagnosis at a glance may be possible because of the perioral cutaneous manifestations. Histopathological examination of follicular atrophoderma revealed proliferation of basaloid cells continuous with the epidermis and coarse collagen fibres, with a decreased density of elastic fibres around the basaloid cells. Two of the eight individuals also showed generalized hypohidrosis. The eight affected persons were the proband, her son, mother, uncle, two younger sisters, cousin and nephew: an autosomal dominant mode of transmission was suggested from this family tree. The patients' symptoms resembled those of Bazex-Dupré-Christol syndrome, except for the different distribution of the follicular atrophoderma and the absence of basal cell carcinoma and hypotrichosis. This disease may be an entirely new syndrome characterized by perioral pigmented follicular atrophoderma associated with milia and epidermoid cysts.

Photo-enhanced modification of human skin elastin in actinic elastosis by N(epsilon)-(carboxymethyl)lysine, one of the glycoxidation products of the Maillard reaction.

Long-term incubation of proteins with glucose leads to the formation of advanced glycation end products (AGEs), which are characterized by fluorescence, brown color, and cross-linking. Formation of AGEs in vitro requires oxygen and is dependent on transition metal-catalyzed oxidation of glucose or Amadori products. AGEs are thought to be involved in aging and age-enhanced diseases such as diabetic complications, atherosclerosis, dialysis-related amyloidosis, and Alzheimer's disease. Chronic exposure of the skin to sunlight induces hyperplasia of the elastic tissue in the upper dermis known as actinic elastosis. Herein we used a monoclonal anti-AGE antibody (6D12) whose epitope is N(epsilon)-(carboxymethyl)lysine (CML), one of the glycoxidation products of AGEs, and demonstrated that the lesions of actinic elastosis were modified by CML. Further immunohistochemical and immunoelectron microscopic examination with 6D12 demonstrated CML accumulates predominantly in elastic fibers especially in the amorphous electron-dense materials corresponding to photo-induced degenerated area rather than the electron-lucent region. Immunochemical analyses with enzyme-linked immunosorbent assay (ELISA) of elastase-soluble fractions demonstrated that the CML levels of the sun-exposed area were significantly higher than those of the sun-unexposed area. We conclude that ultraviolet-induced oxidation may accelerate CML formation in actinic elastosis of photoaged skin.

Weibel-Palade bodies as a storage site of calcitonin gene-related peptide and endothelin-1 in blood vessels of the rat carotid body.

BACKGROUND: The vasculature of the carotid body has been considered to play a role in the regulation of blood flow into this organ. This light and electron microscope immunocytochemistry deals with endothelium-dependent vasomotion by vasodilatory calcitonin gene-related peptide (CGRP) and vasoconstrictive endothelin-1 (ET-1). METHODS: After adult male rats were perfused with a solution of periodate-lysine-paraformaldehyde through the left ventricle, the carotid artery bifurcations were isolated and utilized for light and electron microscope immunolabelings with CGRP and ET-1 primary antisera. RESULTS: By light microscope immunocytochemistry, immunoreactions to CGRP were seen along the endothelium of the carotid body artery (CBA) and its branches, and those of ET-1 were observed along the endothelium of the intralobular capillaries in addition to the above vessels. By immunoelectron microscopy, immunoreactive gold particles of CGRP and ET-1 were identified in the rough endoplasmic reticulum (rER) and in the Weibel-Palade (WP) bodies of endothelial cells of the CBA and its branches. Colocalization of both immunoreactive gold particles was observed in the same WP body. Immunoreactive gold particles of CGRP were also identified in the rER, Golgi apparatus, and specific granules of the dark glomus cells. CONCLUSIONS: Conceivably, CGRP and ET-1 are synthesized in the rER of these endothelial cells and are stored in the WP bodies for the autoregulation of blood flow.

Infantile perianal pyramidal protrusion.

BACKGROUND: The number of reported cases of infantile perianal eruption has been increasing. However, infantile pyramidal protrusion located in the midline anterior to the anus has apparently been reported mainly as acrochordons or skinfolds. OBSERVATIONS: Fifteen infants (14 girls and 1 boy) had a pyramidal protrusion located only in the midline anterior to the anus. Histological examination revealed acanthosis in the epidermis, marked edema in the upper dermis, and mild infiltrates in the dermis. The patients had been brought to the hospital because of swelling of the protrusion. However, all protrusions showed reduction without any treatment. CONCLUSION: Infantile pyramidal protrusion located only in the midline anterior to the anus should be distinguished from acrochordons or skinfolds based on the characteristic clinical features.

[A reliable method for internal jugular vein catheterization for neonates and infants using a small-caliber Doppler probe].

Eighteen neonates and infants scheduled for cardiac surgery, ranging from 1 to 42 months in ages and from 1.9 to 14.6 kg in weight, were placed in supine position under general anesthesia. The neck was moderately extended with the head turned to the left. The pathways of the right carotid artery and the internal jugular vein (IJV) were located with a Doppler probe (2.0 mm in diameter, HAYASHI Electric, TOKYO) using 10 MHz ultrasound wave. The pathways of the vessels were drawn on the skin surface and the IJV was cannulated with a 24-gauge catheter over needle (Neophron, Ohmeda, USA) along the drawn pathway. A 0.015-inch guidewire was passed through the 24-guage catheter, and a central venous catheter was placed in the IJV employing to the Seldinger method. In 16 out of 18 patients the pathways of the IJV was clearly differentiated from that of the carotid artery and the catheterization was successful without any undesirable events. In the remaining 2 patients the left IJV was catherized using the same method. No complications related to the catheterization were observed. Our catheterization method is thought to be highly reliable and safe in small pediatric patients.

Purification and structural analysis of extracellular matrix of a skin tumor from a patient with juvenile hyaline fibromatosis.

Juvenile hyaline fibromatosis is a rare mesenchymal dysplasia that is inherited in an autosomal recessive fashion. The histological features of the tumor-like lesions are characterized by the deposition of amorphous hyaline material in the extracellular spaces of the dermis and soft tissues. We have analyzed the hyaline substance in a specimen of a skin tumor obtained from a 4-year-old Japanese girl with juvenile hyaline fibromatosis. It was found to consist mainly of type VI collagen; a small amount of type I collagen was also present. These components were separated by DEAE-cellulose ion-exchange chromatography under reducing conditions. The ratio of the dry weights of type I and type VI collagen was 1:4. Of the three chains of type VI collagen (alpha 1(VI), alpha 2(VI) and alpha 3(VI)), alpha 3(VI) was the most abundant. Glycosaminoglycans in the tumor tissue comprised dermatan sulfate, chondroitin sulfate and hyaluronan, with dermatan sulfate predominating. In contrast, hyaluronan is the most abundant in normal skin.

The roles of the marginal veins in the differentiation of the rat hind limb bud: an immunocytochemical study.

Rat hind limb buds, aged between prenatal days 14 and 18, were used for electron microscopy and immunocytochemistry of fibronectin, laminin and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining indicative of DNA fragmentation. Fibronectin and laminin were actively synthesized in the rough endoplasmic reticulum of epidermal cells in the apical ectodermal ridge between prenatal days 14 and 15, but most cells underwent apoptosis after prenatal day 15. As the regression of the apical ectodermal ridge progressed, mesenchymal cells associated with the marginal veins were successively incorporated into the endothelium devoid of the basal lamina. No mitotic figures of endothelial cells were recognized either in the marginal vein or in the surrounding growing capillaries. Extracellular matrix components connected the adjacent mesenchymal cells, with the endothelium of such vessels immunoreacting to fibronectin and laminin. In addition, fibronectin-immunoreactive networks among the interstices of the mesenchymal cell cords developed in the avascular zone between the epidermis and the growing capillaries at prenatal day 15, but became inconspicuous at prenatal day 16. These results indicate that the apoptosis of the epidermal cells is the major reason for the regression of the apical ectodermal ridge, and that the capillary ingrowth from the marginal veins to the avascular zone is accelerated by transformation of mesenchymal cells to endothelial ones. Fibronectin and laminin seem to play crucial roles in capillary growth, especially in the adhesion between endothelial cells of the pre-existing vessels and mesenchymal cells.