The effects of a diode laser (810 nm) on pigmented guinea-pig skin.

Various types of lasers, such as the Q-switched ruby laser and the Alexandrite laser, cause selective damage to cutaneous pigmented cells and are currently used in the therapy of pigmented lesions. The aim of this study was to evaluate the effects of a diode laser at a wavelength of 810 nm on pigmented guinea-pig skin. The diode laser was supplied by OcuLight Iris Medical Instruments, Inc. and was used to deliver 0.15, 0.2, 0.3, 0.4, 0.5, 1 and 1.5 J/cm2 laser beams in micropulses of 100 micros. The study was carried out on albino and black-spotted guinea pigs (GP). After irradiation, punch biopsies were taken and analysed by light and electron microscopy. Albino animals developed just a few signs of cutaneous injury. This mostly consisted of spongiotic disarray, after the highest doses were administered (>1 J/cm2). In the black skin of spotted GPs alterations appeared at 0.15 J/cm2 and included melanosome damage and, at doses higher than 0.3 J/cm2, also melanocyte damage. These observations demonstrate that the diode laser at 810 nm selectively affects pigmented structures and that the cellular targets of diode laser radiation are the melanosomes. The diode laser specificity for melanin may provide a biological basis for the treatment of pigmented superficial cutaneous lesions.

Papuloverrucous colloid milium: an occupational variant.

We report a case of adult colloid milium in a 47-year-old mechanic with a long history of professional contact with lubricating oils and of sun exposure. In addition to the typical translucent papules seen on the forehead, there were warty papules on the backs of both hands with unusual histological and ultrastructural features: marked hyperplasia of the epidermis with orthokeratotic hyperkeratosis and papillary deposits of colloid material that were contiguous with the basal layer of the epidermis. The hand lesions caused by occupational exposure to mineral oils and solar radiation represent an occupational variant of adult colloid milium.

Up-modulation of the expression of functional keratinocyte growth factor receptors induced by high cell density in the human keratinocyte HaCaT cell line.

Keratinocyte growth factor (KGF) is involved in the control of proliferation and differentiation of human keratinocytes. It binds to, and activates, the tyrosine kinase KGF receptor (KGFR), a splicing transcript variant of the fibroblast growth factor receptor 2. We have previously shown (C. Marchese et al., Cell Growth Differ., 8: 989-997, 1997) that differentiation of primary cultured keratinocytes triggered by high Ca2+ concentrations in the growing medium induced up-regulation of KGFR expression, which suggested that KGFR may play a crucial role in the control of the proliferative/differentiative program during transition from basal to suprabasal cells. Here we analyzed the process of modulation of the expression of KGFRs in the human keratinocyte cell line HaCaT, widely used as a model to study keratinocyte differentiation. Western blot and double immunofluorescence for KGFR and the K1 differentiation marker showed that cell differentiation and stratification induced by confluence and high cell density correlated with an increase in KGFR expression. KGFRs, present on suprabasal differentiated cells, appeared to be efficiently tyrosine-phosphorylated by KGF, which indicated that the receptors up-regulated by differentiation can be functionally activated by ligand binding. Bromodeoxyuridine incorporation assay revealed that a significant portion of suprabasal differentiated cells expressing KGFR seemed to be still able to synthesize DNA and to proliferate in response to KGF, which suggested that increased KGFR expression may be required for retention of the proliferative activity.

Relationship between theoretical molecular weight and blister fluid/serum ratio of cytokines and five other molecules evaluated in patients with bullous pemphigoid.

Bullous pemphigoid (BP) blisters contain several molecules, some of which spread into the blisters from the interstitial fluid, while others are produced locally and migrate into the circulation. The calculation of the ratios between blister/serum concentrations may help to distinguish between these two types of molecules. The rules regulating the diffusion of the molecules have been described only in suction blisters, where the theoretical molecular weight (MW) represents one of the principal influencing factors. The aim of the present study was to analyse the relationship between theoretical MWs and the ratios of concentrations of several molecules evaluated both in sera and in blister fluids. Eight cytokines (interleukin-2, interleukin-3, interleukin-4, interleukin-5, interleukin-10, tumor necrosis factor-alpha, oncostatin-M and vascular endothelial growth factor), two acute phase reactants (alpha-1 acid glycoprotein, haptoglobin), albumin, one soluble membrane molecule with adhesion functions (sICAM-1) and the eosinophil cathionic protein (ECP) were measured in samples from 15 patients affected with BP by means of commercially available tests. The data suggest that the MW may influence the rate of diffusion throughout the blister, both in input and output directions, despite the discontinuity observed at the basement membrane level on the BP blister floor.

Fibroelastolytic papulosis of the neck: a report of 20 cases.

The clinical and histological features of the entities known as 'white fibrous papulosis of the neck' (WFPN) and 'acquired elastolysis of the papillary dermis simulating pseudoxanthoma elasticum' (PDE) are not clearly defined. This study was conducted to compare our experience of WFPN/PDE with those described in the literature. Twenty patients presented at our institution with papular eruptions involving the neck. The asymptomatic lesions, which ranged in colour from normal skin tones to yellowish, were isolated or coalescent. Microscopically, the papules showed elastolysis and fibrosis of the upper reticular and papillary dermis. A review of the literature shows similar characteristics in cases reported as WFPN and PDE. This study indicates that WFPN and PDE are variants of a single disorder that can be more precisely defined as 'fibroelastolytic papulosis of the neck' and which appears to be a manifestation of intrinsic skin ageing.

Granulomatous slack skin. Report of a case and review of the literature.

We describe a patient with granulomatous slack skin (GSS) who has been followed for 15 years and present clinical, histological, immunohistochemical, ultrastructural, cytogenetic, and molecular findings. The clinical and pathological aspects of the 20 cases of GSS reported in the recent literature are reviewed and compared with those of the present case.

Histology and ultrastructure of arteries, veins, and peripheral nerves during limb lengthening.

The effects of lengthening of the metacarpal bone on peripheral nerves and blood vessels were studied in 8 calves. Specimens for light and electron microscopy were obtained from the palmar neurovascular bundle at 1 cm (8% of the initial length), 2.5 cm (20% of the initial length), and 4 cm (33% of the initial length) of metacarpal lengthening. In 2 calves, specimens were studied 2 months after the end of the lengthening procedure. At 8% of lengthening, myelinated fibers of the palmar nerve showed moderate degenerative changes in the myelin sheath. This became severe at 20% and 33% of lengthening, and affected the axoplasm as well. At 20% of lengthening, the palmar vein started to show fibrous metaplasia of the smooth muscle tissue of the tunica media. This became much thinner than normal. The palmar artery showed moderate alterations of the inner part of the tunica media and the intima. The palmar nerve and blood vessels recovered their normal structure almost completely 2 months after the end of the lengthening procedure. The morphologic alterations of peripheral nerves and vessels may constitute the pathophysiologic basis of the nervous and circulatory disturbance observed in clinical practice.

Ledderhose's disease: case study with histologic and ultrastructural analysis.

Two cases of Ledderhose's disease (plantar fibromatosis) are described with emphasis on histologic features. This disorder has three histologic stages: in the first, or proliferative, stage, numerous fibroblasts associated with a small amount of collagen are observed; in the second, or active, stage, the cells are more mature and the collagen is more evident; in the third, or maturation, stage, the extracellular matrix is composed of dense bundles of collagen fibers with few fibroblasts. Ultrastructural analysis revealed filamentous aggregation of an unknown nature among the collagen fiber bundles. The authors discuss some clinical, physiological, and pathologic aspects of a disorder which is similar to Dupuytren's disease but does not involve retraction of the surrounding anatomical structures.

An ultrastructural study of slipped capital femoral epiphysis: pathogenetic considerations.

Two normal proximal femoral growth plates and core biopsies from six patients with slipped capital femoral epiphysis (SCFE) were studied by electron microscopy. In these SCFE patients, chondrocytes from all the zones of the plate were frequently smaller than normal, more irregular in shape, and many of them were degenerating, with formation of matrix vesicles and cellular debris. Floccular electron-dense material, most likely abnormal proteoglycan, was present in the hypertrophic rough-surfaced endoplasmic reticulum and Golgi apparatus as well as in the extracellular matrix, intermingled with collagen fibrils thinner than normal and loosely arranged. Mineralization of the abnormal matrix of the longitudinal septa of the degenerating zone was either scanty or absent, with scanty formation of irregular and thin bone trabeculae. The abnormalities observed in SCFE seem to be caused by a change in chondrocyte metabolism with consequent altered synthesis and/or extracellular aggregation of both collagen and proteoglycan, and scanty mineralization of the abnormal cartilage matrix.

Establishment, characterization and chemosensitivity of two human glioma derived cell lines.

Two continuous human glioma derived cell lines, LI and DF, were established in our laboratory. Both cell lines showed cytological features and in vitro behavior similar to those of the respective original neoplasms. These two lines were characterized for their main biological properties including in vitro and in vivo growth rate, clonogenic ability and tumorigenicity in nude mice. The plating efficiencies were generally high both during exponential and stationary growth phases and a high tumorigenicity was observed. All injected nude mice developed tumors. The two lines were tested for chemosensitivity to 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and cis-Diamminedichloroplatinum II (DDP). Heterogeneity in biological features and in drug sensitivity was observed. Exposure of the two lines to BCNU and DDP showed that the glioblastoma (LI) was less sensitive than the anaplastic astrocytoma (DF). For both lines BCNU was more effective on cells in plateau than in exponential phase, while the killing effect of DDP was not phase-dependent.

Idiopathic chondrolysis of the hip: an ultrastructural study of the articular cartilage of the femoral head.

Core biopsies obtained from the articular cartilage of the femoral head of a 16-year-old girl with idiopathic chondrolysis of the hip were subjected to an ultrastructural study. Zone I was missing and zone II was the most superficial layer present in the articular cartilage of our patient. Collagen fibrils were thinner than normal and more uniform in diameter, and proteoglycans were normally distributed among them. Degenerating chondrocytes were found, as well as debris of dead cells, but many chondrocytes were still vital and engaged in active synthesis. Most of the chronic degenerative changes observed in ultrastructural studies on osteoarthritis were not observed in this case of idiopathic chondrolysis which seems to be, on the basis of our findings, a pathological process which develops and runs its course very quickly.

Juvenile kyphosis: an ultrastructural study.

Biopsy specimens of the spine, including intervertebral disc, vertebral plate, growth plate, and part of the vertebral body, were obtained from seven patients with juvenile kyphosis treated by anterior spinal arthrodesis. Histological and histochemical studies showed abnormal loose-appearing cartilage in both the vertebral plate and the growth plate. Both plates were missing in some vertebrae. Ultrastructurally the abnormal cartilage had a matrix rich in proteoglycans and very thin collagen fibrils. The mineralization and ossification of the vertebral plates were irregular. Vertebral bone growth was stunted under the areas of abnormal growth plates.

Distribution of laminin and collagen type-IV in benign and malignant lesions of melanocytic origin.

Using antisera for two specific basement membrane (b.m.) antigens such as laminin and collagen type-IV, together with electron microscopy, we have shown that fully antigenic b.m. and morphologically typical basal lamina (b.l.) are associated with normal and transformed cells of the melanocyte lineage in different ways. Thus, while b.m. and b.l. surround individual choroidal melanocytes, intradermal nevus cells and cells of blue nevi are not detectable at the periphery of resting and proliferating epidermal melanocytes. They have a low degree of expression with a heterogeneous pattern of distribution in primary and metastatic melanoma. This heterogeneity is present within single metastases and among autologous metastases. These findings indicate that the presence of b.m. can be an additional marker for cells of the melanocyte lineage and should be considered when applying serological means for the detection and control of neoplasms of melanocytic origin.

De Quervain's disease. An ultrastructural study.

The tendon sheaths of extensor pollicis brevis (EPB) and abductor pollicis longus (APL), obtained from four patients with de Quervain's disease were studied by light and electron microscopy. Three different layers were identified in the sheath which was much thicker than normal. Both the outer and the middle layers had thick bundles of collagen fibres with scattered fibroblasts. The inner layer was mainly formed by chondroid and myxomatous tissue. Collagen fibrils were thicker than normal, reaching 2100 nm in diameter. Numerous cells which resembled "myofibroblasts" were scattered throughout the whole thickness of the sheath. The results seem to indicate that thickening and hardening of the EPB and APL tendon sheaths in de Quervain's disease is caused by increased synthesis of the extracellular matrix, increased thickness of the collagen fibrils and areas of myxomatous and chondroid metaplasia.

The "in vivo" effect of chymopapain on intervertebral disc hernia (an ultrastructural study).

This is an ultrastructural study of disc material obtained from two patients subjected to operation two weeks after an unsuccessful chymonucleolysis. It confirms that the "in vivo" effects of chymopapain are similar to those observed "in vitro", namely that it is ineffective in large contained hernias and in extruded hernias.

Morphologic changes in annulus fibrosus during aging. An ultrastructural study in rats.

Morphologic studies of annulus fibrosus of newborn, young, and old rats were carried out to determine the tissue changes occurring with aging. Newborn annulus consists of an inner portion containing chondroblast-like cells, a middle portion showing cells with intermediate features between chondroblasts and fibroblasts, and an outer portion containing fibroblast-like cells. The cells, arranged in rows, show a prominent ergastoplasm and are surrounded by pericellular lucunae. In the inner portion, the collagen fibers in the matrix are oriented randomly; in the middle and outer portions, the fibers between adjacent cell rows are arranged parallel, forming fibrous lamellae. The collagen fibers have fairly uniform diameters (average 510 nm), are strongly positive to the PASM reaction, and are surrounded by small proteoglycan granules. Both immature and mature elastic fibers are present. Young annulus consists of a transition zone, composed of large cells with no definite arrangement and bundles of collagen fibers oriented in various directions, and a lamellar zone consisting of fibrous lamellae separated by interlamellar septa containing the cells. These show intermediate features between chondrocytes and fibrocytes and well-developed ergastoplasm. The interlamellar septa consist of collagen fibrils and proteoglycans and contain few aggregates of electron dense material. The collagen fibers have highly variable diameters (average 740 nm). The larger fibers are scarcely periodate-silver-methenamine (PASM) positive. The proteoglycans are similar in size to those in the newborn. Few, mature elastic fibers are present. In old annulus, the transition zone is larger than in the young tissue. The cells are less numerous, and most of them show a sparse ergastoplasm.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of gossypol and Lonidamine on electron transport in Ehrlich ascites tumor mitochondria.

The effect of the association of gossypol and Lonidamine on the electron transport in Ehrlich ascites tumor mitochondria has been investigated by addition of drugs to isolated mitochondria. The results may be summarized as follows. (1) Low concentrations of gossypol increase the rate of oxygen consumption at the level of three energy-conserving sites of the respiratory chain. Higher concentrations result in an inhibition of oxygen consumption at (or near) both energy-conserving sites 1 and 2, while energy-conserving site 3 is unaffected. (2) Gossypol, at concentrations at which it exerts its uncoupling effect, stimulates ATPase activity. Higher concentrations inhibit the enzyme activity. (3) The addition of gossypol to mitochondria respiring on pyruvate plus malate or succinate induces a more oxidized state of NAD+ and cytochrome b, respectively. (4) Gossypol enhances the effect of Lonidamine on oxygen consumption. Lonidamine does not affect state 4 respiration, but in the presence of gossypol, it determines a marked decrease in the rate of oxygen consumption. The inhibition of oxidation of NAD-linked substrates is greater than that of FAD-linked substrates. (5) It may be concluded that gossypol is very effective in potentiating the effect of Lonidamine. Moreover, it may be suggested that the antitumor activity of Lonidamine is enhanced if it is used in combination with other drugs and/or treatments, such as hyperthermia, which modify the energy status of mitochondria.

Heterogeneous distribution of actin, myosin, fibronectin and basement membrane antigens in primary and metastatic human breast cancer.

The distribution of actin, myosin, fibronectin and basement membrane antigens has been studied by indirect immunofluorescence in benign and malignant human breast lesions. While benign tumors showed only minor differences from normal mammary tissue, tumors of different histological types displayed a heterogeneous distribution of the antigens studied. Heterogeneity was observed within the same tumor, among different neoplasms and between primary tumors and autologous metastases. As a common characteristic, most of the tumors did not stain for actin and myosin, the pattern being similar to that found in myoepithelial cell distribution. In transformed epithelia there was often a lack of detectable actin with a myosin-positive fluorescence. Staining for both proteins was diffused to most of the cell cytoplasm. Staining for fibronectin was seen in only a minority of the cases, with medullary tumors being the most positive. Basement membrane stain was either absent or decreased and fragmented, except in rare ductal, i.e. papillary, carcinomas. Medullary tumors displayed an almost continuous, though fragmented basement membrane in approximately 70% of cases.

Morphological damage induced in vivo by Lonidamine on human metastatic cancer cells.

Lonidamine was given to several patients affected with different types of neoplasias growing as metastases both in ascites and pleural effusion, and solid cutaneous metastases. The patients with tumor cells in ascites and pleural effusion were treated with Lonidamine per os or in loco injections. In cutaneous metastases, Lonidamine was administered by different routes: (1) per os; (2) local endoarterial; and (3) in association with hyperthermic perfusion with or without antiblastic drugs.