Frequent overexpression of cyclin D1 in sporadic pancreatic endocrine tumours.

Pancreatic endocrine tumours (PETs) occur sporadically or are inherited as part of the multiple endocrine neoplasia type-1 syndrome. Little is known about the molecular events leading to these tumours. Cyclin D1, a key regulator of the G1/S transition of the cell cycle, is overexpressed in a variety of human cancers as well as certain endocrine tumours. We hypothesized that similar to other endocrine tumours, cyclin D1 is overexpressed in human sporadic PETs. Cyclin D1 protein overexpression was found in 20 of 31 PETs (65%) when compared with normal pancreatic tIssue. Furthermore, Northern blot analysis suggests that cyclin D1 up-regulation occurs at the post-transcriptional level in some PETs. Because the key cell growth signalling pathways p42/p44/ERK (extracellular signal-regulated kinase), p38/MAPK (mitogen-activated protein kinase), and Akt/PKB (protein kinase B) can regulate cyclin D1 protein expression in other cell types, pancreatic endocrine tumours were analysed with phospho-specific antibodies against the active forms of these proteins to elucidate a tIssue-specific regulatory mechanism of cyclin D1 in PETs. We found frequent activation of the p38/MAPK and Akt pathways, but down-regulation of the ERK pathway, in cyclin D1 overexpressing PETs. This study demonstrates that cyclin D1 overexpression is associated with human sporadic PET tumorigenesis, and suggests that this up-regulation may occur at the post-transcriptional level. These findings will direct future studies of PETs towards cell cycle dysregulation and the identification of key growth factor pathways involved in the formation of these tumours.

Alternative mRNA splicing in colon cancer causes loss of expression of neural cell adhesion molecule.

BACKGROUND: The neural cell adhesion molecule (NCAM) has numerous isoforms resulting from alternative splicing of mRNA. The 3 major isoforms found in adult tissue are (1) a 120-kDa protein that is linked to the plasma membrane by glycosylphosphatidylinositol; (2) a 140-kDa form that has a transmembrane component and a cytoplasmic tail with unknown function; and (3) a 180-kDa isoform that has an intracellular protein that binds the cytoskeleton. NCAM is capable of homotypic binding and therefore plays a role in cell-cell adhesion for cells expressing the 180-kDa isoform by anchoring groups of cells into epithelial sheets. NCAM-180 is the isoform found in colonocytes, and loss of expression is associated with clinically aggressive colon cancers. METHODS: Western blotting and reverse transcriptase-polymerase chain reaction were used to screen commercially available cell lines for NCAM-180 expression. For cell-line pairs with differential NCAM-180 expression, exon analysis was performed with reverse transcriptase-polymerase chain reaction to determine where the molecule was spliced, culminating in failed expression. These results were confirmed with exon analysis in colon cancers harvested at the time of laparotomy. RESULTS: Analysis of a SW480 cell line (derived from a patient's primary colon cancer lesion) revealed NCAM-180 expression, whereas no expression was found in the SW620 cell line (derived from a metastatic lesion from the same patient). Exon analysis of NCAM mRNA transcripts from SW620 revealed that the transcripts were truncated after exon 12. This region correlates to an area between 2 fibronectin-III domains on the NCAM protein. CONCLUSIONS: The most common site for NCAM alternative splicing is between the 2 fibronectin-III domains corresponding to the border between exons 12 and 13 of the NCAM gene. Loss of NCAM-180 expression in aggressive colon carcinoma results from a splice defect in the same area, which may result in defective intracellular adhesion between colonocytes.

Detection of chromosome 11q13 amplification in head and neck cancer using fluorescence in situ hybridization.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) remains a cancer with one of the lowest five-year survival rates. Despite a better understanding of the disease and recent advances in diagnosis and treatment, survival rates for HNSCC patients have not improved. Chromosomal abnormalities are common in HNSCC, and aberrations of chromosome 11q13 have been correlated with a poor prognosis. MATERIALS AND METHODS: In this study we utilized fluorescence in situ hybridization (FISH) to determine the incidence of 11q13 amplification in twenty primary HNSCC tumors. INT-2 was used as the 11q13 probe, and 9 and 11 centromeric probes were used as controls. RESULTS: Polysomy, greater than two copies of chromosome 11, was found in 2 of 20 tumors. INT2 (11q13) amplification was found in 3 other tumors. CONCLUSIONS: These preliminary studies indicate tht analysis of a larger sample of tumors using FISH may yield important diagnostic and prognostic information about head and neck tumors.

Phosphate depletion in the rat: effect of bisphosphonates and the calcemic response to PTH.

BACKGROUND: The removal of phosphate from the diet of the growing rat rapidly produces hypercalcemia, hypophosphatemia, hypercalciuria, and hypophosphaturia. Increased calcium efflux from bone has been shown to be the important cause of the hypercalcemia and hypercalciuria. It has been proposed that the increased calcium efflux from bone is osteoclast mediated. Because bisphosphonates have been shown to inhibit osteoclast-mediated bone resorption, this study was performed to determine whether bisphosphonate-induced inhibition of osteoclast function changed the biochemical and bone effects induced by phosphate depletion. METHODS: Four groups of pair-fed rats were studied: (a) low-phosphate diet (LPD; phosphate less than 0.05%), (b) LPD plus the administration of the bisphosphonate Pamidronate (APD; LPD + APD), (c) normal diet (ND, 0.6% phosphate), and (d) ND + APD. All diets contained 0.6% calcium. A high dose of APD was administered subcutaneously (0.8 mg/kg) two days before the start of the study diet and on days 2, 6, and 9 during the 11 days of the study diet. On day 10, a 24-hour urine was collected, and on day 11, rats were either sacrificed or received an additional APD dose before a 48-hour parathyroid hormone (PTH) infusion (0.066 microgram/100 g/hr) via a subcutaneously implanted miniosmotic pump. RESULTS: Serum and urinary calcium were greater in the LPD and LPD + APD groups than in the ND and ND + APD groups [serum, 11.12 +/- 0.34 and 11.57 +/- 0.45 vs. 9.49 +/- 0.17 and 9.48 +/- 0.15 mg/dl (mean +/- SE), P < 0.05; and urine, 8.78 +/- 2.74 and 16.30 +/- 4.68 vs. 0.32 +/- 0.09 and 0.67 +/- 0.28 mg/24 hr, P < 0.05]. Serum PTH and serum and urinary phosphorus were less in the LPD and LPD + APD than in the ND and ND + APD groups (P < 0.05). The calcemic response to PTH was less (P < 0.05) in the LPD and LPD + APD groups than in the ND group and was less (P = 0.05) in the LPD + APD than in the ND + APD group. Bone histology showed that phosphate depletion increased the osteoblast and osteoclast surface, and treatment with APD reduced the osteoblast surface (LPD vs. LPD + APD, 38 +/- 4 vs. 4 +/- 2%, P < 0.05, and ND vs. ND + APD, 20 +/- 2 vs. 5 +/- 2%, P < 0.05) and markedly altered osteoclast morphology by inducing cytoplasmic vacuoles. CONCLUSIONS: (a) Phosphate depletion induced hypercalcemia and hypercalciuria that were not reduced by APD administration. (b) The calcemic response to PTH was reduced in phosphate-depleted rats and was unaffected by APD administration in normal and phosphate-depleted rats, and (c) APD administration markedly changed bone histology without affecting the biochemical changes induced by phosphate depletion.

Fractal dimension of the bone marrow in metastatic lesions.

We previously reported that the distribution of the cells in normal bone marrow is fractal and self-similar. The purpose of this study was to determine whether the same is true in metastatic tumors. Thirty-two bone marrow biopsy sections (3 to 5 microm thick) of 28 patients were used to measure the fractal dimensions of the metastatic tumor cells' distribution. Microscopic images were obtained and were used for the fractal measurements. In the two-dimensional images, the fractal dimensions were 1.98 +/- 0.02 (95% +/- 5% cellularity), suggesting a compact nonfractal structure. The dimensions, however, were 1.72 +/- 0.1 (56% +/- 11% cellularity) for the normal components, with a P-value of <.0001 that is in agreement with our previous study. These results suggest that loss of the fractal structure in the metastatic lesions may be attributable to loss or suppression of the regulatory mechanisms maintaining the fractal morphogenesis of the bone marrow. This report provides a novel objective approach in the study of pathophysiology of the bone marrow disorders.

Tumor suppressor activity of neural cell adhesion molecule in colon carcinoma.

BACKGROUND AND AIMS: Neural Cell Adhesion Molecule (NCAM) is a well-characterized member of the immunoglobin superfamily. The structure of NCAM is similar to the tumor suppressor Deleted in Colon Carcinoma (DCC). NCAM has been found in some epithelial tissues and plays a role in tumorigenesis of some cancers. The purpose of the present study was to determine if NCAM is present in normal human colon. Once its presence was established, its function as a tumor suppressor was investigated. METHODS: Colon tumors and normal proximal margins were processed for reverse transcription-polymerase chain reaction (RT-PCR) of the NCAM-180 message. Immunohistochemistry of the tissue was performed to determine the distribution of NCAM. RESULTS: RT-PCR analysis demonstrated the presence of the NCAM-180 kD isoform in normal colonic epithelia. Immunohistochemistry showed NCAM on the basolateral surface of colonic epithelial cells of the villous tips. Tumors from 15 patients followed up for 4 years were studied. All seven tumors expressing NCAM-180 were from patients having a benign clinical course. Seven of eight tumors that lacked NCAM-180 were associated with aggressive clinical behaviors (presenting with obstruction, perforation or metastatic disease, or patient death within 18 months of presentation). The sole exception was in a villous adenoma excised from a patient who has had multiply recurrent polyps on follow-up. CONCLUSION: We conclude that like DCC, NCAM is an important colonic adhesion molecule that functions as a tumor suppressor.

Morphogenesis of the bone marrow: fractal structures and diffusion-limited growth.

Bone marrow (BM) provides a particular spatial organization that allows interaction between its various components. Characterization of the spatial patterns in the BM and understanding the mechanisms that give rise to them may play a role in better understanding of the BM pathologic processes. Morphometric analyses were performed in BM biopsy samples from 30 patients (16 men and 14 women) with an average age of 46 years, ranging from 17 to 77 years. The biopsies were obtained during the course of patient care to rule out BM involvement in a variety of hematologic disorders before or after therapy. Three different, but structurally interrelated, parameters were measured: (A) cellular area, (B) nuclear area, and (C) cell numbers. All three methods, in all cases, showed that the spatial structure of the BM is fractal. The average values of the fractal dimensions (Df) were 1.7 +/- 0.08, 1.64 +/- 0.1, and 1.69 +/- 0.04 for categories A, B, and C, respectively. The overall value of Df for the cellularity in the range of 40% to 60% was about 1.67 +/- 0.09. Fractal dimensions of 1.6 to 1.7 represent configurations that correspond to two-dimensional diffusion limited aggregation structures, suggesting that the structural configuration of hematopoietic cells is dependent on the diffusion of regulatory cytokines in the BM.

Pulmonary reaction to an aspirated balanced liquid diet in a cancer patient.

A 60-year-old man with pharyngeal carcinoma developed spontaneous pneumothorax. A biopsied lung tissue revealed an extensive granulomatous reaction to some yellow material. Because of the patient's frequent aspirations of the liquid diet which was mixed with an artificial lemon flavor, it is considered that the aspirated material provoked the pulmonary reaction and possibly the pneumothorax. An energy dispersive X-ray analysis of the biopsied lung tissue aided in excluding the possibility of the pulmonary reaction and the yellow material being of ubiquitous pneumoconiotic origin.

Fiber contamination of vermiculites: a potential occupational and environmental health hazard.

Vermiculite ores from Montana, Virginia, and South Africa have been analyzed for the presence of amphibole contamination. Fibrous actinolite was found in unexpanded Montana vermiculite ore at a maximum concentration of 2.0%. The fibers persisted in the expanded ore at a maximum concentration of 0.6%. Actinolite was also found in the Virginia vermiculite ore but at a lower concentration and mostly as cleavage fragments with low length-to-width ratios. South African ore contained rare anthophyllite fibers also with low length-to-width ratios. Vermiculite ores have the potential for amphibole contamination and can represent potential health hazards without proper occupational and environmental control measures.

Identification and significance of magnetite in human tissues.

Magnetite or iron oxide has been identified in humans as well as certain animals and bacteria. With the current popularity of magnetic resonance imaging, the presence of these ferromagnetic particles in the tissues may impose biological significance. So far, identification of magnetite in tissue has been mainly based on magnetometry. Hence, a simple technique for direct identification of the magnetic particles in tissues is described. Lung tissues with abundant iron material and particles were digested in 1N sodium hydroxide solution. After rinsing, the sediments were suspended in 95% alcohol and placed on a glass slide located on a strong magnet. The iron-containing particles from the digestion procedure were aligned in a parallel manner along the north-south poles of the magnet and were confirmed to be magnetite by x-ray diffraction. No such effect was observed with hemosiderin-containing granules from the control liver tissues. The results of this experiment show that the "biological magnetite" is distinctly different from hemosiderin and has characteristic properties when subjected to a magnetic field.

Chrysiasis: the role of sun exposure in dermal hyperpigmentation secondary to gold therapy.

To investigate the role of sun exposure in the pathophysiology of chrysiasis, we studied 10 Caucasian female patients with rheumatoid arthritis: 4 with clinically apparent chrysiasis and 6 without apparent pigmentation. Three patients without chrysiasis had received over 4 g of gold and 3 less than one g. The mean melanin score, determined by histological examination of sun exposed and nonsun exposed skin, was significantly higher in the sun exposed skin of the chrysiasis and high dose controls than low dose controls (p less than .05). Concentration of gold measured semiquantitatively by transmission electron microscopy and quantitatively by atomic absorption showed increased gold concentration in sun exposed when compared to nonsun exposed skin of chrysiasis and high dose controls (p = .26). Low dose controls had no gold demonstrated by either method. Our results suggest that gold deposition in the dermis stimulates melain production and that melanin is important in hyperpigmentation of chrysiasis. Furthermore ultraviolet light may induce preferential uptake of gold by the skin.

Zeolite exposure and associated pneumoconiosis.

Naturally occurring zeolite minerals are aluminum silicates widespread in the earth's crust. Several of these minerals have fibrous forms and have been implicated as a possible cause of benign and malignant diseases of the lung and pleura in Turkey. This report describes a patient, living in an area of Nevada rich in zeolites, who presented with idiopathic pleural thickening and pulmonary fibrosis associated with extensive pulmonary deposition of zeolites.

The intracellular degradation of poly(epsilon-caprolactone).

Poly(epsilon-caprolactone) [PEC], a biodegradable aliphatic polyester, undergoes a two-stage degradation process: The first lengthy phase involves nonenzymatic hydrolytic cleavage of ester groups, the second phase beginning when the polymer is more highly crystalline, and of low molecular weight. The cellular events of the second phase were examined by implanting gelatin capsules containing 25 mg of low molecular weight (Mn 3000) PEC powders, 106 to 500 micron, in rats. PEC fragments ultimately were degraded in phagosomes of macrophages and giant cells, the process requiring less than 13 days for completion at some sites. PEC was also identified within fibroblasts. These studies support the intracellular degradation of PEC as the principal pathway of degradation once the molecular weight of the aged polymer is reduced to 3000 or less.

Peri-articular reactions to microscopic erosion of silicone-polymer implants. Light- and scanning electron-microscopic studies with energy-dispersive x-ray analysis.

Two patients are described who underwent implantation of silicone-polymer prostheses: one for an arthroplasty of the left great toe after a bunionectomy, and another for replacement of fractured trapezoid bone of the left wrist. Each patient developed unexplained severe pain at the sites of surgery at 3 and 7 months postoperatively. On removal of the implants and adjacent tissues, there was a microscopic foreign-body reaction to amorphous material in the subsynovial connective tissue. Although both prostheses were grossly intact, scanning electron microscopy revealed multiple erosions on their surfaces. Energy-dispersive x-ray analysis of foreign material in the paraffin-embedded sections revealed a peak for silicon. These two cases indicate that clinically significant pathological reaction may occur as early as 3 months after silicone-polymer implantation and that energy-dispersive x-ray analysis of excised tissue is a useful and specific diagnostic procedure.

Responses of atherosclerotic aorta to argon laser.

Lasers have been advocated to resect atherosclerotic plaques in the cardiovascular system, yet little information is available regarding the effects of laser on the range of occlusive lesions seen in the peripheral arterial tree. This study was conducted to assess the risk of perforation in human cadaveric aorta involved with variable degrees of atherosclerosis. Ten fresh segments of atherosclerotic human aorta were graded for extent of atherosclerosis, then subjected to argon laser energy within 48 hours. Using air as the conduction medium and with the fiber tip 2 or 5 mm from the vessel wall, the argon laser was applied to matched calcified and non-calcified arteries at 3.0-7.0 W and 10.0-13.5 W with energy density identical for matched pairs. Results were compared among segments which were normal in appearance or had only fatty streaks grossly with those with gross regional wall calcification. The mean penetration time (T) for calcified and non-calcified lesions at low and high power outputs was compared. (table; see text) Mean time to perforation and range of time necessary to produce perforation were greater in calcified than non-calcified segments at all power levels employed. These data suggest that atherosclerotic lesions vary in their response to argon laser. The presence of calcium may preclude resection of some plaques and protect against wall perforation.

Acute effects of argon laser on human atherosclerotic plaque.

Although the laser has been demonstrated to vaporize coronary artery plaque, there is little information about its ability to resect or vaporize the range of plaques present in peripheral vessels. This study attempts to determine the ability of the argon laser to resect all grades of atherosclerotic plaque, the risk of perforation during plaque resection, the effects on surrounding arteries, and the effect of different transmission media (air, saline solution, and blood) on the delivery of laser energy to the vessel. Seventy-five adult human cadaveric aortic specimens with a range of atherosclerosis from grossly normal artery to extensive calcification with ulceration were exposed to variable energy densities (200 to greater than 20,000 J/cm2) within 48 hours of harvesting. Specimens were examined grossly for the visual effects of laser and microscopically after preparation with hematoxylin-eosin, trichrome, and/or Verhoeff's elastin stains. Our results indicate that normal arteries and noncalcified plaques absorb laser energy and are vaporized. As the atherosclerosis becomes more complex with calcification, calcified regions are not vaporized and cannot be resected. In normal arteries and noncalcified plaque, perforation times were less than 5 seconds. Where palpable calcification was present in atherosclerotic lesions, average perforation time was doubled. In some vessels areas of calcification prevented wall perforation, but areas of subintimal hemorrhage perforated rapidly because of the selective absorption of laser energy by the red color of hemorrhagic tissue. These results remain the same when saline solution is used as a transmission media, although the amount of energy required to achieve the effects is increased.(ABSTRACT TRUNCATED AT 250 WORDS)

Electron microscopy in small cell lung carcinomas: clinical correlation.

In order to investigate the relationship of subcellular differentiation of small cell lung carcinomas (SCLC) and clinical response, we reviewed the electron microscopic (EM) features of tumor biopsy specimens from 33 patients with SCLC diagnosed by light microscopy (LM). These tumors were divided by EM into four groups according to the ultrastructural features. Group I (13 patients) had tumors with only neurosecretory granules on EM. Group II (11 patients) had tumors with neurosecretory granules and other subcellular features of non-SCLC. Group III (five patients) had tumors that lacked neurosecretory granules but contained subcellular features of non-SCLC. Group IV (four patients) had tumors that lacked all of these features. The complete and partial response rate to systemic chemotherapy with or without radiation therapy was 88% in the total population studied. The response rates were not statistically different in any of the four groups based on EM findings. The results of this study suggest that the LM diagnosis of SCLC alone adequately identifies lung cancer patients with a high response rate to systemic therapy.