Effects of methanol extraction residue of Bacillus calmette-Guérin in humans.

Forty patients with histologically confirmed neoplastic diseases were treated with the methanol extraction residue of Bacillus Calmette-Guérin (MER). Thirty-six received concomitant chemotherapy. MER was initially given intradermally twice a month, 1 week apart, at a dose of 200 mug into each of five sites draining different lymph node-bearing areas on the anterior body surface. Thirty-seven patients developed local ulcerations at least 0.5 cm in diameter at MER injection sites. Typical lesion evolution was characterized by erythema and induration followed by vesicle formation and central necrosis. Either granulation tissue or a thick nonulcerated eschar preceded healing, leaving a linear, flat scar. Systemic toxicity consisted of malaise, fever, and myalgias on the day of MER administration. No hematological or biochemical changes directly attributable to MER were observed. Dose titrations in decreasing 10-fold dilutions in a linear array in a single anatomical region were carried out on 35 occasions. All patients but three developed at least a 5-mm induration to the 1-mug dose within 2 weeks of titration. Dose reductions were necessary in 19 instances. The minimal dose that produced a 1-cm inflammatory lesion with central necrosis was 0.01 mug. Serial biopsies were performed. These indicated a time-related series of changes from a nonspecific inflammatory lesion to an acute inflammatory response with microabscesses, followed by noncaseating granulomata and ultimately fibrosis. MER is a quantifiable nonviable immunostimulant that obeys dose-response relationships in its cutaneous lesions.

Evidence for nonrenal synthesis of 1,25-dihydroxyvitamin D in patients with inflammatory arthritis.

The extrarenal synthesis of 1,25-dihydroxyvitamin D [1,25-(OH)2D] is a characteristic of activated macrophages and has been demonstrated to occur in vitro in synovial fluid macrophages from patients with inflammatory arthritis. To examine whether such synthesis occurs in vivo, 19 patients with rheumatoid arthritis, 5 patient controls, and 5 healthy controls received a challenge oral dose of 250 micrograms 25-hydroxyvitamin D3 (25-OHD3) and the serum 1,25-(OH)2D3 response was measured. The median rise in serum 1,25-(OH)2D3 was significantly greater (22 pg/ml) in the rheumatoid patients compared to either of the control groups (8 pg/ml), although the increase in precursor 25-OHD3 was similar in all groups. The serum 1,25-(OH)2D concentration did not rise above the normal upper limit in any of the control subjects but exceeded the normal range in 8 of the rheumatoid patients. Extrarenal 1,25-(OH)2D synthesis is substrate dependent, unlike renal 1 alpha-hydroxylation, which is homeostatically controlled. Excessive 1,25-(OH)2D3 synthesis in the rheumatoid group on raising the 25-OHD3 concentration is indicative of nonrenal production of the hormonal metabolite. Further evidence for substrate-dependent extrarenal synthesis came from measurements of 25-OHD and 1,25-(OH)2D in paired serum and synovial fluid samples from 19 patients with inflammatory arthritis, including 15 with rheumatoid arthritis. Synovial fluid 1,25-(OH)2D was usually present at a lower concentration than serum 1,25(OH)2D, with which it was strongly correlated (Kendall's R = 0.46, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Antigen localization in immunoperoxidase-stained plastic-embedded soft tissues.

Immunoperoxidase stains were performed on normal and neoplastic tissue from prostate, colon, thyroid, lung, nerve, uterus, and placenta embedded in both plastic (glycolmethacrylate [GMA]) and paraffin. Positive results in plastic section were obtained for carcinoembryonic antigen (CEA), keratin, epithelial membrane antigen (EMA), thyroglobulins, S-100, prostate-specific antigen, human chorionic gonadotrophin (HCG), and beta-HCG. More delicate staining with more precise localization of antigens is noted. Superior (paraformaldehyde) fixation and cold processing followed by GMA polymerization (4 degrees C) allow for optimum antigen survival. After fixation, tissue processing involves a series of 0.1 mol/L phosphate buffer rinses with sucrose and ammonium chloride in a conventional dip-and-dunk processor placed in a 4 degrees C cold room. Acetone dehydrations are used before GMA infiltration, cold polymerization, and sectioning. Before immunoperoxidase staining, the plastic section is digested in .25% bovine trypsin for ten minutes. The immunoperoxidase methods described can be useful when small biopsies are routinely embedded in plastic to obtain improved histologic (hematoxylin-eosin) sections. There may also be research applications in quantifying antigen expression in benign, dysplastic, and neoplastic tissues by examining the stains under high power.

Fat embolism to the cardiac conduction system associated with sudden death.

During a brief hospital stay, a patient under treatment with corticosteroids for lymphoma died suddenly and unexpectedly. At autopsy, fat emboli were identified in the lungs, myocardium, liver, and brain. Examination of the cardiac conduction system revealed fat emboli in the vessels of the bundle of His as well as those accompanying the bundle branches, a finding believed to have resulted in the sudden death. This report is the first of fat embolization to the cardiac conduction system and emphasizes the importance of examination of this system in cases of unexplained or sudden death.

The problems of environmental hazardous substances: an overview.

It was decided to present an overview of problems of environmental hazardous substances by selecting three areas of concern which represent different aspects of the overall subject. All three are topics of great interest at the present time. They provide examples of different causes of the hazards and different approaches to their resolution or containment. Aerial and water contamination by radon gas are a natural hazard only recently recognized. Radon can be contained, but it will require cooperative action and expenditure. Herbicides are a continuing problem demanding increased research activity to establish risk-benefit decisions. Pollution of estuaries and oceans require new development of models for establishing safe levels of some pollutants and prohibition of others.

Plastic embedding as a tool in surgical pathology diagnosis.

Plastic (polymer) embedding is recommended as the method of choice for studying small biopsies such as those obtained from liver, bone marrow, lung and by endoscopy from the gastro-intestinal tract. The method offers the opportunity to examine one to four micron thick sections and, therefore, to evaluate cytologic detail. Multiple levels can be obtained from such fragments, and the reasonably large blocks provide adequate section size for diagnostic purposes.

The mechanism of malignant tumor induction by nickel subsulfide.

Experiments were performed to study the earliest changes and their sequence in the development of malignant tumors following implantation of 10 mg of nickel subsulfide (Ni3S2) into the right quadriceps muscle of seven male Fischer rats. Biopsies were performed when nodules reached one to three mm, and, later, when they were five to seven mm, and, finally, at sacrifice, to confirm the fully developed tumor pattern. Light microscopy of the earliest samples showed groups of cells clumped and scattered among degenerating muscle fibers. Mitoses were seen and inflammatory cells were not a feature. Electron microscopy showed individual degenerating muscle fibers, but also cells with characteristic features of myofibroblasts. In many cells osmium, dense fragments suggestive of crystalline material, were seen in the cytoplasm and nuclei. In the second set of biopsy material, myofibroblasts with well defined and dilated rough endoplasmic reticulum, intracellular membrane-bound collagen, and microfilaments with focal "dense bodies" were numerous. Mitoses were frequent. Immunohistochemistry showed strongly positive reaction to Vimentin and muscle-specific Actin in the tumor cells. In the fully developed tumors, the previously described typical storiform cell pattern with spindle and spheroidal cells with frequent mitoses was seen. Vimentin and muscle-specific Actin stains were strongly positive. The long latent period, the evidence of cell degeneration, necrosis, foreign material (probably of nickel composition), cell invasion, and subsequent rapid myofibroblast-type cell development, proliferating to malignant tumors highly suggestive of malignant fibrous histiocytoma, seem to suggest an epigenetic form of carcinogenicity of cytotoxic variety. Whether the tumor cells derive from transformed myofibrils or from activated pluripotential mesenchymal cells, or from both, remains in doubt.

Sodium diethyldithiocarbamate administration in nickel-induced malignant tumors.

Studies were undertaken to ascertain the possible antitumorigenic effectiveness of sodium diethyldithiocarbamate (dithiocarb) on the development of tumors in rats following the muscular implantations of nickel subsulfide (Ni3S2). These tumors have histologic characteristics which suggest origin from striated muscle and have been classified as rhabdomyosarcoma. Most of the tumors metastasized. In two separate studies over a period of two years, a total of 100 four-month old Fischer rats received muscular implantations of Ni3S2. Of this total, 50 rats (25 males and 25 females) were treated with dithiocarb for a period of four to six weeks. Seventy-eight percent of the untreated rats developed sarcomas as compared to 32 percent of the treated rats. Analyses of the data revealed a striking difference in the sex responsiveness to dithiocarb. Of the 25 female rats with Ni3S2 implantations and treated with dithiocarb, only 12 percent developed sarcomas; of the male rats similarly treated, 52 percent developed sarcomas. The difference in sex response is statistically significant (p = 0.005). Therapeutic implications are suggested.

A modified Brown and Hopps stain for identification of gram-positive and gram-negative microorganisms in glycol methacrylate-embedded tissues.

Plastic embedding is a recently developed technique that has been shown to be superior to conventional paraffin embedding in the histopathologic identification of microorganisms in tissue specimens. This report describes a modification of the Brown and Hopps stain for the differentiation of gram-positive and gram-negative microorganisms in 3-micron-thick plastic sections. This technique is easily performed and has a rapid turnaround time. This staining method permits a more precise histopathologic diagnosis of infectious agents than is possible in paraffin-embedded tissues.

Nickel-induced malignant tumors.

A series of malignant tumors experimentally induced by implanting Ni3S2 in gelatin capsules into the right hind limb of Fischer 344 rats is described. In this series, histology and ultrastructure revealed two major cell types. These are described as fibroblast-like and histiocyte-like. In addition, numerous giant cells, including large bizarre multinucleate giant cells with granular cytoplasm, were also seen. In this preliminary report, no more precise diagnosis will be given for this tumor than "malignant tumor of pluripotential origin". Further work is proposed to define more specifically its histogenesis. The appearances of the experimentally produced tumor are compared with certain tumors found in humans and also in experimentally induced tumors by Maruyama et al designated as malignant fibrous histiocytoma (MFH).

Damage in canine hearts following defibrillator shocks.

Experiments were performed to investigate possible differences in potential myocardial cell damage following the use of two clinically available difibrillators. One had a damped sine wave (DSW), and the other a truncated exponential waveform (TEW). The latter, therefore, had a lower peak current and voltage. After pilot studies to determine damage potential, an energy content of 10 Joules per kg was selected with three transthoracic shocks at 30 second intervals, delivered with paddles, by both defibrillators. Thirteen dogs were shocked with DSW (10 sacrificed at 24 hours and three at 72 hours). Eleven dogs were shocked with TEW (eight sacrificed at 24 hours and three at 72 hours). Three were used as untreated controls. Observations included ECG monitoring, technetium99m pyrophosphate (Tc-PyP) uptake, and gross and microscopic observations including electron microscopy. This report will concentrate on the morphologic changes. Eleven of 13 dogs shocked with DSW developed ventricular tachycardia, transient heart block, Tc-PyP uptake and myocardial cell death with acute necrosis, and aberrant contraction patterns associated with cell death. Progression of the lesions occurred from 24 to 72 hours. Only two (one at 24 hours and one at 72 hours) of the dogs shocked with TEW showed microscopic foci of necrosis. One was a chance finding (24 hours); the other was associated with an overlying "yellow streak". In neither case were arrhythmias or Tc-PyP uptake observed. The results indicate that in dogs at equivalent energy levels, TEW caused significantly less myocardial damage than DSW.

Histogenesis of subcutaneous malignant tumors resulting from nickel subsulfide implantation.

Tumors are described which have been produced by subcutaneous (s.c.) implantation of nickel subsulfide (Ni3S2) in 00 gelatin capsules in the left axillary region of Fischer 344 rats. This is in contradistinction to previous accounts where intramuscular (i.m.) or, in a few cases, subcapsular testicular implantation sites have been used. The advantages of the s.c. route are discussed, including the removal of the difficulties of distinguishing degenerating and regenerating striated muscle cells from tumor cells. The s.c. induced tumors were transplanted as far as the F6 generation, in some cases. Cells from tumor transplanted as far as the F3 generation were grown in tissue culture and then reimplanted s.c. Cytology of the tumors remained remarkably constant in all tumors, even after serial transplants and culture. A characteristic storiform pattern, with two major cell types -myofibroblasts and histiocyte-like cells, was seen. The question of pluripotential cell origin is discussed. The striking similarity of appearance to human malignant fibrous histiocytoma is noted; however, in view of the somewhat controversial origin of the human tumor, the simple designation, "nickel-induced malignant tumor of pluripotential origin", is preferred. The use of this model with its demonstrated ability to reproduce characteristic appearances seems to be valuable for the study of soft tissue sarcomas.

The problem of latency in the development of tumors following exposure to nickel compounds.

Six previously published animal studies of tumor production have been reviewed, in order to relate time interval between exposure to nickel and development of tumor formation. Biopsies at intervals before final tumor formation, in some of these experiments, were reviewed to define interim changes between exposure and tumor diagnosis. Correlation between rat and human life span was used to suggest a latency of human tumor expectancy following exposure to nickel.

Renal osteodystrophy in CAPD.

At start of dialysis most patients have histological bone abnormalities. These can be divided into two groups--high turnover and low turnover bone disease. Low turnover aplastic disease was previously attributed to aluminum accumulation but is now known to occur even in patients with less than 5% surface stainable aluminium. It is characterised by a mineralisation defect, thin osteoid seams, decreased numbers of osteoclasts and osteoblasts and absent aluminium staining. We have avoided aluminium containing phosphate binders (ACPBs) completely, with a combination of oral calcium carbonate and "low calcium" (1.25 mMol/l) dialysis fluid. Phosphate control has been good (mean less than 1.6 mMol/l) and over the first twelve months serum PTH levels have fallen significantly. Transient asymptomatic episodes of hypercalcaemia have occurred but no patient required ACPBs. Bone biopsies at the start of CAPD in 34 patients showed over 50% to have osteitis fibrosa (OF) but in five cases (15.6%) the aplastic lesion was found without aluminium staining. In seven follow-up biopsies OF improved in 3 cases, osteomalacia improved in 1, became aplastic in 1, while aplastic bone worsened in 1 and changed to mild OF in 1. We conclude that the predominant bone lesion in our patients at start of CAPD is OF, but 15% already have aplastic bone. "Low calcium" dialysis fluid enables ACPBs to be avoided in the majority of CAPD patients.