William B. Greene

Effects of prolonged exposure to dietary DDT and PCB on rat liver morphology

Livers from mature female rats exposed for up to 36 weeks to dietary levels of Aroclor® 1242 (75 or 150 ppm) and/or commercial grade DDT (75 or 150 ppm) were compared to those from animals receiving basal diets. In earlier studies, reproductive effects of the test substances were assessed. Moreover, the markedly abnormal gross appearance of the livers led to examination of the hepatic effects of PCB and DDT in more detail, at both the light microscope (LM) and electron microscope (EM) levels.Light microscopy revealed focal liver cell necrosis in rats fed PCB, DDT, and PCB-DDT combinations. Higher levels of PCB (150 ppm) increased the severity of necrosis. Feeding both DDT and PCB produced similar effects at 75 ppm, and caused atypical centrolobular regeneration, occasionally forming nodules resembling small tumors. The experimentally induced injury was associated with the marked accumulation of iron-containing pigment in hepatocytes and Kupffer cells.Electron micrographs demonstrated the presence of whorl structures (myelin figures) within liver cell cytoplasm, and for the first time clearly illustrated the endocytotic expulsion of these membranous whorls from hepatocytes into the bile canaliculi and sinusoids. Other ultrastructural changes were similar to those previously reported in rat livers injured by several hepatotoxic substances. Mitochondria enclosed by or projecting into large non-lipid vacuoles were present in several experimental groups.The electron micrographs provide the most convincing evidence to date to support the hypothesis that myelin figures may be the vehicle whereby the cell rids itself of specific hepatotoxic substances.

Ingested mineral fibers. Do they penetrate tissue or cause cancer

Three different laboratories independently investigated the ability of ingested mineral fibers to penetrate tissues in rats. All concluded that there was no evidence of tissue penetration by ingested mineral fibers. These experimental observations are supported by the findings in coal and hard-rock miners who swallow, during their lifetime, nearly 100 times the amount of dust that is stored in their lungs. The intestinal wall or mesenteric lymph nodes of these people show no evidence of storage of the ingested dust particles. Animals fed asbestos over much of their lifetime and allowed to live to the age of cancer production, failed to provide evidence of a cancerogenic effect.

Elemental analysis and clinical implications of calcification deposits associated with silicone breast implants.

Calcification of the fibrous capsule surrounding silicone breast implants is a well-recognized occurrence that increases with time following implantation. These mineralized deposits potentially confound mammographic breast cancer surveillance already made difficult by the obscuring effects of silicone breast implants. The authors performed elemental analysis of silicone breast implant-associated calcifications to define better their chemical composition as related to mammographic and clinical significance. Electron probe microanalysis and infrared spectroscopy revealed all of the calcification deposits to be calcium complexed with tribasic phosphate. No evidence of calcium oxalate, calcium carbonate, silicone, or talc was observed. Caution must be employed in interpreting mammograms in women with silicone breast implants as well as those who have had their silicone breast implants removed. High-density mammographic calcifications indicative of calcium phosphate associated with a silicone breast implant may represent an accepted consequence of implantation or nearby carcinoma. We recommend baseline mammography on women who have had their silicone breast implants removed to prevent unnecessary fine-needle aspiration or tissue biopsy of retained breast capsule calcifications during subsequent routine surveillance for carcinoma.

Ultrastructural Pathology of Nerve Fibers in Calcium-Induced Myelopathy

Calcium has been proposed as a mediator of nerve fiber degeneration following traumatic injury of the spinal cord. It induces a spongy, necrotizing myelopathy similar in its evolution to that observed in experimental spinal cord trauma. The current study was undertaken to determine the ultrastructural changes in the central nervous system (CNS) nerve fibers associated with calcium-induced myelopathy. A 10% calcium chloride (CaCl2) solution (pH 7.4) was slowly dripped on the dorsal surface of the surgically exposed lower thoracolumbar spinal cord of adult male Sprague-Dawley rats. The posterior and lateral columns of the spinal cords were fixed and processed for electron microscopy. Controls consisted of tissue from normal and sham-operated animals, as well as those receiving equal volumes and osmolalities of sodium chloride (NaCl), magnesium chloride (MgCl2), and potassium chloride (KC1) at the same pH. In the CaCl2 treated animals, spongiosis of increasing severity developed in white matter, as the result of periaxonal, adaxonal and intramyelinic swelling. Vesicular demyelination was consistently observed, beginning within one hour (h) and progressing with increasing severity up to 24–72 h. Axonal changes included pleomorphic spheroids, granular degeneration and intra-axonal calcification. The ultrastructural changes in the nerve fibers provoked by calcium were indistinguishable from those previously reported in experimental spinal cord trauma. These observations strengthen the hypothesis that calcium initiates the nerve fiber degeneration following spinal cord injury.

Silicone Breast Implants: Pathology

Questions as to the bioreactivity of silicone breast implants (SBIs) have recently been intensely scrutinized, most notably by the media and legal system. Pathologists must be aware of the controversy and treat each SBI and associated tissue as a potential lawsuit. Grossly, silicone is a clear, viscous substance that may be observed either within or extruding from a silastic bag. By light microscopy, silicone is a nonstainable, nonpolarizable, refractile substance. Thicker sections, especially when viewed by non-Köhler illumination, phase-contrast, and darkfield microscopy will enhance visualization. Ultrastructurally, silicone is an electron-dense, amorphous substance often located within phagocytic vacuoles or extracellularly within the stroma. Correlating electron probe microanalysis allows for reliable identification. In most cases, a fibrous capsule surrounds the SBI, with the interface lining varying from a virtually acellular to a synovial-like lining composed of phagocytic and secretory cells. Silicone can often be identified within the fibrous capsule and also in distant tissues biopsied for suspected autoimmune disorders, such as synovium, skin, and lymph nodes, often without ultrastructural evidence of cytologic effects. This study has demonstrated that silicone accumulates at distant tissue sites due to preexisting inflammation acting as a stimulus. Thus, silicone is not a primary inducer of inflammatory disease processes. These findings are supported by various large epidemiologic studies.

Caterpillar bodies of porphyria cutanea tarda ultrastructurally represent a unique arrangement of colloid and basement membrane bodies

Caterpillar bodies are eosinophilic, periodic acid-Schiff (PAS)-positive globules arranged in a linear fashion in the epidermis overlying subepidermal blisters of porphyria cutanea tarda (Am J Dermatopathol 1993;15:199-202). We retrospectively studied by transmission electron microscopy nine cases of porphyria cutanea tarda (PCT) that demonstrated caterpillar bodies. We identified three components of the eosinophilic bodies: degenerating keratinocytes, colloid bodies, and basement membrane bodies. The colloid bodies consisted of whorled masses of filaments containing degenerating melanosomes, vacuoles, mitochondria, and desmosomes. Basement membrane bodies were composed of convoluted basement membrane material and associated collagen. Both colloid and basement membrane bodies were often associated with degenerating keratinocytes, were located both intra-and extracellularly, and were occasionally fused to one another. We believe that caterpillar bodies are a combination of degenerating keratinocytes, colloid bodies, and basement membrane bodies formed by repeated blistering and reepithelialization with transepidermal migration. Furthermore, we believe that caterpillar bodies are a diagnostic clue for the diagnosis of PCT.

Silicone deposition in reconstruction scars of women with silicone breast implants

BACKGROUND The possible association of silicone breast implants and disease is a subject of continuous debate and concern. OBJECTIVE Our purpose was to examine microscopically and ultrastructurally the periprosthetic fibrous capsules and reconstruction scars of women with silicone breast implants. METHODS Representative samples from the periprosthetic capsules and reconstruction scars from six women with silicone breast implants were examined by a variety of light microscopy techniques, transmission electron microscopy, and electron probe microanalysis. RESULTS Silicone globules of various sizes were identified in every periprosthetic capsule and reconstruction scar. CONCLUSION Extrusion and seeding of the incision tract during surgery most likely accounts for the presence of silicone in the reconstruction scar specimens. This observation suggests that the identification of silicone in the reconstruction scars of women with silicone breast implants does not necessarily implicate rupture of the silicone breast implant with systemic dissemination of silicone gel.

Synovial metaplasia of a periprosthetic capsule surrounding a polyurethane foam breast prosthesis.

A 43-year-old woman underwent two-stage bilateral reconstruction mammoplasty using silicone-filled, polyurethane foam-covered breast implants. The permanent implants were later removed. The periprosthetic capsule demonstrated synovial metaplasia by light and electron microscopy. Synovial metaplasia is believed to be induced by the physical and chemical characteristics of the prostheses and may have important clinical implications in reducing capsular contracture and increasing host acceptance of implantable biomaterials.

Structural changes in a porcine xenograft after implantation for 105 months

Morphologic studies including light microscopy and transmission and scanning electron microscopy were performed on a formaldehyde-fixed porcine xenograft prosthesis that functioned well for 8 years and had been implanted for 105 months in the tricuspid valve position. It is the oldest implanted valve studied in this manner. Although all leaflets had some gross deterioration, only one had adherent thrombus. The degenerative changes in the connective tissue from all leaflets were similar to those recently reported in glutaraldehyde-fixed porcine valves implanted for shorter periods. The surfaces of the two nonthrombosed leaflets and small portions of the surface of the thrombosed leaflet were endothelized, and the endothelium appeared to have little or no effect on the degenerative changes in the subjacent connective tissue. Further, the endothelium did not extend over the examined portion of the synthetic anulus of the prosthesis, suggesting that its origin was possibly from cells circulating in the blood.

An improved cryo‐jet freezing method

A new cryo‐jet freezing apparatus is described that is easy to use and gives good results using a propane‐butene mixture (3: 1). Our use of the freezer in the study of mouse spinal cord explant cultures is discussed. At the tissue surface, the quality of tissue preservation from freezing, followed by freeze substitution, rivals that of conventional electron microscopic methods. Certain intracellular structures are better visualized using our methods. There is no evidence of the tissue being distorted by the cryogen jet when the freezer is operated correctly. A new freeze substitution device is also discussed.