Theodore F. Beals

Induction of pemphigus in neonatal mice by passive transfer of igg from patients with the disease.

We examined the role of circulating autoantibodies in the pathogenesis of pemphigus vulgaris by passively transferring IgG fractions from five patients with pemphigus vulgaris into neonatal Balb/c mice, in doses of 1.5 to 16 mg per gram of body weight per day. Cutaneous blisters and erosions with the histologic, ultrastructural, and immunofluorescence features of pemphigus occurred in 39 to 55 mice given intraperitoneal injections of IgG from patients with pemphigus and in none of 58 control mice given normal human IgG. IgG fractions with high titers of pemphigus antibodies were most effective in inducing disease, and this effect was dose dependent. Titers of circulating IgG in mouse serum closely correlated with the extent of disease induced (P less than 0.002). This study strongly supports the proposed role of pemphigus autoantibodies in the pathogenesis of pemphigus vulgaris in human beings and demonstrates that pemphigus can be passively transferred to laboratory animals.

Membranous basal cell adenoma of parotid gland, dermal cylindromas, and trichoepitheliomas: Comparative histochemistry and ultrastructure

A basal cell adenoma of parotid, eccrine dermal cylindromas and trichoepitheliomas occurring in the same patient were examined by light and electron microscopy and histochemistry. The eccrine and parotid adenomas were similar both structurally and histochemically except for the presence of Langerhans cells in the cutaneous adenoma and well differentiated mucinous cells in the parotid tumor. The three different hamartomas found in this individual may represent the effect of a single pleiotropic gene acting on ontogenetically related stem cells.

Overexpression of p53 Predicts Organ Preservation Using Induction Chemotherapy and Radiation in Patients with Advanced Laryngeal Cancer

A critical research frontier in head and neck oncology involves defining the use of induction chemotherapy regimens to allow organ preservation and to avoid functionally debilitating surgical resections. Completed clinical trials in laryngeal cancer indicate that such an approach is feasible, but progress thus far has been limited by our inability to predict which patients are likely to respond to chemotherapy and preserve their larynx. Mutation of the p53 tumor-suppressor gene is the most common genetic alteration identified thus far in human cancers, and it may be important in regulation of cell proliferation and chemosensitivity. To determine whether p53 overexpression predicts chemotherapy response, organ preservation, and survival in patients with advanced laryngeal cancer, we analyzed immunohistologic expression of p53 in tissue sections from 178 patients with advanced laryngeal cancer who were entered in the Department of Veterans Affairs Laryngeal Cancer Cooperative Study, a multiinstitutional clinical trial comparing induction chemotherapy (cis-platinum and 5-fluorouracil) plus radiation therapy (94 patients) to surgery plus postoperative radiation therapy (84 patients). Larynx preservation was significantly higher in the group of patients whose tumors overexpressed p53 (74% vs. 52.5%; p = 0.03). The presence of p53 overexpression did not predict survival in either the surgery or the chemotherapy groups (p = 0.82 and p = 0.53).

Actin polymerization in cellular oxidant injury

Microfilaments undergo an ATP-dependent disruption into shortened bundles following cellular exposure to oxidants. This phenomenon does not require a net change in the amount of polymerized actin. However, increased amounts of polymerized actin have been detected in oxidant-injured cells and it was the purpose of this study to determine the conditions under which the actin polymerization may occur. Utilizing the formation of oxidized glutathione (GSSG) as an indicator of cellular sulfhydryl oxidation, conditions were chosen to accentuate sulfhydryl oxidation within the target P388D1 cell line following exposure to the oxidants, H2O2 and diamide. Using the DNase I and flow cytometric assays of actin polymerization, significant polymerization of actin was detected only under conditions in which sulfhydryl oxidation occurred after exposure to the two oxidizing agents. Greater sulfhydryl oxidation early in the course of injury was associated with a greater rate and extent of actin polymerization in the injured cells. Experiments with cells depleted of glutathione (GSH) demonstrated that neither loss of GSH nor absolute levels of GSSG formed during oxidant exposure were responsible for the polymerization of actin. The data presented are consistent with the hypothesis that oxidizing conditions which induce significant sulfhydryl oxidation in target cells are correlated with assembly of polymerized actin and that this represents a process which is distinct and separate from the ATP-dependent gross disruption of microfilaments.

Malignant clear cell acrospiroma.

The histologic features of a malignant clear cell acrospiroma were those of infiltrative local growth, frequent mitoses and angiolymphatic invasion. The histochemical and ultrastructural findings were similar to those reported for benign clear cell acrospriomas. Amputation of the leg and regional node dissection were required for clinical control. Evaluation of this case and review of the literature suggests that the malignant clear cell acrospiroma often behaves in an aggressive manner and frequently metastasizes. As a consequence, therapeutic strategies should be appropriately planned.

Predictive markers for response to chemotherapy, organ preservation, and survival in patients with advanced laryngeal carcinoma.

OBJECTIVE: A systematic retrospective study of the largest randomized trial of induction chemotherapy and radiation for advanced laryngeal cancer was undertaken to determine whether specific tumor or biologic factors were predictive of chemotherapy response, organ preservation, or survival. METHODS: The variables analyzed included clinical and histologic factors, immunohistochemical expression of proliferating cell nuclear antigen and p53, and adjusted DNA index measurements. Variables were evaluated for correlation with outcomes of tumor response, organ preservation, and survival. RESULTS: Multivariate analysis revealed that the best predictor of complete response to induction chemotherapy was low T class. The full multivariate model for predicting larynx preservation in patients treated with induction chemotherapy plus radiation shows that T class, p53 overexpression, and elevated proliferating cell nuclear antigen index were independent predictors of successful organ preservation. CONCLUSIONS: These predictive markers should be included in future clinical trials of advanced laryngeal cancer to determine their usefulness prospectively.

Congenital gingival granular cell tumor with smooth muscle cytodifferentiation.

Congenital epulis of the newborn is a rare benign congenital gingival granular cell tumor (GGCT) of unknown histogenesis which occurs most commonly on the gingiva of the anterior maxillary alveolar ridge in girls. The granular cells in this entity are histologically indistinguishable from those in extragingival granular cell tumors, known historically as granular cell myoblastoma (GCM), which occur at any age and appear to be of Schwann cell origin. Ultrastructural, histochemical, and immunohistochemical features of three GGCT were examined and compared to three GCM and a granular cell ameloblastoma. This is the first instance in which the ultrastructure of granular cells in a congenital epulis showed evidence of smooth muscle differentiation. Carcinoembryonic antigen-like immunoreactivity was localized in granular cells from all granular cell tumors studied, but S-100 protein was present only in GCM. The smooth muscle ultrastructural features and the lack of S-100 protein in GGCT strongly suggest a different histogenesis from that of GCM. The GGCT is likely derived from a primitive gingival perivascular mesenchymal cell with the potential for smooth muscle cytodifferentiation.

A cellular model of endothelial cell ischemia

Endothelial cell dysfunction in ischemia may cause increased capillary permeability. We examined the effect of failing ATP synthesis, a major consequence of ischemia, on microfilaments--important structural determinants of the endothelial cell. Glycolytic and mitochondrial ATP synthesis in bovine pulmonary artery endothelial cells was inhibited by glucose depletion and 650 picomole (pmole) oligomycin/micrograms DNA, respectively. ATP levels were monitored with the luciferase-luciferin assay over a 2-hr time course followed by recovery for 1 hr after removal of the oligomycin and addition of 5.5 mM glucose. ATP levels fell to 83.6 +/- 63.8 pmole/micrograms DNA (n = 11) by 30 min, 26.9 +/- 13.8 pmole/micrograms DNA (n = 11) by 60 min, and 17.2 +/- 3.8 pmole/micrograms DNA (n = 6) by 120 min, whereas control uninjured cells had 541.3 +/- 196.8 pmole/micrograms DNA (n = 6) at 120 min. Fluorescence microscopy of microfilaments stained with rhodamine-phalloidin revealed progressive disassembly and shortening of the microfilaments in greater than 90% of cells over 60 min which correlated with the fall in ATP. Ultrastructural examination revealed that side to side aggregation of microfilaments had occurred over the 120-min time course. Two hours of glucose depletion (305.5 +/- 130.8 pmole ATP/micrograms DNA, n = 6) or oligomycin alone (480.0 +/- 90.1 pmole ATP/micrograms DNA, n = 6) failed to produce the dramatic fall in ATP or the microfilament changes. During cell recovery, there was a rapid reassembly of microfilaments, detected by fluorescence microscopy, which was nearly complete in 85-90% of cells by 45-60 min. ATP levels increased significantly (P = 0.002) to 96.1 +/- 36.8 pmole/micrograms DNA (n = 6) by 30 min. This model should provide insight into the pathogenesis and treatment of the capillary leak seen with ischemia.

Neuroblastoma-like neurilemoma

We report three cases of neurilemoma occurring in the superficial soft tissue of the palm, posterior neck, and flank that closely resembled neuroblastoma by virtue of a predominance of small, round hyperchromatic Schwann cells with scant cytoplasm which, in one case formed perivascular rosettes and, in the others, giant rosettes with central collagen cores. Although the diagnosis of neuroblastic tumor was seriously entertained in all cases, the diagnosis of a schwannoma was substantiated by the finding of focal areas of conventional schwannoma and by the typical immunophenotypic profile. All tumors strongly and diffusely expressed S-100 protein but lacked neurofilament protein, protein gene product (PGP), and synaptophysin. Ultrastructurally the cells contained slender cytoplasmic processes invested with basal lamina, but no dense core granules. All were excised conservatively, and none has recurred during the 2, 12, and 27 month follow-up periods.

A cellular model of oxidant-mediated neuronal injury

Oxidants derived from the partial reduction of oxygen are thought to play a significant role in neuronal injury. We present here a cellular model of neuronal injury mediated by hydrogen peroxide (H2O2) using the PC 12 rat pheochromocytoma cell line. The organization of microtubules and microfilaments within neurites of PC 12 cells differentiated by exposure to nerve growth factor was examined after H2O2 injury using fluorescence microscopy. Concentrations of H2O2 as low as 100 microM produced an initial periodic pattern of microtubule depolymerization over 3-4 h which later progressed to complete depolymerization. Neuritic microspikes containing actin filaments were relatively more resistant to injury by H2O2 than microtubules. Blebbing of PC 12 cell bodies and neurites also was seen after H2O2 injury and the blebs appeared to contain microtubules. The destructive changes affecting neuritic structure preceded but were not essential for PC 12 cell lysis. Exposure of the cells to the Ca2+ ionophore, ionomycin (25 microM) also produced the same pattern of microtubule depolymerization in PC 12 neurites as was seen after H2O2 injury suggesting that H2O2 may mediate its destructive effect on the neurites via elevation of intracellular Ca2+.