Steam heat with an EDTA buffer and protease digestion optimizes immunohistochemical expression of basal cell-specific antikeratin 34betaE12 to discriminate cancer in prostatic epithelium.

In select cases of prostatic carcinoma, antikeratin 34betaE12 immunohistochemical analysis is diagnostically useful for specific labeling of basal cells. This antibody, however, is prone to variability in staining, and the optimal conditions were not, to our knowledge, previously defined. We combined steam heat with EDTA buffer (steam-EDTA) and protease digestion (steam-EDTA + protease) to optimize epitope retrieval of antikeratin 34betaE12 in 42 cases of prostatic cancer. Results were judged by the percentage of cells staining and by staining intensity. In benign epithelium, steam-EDTA + protease significantly increased the percentage of immunoreactive cells (from 74 to 93%) and the intensity of staining (from 2.1 to 3.0 on a scale of 0-3+) by comparison with protease alone (all P<.001). In high-grade prostatic intraepithelial neoplasia, the percentage of cells staining increased from 55 to 73% and intensity increased from 1.7 to 2.8 (both P<.001). Steam-EDTA + protease also minimized variability in results between cases, with essentially no background stromal staining. Cancer was negative in all of our cases by both methods. We conclude that steam-EDTA + protease significantly enhances basal cell immunoreactivity compared with protease treatment alone in noncancerous prostatic epithelium. This helps to prevent misinterpretation of histologic mimics of cancer, such as atrophic acini and high-grade prostatic intraepithelial neoplasia, that result from false-negative staining.

Neuroendocrine differentiation in prostatic intraepithelial neoplasia and adenocarcinoma.

Neuroendocrine cells are thought to have a regulatory role in prostatic epithelial growth and may be prognostically useful in prostatic adenocarcinoma. To determine the extent of neuroendocrine differentiation in high-grade prostatic intraepithelial neoplasia (PIN), a putative precursor of cancer, we studied the immunohistochemical expression of 10 markers in 26 radical prostatectomy specimens with PIN and adenocarcinoma. Expression was measured as mean percent of positive cases and positive high-power (x40) fields. The highest percentage of cases showed immunoreactivity for serotonin (73%, PIN; 54%, carcinoma), neuron-specific enolase (NSE) (67%, PIN; 46%, carcinoma), chromogranin (62%, PIN; 65%, carcinoma), and human chorionic gonadotropin (hCG) (30%, PIN; 22%, carcinoma); the remaining markers showed immunoreactivity in fewer than 5% of cases (somatostatin, calcitonin, corticotropin) or in no cases (thyrotropin, prolactin, and glucagon). At least one of the markers was present in 88% of cases of PIN and 92% of carcinoma. Non-neoplastic epithelial cells expressed serotonin, NSE, chromogranin, and hCG in every case, and the expression was significantly greater than in PIN and cancer. Stepwise regression analysis revealed the following positive correlations: chromogranin expression in PIN and patient age, NSE expression in cancer and number of lymph node metastases, and hCG expression in cancer and percentage of Gleason pattern 5; serotonin expression in PIN and cancer did not correlate with any of the clinical and pathologic factors. Neuroendocrine differentiation is downregulated in prostatic carcinogenesis, with intermediate levels of expression in PIN compared with normal cells and carcinoma.

Duodenal gangliocytic paraganglioma. Clinicopathologic and immunocytochemical study of 11 cases.

The clinicopathologic features of 11 cases (8 in men) of duodenal gangliocytic paraganglioma are presented. The patients averaged 56 years of age; none showed evidence of phakomatosis. Ten tumors occurred in the second portion of the duodenum, and one arose in the third portion. All tumors were polypoid, and half presented with gastrointestinal bleeding. The neoplasms were composed of paraganglioma and carcinoid-like elements, neurons, and Schwann as well as sustentacular cells. All tumors behaved in a benign fashion after local resection or snare polypectomy; long-term follow-up (1-25 years; mean, 8.3 years) showed no recurrence in any case. Immunocytochemical examination demonstrated the presence of somatostatin, serotonin, and human pancreatic polypeptide within endocrine cells and neurons.

Paraganglioma of the cauda equina region. Clinicopathologic study of 31 cases with special reference to immunocytology and ultrastructure.

Thirty-one paragangliomas of the cauda equina region were studied (18 men and 13 women, ages 30-71 years [mean, 51 years]). Symptoms (1 day to 15 years in duration; mean, 48 months) included low back pain (87%), sensory/motor deficits (35%), urinary/fecal incontinence (13%), and paraplegia (6%). All patients studied had some myelographic block. Cerebrospinal fluid protein level ranged from 56 to 7000 mg/dl (mean, 1109 mg/dl). Most tumors were limited to the filum terminale, although one also involved the conus medullaris and two clearly arose from a caudal nerve root. All but one were entirely intradural. The tumor was totally excised in 26 cases; these patients remain disease-free. Of three patients whose tumors were excised subtotally, two received radiotherapy; the one non-radiated patient died of tumor-related complications. No autopsy was performed. One partially encapsulated tumor that had been subjected to biopsy and irradiation presented 1 year later with osseous invasion and retroperitoneal extension; 20 years after subtotal excision, this patient is alive but paraplegic. Morphologically, all tumors resembled paraganglioma at other sites. Cytologic atypia and mitotic activity generally were absent to mild. Fourteen (45%) cases showed ganglionic differentiation. All tumors tested were immunoreactive for neuron-specific enolase and neurofilament protein, and most showed somatostatin or serotonin reactivity. S-100 protein immunoreactivity was noted in sustentacular cells and, to a lesser extent, within chief cells and neurons. The authors conclude that paragangliomas are largely benign and encapsulated and respond to simple resection. When surgically feasible, gross total removal should be the goal of surgery. When subtotal resection is necessary or when local invasion leaves a question as to completeness of tumor removal, irradiation seems mandatory although far from guaranteeing prevention of recurrence. Biopsy alone is undesirable.

A study of the techniques of cardiac massage with the abdomen open.

The present study compares the hemodynamic effectiveness of closed-chest cardiac massage (CCCM) with closed subdiaphragmatic massage (CSDM) and four open transdiaphragmatic cardiac massage techniques during cardiac arrest with an open abdomen. In 10 dogs CCCM resulted in the lowest cardiac index (CI), mean arterial pressure (MBP), and carotid blood flow (CBF) of all cardiac massage techniques tested. CSDM was not statistically superior to CCCM in the dog (p greater than 0.05) but did result in a 23% increase in CI and a 54% increase in CBF. Transdiaphragmatic retrocardiac massage through an incision in the diaphragm resulted in the highest CI, MBP, and CBF of all the four open transdiaphragmatic techniques and had significantly higher values than those for CCCM in the dog (p less than 0.05). In three cadaveric renal donors, all four open transdiaphragmatic techniques and CSDM were noted to be equal to or superior to CCCM. Three patients have been successfully resuscitated with diaphragmatic cardiac massage techniques for cardiac arrest while undergoing abdominal operations. These studies reveal that all subdiaphragmatic or transdiaphragmatic techniques for cardiac massage are hemodynamically equivalent to or superior to the standard CCCM without such complications as fractured ribs and should be considered the treatment of choice for cardiac arrest in the patient with an open abdomen.

Immunocytochemical localization of a granuliberin-like peptide in Rana pipiens brain.

The histamine-releasing peptide "granuliberin", originally isolated from the skin of Rana rugosa, was localized by immunofluorescence within nerve cell bodies and fibers in the brain of Rana pipiens. The granuliberin-positive neurons were characterized ultrastructurally by electron microscopic observation of ultrathin sections stained either with immunoperoxidase or with conventional stains. Granuliberin-positive nerve cell bodies were seen throughout the hypothalamus, from the suprachiasmatic area rostrally to the full length of the periinfundibular grey matter caudally. Similarly positive nerve fibers were localized in the hypothalamus radiating upwards to the optic vesicles in the midbrain, extending through the preoptic area into the subpallium in the forebrain, and throughout the white matter surrounding the floor and lateral walls of the fourth ventricle in the brainstem. The granuliberin-positive nerve cells showed the presence of variable numbers of small cytoplasmic neurosecretory granules, possessing an electron dense elongated core, and measuring 250-350 nm in their largest diameter. The functional significance of a granuliberin-like peptide in neurons of the frog brain is not known.

Renal microembolization syndrome. A cause for renal dysfunction after abdominal aortic reconstruction.

Moderate renal dysfunction due to renal microembolization developed in eight patients who underwent abdominal aortic reconstruction for aneurysmal or occlusive disease. In each patient, the aorta around the renal arteries was thrombus-lined or severely ulcerated and was therefore the source of embolization, and aortic clamping near the renal arteries was required and provided the mechanism for embolization. Renal failure was moderate and did not require dialysis. Renal dysfunction appeared to be largely reversible, although some degree of permanent damage did occur. Similar changes in renal function were noted in a dog model of renal microembolization. Prevention of this complication depends on awareness of aortic lesions that increase the risk of renal embolization.

Recovery of pancreas from mild puromycin-induced injury. A histologic and ultrastructure study in rats.

Pancreatic acinar cells undergo degeneration or necrosis following injection of puromycin intraperitoneally in rats. The purpose of this study was to characterize recovery following injection of four hourly doses of puromycin, 40mg/kg of body weight, examining the pancreas histologically and by electron microscopy. The number of dividing acinar cells increased following injury. By 12 to 24 hours following treatment, electron microscopy showed numerous autophagic vacuoles and intracisternal granules in the cells. By 48 hours, these were largely cleared from surviving cells although the intracisternal granules persisted in isolated acinar cells as long as 144 hours. At 24 hours, there was debris in the acinar lumens and interstitial space. We conclude that some acinar cells injured by puromycin may survive and be restored to normal structure; that surviving acinar cells can extrude autophagic vacuoles; and that necrotic acinar cells are replaced by regeneration following puromycin-induced injury in rats.