Preparation of thyrotroph cells from adult male rat pituitary glands by centrifugal elutriation.

To study the metabolism of thyrotrophs and dynamics of TSH secretion in vitro, it is desirable to have a highly enriched population of thyrotrophs. For that purpose, centrifugal elutriation, a recently developed cell isolation method based on the size and density of cells, was used to prepare thyrotrophs from a cell suspension of adult male rat pituitary cells. Trypsin-dispersed cells (4-8 X 10(7] were loaded into the elutriation rotor (Beckman, JE-6) operating at 2800 rpm. Twelve cell fractions were collected at variable rotor speed (2000-2800 rpm) and increasing medium flow rate (10-103 ml/min). Cell recovery was 77-98%. The viability of the cells after elutriation was 90-95% based on trypan blue exclusion. Each fraction was analyzed for TSH, GH, and PRL content and for TRH-stimulated TSH release by RIA. Thyrotrophs were found predominantly in fractions 8-11 (flow rate 38-75 ml/min) based on TSH RIA. The mean TSH concentration in these fractions was 56 +/- 13.6 (+/- SD) microU/10(3) cells compared with 7.6 +/- 3.8 microU/10(3) cells in the initial cell suspension, representing a 7- to 8-fold enrichment of the thyrotrophs. Incubation with 20 nM TRH for 3 h increased the TSH release of cells eluted in fractions 8-11 by 3- to 5-fold; there was no significant increase in TSH release in fractions 3-6. Centrifugal elutriation may be used to prepare a uniform highly enriched thyrotroph fraction with excellent recovery from a suspension of rat pituitary cells. This technique should be valuable for study of the metabolism of thyrotrophs.

Postinfectious glomerulonephritis in a renal allograft associated with a mycotic aneurysm of a coronary artery.

Postinfectious glomerulonephritis is well known to occur after various infections but is rare in renal allografts, perhaps as a result of immunosuppressive therapy. This report describes a 47-year-old man who, seven years after receiving a cadaveric renal transplant, had biopsy-proved crescentic glomerulonephritis presenting with gross hematuria and rapidly progressive renal failure. The patient underwent cardiac surgery to define an abnormal structure shown on echocardiography, and a mycotic aneurysm of the left circumflex artery was discovered. Such aneurysms are uncommon and rarely diagnosed during life. This case appears to be the first report of glomerulonephritis associated with a mycotic aneurysm of a coronary artery and one of the few reports of postinfectious glomerulonephritis in a renal allograft.

Acute interstitial nephritis due to methicillin.

Fourteen patients are described with a syndrome of methicillin-induced interstitial nephritis. In all patients severe renal dysfunction developed with an average peak serum creatinine of 8 mg/100 ml. An increased total peripheral eosinophil count was found in all patients. All patients had sterile pyuria and each of nine patients studied by Wright's stain of urine sediment had marked eosinophiluria. These findings are suggestive of methicillin-induced interstitial nephritis, although proteinura was a variable finding in our patients. Eight of 14 patients in our study received prednisone therapy for their interstitial nephritis, and the time lapse between maximal and final base line serum creatinine levels was statistically less in the prednisone-treated compared to the nontreated groups. Clinical manifestations of this syndrome are discussed, and the light and electron microscopic and immunofluorescent findings on renal biospy are described.

Primary leiomyosarcoma of the aorta.

A case of primary leiomyosarcoma of the abdominal aorta is reported. The tumor replaced the full thickness of the aortic wall with formation of a large saccular aortic aneurysm filled with an admixture of tumor and mural thrombus. Electron microscopy of the surgical biopsy of the tumor revealed neoplastic smooth muscle cells with myofilaments and a discontinuous basal lamina. To our knowledge, leiomyosarcoma of the aorta has never been reported.

Legionnaires' disease. Postmortem pathologic findings of 20 cases.

From March 1977 to December 1978, postmortem examination was performed at Wadsworth Veterans Administration Medical Center for 20 patients who had had nosocomially acquired Legionnaires' disease. Seventeen patients died during the acute illness due to Legionnaires' disease, and three patients died after clinical resolution of the acute process. The only consistent postmortem findings were limited to the lungs. Confluent bronchopneumonia, and less frequently lobar pneumonia, was present in most cases. Although a spectrum of microscopic pulmonary findings was observed, the characteristic histologic features of acute Legionnaires' disease were an extensive intra-alveolar exudation of macrophages and neutrophils in varying proportions, erythrocytes, and fibrin. Lysis of the inflammatory cells was frequently found. Areas of coagulative necrosis of the lung parenchyma and edematous thickening of the alveolar septa were typically seen. Microscopy of lung tissue from the three patients who died after clinical resolution of the acute process revealed organized pneumonia, with patchy organization of the intra-alveolar exudate and focal obliteration of the alveolar septal framework. Associated postmortem findings were fibrinous endocarditis in one case and hemorrhagic infarction of the adrenal glands in two cases. Electron-microscopic examination of the lungs revealed as many as 23 separate bacillary profiles within a single macrophage. Septate binary fission or spore-like structures were not observed.

Patency of biologic and prosthetic inferior vena cava grafts with distal limb fistula.

A 5-cm segment of the infrarenal inferior vena cava (IVC) in 27 dogs was excised and replaced with a prosthesis of bovine heterograft, venous allograft, or polytetrafluoroethylene (ePTFE). On completion of the IVC graft, a side-to-side femoral arteriovenous fistula was constructed in the thigh. At six weeks an IVC venogram was obtained to assess patency and the femoral arteriovenous fistula closed. After three months, the dogs were reexplored, the grafts were removed, and patency was determined. Patency of the venous allograft and bovine heterografts were maintained with the arteriovenous fistula in only two dogs and further deteriorated after closure of the fistula so that only one dog in each group had a patent IVC graft at the end of three months. Eight of the 13 e-PTFE prostheses were patent at six weeks and remained patent for three months after fistula closure. Light and electron microscopy of the patent e-PTFE specimens excised at three months showed regenerative endothelium. Expanded e-PTFE was the most suitable prostheses for replacement of the IVC, resulting in 61% patency three months after closure of a distal arteriovenous femoral fistula.

Intestinal bypass complications involving the excluded small bowel segment.

We have examined complications involving the defunctionalized bowel in 119 intestinal bypass patients. In this group, we found a 66% of incidence of bypass enteropathy. Pneumatosis cystoides intestinalis was present in three patients, severe blood loss in three, localized ulcerations in two, intermittent or chronic intussusception of the proximal jejunal stump in 10, and extensive stenosis relating to tight fibrous adhesions in one patient. The stenosis may become manifest as an obstructive process only after reconstitution of normal bowel continuity. Bacterial overgrowth in the bypassed small bowel was the primary cause for most of the lesions. A consistent diagnostic finding, suggesting disease in the excluded bowel, was ileal distention and the presence of gas-fluid levels on upright abdominal x-rays. Definitive diagnoses of ulceration, intussusception, and/or obstruction were sometimes possible only during laparotomy. Because the bypassed bowel cannot be examined with conventional techniques, these various abnormalities must be suspected when ill-defined abdominal complaints are observed in bypass patients. Metronidazole, to suppress anaerobic organisms, or suitable broad spectrum antibiotics can relieve the various lesions of the inflammatory process, whereas appropriate surgical procedures may be required for some of the chronic or recurrent complications.

The vascular dementia of Fabry's disease.

Fabry's disease, a rare X-linked disorder of glycosphingolipid metabolism, can present as an insidious dementia in middle or later life. This genetic disorder produces a deficiency of alpha-galactosidase A which results in the deposition of glycosphingolipids in blood vessel walls in the brain as well as in the kidney, heart, peripheral nerves, and other organs. Among the cerebrovascular manifestations of this disorder is a vascular dementia from involvement of multiple small penetrating blood vessels. Fabry's disease is a consideration in the workup of an otherwise unexplained vascular dementia, particularly in males less than 65 years of age.

Papillary serous carcinoma of the peritoneum in a man: a case report.

BACKGROUND: Papillary serous carcinoma of the peritoneum (PSCP) is a rare primary peritoneal tumor, described exclusively in women. It is believed to arise from the secondary müllerian system, which is comprised of the pelvic and lower abdominal mesothelial lining and subjacent (subcoelomic) mesenchyme in women. Both mesotheliomas and PSCP arise from the coelomic epithelium, but are clinicopathologically and biologically distinct entities. METHODS: The authors report clinicopathologic findings in a man, age 74 years, who died 3 months after the diagnosis of an extensive malignant abdominal disease. RESULTS: The routine histologic and immunocytochemical studies of tumor tissue, obtained during the patient's lifetime and at autopsy, validated the unique occurrence of PSCP in a man. CONCLUSIONS: This case illustrates that PSCP can occur in a man and that this diagnosis may be considered in the differential diagnosis of papillary serous tumors of the peritoneum in male patients. Although rare, PSCP is a diagnostically distinct entity the treatment of which is similar to ovarian serous tumors rather than mesotheliomas.

Tumor necrosis factor-alpha activates nuclear factor kappa B and induces manganous superoxide dismutase and phosphodiesterase mRNA in human papillary thyroid carcinoma cells.

Human papillary thyroid carcinoma (PTC) has a relatively benign prognosis despite a high frequency of lymphatic metastasis. This suggests that local anticancer factors, generated in lymph nodes, control PTC progression. The cytokine, tumor necrosis factor-alpha (TNF-alpha), may be one such factor. We have previously shown that a human PTC cell line (NP-PTC) has high affinity TNF-alpha receptors. We now report on the action of TNF-alpha in these cells. TNF-alpha decreased [3H]thymidine incorporation as well as cellular DNA content and cell number in a dose-dependent manner. The abundance of phosphodiesterase and manganous superoxide dismutase mRNA species was increased in a time- and dose-dependent manner in the NP-PTC cells after TNF-alpha treatment. TNF-alpha activated NF-kappa B, a nuclear factor thought to mediate multiple actions of TNF-alpha, in these cells with a maximum effect observed after 30 min of treatment. Thus, TNF-alpha has an antiproliferative action on NP-PTC cells, despite its ability to induce the accumulation of mRNA that encodes an enzyme (manganous superoxide dismutase), thought to be cytoprotective. The net antiproliferative effect must therefore be explained by a balance of protective and tumoricidal or static effects that ultimately result in control of tumor spread. These antiproliferative effects may be in part mediated by NF-kappa B and PDE.

Expression and stereochemical and isotope effect studies of active 4-oxalocrotonate decarboxylase.

4-Oxalocrotonate decarboxylase (4-OD) and vinylpyruvate hydratase (VPH) from Pseudomonas putida mt-2 form a complex that converts 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate in the catechol meta fission pathway. To facilitate mechanistic and structural studies of the complex, the two enzymes have been coexpressed and the complex has been purified to homogeneity. In addition, Glu-106, a potential catalytic residue in VPH, has been changed to glutamine, and the resulting E106QVPH mutant has been coexpressed with 4-OD and purified to homogeneity. The 4-OD/E106QVPH complex retains full decarboxylase activity, with comparable kinetic parameters to those observed for 4-OD in the wild-type complex, but is devoid of any detectable hydratase activity. Decarboxylation of (5S)-2-oxo-3-[5-D]hexenedioate by either the 4-OD/VPH complex or the mutant complex generates 2-hydroxy-2,4E-[5-D]pentadienoate in D(2)O. Ketonization of 2-hydroxy-2,4-pentadienoate by the wild-type complex is highly stereoselective and results in the formation of 2-oxo-(3S)-[3-D]-4-pentenoate, while the mutant complex generates a racemic mixture. These results indicate that 2-hydroxy-2, 4-pentadienoate is the product of 4-OD and that 2-oxo-4-pentenoate results from a VPH-catalyzed process. On this basis, the previously proposed hypothesis for the conversion of 2-oxo-3-hexenedioate to 2-oxo-4-hydroxypentanoate has been revised [Lian, H., and Whitman, C. P. (1994) J. Am. Chem. Soc. 116, 10403-10411]. Finally, the observed (13)C kinetic isotope effect on the decarboxylation of 2-oxo-3-hexenedioate by the 4-OD/VPH complex suggests that the decarboxylation step is nearly rate-limiting. Because the value is not sensitive to either magnesium or manganese, it is likely that the transition state for carbon-carbon bond cleavage is late and that the metal positions the substrate and polarizes the carbonyl group, analogous to its role in oxalacetate decarboxylase.

Renal damage with intestinal bypass.

Renal function and biopsies were studied in 18 patients, 7 to 108 months after intestinal bypass. Enteropathy was found in 12 and hyperoxaluria in 16. Every biopsy showed a type of focal interstitial nephritis, tubular atrophy, fibrosis, and glomerular hyalinization. Damage ranged from minimal to extensive and renal function from normal to end-stage failure. Tubular injury had resulted partly from oxalate deposits. However, in 10 patients no oxalate crystals were seen. In eight others, most of the damaged areas were remote from crystal deposits. Immunoglobulin M and C3 deposits, found in glomerular capillaries and the messangium in six of 11 specimens, and the presence of circulating immune complexes in five of 10 patients, in addition to the extraintestinal organ involvement, suggested immune complex mesangial injury as one factor in bypass nephropathy. With progressive impairment of renal function, a biopsy appears justified. If damage is significant, the bypass should be dismantled.

Microvasculature in brain biopsy specimens from patients with Alzheimer's disease: an immunohistochemical and ultrastructural study.

Brain biopsy specimens from five patients with Alzheimer's disease obtained in the course of a trial of intracerebroventricular bethanechol were studied by immunohistochemical (antibody to A4 peptide) and ultrastructural techniques, with particular emphasis on the microvessels. In some cases, numbers of A4-immunoreactive lesions (senile plaques) correlated well with numbers of plaques demonstrable by silver stains. Prominent A4-immunoreactive amyloid angiopathy was seen in one patient. The patient with severe cerebral amyloid angiopathy (CAA) showed extensive arteriolar deposition of amyloid filaments with apparent destruction of the media but remarkably intact endothelium. A cell of origin for amyloid filaments was not apparent, although close proximity to smooth muscle cell remnants in the arteriolar media suggested this as one possible cell of origin. Frequent vessels showed medial or adventitial collagen deposition, even when the amount of amyloid was minimal or negligible. Thus relatively severe CAA can exist in the absence of overt endothelial injury, although related studies on this tissue indicate definite abnormalities of the blood-brain barrier. Conversely, destruction of smooth muscle cells and collagen deposition in vessel walls may be the cellular correlates of arteriolar weakening that can lead to CAA-related brain hemorrhage.

Parenteral aluminum administration in the dog: I. Plasma kinetics, tissue levels, calcium metabolism, and parathyroid hormone.

Aluminum (Al) may cause both osteomalacia and encephalopathy in dialysis patients. Little is known about the biology of Al. This study examined the initial distribution kinetics of Al and its biological effects after injections of 1 mg/kg/day into dogs for 3 to 5 weeks. Following one intravenous dose, the plasma half-life (x +/- SE) was 276 +/- 51.8 min, with an apparent volume of distribution of 1.30 +/- 0.17 liters or 5.90 +/- 0.30% body wt; 10 to 21% of administered Al was excreted in the urine over 150 min, and the renal contribution to plasma clearance of Al correlated with GFR (r = 0.77, P less than 0.05). The total plasma clearance of Al (4.43 +/- 2.83 ml/min) exceeded the renal contribution to plasma clearance (1.94 +/- 0.36 ml/min) in each dog, and in only two instances did the renal contribution reach 50% of total plasma clearance. Serum calcium rose from 9.4 +/- 0.2 to 11.1 +/- 0.3 mg/dl and immunoreactive parathyroid hormone (iPTH) fell by 27 +/- 4% following one Al injection. With repeated Al injections, serum calcium increased from baseline levels of 10.2 +/- 0.07 mg/dl to 11.1 +/- 0.22 and 11.3 +/- 0.46 mg/dl after 1 and 2 weeks, respectively. Renal function declined in all dogs, and serum creatinine exceeded 3.5 mg/dl in four; over the 5 weeks of study, serum calcium correlated with serum creatinine (r = 0.91, P less than 0.001). Liver, kidney, and spleen showed the highest tissue content of Al, and there was substantial uptake by bone; the parathyroid content of Al was modest.(ABSTRACT TRUNCATED AT 250 WORDS)