Identification of conserved lentiviral sequences as landmarks of genomic flexibility.

Considering that recombinations produce quasispecies in lentivirus spreading, we identified and localized highly conserved sequences that may play an important role in viral ontology. Comparison of entire genomes, including 237 human, simian and non-primate mammal lentiviruses and 103 negative control viruses, led to identify 28 Conserved Lentiviral Sequences (CLSs). They were located mainly in the structural genes forming hot spots particularly in the gag and pol genes and to a lesser extent in LTRs and regulatory genes. The CLS pattern was the same throughout the different HIV-1 subtypes, except for some HIV-1-O strains. Only CLS 3 and 4 were detected in both negative control HTLV-1 oncornaviruses and D-particle-forming simian viruses, which are not immunodeficiency inducers and display a genetic stability. CLSs divided the virus genomes into domains allowing us to distinguish sequence families leading to the notion of 'species self' besides that of 'lentiviral self'. Most of acutely localized CLSs in HIV-1s (82%) corresponded to wide recombination segments being currently reported.

Elution of tumor-directed antibody from kidneys of mammary tumor-bearing mice.

Immunoglobulin-containing eluates, prepared from kidneys of Paris RIII mammary tumor-bearing mice approximately 10 months old, were incubated with frozen sections of mouse mammary tumor and normal lactating mammary gland and examined by immunofluorescence. The eluates diffusely strained the tumor but did not stain the nonneoplastic lactating mammary gland. Mammary tumor and normal lactating mammary gland were both stained by an antiserum to mammary tumor virus (MuMTV). The euglobulin portion of the eluates, when fractionated on a Sephadex G-200 column, yielded an IgG fraction which was shown by immunodiffusion to react with Paris RIII mammary tumor extract and with anti-mouse globulin but not with nonneoplastic mouse tissue extracts. Since the lactating mammary glands of these animals contain MuMTV, the antibody-containing fraction eluted from the kidneys appeared to be tumor directed rather than virus directed.

Glomerular immune complex deposits associated with mouse mammary tumor.

Adult Paris R-III mice with localized mamary tumors were found to have subendothelial glomerular immune complex deposits by electron microscopy and immunofluorescence. In addition to IgG adn B-1-C, the glomeruli contained antigenic material that reacted with an antiserum to mouse mamary tumor virus. These findings support the hypothesis that many animals with congenitally or neonatally induced virus-related tumors are not immunologically tolerant to the oncogenic virus.

Electron microscopy and immunofluorescence of glomerular immune complex deposits in cancer patients.

Kidneys of 29 patients without clinical renal disease were studied by electron microscopy for the presence of glomerular basement membrane deposits: those from 11 of 20 patients with cancers of various sites, but only 1 of 9 patients without cancer contained electron-dense subendothelial deposits. The majority of these kidneys gave positive immunofluorescent reactions for immunoglobulin and complement. However, among a number of the cases studied, a lack of correlation between electron microscopic and immunofluorescence findings has yet to be investigated. Although the number of patients in this study is small because of the difficulty in obtaining tissue for electron microscopy, it is postulated that the deposition of immune complexes in the kidney occurs with a high frequency among cancer patients. The kidneys may thus be a valuable source for isolating tumor-associated antigens and corresponding antibodies.

Demonstration of an antibody to tubular epithelium in glomerulonephritis associated with obstructive uropathy.

It has been postulated that in some patients with obstructive and reflux uropathy proteinuria develops through an intermediate mechanism of immune complex glomerulonephritis involving antigenic material of renal tubular epithelium. A patient with a unilateral ureterocele and nephrotic syndrome underwent bilateral renal biopsies during surgical correction of the obstruction. The obstructed kidney showed mild pyelonephritis, but both kidneys showed a glomerulopathy with electron-dense deposits in the mesangial and paramesangial regions associated with positive immunofluorescence for immunoglobulin M (IgM) and the third component of complement (C3). An IgM antibody was eluted from the biopsy specimens and it reacted by indirect immunofluorescence with normal renal tubular epithelium and with the patient's renal tubular epithelium. The eluate also reacted with pre-eluted glomeruli of the patient, but not with normal glomeruli. All antibody activity could be removed from the eluate by pre-incubation with normal kidney. It is concluded that the unilateral renal obstruction produced tubular injury so that as yet unidentified antigens were recognized by the immune system. The resultant antibody response gave rise to circulating immune complexes which were then deposited in glomeruli with subsequent glomerular damage and nephrotic syndrome.

Pseudoinvasion with high-grade dysplasia in a colonic adenoma. Distinction from adenocarcinoma.

High-grade dysplasia was found to extend to an area of pseudoinvasion in the submucosa of a colonic adenoma mimicking invasive carcinoma. The presence of both benign and cytologically malignant epithelium and residual foci of lamina propria among the submucosal glands distinguishes this entity from adenocarcinoma arising in an adenomatous polyp.

Dysplasia and early carcinoma in inflammatory bowel disease and colorectal adenomas.

The control of colorectal cancer is currently dependent on early detection, and its prevention requires the recognition and treatment of its precursor lesions. The adenoma has been established as the precursor of colorectal carcinoma in the general population. Among patients with inflammatory bowel disease (IBD), dysplasia is associated with, and precedes, invasive carcinoma. In this section criteria are described for the histological detection of preinvasive and early invasive neoplasia in the large intestine of patients with and without IBD. The therapeutic implications of these diagnoses are stressed. A brief review of subcellular changes, including genetic alterations, in colorectal neoplasia is included.

Renal manifestations of extrarenal neoplasms.

In a study of 1270 consecutive autopsies there were 314 patients with malignant neoplasms arising in sites other than the kidney and central nervous system. In over 50 per cent of these there was significant renal damage related to cancer. Renal damage was produced by direct involvement of one or both kidneys by the neoplasm or by indirect effects. The latter included ischemic damage, metabolic injury, immunologic injury, and effects of therapy directed at the malignant tumor. In patients with cancer, tumor bulk and invasion of vital organs do not always explain the clinical deterioration and cause of death. Recognition of the indirect effects of tumors on the kidney and other organ systems is essential to the understanding of the generalized host response to malignant disease.

Mucinous cystadenoma of the ovary with stromal luteinization and hilar cell hyperplasia during pregnancy.

A 32-year-old woman was delivered of a healthy, full-term infant by cesarean section, at which time a large ovarian cyst was removed. The cyst proved to be a mucinous cystadenoma with prominent luteinization of the stroma subtending the epithelium and with numerous foci of hyperplastic Leydig cells in the cyst wall and ovarian hilum. These hormonally induced changes must be recognized in order to avoid mistaking them for invasive epithelial components.

Malignant giant cell tumor of bone: ultrastructural and immunohistologic evidence of histiocytic origin.

A malignant giant cell tumor of the carpal bones metastatic to the lung in a 23-year-old woman is described. Findings from ultrastructural and cell marker studies support the concept that this tumor is of histiocytic origin.

Invasive and metastasizing carcinoma in a small adenomatous polyp of the colon: report of a case with demonstration of a tumor associated antigen.

An unusual case of metastasis to two lymph nodes from an adenocarcinoma arising in an adenomatous polyp of the sigmoid colon is presented. Invasion of the stalk could not be detected, but penetration of the muscularis mucosae was observed. Immunoperoxidase staining was used to detect an antigen in the invasive and metastatic components of the tumor that reacted with an antiserum to a tumor associated antigen. A focus of intraepithelial carcinoma was also detected by this method. The relevance of these findings to the adenoma-carcinoma sequence is discussed.

Tumor directed antibody and carcinoembryonic antigen in the glomeruli of a patient with gastric carcinoma.

A patient dying of gastric carcinoma was one of a group of cancer patients examined for the presence of glomerular immune complex deposits not associated with the nephrotic syndrome. The deposits were distributed in the mesangial and subendothelial regions. This distribution is found in experimental animals with neoplasia and glomerulopathy as well as in over 30 per cent of humans with cancer. Immunofluorescence showed IgG and C3 in the patient's glomeruli. Carcinoembryonic antigen was identified in the patient's glomeruli by the immunoperoxidase staining method. An IgG antibody was eluted from the kidney and found to be reactive with the patient's tumor, as well as another patient's colonic carcinoma. This reactivity was blocked by preincubation of the tumor substrate with anticarcinoembryonic antigen. Thus, both a tumor associated antigen and a corresponding antibody were shown to be contained in the glomerular deposits. It is concluded that circulating immune complexes of high molecular weight containing carcinoembryonic antigen produced by the gastric carcinoma led to the formation of subendothelial deposits without significant renal damage. This is in contrast to the usual finding of membranous glomerulonephritis among cancer patient with the nephrotic syndrome and more closely resembles the animal models. Whereas tumor reactive antibodies can be found in the glomeruli of patients with cancer, a specific tumor associated antigen to which the antibody is reactive has only occasionally been demonstrated.

Immunohistologic detection of estrogen receptors in paraffin-embedded breast cancers: correlation with cytosol measurements.

Paraffin-embedded sections of 76 human breast tissue specimens were analyzed for estrogen receptors (ER) and endogenous bound estrogen (ER-E). Preincubation of sections with polyestradiol phosphate was followed by stabilization of the complex with glutaraldehyde. The bound hormone was then visualized by the peroxidase-antiperoxidase (PAP) technique with antiestradiol as the primary antiserum. Normal breast tissue and benign proliferations were consistently positive for ER and ER-E. All specimens were examined for free and bound receptors in cytoplasm and nuclei. Among the carcinomas examined, a high correlation was found between the presence of ER by the PAP method and by the biochemical analysis of cytosol preparations. The PAP method, requiring no special preparation of surgical specimens, overcomes many of the disadvantages of the cytosol method and adds the advantage of independent evaluation of nuclear and cytoplasmic estrogen binding sites.

Primary malignant fibrous histiocytoma of the pancreas.

A patient with a large tumor of the head and body of the pancreas had a Whipple procedure performed after the intraoperative diagnosis of "mesenchymal tumor" was made. The final histopathologic diagnosis was malignant fibrous histiocytoma arising in the pancreas. The patient died of postoperative complications, and no metastatic disease was found at autopsy. This is the first detailed description of a primary pancreatic malignant fibrous histiocytoma.

Immunoperoxidase: a sensitive immunohistochemical technique as a "special stain" in the diagnostic pathology laboratory.

The present communication is a brief review of the origins, theory, and applications of a relatively new immunohistochemical technique that can be performed on routinely fixed, paraffin embedded tissues in which visualization by bright field light microscopy is feasible and which thus can be readily adapted to routine diagnostic work. The chief concern of this presentation is the practical diagnostic application of the immunoperoxidase technique, a method whose reputation as a sensitive investigative tool is well established.

Glomerulonephritis and virus-like particles associated with prostatic cancer.

A patient with carcinoma of the prostate and metastases developed the nephrotic syndrome following hormonal therapy. A decrease in the dose of estrogens was associated with decreased proteinuria, but when therapy was increased the nephrotic syndrome became more severe. Renal biopsy performed when proteinuria was present showed subendothelial electron dense deposits, complement by immunofluorescence, and a morphologic pattern of membranoproliferative glomerulonephritis. There was evidence of resorption of the deposits by endothelial cells. Microspherical particles resembling those described in prostatic cancer were found in the glomeruli. On the basis of previous reports it is concluded that circulating tumor antigen-antibody complexes produced glomerulonephritis, the severity of which was related to the amount of soluble antigen released by tumor cell destruction. Apparent phagocytosis of immune complexes by glomerular endothelial cells was believed to account for the reversibility of the nephrotic syndrome. The role of the virus-like particles in the process is as yet unclear.

Hemorrhagic pancreatitis: a complication of transcatheter embolization treated successfully by total pancreatectomy.

Since pancreatitis can be produced experimentally in dogs by embolization of microspheres into the pancreatic arterial circulation, there has been speculation that intentional or inadvertent embolization of the pancreas in human subjects could also produce pancreatitis. Although such therapeutic embolization has increased, no pathologically documented case of this complication has been recorded. We have reported the first such case occurring in a patient with a large, highly vascular, nonfunctioning islet cell carcinoma of the tail of the pancreas preoperatively embolized with Gianturco coils and Gelfoam particles suspended in sodium tetradecylsulfate solution to facilitate distal pancreatectomy. The resultant hemorrhagic pancreatitis and duodenal necrosis required a total pancreatectomy. We conclude that, by itself, occlusion of the origin of the splenic and gastroduodenal arteries with coils would have been effective and without complication; however, the addition of Gelfoam particles in a sclerosing solution reduced the microscopic pancreatic circulation to a critical point and resulted in hemorrhagic pancreatitis.

Gut-trophic effects of keratinocyte growth factor in rat small intestine and colon during enteral refeeding.

BACKGROUND: Keratinocyte growth factor (KGF) induces proliferation of gut epithelium in rat models, but KGF-nutrient interactions have not been studied. An experimental model of fasting-induced gut atrophy followed by different levels of enteral refeeding was used to investigate the influence of nutrient availability on the gut-trophic effects of exogenous KGF. METHODS: After a 3-day fast, rats were enterally refed either ad libitum or at 25% of ad libitum intake for 3 subsequent days. Either intraperitoneal KGF (5 mg/kg/d) or saline was given in each dietary regimen. Wet weight, DNA, and protein content were measured as indices of full-thickness cellularity in duodenum, jejunum, ileum, and colon. Villus height in small bowel segments and crypt depth in all gut tissues were measured as specific indices of mucosal growth. RESULTS: Refeeding at 25% of ad libitum intake significantly decreased full-thickness cellularity and mucosal growth indices in duodenum, jejunum, and ileum. In the colon, only protein content fell significantly and crypt depth was maintained. KGF administration during 25% refeeding did not alter full-thickness indices in any small bowel segment or affect jejunal mucosal growth. In contrast, KGF normalized duodenal villus height (p < .01) and duodenal and ileal crypt depth (p < .05) only in the 25%-refed model. KGF significantly increased ileal villus height in both ad libitum and 25%-refed rats (by 43% and 48%, respectively, p < .05) and markedly increased colonic cellularity and mucosal crypt depth with both levels of refeeding (p < .01). CONCLUSIONS: Rat small bowel growth is more sensitive than colon to the level of enteral refeeding after a 3-day fast. KGF administration does not affect jejunal growth, but specifically prevents atrophy of duodenal and ileal mucosa during hypocaloric, hyponitrogenous refeeding. In ileum and colon, some KGF-mediated growth responses are independent of the level of enteral refeeding. Thus gut-trophic effects of KGF and KGF interactions with the level of nutrient intake are tissue-specific.

Metaplastic breast carcinoma with epithelial phenotype of pseudosarcomatous components.

A case of metaplastic breast carcinoma was subjected to an immunohistochemical study to characterize its apparent fibrosarcomatous and chondrosarcomatous elements. Polyclonal and monoclonal antibodies against epithelial cell components such as keratin proteins, epithelial membrane antigen, membrane and cytoplasmic antigens of human mammary carcinoma cells, and carcinoembryonic antigen were used, as well as antibodies against the mesenchymal antigens, desmin, and vimentin. The cells with a mesenchymal appearance had an epithelial derivation, as shown by the presence of epithelial cell markers and absence of mesenchymal cell markers.

Carcinoembryonic antigen: immunohistologic identification in invasive and intraepithelial carcinomas of the lung.

A total of 58 pulmonary lesions from 48 patients were examined for carcinoembryonic antigen (CEA). The three-layer immunoperoxidase procedure for antigen detection was used with a monospecific anti-CEA antiserum. The control serum was the same antiserum with its specificity removed by affinity chromatography. Normal goat serum was also used as a control. Carcinoembryonic antigen was present in the majority of pulmonary adenocarcinomas and generally absent in the squamous cancers. The major exception was in the well-differentiated squamous lesions where CEA was occasionally found in the keratinizing areas. Of special interest was the finding of CEA in all areas of intraepithelial squamous neoplasia studied.