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Background: The molecular basis of colorectal dysplasia and carcinoma arising in ulcerative colitis is poorly understood. Loss of heterozygosity involving the tumor suppressor gene p53 occurs frequently in neoplastic ulcerative colitis lesions. Point mutation affecting p53 is associated with loss of heterozygosity in other cancers. Therefore, it was determined whether p53 point mutation occurs in ulcerative colitis-associated neoplasia., Methods: Single-strand conformation polymorphism analysis, DNA sequencing, and loss of heterozygosity studies were performed on 45 patients with ulcerative colitis-associated dysplasia and carcinoma., Results: Point mutations were detected in 26 lesions from 20 patients, including 18 carcinomas, 6 dysplasia-associated masses, 1 flat dysplasia, and 1 lymph node metastasis. In two cases, identical p53 mutations were observed in both carcinoma and adjacent dysplasia. Missense mutations causing amino acid substitutions as well as nonsense mutations resulting in premature stop codons were seen. Tandem mutations, in which more than 1 sequence alteration occurred on the same allele of p53, were also detected. Point mutation was accompanied by loss of the other p53 allele in 8 of 10 patients informative for both loss of heterozygosity and mutation assays., Conclusions: These findings suggest that inactivation of p53 by mutation and loss of heterozygosity is a common mechanism of malignant transformation in ulcerative colitis. They also imply that in contrast to sporadic colorectal carcinoma, ulcerative colitis-associated neoplastic progression may involve p53 inactivation at relatively early, noninvasive stages.

One hundred two women admitted for evaluation of an adnexal mass were tested prior to surgery by the computerized tube leukocyte adherence inhibition (LAI) assay. The test was positive in 24/35 (69%) patients, diagnosed after surgery as epithelial ovarian cancer while 50/67 (75%) of those diagnosed as benign masses were negative. Only 2/38 (5%) of normal control subjects showed a false-positive result. The LAI test described in this report showed a sensitivity of 88% and a specificity of 75% in stage I ovarian cancer; thus it may be useful for early diagnosis of ovarian malignancy.

Twenty consecutive patients with unresectable, locally advanced pancreatic cancer were treated with split courses of radiotherapy (RT) and simultaneous multidrug chemotherapy consisting of 5 fluorouracil, continuous infusion, streptozotocin, and cisplatin. A separate, retrospective study identified a group of 28 contemporary patients with less advanced pancreatic cancers, all of which were successfully resected. The survival rate of the two groups were similar over the first 2 years, although it initially favored the unresectable group. This pattern of survival among patients treated with combined modality therapy provides a basis for new studies. At the two clinical extremes, these include treatment of unresectable tumors previously considered ineligible for this treatment and initial treatment before resection of stage I tumors.

Serum levels of the soluble form of tumour necrosis factor receptor type II (p75) (sTNF-R) were determined in HIV-infected individuals and risk groups and were then correlated with the course of infection and prognosis. sTNF-R levels were determined by an ELISA with MoAbs and polyclonal antibodies to urine-derived sTNF-R proteins. The mean +/- s.e. levels of sTNF-R in the sera of 49 HIV+ male homosexuals, 34 HIV- male homosexuals and 44 matched controls were 6.1 +/- 0.3 ng/ml, 4.4 +/- 0.3 ng/ml and 3.4 +/- 0.2 ng/ml, respectively. All these values were significantly different between each of the groups (P less than 0.001-0.05). Sequential studies of sTNF-R revealed higher levels following seroconversion in 5/8 individuals, remained persistently high during the asymptomatic phase of the infection and became even more elevated in some ARC and AIDS patients. At the same time TNF-alpha was undetectable in sera obtained from HIV+ male homosexuals and from healthy controls. This was independent of stage of HIV infection, serum sTNF-R level and type of ELISA kit used. These findings suggest that TNF-alpha/TNF-R system is turned on before and during HIV infection and raise the possibility that sTNF-R, the natural inhibitor of TNF, may be of importance in determining the course and probably prognosis of the disease.

There is an extensive literature documenting the increased or deregulated expression of the c-myc oncogene in human malignancies. The authors have recently devised a sensitive immunocytochemical method for studying the tissue localization of c-myc protein in tissue sections of human colon. We have compared nuclear c-myc staining using a polyclonal rabbit anti-c-myc antibody and a mouse monoclonal myc antibody NCM II 274. Microscopic observation of the tissue specific pattern of c-myc protein distribution shows that nuclear staining intensity varies in normal and neoplastic crypt cell nuclei in parallel with morphologic criteria of neoplasia. These studies yield further information on the usefulness of c-myc protein as a prognostic indicator.

Whether borderline serous tumors of the ovary can be differentiated from invasive serous cystadenocarcinomas by morphometric analysis of nuclear features of the neoplastic epithelium was examined. Multiple descriptors extracted from nuclear tracings and intranuclear gray-level analysis of argyrophilic nucleolar organizer regions were evaluated in 11 borderline and 18 invasive tumors using computerized interactive morphometric analysis. These descriptors included: nuclear area and perimeter; number, area, and perimeter of argyrophilic nucleolar organizer regions; standard deviations of these findings; and a size distribution of nuclear areas in discrete classes. Multivariate statistical analysis showed the internal consistency of the two groups in terms of physical descriptors and the discriminating value of the parameters of nuclear size and pleomorphism (independently of nucleolar organizer region parameters). The results indicated that interactive morphometric analysis of nuclear features, combined with appropriate statistical methods, could be used to distinguish between these tumors.

Allelic deletions of tumor suppressor genes have been observed frequently in a variety of human tumors. These losses are believed to contribute to the development of human cancer. Three of the most frequently deleted chromosomal loci contain the tumor suppressor genes p53, retinoblastoma (Rb), and mcc/apc. In order to detect loss of heterozygosity (LOH) within these genes in dysplastic and cancerous ulcerative colitis, we used an application of the polymerase chain reaction. LOH affecting p53 was observed in 8 of 17 (47%) of heterozygous patients, while LOH of Rb and the mcc/apc locus was observed in 9 of 27 (33%) and 13 of 39 (33%) of heterozygotes, respectively. Among 35 patients heterozygous at 2 or more loci, LOH of p53, Rb, and/or mcc/apc was observed in 18 (51%). LOH was more common in left-sided neoplasms. These data suggest that allelic deletion of p53, Rb, mcc, and/or apc is involved in the pathogenesis and/or progression of at least a subset of colonic dysplasias and carcinomas occurring in the setting of ulcerative colitis.

Immunologic methods for detection of colorectal neoplasia based on examination of stool or colonic effluent are being developed. Most current oral lavage preparations contain polyethylene glycol (PEG), and if PEG adversely interferes with immunologic testing these tests may become less useful. We describe a decrease in sensitivity of ELISA for tumor-associated antigens (TAA) when effluent samples are diluted in PEG-electrolyte lavage solution, equivalent to a commonly used oral lavage solution based on PEG. Radioisotope-labeled antigen binding to plastic plates was decreased by dilution in the PEG lavage solution. Antigen binding, present in colonic effluent collected by the laxative purge method, was absent in effluent collected by PEG oral lavage from the same patient. We conclude that PEG and PEG-containing lavage solutions interfere with ELISA detection of TAA in colonic effluents. We speculate that the in vitro, and possibly the in vivo, effect occurs at the level of antigen binding to the plate either by a steric effect or alteration of charge by the nonpolar properties of PEG.

The PiZ mutation of the gene coding for alpha 1-antitrypsin results in a serum deficiency of this protein leading to early onset emphysema and liver disease. The PiZ gene has a Z-specific point mutation in exon V together with a point mutation in exon III which is also present in some normal (PiM) individuals. There has thus far been no system to study the effects of PiZ point mutations in tissue culture. We constructed plasmids containing alpha 1-antitrypsin cDNA synthetically altered at either exon III or exon V mutation sites and linked to simian virus 40 promoter sequences. Such constructs with the exon V mutation were transfected into monkey COS1 cells followed by analysis of expression of alpha 1-antitrypsin gene products. COS1 cells normally synthesize virtually no alpha 1-antitrypsin mRNA or protein. alpha 1-Antitrypsin mRNA is transcribed at high levels in cells transfected with either M or Z plasmids. Immunologic staining of COS1 cells within 48 h of transfection localizes alpha 1-antitrypsin protein to specific regions of the cytoplasm. This extranuclear localization is also observed with human HepG2 hepatoma cells, which synthesize alpha 1-antitrypsin at high levels, and with human SK-Hep1 hepatoma cells transfected with an M plasmid. The cloned synthetically altered alpha 1-antitrypsin genes provide a system for dissecting contributions of distinct point mutations to the pathological effects of the PiZ protein.

Accessory and supernumerary ovaries are uncommon congenital anomalies of the female reproductive tract. Tumors arising in ectopic ovaries are extremely rare. We present a case of a Brenner tumor that arose in an accessory ovary and discuss this case in relation to the literature concerning neoplasms in ectopic ovaries.

Skh/hr-1 hairless albino mice were irradiated with photocarcinogenic dosages of ultraviolet light for periods of 30 weeks or longer. A high proportion of mice developed pleomorphic spindle cell tumors and epidermal neoplasms of various types. These spindle cell tumors were studied by immunofluorescence and immunoperoxidase techniques and by electron microscopy. Freshly isolated tumor cells were grown in tissue culture. Immunocytochemical analysis showed varying expression of markers of mesenchymal differentiation: vimentin, procollagens I and III, type I collagen, and lysozyme. Electron microscopy showed spindled and cuboidal cells with abundant endoplasmic reticulum, filopodia, and lysosomes, but no intercellular connections. The cells grown in vitro were cuboidal and stellate and also showed mesenchymal differentiation by electron microscopy. These results are perhaps similar to those described for a human actinically produced fibrohistiocytic neoplasm, atypical fibroxanthoma, and this system may provide a useful model of ultraviolet-induced dermal neoplasia.

The LAI reactivity of a colon organ specific neoantigen (OSN) was recovered from the urine of advanced colon cancer patients. In this study three physico-chemical steps were employed; precipitation by 80% ammonium sulfate, ion exchange, and molecular sieve chromatography. Each isolate was tested for activity and specificity by the direct tube leukocyte adherence inhibition (LAI) assay employing leukocytes from colon and breast cancer patients. The OSN enriched isolate was then used to generate monoclonal antibodies (Mab). Hybridomas were screened by ELISA. One hybridoma designated Bac 18.1 reacted preferentially with colon OSN and not with the urine from normals or patients with breast cancer. Affinity purification of colon OSN was achieved and it was shown to consist of a single band in SDS-PAGE with an apparent molecular weight of 30,000. The eluted polypeptide was specifically reactive in ELISA as well as in the LAI assay.

A monoclonal antibody (mAb) was prepared against a semipurified preparation of an organ specific neoantigen (OSN) reactive in the leukocyte adherence inhibition (LAI) assay. The mAb (LC20.1) induces a positive LAI response when incubated with leukocytes of normal individuals in the presence of OSN derived from either human colon or lung carcinoma cell lines. Absorption of crude OSN preparations from these cell lines on immobilized LC20.1 mAb eliminates all the LAI reactive material suggesting that the mAb recognizes a common determinant on OSN from both colon and lung carcinomas. The LC20 mAb was used to affinity purify the colon cancer OSN as well as a cross-reactive normal protein from the urine of colon cancer patients and healthy donors, respectively. The colon cancer OSN and normal cross-reactive protein display an apparent molecular weight of 29,000, have a similar linear tryptic peptide map, and are indistinguishable by isoelectric focusing analysis. Regardless of the molecular similarity, only the colon cancer OSN preparation could induce a positive LAI when incubated with leukocytes of colon cancer patients. Seven additional anti-colon cancer OSN mAbs were prepared against purified material. These mAbs can be divided into three groups, each of which recognizes a distinct antigenic determinant that is shared by the colon cancer neoantigen and its cross-reactive normal protein.

Caroli's disease is a rare form of fibropolycystic disease of the hepatobiliary system characterized by segmental cystic dilatation of intrahepatic ducts. It is associated with intrahepatic cholelithiasis, cholangitis and hepatic abscesses. Like other forms of fibropolycystic disease of the liver, Caroli's disease is often accompanied by cystic renal disease, specifically renal tubular ectasia or medullary sponge kidney. Adult-type polycystic kidney disease associated with Caroli's disease is rare, only one such case having appeared in the literature to our knowledge. We present a second instance of this association.

Lipoadenoma of the parathyroid gland is a rare histologic variant of parathyroid adenoma that is usually functional and associated with clinical hyperparathyroidism. We report a case in which a radiolabeled thallium scan failed to demonstrate evidence of an adenoma, presumably because of the tumor's high fat content. The literature concerning this entity is reviewed. To our knowledge there are no other reported cases in which parathyroid scanning was used in diagnostic studies of parathyroid lipoadenoma.

Immunity to colorectal cancer antigen was tested by the tube leukocyte adherence inhibition (LAI) assay, using spent medium of human carcinoma cell lines as a source of antigen. The assay was positive in 18 out of 43 patients (41%) with colorectal tumor in comparison to only 2 out of 34 (5.8%) of colorectal cancer patients clinically free of disease, and to 4 out of 150 (2.6%) healthy subjects. The frequency of positive results correlated negatively with the tumor burden, 16 out of 29 (55.1%) patients with Dukes' A, B and C being positive compared to only 2 out of 14 (14.2%) patients with Dukes' stage D disease. Addition of PGE2 enhanced the sensitivity of the assay without affecting its specificity. The number of positive assays increased from 18 to 32 (41.8% to 74.4%) in the whole group of 43 colorectal cancer patients without altering significantly the frequency of positive results in the control group. The results with all groups of patients were influenced similarly by the addition of prostaglandins, the frequency of positive assays increasing from 55.1% to 82.7% and from 14.1% to 57.1% in early and advanced disease, respectively. These results lend further support to the value of the LAI assay in diagnosis and monitoring of the immune response in human colorectal cancer.

Highly glycosylated, water-soluble ABH-specific sphingolipids, designated macroglycolipids, were isolated in high yield, up to 5 mg per unit of blood, from the crude human-erythrocyte-membrane glycoprotein fraction which is obtained by extraction of the membranes with chloroform/methanol/water. Both serological tests and radioactive labelling experiments indicated that these substances, rather than the glycoproteins, are the principal ABH-components in this fraction. The activities of A-specific, B-specific and H-specific macroglycolipids were very high, approximately 0.1 microgram inhibiting four hemagglutinating doses of the respective agglutinating reagents, and were thus comparable to those of secreted blood-group ABH-specific glycoproteins. The substances were stable to mild alkaline conditions. They contained fucose, galactose, glucosamine, glucose, sialic acid, sphingosine and fatty acids; blood-group-A-specific substances contained, in addition, galactosamine. No amino acids were detected. Assuming one glycosyl residue per molecule, the average number of sugars in A and B macroglycolipids was 31, and their molecular weights approximately 6100. The presence of beta-D-galactosidase-labile and sialic acid residues indicated that these substances contain nonreducing termini additional to the ABH immunodeterminants. In the B macroglycolipid, the ratio between nonreducing terminal alpha-D-galactopyranosyl and beta-D-galactopyranosyl residues was 1.7:1.0. The macroglycolipids formed clear aqueous solutions at concentrations as high as 30 mg/ml, were insoluble in 60--70% aqueous ethanol, and did not migrate on thin-layer chromatography unless they were acetylated. Polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate showed the macroglycolipids to be a heterogeneous mixture migrating throughout most of the region in which the periodic acid/Schiff-positive membrane glycoproteins are found. On the basis of the evidence presented, it is concluded that macroglycolipids are the predominant ABH-specific component in human erythrocyte membranes, and that they most likely account for previous observations of ABH activity in membrane glycoprotein fractions.

Spleen cells of BALB/c mice immunized with human chorionic gonadotropin (HCG), a glycopeptide hormone composed of two nonidentical alpha and beta subunits, were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, cone PC2 and clone PD3 secreting monoclonal antibodies (McAb) to HCG were isolated. With the aid of a double antibody radioimmunoassay it was established that PC2-McAb recognizes an immunodeterminant associated with beta-HCG subunit, whereas, PD3-McAb recognizes an epitope present on the alpha-HCG subunit. Both monoclonal antibodies showed an affinity constant of approximately 5 X 10(10) L/M. Because of these immunological characteristics, only PC2-McAb specifically reacts with HCG and shows cross-reactivities of less than 0.1% to other related human glycopeptide hormones. On the other hand, using a hemagglutination test based on antibody induced agglutination of sheep red blood cells coated with HCG, it was shown that only PD3-McAb is capable of inducing a positive reaction, although both monoclonal antibodies have a similar binding capacity to the coated cells. The potential of these two new monoclonal reagents for the qualitative detection and quantitative determination of HCG in human biological fluids is discussed.

A 44-year-old man with inflammatory pseudotumor of the thymus is reported. The patient was seen with fever, myalgia, and dyspnea and was found to have an anterior mediastinal mass and bilateral pleural effusions. The resected lesion consisted of a well-circumscribed mass of chronic inflammatory and fibrous tissue that virtually replaced the thymus. There was no morphological evidence of neoplasia. The patient's symptoms and roentgenographic abnormalities resolved with excision of most of the mass. To our knowledge, this report is the first instance of inflammatory pseudotumor of the thymus.

Urogenital cysts are retroperitoneal or mesenteric cysts that are derived from vestigial remnants of the embryonic urogenital apparatus. Although their precise embryonic origins are usually obscure, occasionally they display histologic features indicative of a mesonephric or metanephric origin. This report describes an unusual mesenteric müllerian cyst consisting of an endosalpingial lining and two-layered smooth muscle walls, probably representing a müllerian duct duplication cyst.

A computerized tube LAI assay was used to determine the antitumor immune response in lung cancer patients. A standardized assay was established by the use of tumor antigen recovered from the spent medium of lung and colon adenocarcinoma cell lines. Furthermore, modulation of the assay by prostaglandin E2 (PGE2) was used to increase its sensitivity. Of 65 patients with epidermoid lung cancer, 34 (52.3%) had a positive nonadherence index (NAI). The NAI was inversely related to tumor burden, with 31 of 44 (70.6%) patients with lung cancer stages I and II and only 3 of 21 (14%.2%) patients with stage III having positive assays (greater than 30%). Addition of PGE2 to the as say increased the overall sensitivity so that a positive NAI was obtained in 88.6% of patients with lung cancer stages I and II, and 88.6% with stage III, respectively. Of 174 individuals from the control group, positive NAI was found in only 9 and 3.8% of patients with chronic obstructive pulmonary disease and normal controls, respectively. These results support previous observations on the specificity and sensitivity of the LAI assay in the diagnosis and study of human cancer.

Polymorphous low-grade adenocarcinoma (PLGA) is a minor salivary gland neoplasm that is characterized by morphologic variability, cytologic uniformity, and an infiltrating growth pattern. To date, this entity has been identified either within the confines of the oral cavity or, rarely, in the nasal cavity. The authors report a case of PLGA that arose in the nasopharynx of a 44-year-old female, representing the first documented occurrence of this tumor outside the oral cavity or the sinonasal tract. The histopathologic and immunohistochemical differentiation from pleomorphic adenoma and adenoid cystic carcinoma is discussed.

Histologic, histochemical, immunocytochemical, and ultrastructural features of two cardiac myxomas containing glandular elements are reported. Glandular elements in both cases stained positively with both mucicarmine and periodic acid-Schiff reagent with diastase pretreatment (DPAS). Immunoperoxidase studies demonstrated positivity of the glandular cells for carcinoembryonic antigen (CEA), epithelial membrane antigen (EMA), and keratin. Factor VIII-related antigen (FVIIIAg) was identified only in cells lining vascular spaces. Electron microscopic study of one tumor demonstrated well-formed glands having basement membranes, junctional complexes, and apical secretory granules. These findings indicate the capacity for true epithelial differentiation of cardiac myxomas and have implications both as regards the histologic diagnosis of these tumors and their histogenesis.

Seventy-four women admitted for breast biopsy were monitored before surgery for anti-tumor cell-mediated immunity using a computerized tube leukocyte adherence inhibition (LAI) assay. Spent medium from breast, lung and colon adenocarcinoma cell lines was used as the source of organ-specific neoantigens for standardization of the assay. Peripheral blood leukocytes from 25/40 (62.5%) patients diagnosed after surgery as breast cancer responded to spent medium with a positive non-adherence index (NAI). A positive NAI was inversely related to tumor mass because only 7/18 (38.8%) of those with Stage III or IV had a positive NAI; while 18/22 (81.8%) of those with Stage I or II were positive. Cross-reactive antigenicity was not observed when spent medium from breast cancer was incubated with leukocytes from patients with several other solid tumors nor when leukocytes from breast cancer patients were incubated with spent medium from lung or colon carcinoma cell lines. The antigenic material in the spent medium appears to be an organ-specific neoantigen because only 1/34 patients with benign breast disease had a positive NAI and all normal healthy control individuals were negative. The results of this study show that spent medium of a breast carcinoma cell line is more reliable than crude cancer extracts for use in LAI to detect specific anti-tumor cellular immune responses. The improved method presented in this report can be a useful tool in the early diagnosis of breast cancer and for monitoring of patients with this disease.

The heterogeneity of glycopeptides prepared from pronase digests of various glycoproteins has previously been demonstrated primarily by lengthy paper chromatography of oligosaccharides released from the glycopeptides by enzymatic or chemical means. We describe the use of high voltage paper electrophoresis in borate for the resolution of seven glycopeptides from ovalbumin and seven glycopeptides from human immunoglobulin G. This technique has been used in the preparation of glycopeptide substrates required for the study of certain glycosyltransferases and glycosidases.

alpha-D-Galactosidase was isolated from untoasted soybean meal and purified to homogeneity by affinity chromatography on N-epsilon-aminoacaproyl alpha-D-galactopyranosylamine-Sepharose. The purified enzyme destroyed the B-specificity of human ovarian cyst B-glycoprotein with an accompanying increase in H-specificity, and converted human type-B erythrocytes to type O. The enzyme consists primarily of a tetramer, molecular weight 150 000 +/- 5 000 at pH 4.0 and of a monomer, molecular weight 40 000 +/- 3 000 at pH 8.0. Polyacrylamide gel electrophoresis in dodecyl sulfate at pH 7.2 distinguished between two types of monomeric unit of similar molecular weight. N-terminal alanine was identified as the sole N-terminal amino acid residue. The enzyme was shown to be devoid of carbohydrate.