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The neurologic symptoms in human shigellosis have often been attributed to Shiga toxin, although its exact role has not been determined. By use of a [3H] thymidine-labeled HeLa cell assay, cytotoxic activity was demonstrated in stool but not cerebrospinal fluid or serum from five patients with shigellosis presenting with seizures or encephalopathy. Bacterial isolates produced 16.0-88.2 CD50 (50% cytotoxic dose) of cytotoxin/mg of protein. The toxin activity in stool and the cytotoxic activity of the isolates were not neutralized by antiserum to purified Shiga toxin. DNA hybridization studies showed that Shigella isolates from these patients lacked the structural genes for Shiga toxin. The cytotoxin produced was also distinct from Shiga-like toxins I and II. Sonicates of the Shigella strains injected intraperitoneally into mice caused lethargy and lethality. The toxin activity was heat-labile and sensitive to trypsin, indicating that its active component is protein. Ultrafiltration and gel filtration chromatography showed a molecular mass of 100-125 kDa. Thus Shiga toxin production is not essential for the development of neurologic manifestations of shigellosis; other toxic products may play a role.

Stage B adenocarcinomas of colon (61 carcinomas) and rectum (44 carcinomas) were retrospectively reviewed in order to define prognostic indicators and to determine if they are different for tumors arising at the two sites. Parameters evaluated included the substage of the tumor, histologic grade, presence of vascular/lymphatic invasion, residual adenoma or extracellular mucin, and tumor cell DNA content as determined by flow cytometry of paraffin-embedded tissues. Five-year survival was 81.0% for patients with colonic and 87.5% for those with rectal tumors. DNA ploidy was a significant predictor of overall and disease-free survival in patients with rectal adenocarcinomas (improved survival for those with diploid tumors) but not in those with colonic tumors and not in the two groups combined. The degree of differentiation of the tumor was prognostically significant in the colonic group and for the groups combined but not in the rectal group. There was a trend toward better survival in substage B1 tumors, as compared with substages B2 and B3, particularly in rectal neoplasms. Other histologic parameters did not predict survival. We conclude that the prognostic indicators in colonic adenocarcinoma seem to differ from those of adenocarcinoma arising in the rectum and that DNA ploidy has a significant impact upon outcome in stage B rectal adenocarcinomas.

The degree of focal chronic sialadenitis in a labial salivary gland biopsy specimen, expressed in terms of a semiquantitative focus score, is the best contemporary assessment of the salivary component in Sjögren's syndrome. A labial salivary gland focus score of greater than 1 focus per 4 mm2 serves as an acceptable threshold for the diagnosis.

The authors report three cases of sarcomatoid carcinoma arising in the stomach. This uncommon tumor is characterized by a mixture of malignant epithelial and spindle cell elements. All three tumors were large (average diameter, 5 cm) and infiltrated deep into the stomach wall. Two of the tumors had a polypoid configuration; the third was ulcerated and endophytic. Intestinal metaplasia was present adjacent to the tumor in all cases, with dysplasia in two. Immunohistochemical studies showed positivity for cytokeratin, carcinoembryonic antigen, and epithelial membrane antigen in the epithelial component of all tumors, and Leu-M1 was positive in the epithelial component of one. The spindle cell components contained vimentin, and in tumor 2, the spindle cell component was also positive for desmin. Two tumors showed focal positivity for cytokeratin in the spindle cells immediately adjacent to the epithelial component. Ultrastructurally, the spindle cell component of two tumors was composed of undifferentiated cells without specific epithelial or mesenchymal features. The third tumor contained occasional cells with features of myofibroblasts.

Undifferentiated carcinoma with lymphoid stroma or lymphoepithelial carcinoma of the major salivary glands is a demographically and histopathologically unique malignancy. Although whites may have the disease, it is preponderantly a carcinoma of North American Eskimos and native Greenlanders. The carcinoma shares many features with undifferentiated nasopharyngeal carcinomas, from which it must be distinguished: histologic appearance, putative relationship with Epstein-Barr virus, predilection for mongoloid races, and response to therapy. In some cases, the carcinoma appears to have evolved from a lymphoepithelial lesion.

Patients with human colorectal carcinoma have a poor prognosis when serum carcinoembryonic antigen level exceeds 5 ng/mL. The hypothesis that carcinoembryonic antigen enhances metastasis by promoting the attachment of tumor cells to Kupffer cells and hepatocytes was tested in an experimental metastasis model in which colorectal carcinoma cells were injected into the spleens of BALB/c athymic nude mice and liver colonies counted 5 weeks later. Pretreatment with systemic injections of carcinoembryonic antigen significantly increased the metastatic potential of a poorly metastatic colorectal carcinoma cell line KM-12c, but did not induce the nonmetastatic colorectal carcinoma cell line HC 2998 to produce metastases, nor did carcinoembryonic antigen make the highly metastatic colorectal carcinoma cell line mHC 1410 more metastatic. Carcinoembryonic antigen did not stimulate proliferation of colorectal carcinoma but appeared to be a cofactor for metastasis possibly as an adhesion factor.

Four monoclonal antibodies specific to various Lewisx-related antigens were tested for binding to monolayers and detergent extracts of the human colon carcinoma cell line HT-29 P and its metastatic variant HT-29 LMM. Only monoclonal antibody FH6 (antisialyl-dimeric Lex) bound more to the metastatic variant compared with HT-29 P. Detergent extracts of human tissues were then assayed for their FH6-binding activity. Normal colonic mucosa displayed less FH6 binding than either Dukes stage B or D primary colon carcinomas. Liver metastases from colorectal carcinoma demonstrated greater binding than stage B and stage D primary tumors, and more than normal liver. Expression of sialyl-dimeric Lex increases as the metastatic potential of colorectal carcinomas increases, and is related to the progression of colorectal cancer.

Lymphoepithelial cysts of the parotid gland and its lymph nodes have been known for nearly 100 years, but only in the past half-decade have they gained clinical prominence, if not significance. The lymphoepithelial cysts can now be added to the growing list of clinicopathologic manifestations of human immunodeficiency virus-associated diseases.

We studied alterations in the transitional mucosa of the colon in 18 cases of primary colonic adenocarcinoma (4 from the cecum; 4, the ascending colon; 5, the descending colon; 2, the sigmoid colon; and 3, the rectum). One patient had 2 separate primary tumors. Formalin-fixed, paraffin-embedded sections were studied in all cases, and tissues fixed in methacarn fixative were available in 9. Sections of tumor and transitional mucosa were compared with those of normal mucosa distant from the tumor. Sections were stained with hematoxylin-eosin, Masson's trichrome, and alcian blue at a pH of 0.9 (sulfomucin) and at a pH of 2.5 (sialomucin and sulfomucin). Immunoperoxidase stains for collagen type IV (basal lamina) were also performed. Transitional mucosa showed morphological alterations, including increase in mucosal thickness and crypt distortion. An increase in lamina propria fibrosis was noted in transitional mucosa in 3 cases. Abnormal mucin content, consisting of a predominance of sialomucin, was noted in transitional mucosa in 12 cases. In one of these, sialomucin was predominant in both transitional and normal mucosa. No alterations in the thickness of the subepithelial collagen table were noted. Collagen type IV staining was effective in demonstrating an increase in lamina propria vascularity in transitional mucosa in 11 specimens.

Pneumocystis carinii is the most common protozoan organism causing infection in immunosuppressed patients. This study, based on a review of 32 lung biopsies and 13 autopsies of patients with neoplastic diseases who developed P carinii pneumonia, emphasizes the morphologic variation of this disease. Excluded from this study are those patients with PCP secondary to bone marrow transplantation and AIDS. P carinii pneumonia occurred predominately in patients with malignant lymphoreticular neoplasms, 73% of those in our series, and the remaining 27% had solid tumors. There were 27 males and 18 females and their ages ranged from 1 to 73 years, with a median age of 30 years. Microscopically, the diagnostic intraalveolar foamy exudate of P carinii pneumonia was present in only 58% of the cases. Furthermore, diffuse alveolar damage alone was present in 26% of cases. This change was especially prominent when the characteristic foamy material was scanty. Other tissue reactions to P carinii included the presence of giant cells, epithelioid granulomas, desquamative interstitial-like pneumonitis, and interstitial lymphoid infiltrate. Extrapulmonary dissemination was not observed in this patient population. The pathologist should be aware of the marked variations that occur in the morphologic appearance of PCP, and the diagnosis of Pneumocystis infection should not be discarded until a careful search for the organisms using silver stains is rendered negative.

Sulfated macromolecules synthesized in tumor and mucosa tissues derived from colorectal cancer patients were labeled with [35S]sulfate and separated into two fractions on DEAE-Sephacel: the slightly acidic peak (peak I) was eluted with 0.2 M NaCl and the highly acidic peak (peak II) was eluted with 0.5 M NaCl. A total of 40 specimens, which included primary colon cancer, liver metastases, and normal mucosa obtained at surgery (16 patients), were examined regarding the amount of peak I and peak II. The amount of peak I significantly decreased in the order of normal mucosa greater than primary tumors greater than metastases, while the amount of peak II did not significantly change among the tissues. Peak I was mostly resistant to chondroitinase ABC and nitrous acid treatment under acidic conditions, whereas combined chondroitinase-sensitive materials and nitrous acid-sensitive materials were greater than 80% of the radioactivity in peak II. The major radioactive component of peak I migrated at a position corresponding to Mr greater than 300,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and became Mr less than 40,000 after alkaline borohydride treatment. The major component of peak I was likely to be a sulfated glycoprotein containing sulfate groups on alkaline labile carbohydrate chains. Peak II consisted of a mixture of heparan sulfate proteoglycans and chondroitin sulfate proteoglycans. Differential incorporation of [35S]sulfate into peak I among normal mucosa, primary colon carcinoma, and colon carcinoma metastasis was observed. Therefore, decreased peak I production may be a biochemical change associated with colorectal cancer progression and metastasis.

Previous studies using metabolic labeling of fresh colonic mucosa and colorectal carcinoma with [35S]sulfate followed by biochemical analysis demonstrated that the amount of a sulfated high-molecular-weight glycoprotein expressed in primary colorectal carcinoma was lower than that in normal mucosa, and that the amount further decreased in liver metastases. This suggested that this sulfated molecule represented a sulfomucin previously defined by histochemical reactivity with a cationic dye. We have extracted and partially purified this high-molecular-weight sulfated glycoprotein from normal human colonic mucosa. We immunized mice with the partially purified sulfomucin and generated hybridomas. One cloned hybridoma, designated as 91.9H, produced a monoclonal antibody strongly reactive with a component which migrated at an identical position as the metabolically [35S]sulfate-labeled high-molecular-weight glycoprotein after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The reactive molecules appeared to have a polydisperse nature with a molecular weight ranging between 400,000 and 900,000. The [35S]sulfate-labeled high-molecular-weight glycoprotein was bound to Staphylococcus Protein A-agarose coated with this monoclonal antibody but did not bind to unconjugated Protein A-agarose. The immunoprecipitated substance also migrated at an apparent molecular weight range of 400,000 to 900,000. The reactivity of monoclonal antibody 91.9H with the extracts of normal mucosa, colorectal primary carcinoma, and metastasis was compared by dot blot assay on a nitrocellulose membrane. This antibody was more reactive with the extracts of mucosa adjacent to carcinoma tissues than with the carcinoma extracts. Primary tumors showed higher reactivity than metastases in most of the cases. These results strongly suggest that this antibody is specific to colonic sulfomucins or at least to mucins closely related to colonic mucins previously identified by metabolic labeling with [35S]sulfate.

Basal cell hyperplasia (BCH) is an uncommon proliferative lesion of the prostate gland. We studied ten cases of BCH, one case of an unusual adenoid basal cell tumor (ABT), and one case of a prostatic adenoid cystic carcinoma (ACC), using a panel of antibodies to define the histogenesis of these lesions. Monoclonal antibodies (MoAb) directed against a cytokeratin, which selectively stains basal cells (34 beta E12), and against muscle-specific actin, which stains myoepithelial cells (HHF35), were used. In addition, antibodies directed against prostatic acid phosphatase (PAP), prostate-specific antigen (PSA), S-100 protein, and vimentin were used. In the normal prostate, epithelial cells reacted positively with 34 beta E12, PAP, and PSA, and negatively with the actin, S-100 protein, and vimentin antibodies. In BCH, positive staining was seen for 34 beta E12, PSA, and PAP, with no reactivity for actin, S-100 protein, and vimentin. In ABT and ACC, positive reactivity was demonstrated for all antibodies except actin and vimentin. These findings indicate that the basaloid cells of BCH, ABT, and ACC are derived from basal cells of the normal prostate gland and suggest a continuum among the three lesions. The presence of S-100 protein in ABT and ACC may be related to the lack of this antigen's specificity for myoepithelial cells. The absence of reactivity with the HHF35 MoAb supports our belief that the S-100 positivity does not necessarily indicate myoepithelial cell differentiation.

The prevalence of Campylobacter pylori (CP) in gastric biopsies from cancer patients has not been previously studied. We reviewed 112 gastric biopsies from 78 adult patients at a cancer hospital. The patients had a previous history or current diagnosis of gastric or esophageal carcinoma (26 patients), gastric lymphoma (15 patients), gastric stromal tumor (three patients), carcinoid tumor (two patients), nongastroesophageal malignancy (29 patients), or atrophic gastritis with nonhealing ulcer (three patients). The biopsies were stained with hematoxylin-eosin (H&E), Warthin-Starry, Giemsa, and Brown-Hopps. CP was identified in 30 biopsies from 26 patients. Eighteen of the biopsies showed active gastritis, nine showed chronic inflammation/intestinal metaplasia, and one contained an ulcer. CP was also identified in two stomachs bearing tumors (one adenocarcinoma, one carcinoid tumor). Active gastritis was present without CP in nine biopsies, including three from patients on chemotherapy and one from a patient with bile reflux. The organism was detected by the Warthin-Starry stain in 30 biopsies. The Giemsa stain was positive in 27, Brown-Hopps in 27, and H&E in 24. Non-CP bacteria were identified in the Brown-Hopps in seven cases and were mistaken for CP in H&E stains in two. We conclude that CP is frequently found in the cancer patient with active gastritis and is an occasional finding in tumor-bearing stomachs. Special stains are useful in identifying CP, and the Brown-Hopps is helpful in distinguishing CP from other bacteria.

Basal cell hyperplasia of the prostate is a rare, benign lesion that often has been misdiagnosed as adenocarcinoma. Thirteen cases of basal cell hyperplasia were reviewed. All of the patients were over 60 years of age (range 63-83) and all had benign prostatic hypertrophy in addition to basal cell hyperplasia. Histologic features consisted of nests of uniform small cells with scant cytoplasm forming solid nests and acinar structures. Some foci had nodular configurations and most were located within larger nodules of "typical" glandular hyperplasia. In some instances there was a merging of the two types of hyperplasia. There was neither nuclear atypia nor pleomorphism, and the adjacent stroma was hyperplastic. Small elongated cells with desmosomes and nonspecific, dense bodies were seen on electron microscopic studies of formalin-fixed material. The architectural and cytologic features of the basal cell proliferations were not those of carcinoma. However, synchronously, two patients demonstrated foci of prostate carcinoma unrelated to the basal cell hyperplasia.

Carcinomas involving the jejunum and ileum are rare tumors. During a review of small intestinal neoplasms, six primary carcinomas of jejunum or ileum with an anaplastic and sarcomatoid histology were identified. At presentation, three of the patients had symptoms related to metastatic disease and three had symptoms referable to the local tumor. The tumors were large (greater than 4.5 cm in diameter), usually endophytic masses composed of large cells with eosinophilic cytoplasm, anaplastic nuclei, and prominent nucleoli. In many areas, the cells had a spindled configuration. Mucin positivity was identified in all six cases. Electron microscopic findings in two cases were indicative of epithelial differentiation. The tumors behaved aggressively; all five patients for whom there was clinical follow-up died of metastases within 40 months. The six anaplastic and sarcomatoid carcinomas were compared with 29 typical adenocarcinomas arising in the jejunum or ileum. Only two of the latter group had symptoms referable to distant metastases at presentation. These tumors also tended to be smaller at presentation (11 tumors were less than 4 cm in greatest dimension). Of 25 patients with typical adenocarcinomas who had acceptable follow-up, 18 (72%) died of disease and five (20%) were alive with no evidence of disease after 5 years. We conclude that anaplastic and sarcomatoid carcinoma is a rare variant of small intestinal carcinoma with an aggressive clinical course.

To determine if the amount of chondroitin sulfate proteoglycan (CSPG) in human colorectal tumor tissue correlates with the tumor's aggressiveness we immunochemically determined the CSPG levels in colorectal carcinomas at different stages. A total of 50 specimens--4 polyps, 15 stage B tumors, 9 stage C tumors, 12 stage D tumors, 7 liver metastases, and 3 lymph node metastases--were examined. Tumor tissues were extracted with 4 M guanidine hydrochloride containing protease inhibitors. The extracts were serially diluted and blotted onto nitrocellulose membranes. Reactivity of a chondroitin sulfate-specific mouse monoclonal antibody (CS-56) was determined by biotinylated goat antimouse Ig and avidin-biotin-peroxidase complex. After comparing tissues from tumors at different stages (classified by the presence or absence of metastasis), we could not find a positive or negative correlation between the amount of CSPG in primary colorectal carcinoma tissues and the tumor's metastatic potential. However, the metastatic foci in the liver or lymph node contained higher amounts of CSPG than the primary tumors did. Immunohistochemical staining of colon carcinoma tissue with CS-56 revealed that CSPG is predominantly localized in fibrotic portions in the tumor tissues. Two-year follow-up studies indicated that a high level of CSPG in primary tumors was not predictive of recurrence.

We collected a total of 78 tissue specimens, including primary colorectal carcinoma, normal colonic mucosa, and liver metastases of colon carcinoma, to examine whether the extracts of these tissues inhibited the binding of a monoclonal antibody FH6, specific for sialyl-dimeric LeX antigen. The results of inhibition assays demonstrated that: (a) contents of FH6-reactive molecules were greater in carcinoma tissues than in normal colonic mucosa; (b) metastatic foci in livers contained more FH6-reactive molecules than primary tumors; (c) primary tumors from Dukes' stage B1 patients contained less FH6-reactive molecules than primary tumors from Dukes' stage D patients. The inhibitory activity of these tumor tissue extracts against the binding of a monoclonal antibody FH6 to cultured colon carcinoma cells was eliminated by prior treatment of the extracts with sialidase, confirming that the FH6-reactive materials were sialyl-dimeric LeX antigen. Electrophoretic separation of tumor tissue extracts on 3% polyacrylamide gels followed by direct staining with monoclonal antibody FH6 revealed that very high molecular weight glycoproteins, presumably mucins, contained sialyl-dimeric LeX antigen.

Electron microscopy can be of limited value in distinguishing among poorly differentiated hepatocellular carcinomas, bile duct carcinomas, and other adenocarcinomas, but may be very helpful in identifying tumors that are metastatic to the liver. The characteristic ultrastructural features of endocrine and exocrine tumors of the pancreas are readily distinguished by electron microscopy when the tissue is adequately preserved, and this technique can be very useful in establishing a diagnosis.

The condition of a 6-year-old boy was diagnosed as mixed-cellularity Hodgkin's disease that involved the right side of the neck. Five years after the completion of radiation therapy for the involved area, followed by six courses of chemotherapy with mechlorethamine hydrochloride (nitrogen mustard), vincristine sulfate, procarbazine hydrochloride, and prednisone, he developed cerebral gliosarcoma. Numerous second malignant neoplasms have been reported in adults following treatment for Hodgkin's disease; however, the sequence of events in our patient is a new finding that has not, to our knowledge, been reported previously. While the second malignant tumor may have been induced by prior treatment, direct evidence is lacking.

Antibodies to specific human immunodeficiency virus (HIV) polypeptides are important laboratory markers of HIV infection. We have used an antibody to the major structural gag protein p24 of HIV-1 virus to immunochemically localize this capsid antigen in lymphoid cells from seven of eight patients at risk for HIV infection and who presented with parotid lymphadenopathy and lymphoepithelial cysts of the parotid gland. A clinicopathological assessment of these two manifestations as they relate to HIV infection is also presented.

To determine whether the production of experimental hepatic metastases in athymic nude mice by human colorectal carcinomas (HCC) correlated with the clinical outcome in patients, we harvested colorectal carcinomas from 82 patients, dissociated the tumors with collagenase and DNase, and injected them into groups of nude mice, either in the flank to assess experimental tumorigenicity or into the spleen to produce experimental metastasis in the liver. Growth in mice was then associated with clinicopathological factors and clinical outcome. Growth of HCC in either the flanks or the livers of nude mice was associated with the time to recurrence in a Wilcoxon analysis. Analysis of the outcome data in a Cox proportional hazards model suggested that there was an interaction between tumorigenicity and metastatic potential of HCC in nude mice and serum CEA concentration in the patient and stage of disease. A univariate analysis indicated that both tumorigenicity and metastatic potential of HCC in nude mice were significantly associated with the serum CEA concentration of the patient but not with the other variables of stage of disease, mucin production, local tissue invasion, state of differentiation, or sex. A subset of 57 patients was operated upon for cure and followed prospectively for up to 61 months. Tumorigenicity and, to a lesser extent, experimental metastatic potential were associated with disease recurrence in 23 of these patients. Seventy-eight % of the subset of patients who were operated upon for cure developed liver metastasis as one site of their progressive disease. Thus, the ability of HCC cells isolated from surgical specimens to grow in athymic nude mice correlates with the development of advanced disease in patients.

A 29-yr-old woman developed severe, progressive cholestasis 5 months after allogeneic bone marrow transplantation. Extrahepatic graft-versus-host disease (GVHD) was absent. Skin biopsy was equivocal 2 months after transplant, rash was absent during the period of cholestasis, and cholangiographic abnormalities were absent. Liver biopsy 7.5 months posttransplant revealed chronic hepatic GVHD. Cholestasis dramatically resolved with high dose corticosteroid therapy. Chronic hepatic GVHD occurs in the absence of overt extraintestinal GVHD and respond promptly to therapy. This underscores the importance of aggressive diagnostic evaluation of posttransplant cholestasis.

The antibody response of patients was used to characterize the autoantigens in human colorectal carcinoma. Twenty-seven primary and 13 metastatic carcinomas with paired normal tissues were extracted and transferred onto nitrocellulose membranes by the Western transfer technique. After the transfers were incubated with the serum of the patient from whom the tumor was derived, autoantigens were identified by indirect immunoperoxidase staining. All tumors contained at least one autoantigen. Six tumor-associated autoantigens, ranging in molecular weight from 26 to 58 kilodaltons (kD), were identified by antibodies in 25% or more of the sera. Eleven metastases expressed a 41-kD autoantigen that was present in only a third of the extracts of normal liver or lung. Thus, the number of dominant polypeptide autoantigens in colorectal carcinoma is restricted to six molecules. These autoantigens may be organ-associated antigens that are expressed by neoplastic cells. The 41-kD autoantigen may be a potential marker for metastases. A generic vaccine appears to be feasible for colorectal carcinoma since the number of dominant antigens is limited.

A Ulex europeus agglutinin I (UEAI)-reactive glycoprotein(s) with molecular weight higher than 300,000 was detected by direct binding of 125I-labeled UEAI to lysates of rectal or sigmoid colon cancer tissues separated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. This was the major UEAI-reactive molecule in tumor tissues and different tumors possessed varying reactivities. Very little UEAI binding was detected to lower molecular weight components. Histochemical localization of UEAI confirmed that the UEAI-reactive molecules were mostly localized to the surface of carcinoma cells. A total of 135 fresh tissue samples, including those from adenocarcinoma, villous adenoma, and normal mucosa in surgical specimens (69 patients), were examined to determine the reactivities of UEAI to the high molecular weight component in the tissue extracts. The quantitative binding of 125I-UEAI was compared according to the stage of colorectal cancer at the time of surgery. The relative amount of UEAI-reactive high molecular weight substance was significantly higher in the carcinomas than in normal mucosa. UEAI binding to high molecular weight regions of the polyacrylamide gel was significantly lower in primary colorectal adenocarcinomas from stage C or D patients than in those from stage B1 patients. Therefore, increased expression of the UEAI-reactive molecule was related to transformation of colorectal epithelial cells and decreased expression appeared to be associated with progression and metastatic potential.

Electron microscopy offers substantial aid in the diagnosis of gastrointestinal tract tumors, especially small-cell carcinoma and enterochromaffin-cell tumors. Tumors that are most difficult to classify by light microscopy--that is, poorly differentiated ones--may also have ultrastructural features by which the cell of origin can be determined. As always, the well-preserved tissue is more likely to retain diagnostic features than is the inadequately preserved.

Wheat germ agglutinin (WGA)-binding cellular glycoproteins produced by HT-29 human colon carcinoma and its variant cells established from liver metastases in nude mice after intrasplenic injection were analyzed by polyacrylamide gel electrophoresis. On 5.5% polyacrylamide gels five major sialoglycoproteins (approximate Mr 115,000, 145,000, 190,000, 450,000, and 740,000) reactive with WGA were common to the parental and metastatic sublines. There was an additional component of Mr approximately 900,000 that was prominent in cells established from liver metastases. Specific removal of sialic acid from the glycoproteins eliminated WGA binding, indicating that all the WGA-binding glycoproteins including the Mr 900,000 component were sialoglycoproteins. Smith degradation following mild acid hydrolysis resulted in formation of WGA-binding carbohydrate chains on Mr 115,000, 145,000, 190,000, and 900,000 components, but not on Mr 450,000 and 740,000 components, which indicated that these two sialoglycoproteins bore different oligosaccharides from the other sialoglycoproteins. The Mr 900,000 component was more prominent with HT-29 cells growing in nude mice than those growing in vitro. WGA binding to the Mr 900,000 component of metastasis-derived HT-29 cells growing in a nude mouse was higher than that of parental cells growing in nude mice. The expression in liver metastases derived from parental as well as metastatic cells was higher than the primary tumor growing in the spleen of the same mouse, indicating that the levels of Mr 900,000 sialoglycoprotein (SGP = 900) were regulated by intrinsic and environmental factors. The influence of organ microenvironmental factors was confirmed by analyzing sialoglycoproteins of HT-29 cells growing in the liver of a nude mouse following intrahepatic injection. Analyses of human colorectal carcinoma tissues and liver metastases revealed a polydisperse WGA-reactive high-molecular-weight component similar to that seen in tumors growing in nude mice. The mean value of WGA binding to high-molecular-weight glycoproteins in the primary tumors of stage B1 patients was smaller than that of all other primary tumors. Comparison of primary tumors with liver metastases from the same patients indicated that the level of SGP-900-like high-molecular-weight glycoproteins in metastases was not always higher than those in primary tumors.

Collagenolytic enzymes produced by tumor cells are believed to play a significant role in the destruction of surrounding normal tissue and, in certain experimental animal systems, the ability of tumor cells to degrade type-IV collagen (basement membrane collagen) correlates positively with those cells' metastatic capacity. We measured collagenolytic activity levels of extracts from freshly excised colorectal carcinoma tissues and of conditioned media from primary organ culture (total of 114 tissues from 53 patients) by using purified radiolabelled type-I (rat tail) and type-IV (mouse Engelbreth-Holm-Swarm [EHS] sarcoma) collagens. Both type-IV and type-I collagenolytic activity levels of extracts from tumor and adjacent mucosa ranged from less than 1 to 80 ng/hr/mg wet tissue, and no significant differences between mucosa and carcinoma tissues were observed. In conditioned media, the type-IV collagenolytic activity was low for normal mucosa and benign tumors and slightly higher for carcinoma than for mucosa. In 5 of 32 primary tumors, collagenolytic activity levels were 2-5 times higher than in the rest of the tumors and mucosal tissues. There were no significant differences in collagenolytic activity levels of conditioned media and tissue extract from colorectal carcinoma of different Dukes' stages. Deep and superficial areas of primary tumors released similar type-IV collagenolytic activity levels, suggesting that there was little intratumoral heterogeneity in the release of this enzyme.

Twenty patients with colon cancer metastatic to the liver underwent successful hepatic resection and adjuvant perioperative therapy that included hepatic arterial mitomycin C and floxuridine (FUDR). The median survival for all 20 patients was 51 months: 10 are still alive with a median postoperative follow-up of 49 months; 6 are disease-free with a median postoperative follow-up of 43 months. Among 10 patients in whom the surgical margins of the specimen contained tumor cells, the median survival was 52 months. This survival was comparable to that among 10 patients in whom the surgical margins were tumor free (P = 0.22). Neither the number of metastatic liver deposits nor the disease-free interval between the primary diagnosis of colorectal carcinoma and the development of liver metastases significantly affected survival. A transient chemical hepatitis which resolved before the next scheduled treatment was associated with 50% of arterial chemotherapy cycles (approximately 70% of the patients). Gastric or duodenal ulcerations occurred in 23% of the patients. Surgical complications were either pulmonary such as pleural effusion or atelectasis, or wound infections and subphrenic abscesses. Although these results compare favorably with the results in previously published series, this aggressive adjuvant chemotherapy appears to be particularly justified in patients with tumor positive surgical margins or those with multiple tumor masses and, therefore, are characterized by a poor prognosis.