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Twelve partial nephrectomies were performed in 12 piglets using either the combination (contact and non-contact) Nd:YAG laser technique or a steel scalpel. Additional haemostasis was attempted with ligatures. The renal artery was not clamped and renal cooling was not attempted. Total nephrectomy was performed on the contralateral side. Serum creatinine and urea levels were measured, and 99mTc-DTPA renography was performed pre-operatively and 1 and 2 weeks post-operatively. One week post-operatively the mean serum creatinine level was 35% higher than the pre-operative level in the laser group and 30% higher in the steel scalpel group. Two weeks post-operatively the respective differences were 34 and 24%. The mean urea level 1 week after operation was 50% higher than the pre-operative level in the laser group and 17% higher in the steel scalpel group. Two weeks post-operatively the respective differences were 38% in the laser group and 20% in the steel scalpel group. The mean DTPA disappearance rate was 34% lower 1 week after operation in the laser group and 23% lower in the steel scalpel group when compared with the preoperative state. Two weeks post-operatively the respective changes were 48 and 25%. These data indicate that there is no significant difference in renal function when the Nd: YAG combination laser technique is used in partial nephrectomy as compared with the steel scapel.

We have analysed the relationship between carcinoembryonic antigen (CEA) and DNA ploidy prospectively in 130 colorectal carcinoma patients. CEA was elevated preoperatively significantly more often in patients with DNA-aneuploid tumours than in DNA-diploid or DNA-tetraploid tumours--that is, in 48% (36 of 75) of patients with aneuploid tumours, in 34% (14 of 41) of patients with diploid tumours, but only in 14% (2 of 14) of patients with tetraploid tumours (p less than 0.05). Aneuploid tumours had an elevated CEA level in 38% of stage A-B disease and in 61% of stage C-D disease. The elevated CEA values (greater than or equal to 5.0 micrograms/l) correlated with tumour stage in patients with aneuploid tumours but not in patients with diploid tumours. Whereas CEA is a suitable marker for aneuploid carcinomas, other more sensitive tumour markers should be sought for diploid and also for tetraploid tumours. If such markers are found, flow cytometry could provide the most important information in selecting individual follow-up programmes for colorectal cancer patients.

The expression of tumour markers CA 19-9 and CA 50, defined by the monoclonal antibodies 1116 NS 19-9 (19-9 antibody) and C 50, was studied by the immunoperoxidase technique in formalin-fixed, paraffin-embedded tissue sections from 11 hepatocellular carcinomas and 10 cholangiocarcinomas of the liver, and from specimens of normal liver and liver cirrhosis. The 19-9 and C 50 antibodies react with sialosylfucosyllactotetraose, corresponding to sialylated blood group antigen Lewis, and the C 50 antibody also with another sugar moiety, sialosyllactotetraose. Neither marker was cancer specific. The CA 19-9 and CA 50 antigens are normal constituents of bile ducts. Nine out of 10 cholangiocarcinomas stained for CA 50, and eight out of 10 for CA 19-9. There was no apparent difference between the staining pattern of CA 19-9 and CA 50. Hepatocellular carcinomas were consistently negative for both markers. Thus, hepatocellular carcinomas and cholangiocarcinomas showed a clear difference in the reactivity for tumour marker antigens CA 19-9 and CA 50. This difference might be of clinical importance in the differential diagnosis between hepatocellular carcinoma and cholangiocarcinoma.

One hundred and eighty-seven patients with a transitional cell bladder cancer were followed by retrospectively for a mean of 9.5 years. Clinical stage (p less than 0.0001), histological grade (p less than 0.0001), papillary (p less than 0.0001), SPF (p less than 0.0001), M/V index (p less than 0.0001), MAI (p less than 0.0001), DNA index (p = 0.0001) SDNA (p = 0.0004), NA10 (p = 0.0023), NA (p = 0.0044) and G2% (p = 0.0158) were significantly correlated with survival in univariate analysis. In Cox's analysis T-category, papillarity and MAI had independent prognostic value and in combination they predicted bladder cancer-related survival significantly (X2 = 117.5, p less than 0.0001). When histological parameters only were analysed, WHO grade, SPF and papillarity were independent predictors and their combined prognostic significance was high (X2 = 76.6, p less than 0.0001). In papillary tumours SPF (p v 0.0005), MAI (p = 0.0009), DNA index (p = 0.0010) and M/V index (p = 0.0021) predicted survival significantly in addition to stage (P less than 0.0001) and grade (p = 0.0003) in univariate analysis. In Cox's analysis T-category, MAI and M/V index were independent predictors and their combined prognostic value was high (X2 = 54.1, p less than 0.0001). In Ta-T1 and in T2-T3 tumours, WHO grade, papillarity, SPF, mitotic indexes and DI were significant predictors in univariate analysis. In Cox's analysis of Ta-T1 tumours papillarity had independent prognostic value, whereas in papillary Ta-T1 tumours mitotic activity included all prognostic information. In T2-T3 tumours WHO grade and SPF were independent predictors. In conclusion, in papillary bladder tumours T-category is the most important predictor of survival followed by parameters reflecting proliferative activity of cancer cells.

DNA flow cytometric (FCM) data, histological features and clinical stage of bladder tumours in 222 patients were related to outcome during a mean follow-up of 10 years. Aneuploidy was detected in 82 (37%) of tumours and 140 (63%) were diploid. Intratumour heterogenity of DNA ploidy was found in 27% of 30 cases. Aneuploidy and high S-phase fraction (SPF) were related to clinical stage, histological grade and papillarity (P less than 0.0001). Aneuploidy (P less than 0.0001) and high SPF (P less than 0.0001) predicted metastasis to pelvic lymph-nodes and to distant sites. T category (P less than 0.0001), SPF (P less than 0.0001), histological grade (P less than 0.0001), papillarity (P = 0.0021), DNA aneuploidy (P = 0.0094) and G2 fraction (P = 0.0340) predicted cancer-related survival. Multivariate analysis showed DNA aneuploidy as the most important predictor of progression in T category (P = 0.0003) and T category was the most important predictor of lymph-node involvement (P = 0.0083) and metastasis (P = 0.0015), followed by FCM parameters. In Ta-T1 tumours SPF predicted progression independently (P = 0.0153). FCM offers more accurate prognostic information in Ta-T1 tumours than conventional methods. In invasive tumours, FCM offers quantitative prognostic information in terms of pelvic lymph-node metastasis and metastasis to distant sites.

The Nd: YAG contact laser technique with 12 W power output was used in partial nephrectomies in six piglets. Blood loss, total time and number of ligatures needed for haemostasis were compared in six partial nephrectomies performed with a steel scalpel. Mean blood loss was 53 g with the laser and 108 g with the steel scalpel (p less than 0.04). The mean time needed for haemostasis were 9.5 and 16.2 min respectively (p = 0.2) and the average number of ligatures required 1.5 and 8.8, respectively. The mean temperature rise at 5, 10 and 20 mm distance from the resection line was 5.3, 1.2 and 0.3 degrees C, respectively. The use of laser reduced blood loss and shortened the operation time. Further studies are needed concerning renal function after kidney surgery with a laser.

The relationship between the mean nuclear area, standard deviation of the nuclear area, the mean area of the 10 largest nuclei in a section (NA10), volume-corrected mitotic index (M/V index), DNA index (DI) and S-phase fraction (SPF) were assessed in a panel of 222 transitional cell bladder tumors. The relations between the morphometric variables and DI (p less than 0.0001), and between the morphometric variables and SPF (p less than 0.0001) were highly significant. Pearson's correlation coefficients between NA10 and DI (r = 0.413) and between SPF and the M/V index (r = 0.536) were also highly significant (p less than 0.0001). The SPF could be estimated reliably by morphometric methods, whereas the estimation of DI is not perfect. The combination of morphometric variables allowed a more reliable estimation of flow cytometric parameters. The results suggest that DI and SPF can be semiquantitatively estimated in routine histopathological sections by simple morphometric methods.

DNA flow cytometry (FCM) was performed from multiple samples in 60 transitional cell bladder carcinomas (TCC) to detect intratumoral heterogeneity of DNA index (DI). The samples for analysis were selected randomly and they were paraffin-embedded peroperative biopsy specimens from the primary TCCs. Intratumoral heterogeneity of DI was found in 12 of 60 (20%) samples. All WHO grade I tumors were diploid whereas 12 of 41 (29%) WHO grade II-III tumors showed a heterogeneous DI. DI heterogeneity was related to WHO grade (p = 0.004), mitotic frequency (p = 0.002), inflammatory cell infiltration (ICI; p = 0.003) and nonpapillar growth pattern (p = 0.06). In multivariate analysis, the WHO grade (p = 0.027) and the degree of ICI (p = 0.041) were independently related to heterogeneous DI. The results show that (a) many of the WHO grade II-III TCCs are composed of diploid and aneuploid cell populations; (b) the presence of dense ICI and TCCs may lead to false diploid DI. Consequently, the analysis of multiple samples is recommended in WHO grade II-III tumors to detect aneuploidy particularly when dense ICIs are present.

Five patients with severe acute pancreatitis (AP) underwent subtotal pancreatectomy, and six patients with advanced chronic pancreatitis (CP) were subjected to pancreatic resection. Microangiography and histological studies were performed on the resected pancreata. All patients with AP had histologically verified necrotizing pancreatitis. Pancreatic ducts in the necrotic areas had severe inflammation in their walls and a decrease in their vascularity. The ductal walls of CP patients were indistinguishable from the surrounding fibrosis and the vascular supply of the ducts was markedly diminished. The vessels were reduced in number, and their calibers varied considerably. Ductal ischemia in connection with AP and CP is discussed.

The results of DNA flow cytometry (FCM), histological features and clinical stage of prostatic adenocarcinoma were correlated to outcome in 91 patients during a mean follow-up period of 15.6 years. Aneuploidy was detected in 33 (36%) tumours, and 59 (64%) tumours were diploid. Eighteen (20%) tumours showed a tetraploid DNA index. The frequency of tetraploidy increased towards high-stage and high-grade tumours. Aneuploidy, high S phase fraction (SPF) and G2 fraction were significantly related to clinical stage, histological grade and perineural infiltration. Progressing tumours (T category) had higher SPF values (p = 0.0248), and progression in N (p = 0.0122) and M categories (p = 0.0021) was related to high G2 fraction as well. In T1-2 tumours, DNA ploidy (p = 0.0280) and SPF (p = 0.0230) predicted progression, whereas histological grade had no significant predictive value. The clinical stage (T) predicted crude survival (p = 0.0005). Results show that FCM gives prognostic information in prostatic adenocarcinoma beyond that of histological grading.

The DNA ploidy of pancreatic cancer tissue from paraffin blocks was measured by flow cytometry in 46 patients whose disease had been detected and treated with surgery. Lymph node involvement was observed at the time of diagnosis in 36% of patients with diploid tumors and in 79% of patients with aneuploid tumors (p = 0.017), but no clear relation to metastasis could be observed (p = 0.201). The S-phase fraction (SPF) was significantly higher in aneuploid than in diploid tumors (p = 0.007). All patients who underwent radical surgery had diploid DNA content and SPF below the median (11.5%). Seven patients with a diploid tumor (32%) and none of the aneuploid cases survived 1 year. Over the 1-year period, in order of importance, the type of treatment (p less than 0.001), DNA ploidy (p = 0.004), tumor size (p = 0.0046), and lymph node status (p = 0.027) predicted survival. Aneuploidy showed a significant association with decreased cumulative survival (p = 0.015), and a suggestive relationship with SPF was found. The results suggest that DNA ploidy of pancreatic cancer can be used in dividing the patients into different prognostic groups. The value of the detection of aneuploidy, however, is limited, because diploid pancreatic cancers are also generally rapidly fatal.

One hundred and fifty-seven patients with usual colorectal cancer were analysed prospectively for DNA-ploidy, DNA-index and S-phase fraction (SPF) using flow cytometry. An abnormal DNA-stemline was observed in 68% of tumours. The patients have been followed for a median of 36 months. In univariate analysis, tumour stage was the most significant prognostic factor. After excluding patients with stage D disease, DNA-aneuploidy was significantly associated with a shorter survival and a shorter disease free survival. SPF, however, did not correlate with prognosis. In multiple samples from the same tumour there was on average a 29% difference between the highest and the lowest SPF indicating considerable heterogeneity in proliferative activity within the tumours. In diploid tumours the variation was even higher. Patients with proximal tumours as well as female patients had DNA-diploid tumours more often than the others. This may indicate that there are different, so far unknown, aetiological factors leading to different types of ploidy pattern.

A multivariate prognostic index based on clinical data and the results of flow cytometry for the grading of breast cancer was evaluated in 117 patients whose disease had been detected and treated by mastectomy with axillary clearance between 1974 and 1976. Survival analysis with Cox's regression model pointed to three important prognostic factors: lymph node involvement (p less than 0.001), DNA ploidy (p less than 0.01) and tumour size (p less than 0.01). These factors were incorporated into a prognostic index, in which the lymph node involvement, DNA ploidy, and tumour size contributed to the index in that order. Logistic discriminant analysis with five year follow-up as the fixed end point (70 alive, 47 dead) gave the same result; lymph node involvement, tumour size, and DNA ploidy were the best prognostic indicators of survival. The result showed that our multivariate prognostic index was more powerful than lymph node involvement alone. The use of this prognostic index is recommended for selecting patients for different treatments.

In 83 bladder cancer patients with adequate follow-up (mean 13 years, range 9-22) the prognostic value of morphometric, DNA flow cytometric and clinical parameters was assessed. Paraffin embedded material was used in flow cytometry. Univariate life-table analysis showed the statistically significant relation of clinical stage, histological grade, mean nuclear area, the Standard Deviation (SD) of nuclear area, mean maximal nuclear diameter, mean nuclear perimeter and the volume corrected mitotic index (M/V index) to survival when bladder cancer deaths alone were used in the analysis. The recurrence of bladder cancer could be predicted with the M/V index. Survival analysis with Cox's regression model pointed to primary tumour clinical stage as the most important prognostic factor of crude survival. Histologically, grade and the SD of nuclear area were the best prognostic factors. Primary tumour stage and histological grade were the best predictors of death from bladder cancer. In Cox's model, histoquantitative methods are almost as good as clinical staging in predicting prognosis. DNA flow cytometry of paraffin embedded material offered no advantage over clinical stage, histological grade or morphology in assessing prognosis.

The nuclear DNA content of archival paraffin-embedded bladder cancer samples (70 patients) of WHO grades I-III has been measured by flow cytometry. The female/male ratio was 15/55. The mean follow-up time was 13 years (range 9.6-22.0 years). 37 of 70 (53%) patients had DNA index 1.0 (diploid DNA content), and the remaining 33 (47%) patients had an aneuploid tumor. There was no significant difference in the age (mean +/- SD) of the patients having a diploid (66 +/- 9 years) or an aneuploid tumor (68 +/- 11 years) at the time of diagnosis. 47 deaths occurred during the follow-up period; 24 (51%) of these were due to bladder cancer (12 diploid, 12 aneuploid tumors). No significant difference was found after radical treatment during the disease-free interval (mean +/- SD) between diploid (48 +/- 45 months) and aneuploid (35.5 +/- 35 months) groups of patients. Recurrences during the follow-up period were equally common among aneuploid and diploid tumors. A statistically significant relation between histological grade and survival could be demonstrated, but DNA ploidy and S phase fraction had little prognostic value in this respect. There was no statistically significant difference in survival between aneuploid (30%) and diploid (35%) groups of tumors during the follow-up period. The study suggests that flow cytometric determination of nuclear DNA ploidy from paraffin-embedded samples in bladder tumors does not add to the prognostic power of subjective histological grading.

The expression of the tumor marker antigen CA 50, defined by the monoclonal antibody (MAb) C 50, was studied by the immunoperoxidase technique in formalin-fixed, paraffin-embedded tissue sections from normal pancreata, from pancreata with pancreatitis and from benign and malignant pancreatic neoplasms. The results were compared with those obtained with Mab 1116 NS 19-9. The C 50 antibody reacts, like the 1116 NS 19-9 antibody, with sialosylfucosyllactotetraose (corresponding to sialylated blood group antigen Lewisa), but also with another sugar moiety, sialosyllactotetraose. Thirty-two of 37 well- to moderately-differentiated adenocarcinomas and all cystadenocarcinomas were positive for CA 50. The staining was most intense in the apical border of the cells, and in the intraluminal mucus. The number of positive cells was smaller in poorly differentiated adenocarcinomas and only occasional cells were stained in anaplastic carcinomas. In acute and chronic pancreatitis small terminal ducts, centro-acinar cells and some large ducts stained for CA 50. In normal pancreas only a few small terminal ducts were CA-19-9-positive, whereas both ducts and centro-acinar cells were C-50-positive. Normal pancreatic tissue adjacent to carcinoma usually stained more strongly for CA 50 than the carcinoma, whereas the opposite was true for CA 19-9. Eight out of 11 CA-19-9-negative carcinomas were CA-50-positive. Serous cystadenomas and malignant islet-cell tumors were focally positive for CA 50, but negative for CA 19-9. It seems apparent that the C 50 antibody reacts with another determinant than sialylated Lewisa in CA-19-9-negative specimens, serous cystadenomas and malignant islet-cell tumors. Serum CA 50 and CA 19-9 levels were determined in 29 patients with pancreatic cancer. The sensitivity was similar for both markers (76%), and there was a positive correlation between the serum levels. However, there was no correlation between the serum levels and the histological expression of the CA 50 and CA 19-9 antigens.

Five cases with multiple jaw cysts from two families, and two other unrelated patients are presented. Multiple keratocysts of the jaws were found in all patinets whose cysts had been examined histologically. There were also several follicular cysts without keratinization. In some cysts there was even a pronounced proliferation of the basal cells, which resembled basal cell carcinoma. Basal cell carcinomas were seen in only three patients, apparently because the series had been selected for the presence of multiple jaw cysts. However, two patients with only pigmented nevi showed a marked proliferation of the basal cells. An elevation of serum alkaline phosphatase was noted in four out of five patients. The sella turcica was bridged in all but one patient and fusion of the vertebrae was seen in all but two patients. In addition, a high frequency of rib anomalies, frontal and parietal bossing, mild mandibular prognathism and dural calcification was seen. Therefore, what is called the basal cell nevus syndrome may be regarded as a skeletal dystrophy syndrome. Based on the family pedigrees, an autosomal dominant inheritance with poor penetration seems probable.

The lysosomal storage material in cultured fibroblasts from patients with various lysosomal storage diseases was characterized by fluorescence microscopy using lectins specific for different saccharide moieties. In normal fibroblasts and cultured amniotic fluid cells lectins specific for mannosyl and glucosyl moieties, Con A and LcA gave a bright perinuclear cytoplasmic staining corresponding to the localization of endoplasmic reticulum in the cells. All other lectins stained the Golgi apparatus as a juxtanuclear reticular structure. In fucosidosis fibroblasts, only lectins specific for fucosyl groups LTA and UEA, distinctly stained the lysosomal inclusions. The lysosomes in mannosidosis fibroblasts did not react with Con A and LcA, both specific for mannosyl moieties of glycoconjugates, but were brightly labeled with WGA, a lectin specific for N-acetyl glucosaminyl moieties. In I-cell fibroblasts, the numerous perinuclear phase-dense granules, representing abnormal lysosomes, were labeled with every lectin used. In fibroblasts from patients with Salla disease, a newly discovered lysosomal storage disorder, the lysosomes were brightly stained only with LPA, indicating the presence of increased amounts of sialic acid residues in the lysosomal inclusions.

The expression of the gastrointestinal cancer associated antigen CA 19-9, defined by the monoclonal antibody 1116 NS 19-9, was studied by immunoperoxidase staining in routine formalin-fixed, paraffin-embedded tissue sections from normal pancreata, pancreata with pancreatitis and from benign and malignant pancreatic neoplasms. The formalin-fixed specimens were treated with pepsin, which enhanced the staining intensity. Eighty-five per cent of well to moderately differentiated adenocarcinomas were positive. The staining was most intense in the apical border of cells lining the lumina of malignant glands, and in mucus inside the lumina, but cytoplasmic staining was also seen. In poorly differentiated adenocarcinomas the number of positive cells was smaller and in anaplastic carcinomas only occasional cells were stained. All mucinous cystadenomas and cystadenocarcinomas stained intensely, whereas serous cystadenomas, and all benign and malignant islet cell tumours were negative. Ducts in chronic pancreatitis and in normal pancreata were positive in 96% and 79%, respectively, but the staining was focal and usually weaker than in carcinomas. In acute pancreatitis (92% positive) the staining was more intense, and the CA 19-9 expression was seen predominantly in small terminal ducts and in centroacinar cells. There was an apparent correlation between the degree of differentiation of the ductal adenocarcinomas and the expression of CA 19-9, whereas the correlation between tissue expression and serum levels of CA 19-9 was poor.

Guinea pig lymph node and blood lymphocytes have been physically fractionated in preparative cell electrophoresis into two functionally viable populations, the high mobility cell population (HMC) and the low mobility cell population (LMC). By using cell surface markers and functional tests known to be specific for T and B lymphocytes, respectively, it is shown that the T lymphocytes localize in the HMC population and the B lymphocytes in the LMC population. The spleen lymphocytes do not separate into the two populations. They move into one single broad peak containing both T and B cells. This finding indicates the presence of electrokinetically different subsets of T and B lymphocytes in the spleen on one hand and in the lymph node and blood on the other hand.

The correlation between the histopathological diagnosis of biopsies from colorectal villous tumours and that of the excised tumour (in 16 cases of carcinoma only clinical diagnosis, autopsy or death certificate) was investigated. In the persent retrospective study 235 biopsies were taken from 134 tumours where the final diagnosis had been established. In the benign groups the grade of the biopsy was the same as that of the surgically excised tumour in about 50%. However, 25% of the biopsies with a benign diagnosis were in fact taken from carcinomas. Not even multiple biopsies exclude the presence of cancer in a villous papilloma. The tumour should be totally excised and histologically examined.

The distribution of laminin, a basement membrane glycoprotein, was studied by immunohistological techniques in 10 samples of normal pancreatic tissue, in 15 samples of chronic pancreatitis, and in 33 pancreatic neoplasms. Sections of formalin-fixed, paraffin-embedded specimens were pretreated with pepsin and immunostained for laminin. As judged by the expression of laminin, normal pancreatic glands were surrounded by a continuous, intact basement membrane. In chronic pancreatitis the basement membrane was also mainly continuous, but focally weaker and thinner than around normal glands. In pancreatic adenocarcinomas laminin was irregularly distributed and in large areas totally absent. In anaplastic carcinomas no extracellular laminin was seen, but two cases showed some intracellular laminin in a punctate pattern. The findings suggest that these cancers have defects in the deposition of a basement membrane or that it is degraded. Our data suggest that the integrity of the basement membrane correlates with the degree of malignancy in ductal adenocarcinomas, but this is not the case for mucinous cystic neoplasms or for islet cell tumors. In these neoplasms a nearly intact basement membrane was seen both in malignant tumors and in their benign counterparts.

Changes in peritoneal morphology were examined histologically and by scanning electron microscopy during porcine acute hemorrhagic (n = 8) and edematous (n = 9) pancreatitis and after intraperitoneal installation of hemorrhagic pancreatitis-associated peritoneal exudate in healthy piglets (n = 3). In all experimental groups peritoneal inflammatory changes with mesothelial damage were evident already 1 h after the induction of the disease, and increased with time. Hemorrhagic pancreatitis caused desquamation of mesothelial cells and denudation of the basal membrane. Intraperitoneal installation of hemorrhagic pancreatitis-associated peritoneal exudate in healthy piglets caused similar changes, whereas the changes in edematous pancreatitis were much less extensive. Peritoneal exudate accumulating in the peritoneal cavity during hemorrhagic pancreatitis caused early chemical peritonitis characterized by severe inflammation of the peritoneum with destruction of the mesothelial cell layer, leading to denudation of the underlying connective tissue. The significance of these changes in the pathophysiology of acute fulminant pancreatitis remains to be further studied.

Isolated metanephrogenic mesenchyme can be induced for epithelial differentiation and tubule formation by an embryonic spinal cord that is cultivated in a transfilter position to the target cells. Under such conditions, determination of the cells starts at 12 hr and is completed around 24 hr. To correlate this event to DNA synthesis, thymidine incorporation was followed in such cultures from 2 to 96 hr after setting up the transfilter explants. Controls were cultured under conditions which did not result in induction. Incorporation into both induced and noninduced mesenchymes dropped rapidly during the first 12 hr when the tissues were transferred to the in vitro conditions. During the subsequent critical 12- to 24-hr period, mesenchymes continuously exposed to an inductor showed a marked increase in their incorporation of thymidine. If induction was interrupted during the first 10 hr there was no stimulation of incorporation when measured at 24 hr. Similarly, isolated mesenchymes and those exposed to a tissue devoid of inducing capacity did not show stimulation, and their incorporation remained at a low level when measured at 24 hr. Parallel DNA measurements were made by flow cytometry of individual nuclei from mesenchymes exposed to the inductor from 0 to 46 hr. No major changes in the distribution of the various phases of cell generation cycle were detected nor could any synchronization be shown. Hence the changes in the incorporation rate of thymidine seem to be due to variations in the total length of the cell cycle. We conclude that induction of differentiation of the metanephric mesenchyme is accompanied by a contact-mediated stimulation of DNA synthesis. The stimulation may represent a permissive effect on a predetermined cell population in the metanephrogenic mesenchyme.

Glomerular sialic acid was chemically measured in rats with experimental proteinuria induced by N,N'-diacetylbenzidine (DAB) or with autoimmune Heymann nephritis. In DAB nephrosis and in Heymann nephritis the relative amount of glomerular protein was increased. In DAB nephrosis the quantity of sialic acid expressed per amount of protein was decreased, but expressed per amount of DNA, which reflects the number of cells, there was no significant change. In Heymann nephritis the amount of sialic acid was not significantly altered when expressed per amount of protein or per amount of DNA. In individual animals, the amount of glomerular sialic acid expressed per amount of protein or per amount of DNA did not correlate with the severity of proteinuria. An increase in the total number of glomerular cells was found only in Heymann nephritis. The results suggest that proteinuria in the reported models is not the result of changes in glomerular sialic acid.

The serum and urine concentrations of a tumour-associated trypsin inhibitor, TATI, were determined by radioimmunoassay in patients with pancreatic cancer and with benign pancreatic and biliary diseases. Elevated serum levels (greater than 20 micrograms l-1) were found in 85% of the patients with pancreatic cancer, and elevated urine levels (greater than 50 micrograms g-1 creatinine) in 96% of the patients. Thus low TATI level, especially in urine, makes the possibility of pancreatic cancer less likely. Serial assay of TATI in serum from three patients with surgically removed pancreatic cancer showed elevation of the TATI level at the time of detection of recurrence. However, high serum and urine levels were also seen in pancreatitis and in benign extrahepatic cholestasis. Thus TATI is a sensitive, although not specific, indicator of pancreatic and biliary disease, but the use of TATI as a tumour marker in the primary diagnosis of pancreatic cancer is limited. Immunohistochemical staining of pancreatic lesions showed that half of the pancreatic tumours expressed TATI, but the pancreatic tissue adjacent to a carcinoma always stained stronger than the carcinoma. It therefore seems that the main source of TATI in serum and urine of patients with pancreatic cancer are the normal acini and not the tumour tissue. In pancreatitis the staining was intense and clearly stronger than in normal pancreas.

With monoclonal antibody reacting with both human and porcine Ia antigens, we demonstrate with immunoperoxidase staining that Ia expressing endothelial cells are found in Langerhans' islets as well as in exocrine tissue of the pig and human pancreas. However, neither endocrine cells in Langerhans' islets nor acinar or ductal cells do express the Ia antigen.

The human En(a-) blood group is a rare recessive trait. These erythrocytes lack the major membrane sialoglycoprotein (PAS1, MN protein, or glycophorin) and contain a decreased amount of PAS2 as demonstrated by radiolabeling of surface porteins and chemical techniques. A third glycoprotein, Band 3, contains two labeled oligosaccharide chains; the more complex oligosaccharide has a higher molecular weight in En(1-) cells than in normal cells. A fourth glycoprotein, PAS3, is present in usual amounts in En(a-) cells. Cells heterozygous for En(a) are intermediate in these respects. The sialic acid is decreased in En(a-) cells, but the total carbohydrate is similar in the different membranes. The glycolipids are present in normal amounts but are much more exposed to galactose oxidase in En(a-) cells than in normal cells.

The levels of tumour-associated trypsin inhibitor (TATI), CA 125 and CEA were measured in ovarian cyst fluids from 21 patients. TATI in cyst fluid was immunologically and physicochemically similar to the peptide originally isolated from the urine of a patient with ovarian cancer. Mucinous cysts contained significantly higher levels of TATI than did serous cysts. Immunohistochemically TATI was localized in the apical parts of cells of mucinous ovarian cysts. These results suggest that this tumour-associated peptide is actually produced by a tumour. Like TATI, CEA occurred at higher concentrations in mucinous than in serous cyst fluids, whereas CA 125 was found in higher concentrations in serous than in mucinous cyst fluids. The concentrations of these tumours markers in cyst fluids did not correlate with circulating levels of the same markers. In spite of the very high levels of all these tumour markers in benign cyst fluids, serum levels were normal or only slightly elevated. Clearly elevated serum levels occurred only in patients with malignant tumours. Cyst fluid levels of these tumour markers could not be used to distinguish between benign and malignant tumours.

Acute hemorrhagic pancreatitis was induced in 15 piglets, after which 8 of the piglets were treated with saline infusion only and 7 were treated with saline infusion and peritoneal lavage. Hemodynamic variables were measured hourly. Organ and peritoneal blood flow was determined at 0, 1, and 5 h with radioactive microspheres. Peritoneal morphology was studied at 0, 1, 3, and 5 h with light microscopy and scanning electron microscopy. According to the results, changes in cardiac output, mean blood pressure, and peritoneal blood flow and the peritoneal inflammatory reaction were similar in the two groups. However, a significant increase in heart rate and a significant decrease in the blood flow to the pancreas, liver, and spleen were observed in the saline group, in contrast to the lavage group. In addition, blood flow to the adrenal glands was significantly higher in the saline group after 1 h. In conclusion, peritoneal lavage prevented the increase in heart rate and the development of a significant decrease in pancreatic blood flow in experimental hemorrhagic pancreatitis. The significance of these findings remains to be further evaluated.

We evaluated 58 patients who were still alive more than ten years after operative treatment of gastric cancer. We reexamined their histologic specimens and compared them with those of matched paired controls of the same sex and age who had died of gastric cancer. Forty-two patients consented to a follow-up study. The age of the patients did not affect survival. For patients with gastric cancer, those with distal cancer or an ulcer simulating cancer had had a better prognosis. Forty percent of the patients had had an early gastric cancer. Only two patients had had lymph node metastases in regional lymph nodes, and macroscopic tumor growth through the serosa had been recorded in only four cases. In 23 cases, a distal resection had proved successful. No significant correlation between intestinal or diffuse types of cancer and prognosis was observed. One recurrence after ten years was found; in one case, there was a new cancer in the gastric remnant. In addition, biopsy specimens from two patients showed grave dysplasia. We suggest that throughout their lives annual follow-up examinations be performed in patients who have undergone radical operations for gastric cancer.

The experience with 35 patients with unilateral condylar hyperplasia is described. Possibilities of distinction between active and non-active cases are discussed as well as the histological findings. Different surgical procedures were used for treatment. Pain in the joint area is a prominent feature in this group. The treatment was effective in this regard in all patients.

Charged polymers, known to alter cell surface characteristics, reversibly inhibit the intercellular communication required in kidney tubule induction. This effect seems to be a function of their charge density. It is suggested that this effect is due to the formation of a molecular barrier between the interacting cell surfaces.

The DNA ploidy of breast cancer tissue from paraffin blocks was measured by flow cytometry in 117 patients whose disease had been detected and treated with surgery between 1974 and 1976. Patients with aneuploid tumours had positive axillary nodes and distant metastases more often than those with diploid tumours. Aneuploid tumours were more common in postmenopausal than premenopausal women. The S-phase fraction (SPF) was significantly higher in aneuploid than in diploid tumours and positive axillary lymph nodes were found in 26 per cent of the patients who had a tumour with a SPF below the median (4.8 per cent) and in 48 per cent of those with tumours with SPF values above the median. At the primary clinical investigation 2 per cent of the patients with diploid tumours and 6 per cent of those with aneuploid tumours had distant metastases. During the follow-up, the proportion of patients with distant metastases increased to 42 and 72 per cent, respectively. With a follow-up of 11.5 years, the DNA aneuploidy of the tumour showed a significant association with decreased survival. Thirty-three per cent of patients with diploid and 65 per cent of patients with aneuploid tumours had died from breast cancer during the follow-up (P less than 0.001). All patients with hypertetraploid or multiploid tumours died from breast cancer. High SPF values were associated more closely with distant metastases or death during the follow-up than low SPF values. Our results suggest that DNA ploidy measured by flow cytometry from paraffin embedded tissue blocks of human breast cancer can be used to predict the aggressiveness of the tumour and the survival of the patients.

The nuclear DNA content of 38 malignant and 25 benign bone tumours was measured by flow cytometry. The specimens were taken either from biopsies or from surgical specimens. Seventeen of 26 primary malignant bone tumours were aneuploid, 15 had a single aneuploid DNA content, and 2 had a biclonal abnormality. Thirteen of 15 osteosarcomas were aneuploid, but only 2 of 6 chondrosarcomas showed an aneuploid DNA content. Six of 12 metastatic malignant bone tumours were also aneuploid. All 25 benign tumours had a diploid DNA content. Cell cycle analysis showed that the proportion of cells in S- and G2M-phases was higher in the malignant compared to benign tumours, indicating a higher proliferative activity. The increase was statistically significant (p less than 0.05) both in diploid and in aneuploid tumours. Among five tumours studied after chemotherapy, four displayed a marked hyperdiploid abnormality. Aneuploidy and high proliferative activity both were highly associated with malignant bone tumours, suggesting that DNA flow cytometry may be an adjunct in the assessment of malignancy of bone tumours.

The induction of kidney tubules in metanephric mesenchyme has previously been shown to require close contact between the interacting tissues. In our study we show that low concentrations of inhibitors of RNA, DNA and protein synthesis inhibit tubule induction, although they do not seem to prevent the formation of contacts between the interacting tissues. The effective concentrations were about the same as those which inhibited the synthesis of macromolecules. Cycloheximide caused an increased synthesis of RNA. Low concentrations of Mitomycin C inhibited DNA synthesis but not tubule formation. A concentration of the inhibitors which caused weakened induction also caused a marked decrease in leucine incorporation. We concluded therefore, that a decrease in protein synthesis in the inducing tissue is responsible for the inhibition of induction.

Chronic pancreatitis was induced in 22 piglets by dividing all pancreatic attachments to the duodenum; five sham-operated piglets served as controls. Two piglets died of postoperative complications. The animals were autopsied 2, 4, or 6 weeks postoperatively. All operated animals developed chronic pancreatitis. Concomitant with the development of interstitial fibrosis, an increasing progressive atrophy of the exocrine parenchyma occurred, with preservation of the islets of Langerhans. This atrophy and fibrosis were considerable already after 2 weeks. In one piglet only there was some acute inflammation and fat necrosis, whereas all showed at least moderate chronic inflammation, which did not change with time. The growth of the piglets stopped, and all had diarrhoea, which was thought to reflect exocrine insufficiency. Two animals (9%) developed a large pancreatic pseudocyst, and all animals had wide pancreatic ducts. The endocrine function was undisturbed. Intravenous glucose tolerance tests showed that the animals did not become diabetic. This model is appropriate for the study of experimental pancreatitis.

A 34-year-old woman with xanthogranulomatous pyelonephritis mimicking a renal tumour is presented. Preoperative examinations including renal ultrasonography, computed tomography, renal angiography and fine needle biopsy could not exclude malignancy. The diseased kidney had good function and there was no history or signs of urinary tract infection or calculi. During operation a frozen section biopsy showed xanthogranulomatous pyelonephritis and a kidney-saving enucleation of the tumour was performed. The patient recovered uneventfully with good kidney function on the operated side.

The effects were studied of two antipsoriatic drugs, dithranol (anthralin) and butantrone on the proliferation of HeLa cells. Non-confluent monolayers of HeLa cells were treated with various concentrations (10(-5)-10(-8) M) of dithranol or butantrone for 45 min. The cells were then cultivated for up to 48 h and treated with 10 microM bromodeoxyuridine (BrdU) and 10 microM fluorodeoxyuridine for 30 min before harvesting. Both drugs at concentrations of 10(-6) and 10(-5) M inhibited the BrdU uptake of HeLa cells, but did not cause any significant change in the proportion of cells in S-phase. The inhibitory effect of dithranol on cell number was linear with concentration 24 h after incubation. The highest (10(-5) M) concentration of butantrone caused an abrupt decrease in cell number.

The presence of cytoplasmic material inside thin Millipore filters between interacting mouse metanephric mesenchyme and spinal cord was investigated using different fixation methods. The transmission of induction was studied from sections of Zenker-fixed paraffin-embedded explants. Formation of kidney tubules was taken as evidence for induction. Filters with 0.8 and 0.22 mum pores permitted induction, whereas only 6 out of 31 filters with 0.1 mum pores did so. Glutaraldehyde-fixed Epon-embedded explants were used for study of cytoplasmic penetration into filters. In thick sections, filters with large pores were seen to contain cytoplasmic material at all levels. Filters with 0.1 mum pores usually showed only shallow ingrowth, but those which had permitted passage of induction contained material at least half way from the spinal cord and shallow ingrowth from the mesenchyme. With 0.8 mum filters the ingrowths from both sides met first after 18 h of transfilter cultivation. This has previously been shown to be the minimum time needed for induction to take place in this system. In electron microscopy cytoplasmic processes were seen deep inside the 0.8 and 0.22 mum filters regularly permitting induction. In small pores such material was only preserved by certain glutaraldehyde fixatives. Diffusion studies did not reveal major differences between induction-permitting 0.22 mum filters and induction-preventing 0.1 mum filters. Thus in the kidney tubule induction system this and our previous work speak in favor of a mechanism based on close apposition of cells rather than on long-range diffusion of inductive substances or on matrix interaction.

The distribution of laminin, a basement membrane glycoprotein, was studied by an immunohistochemical technique in six samples of benign sclerosing lesions of extrahepatic bile ducts and in 11 sclerosing cholangiocarcinomas. The expression of laminin showed that benign glandular structures were surrounded by a mainly intact basement membrane. In sclerosing cholangiocarcinomas laminin was irregularly distributed, and in large areas totally absent. The findings suggest that deposition of basement membrane material is defective. Sclerosing cholangitis may be extremely difficult to distinguish from well differentiated sclerosing cholangiocarcinoma. This sample was small, but the diseases studied were relatively rare. Staining for laminin might be of additional use to surgical pathologists in the differential diagnosis of benign sclerosing lesions and sclerosing cholangiocarcinomas.

A change in cell surface charge density during early avian development is shown with a free-flow electrophoresis apparatus. The blastoderms of freshly laid eggs consist of two electrophoretically distinct cell populations. After the onset of gastrulation a third cell population with an intermediate electrophoretic mobility appears. With increasing time of incubation there is a shift in the proportion of these populations and an increase in the mobility of the fastest fraction.

The thick sialoglycoprotein coat of the glomerular epithelial cells is a part of the glomerular filtration barrier. It can be demonstrated in paraffin histoslogy by means of the colloidal iron procedure. We have studied the special requirements of tissue processing to obtain reproducible results by this staining method. The requirements include proper fixation, avoiding heat during sectioning and ample rehydration. We have further tested a colloidal iron-PAS procedure for simultaneous demonstration of the glomerular epithelial coat and the basement membrane.

The DNA ploidy of breast cancer tissue from paraffin blocks was measured by flow cytometry in 122 patients. In this material there was a difference in lymph node involvement and in the presence of distant metastases between diploid and aneuploid tumors. Diploid tumors were smaller than aneuploid tumors. Aneuploid tumors were more common in postmenopausal than in premenopausal women. Near tetraploid tumors were found in older patients rather than tumors with other ploidy patterns. The ploidy pattern was not associated with survival during the mean follow-up of 4.1 years. We specially studied the S-phase fraction (SPF) which was distinctly higher in aneuploid tumors than in diploid tumors. Also near tetraploid, hypertetraploid and multiploid tumors showed higher SPF than diploid tumors. The median of SPF in our material was 8.5%. Positive axillary lymph nodes were found in 32% of the patients who had tumors with an SPF below the median and in 46% of those with tumors above the median (SPF greater than 8.5%). The difference, however, was not statistically significant. In our material the SPF of the tumor did not show a significant association with survival. However, longer follow up time is needed for firm conclusions on the predictive value of cell DNA on survival.