Search

Human papillomavirus (HPV) detection in biopsies from the lower genital tract may be requested by clinicians as a complement to ordinary histopathological diagnosis. In the present study, two commercial kits, (Enzo Diagnostics Inc., New York, USA and Biohit, Helsinki, Finland) used for in situ hybridization with biotinylated c-DNA probes were compared and the HPV-expression was evaluated in relation to histopathological findings. The Enzo kit identifies HPV-types 6/11, 16/18, 18 and 31/33/51, whereas the Biohit kit has separate probes for HPV 6, 11, 16, 18, 31 and 33, but none for HPV 51. The usefulness of a general probe (probemix) for the visualization of HPV irrespective of type (Enzo Diagnostics Inc.) was also studied Altogether 226 biopsies from the lower female genital tract were formalin-fixed, paraffin-embedded and processed for routine histopathological grading. Consecutive sections were employed for in situ hybridization. 50 biopsies were subject to double-testing with Enzo and Biohit, whereas 176 were tested with Enzo only. Of the double-tested biopsies, 30% displayed a nuclear staining with the Enzo kit and 28% with the Biohit kit. It is concluded that the probes of these two kits have the same sensitivity in detecting HPV in tissue sections. Condylomata acuminata were HPV-positive in 81%, mostly for types 6/11. Flat condylomas were HPV-positive in 35%. The HPV-positivity of biopsies with low grade SIL (I) was 50% and that of high grade SIL (II and III) was 37%. High grade SIL contained either HPV-types 16/18 or 31/33/51. A correlation was found between the occurrence of koilocytosis and the presence of HPV-DNA. HPV-expression was most easily visualized in condylomata acuminata. In epithelium of normal appearance or with inflammatory alterations HPV-DNA was not seen.

Among 33 patients with endocrine pancreatic tumors due to multiple endocrine neoplasia type 1 (MEN-1), 19 (58%) patients had hypergastrinemia, 7 (21%) patients had hyperinsulinism, and 7 (21%) patients had clinically non-functioning lesions. At least one gross tumor was found in all patients undergoing pancreatic surgery, including those with negative localization studies prior to operation. The patients also had additional macroscopic tumors as well as numerous microadenomas, and the lesions frequently were positive for immunostaining with multiple hormones, mainly pancreatic polypeptide, insulin, glucagon, and somatostatin. Duodenal endocrine lesions were found in 4 of 5 investigated patients and stained with gastrin and somatostatin antibodies. Distal, mainly subtotal pancreatic resection, was performed in 18 patients, eventually combined with caput tumor enucleation or duodenotomy, while a few patients underwent only tumor enucleation or a Whipple procedure. The long-term outcome of operation was most favorable in patients with hyperinsulinism; only 1 patient had clinical recurrence. Patients with hypergastrinemia experienced only transitory lowering of serum gastrin values after pancreatic surgery and 47% of them had or developed metastases. Such tumor spread was seen in 57% of the patients with non-functioning lesions. Nine patients died from progressive tumor disease during follow-up. Consistent with previous studies, we found that surgery is indicated in MEN-1 patients with hyperinsulinism even if a lesion is not visualized by radiology. In addition, these indications should be extended to also include patients with only biochemical markers of disease, including elevations of gastrin, as these indicate the presence of gross tumors.(ABSTRACT TRUNCATED AT 250 WORDS)

Applying the WHO criteria for the histopathological diagnosis of medullary thyroid carcinoma (MTC)--as well as the criterion that a significant amount of argyrophil cells, amyloid deposits, or calcitonin (CT) immunoreactive cells shall be present--122 cases were identified from the files of the Swedish Cancer Registry. Both non-occult (n = 110) and "occult" (less than 1 cm in diameter) (n = 12) MTCs were included. Both primary tumours (n = 91) and metastatic lesions (n = 31) were investigated. The specimens available were all only conventionally formalin-fixed and paraffin-embedded. The presence of neoplastic cells immunoreactive with antisera against chromogranin A (Chr A) was compared with that of the other three MTC markers. Chr A immunoreactive cells were present in practically all the cases. Similar results were obtained when the argyrophil reaction alone and CT immunoreactivity alone were used as markers. When two of the three MTC markers were combined, it was found that virtually everyone of the 122 tumours could be identified as a MTC. In contrast, the presence of amyloid deposits was found to be a less constant MTC marker; whereas 94% of the primary tumours had amyloid deposits, they were present in only approximately 70% and 60% of the metastatic and "occult" tumours respectively. No differences in the staining reaction patterns were found between familial (n = 18) and the sporadic (n = 104) types of MTC.(ABSTRACT TRUNCATED AT 250 WORDS)

The expression of a proliferating antigen by Ki-67 immunohistochemistry was evaluated in 32 gastrointestinal carcinoids and in 5 pancreatic islet cell tumors. In the tissue sections the number of labelled nuclei was calculated per tumor area. The tumors were classified as low proliferating (less than 0.3 labelled cells/mm2), medium proliferating (0.3-1 labelled cells/mm2), and high proliferating (greater than 1 labelled cell/mm2). In 26 tumors obtained from patients receiving antitumor therapy (alpha-interferon) the proliferative activity was decreased. In treated midgut carcinoids the proliferative activity in metastatic tissue was significantly reduced (p less than 0.05). Though not statistically significant, primary midgut carcinoids collected from untreated patients displayed a lower proliferative activity than liver metastases. A survival analysis revealed that patients with tumors displaying low proliferative activity had a better survival than those with high proliferative activity (p less than 0.05). Single cell cytofluorometric DNA analyses showed regular diploid stem cell lines in the majority of tumors from untreated patients (9/11 cases). No correlation was found between the calculated proliferative activity and the DNA profile. The obtained results indicate that the expression of a proliferation antigen by Ki-67 immunohistochemistry can be used to evaluate the biological behavior of neuroendocrine tumors of the digestive system and predict survival.

A 50-year-old man developed a pancreatic islet tumor with liver metastases. High levels of islet amyloid polypeptide (IAPP) were recorded in plasma-35,000 pmol/L-concomitant with the occurrence of type II diabetes mellitus (the clinical syndrome has recently been described in detail) [25]. Light microscopically, the tumor contained considerable amounts of amyloid and displayed IAPP immunoreactivity both in the tumor cells and in the amyloid stroma. Electron-microscopical examination of the liver metastases showed presence of round secretory granules in the tumor cells. The granules were immunoreactive to chromogranin A and B and IAPP but not to insulin. The amyloid deposits were mainly accumulated in the extracellular spaces but were also present in the tumor cell cytoplasm. The intracellufar amyloid fibrils were, as revealed by immunogold labeling, IAPP immunoreactive and seemed to emerge from the secretory granules in the shape of radiating threads. The results show that in this particular case, the amyloid formation started already at the intracellular level and in close proximity to the lAPP-storing secretory granules. The findings may have some significance for understanding the development of pancreatic islet B-cell-related amyloidosis in type II diabetes mellitus.

Time trends in the incidence of malignant neoplasms of the thyroid were examined for Sweden for the period 1958-81, 5,838 cases being studied. Autopsy-diagnosed cases were excluded. Mean annual changes in the age-standardized rates over this period were 1.9% and 1.2% for women and men, respectively, for all thyroid carcinomas combined; 4.9% and 2.1% for papillary carcinoma; 0.9% and 2.1% for follicular carcinoma; and -1.0% and -2.1% for anaplastic carcinoma. Age, period and cohort models were fitted. There was a progressive increase in papillary cancer incidence for cohorts born since 1919. This increase cannot be satisfactorily explained as an artefact of trends in classification, and probably reflects a real increase in incidence. A less marked increase was seen for follicular cancer, and there was a decline for cohorts born since 1939. While the risk of papillary and anaplastic carcinoma was lower in iodine-deficient areas, follicular cancer risk was twice as high in these areas, for men only. There was a steady decline of anaplastic cancer in both sexes after 1965, largely attributable to a decline in cohorts born since 1924.

Abnormally expressed oncogenes are implicated in neoplastic transformation. We have investigated a series of endocrine tumours using immunocytochemistry as a first screening tool to detect oncogene expression. Paraffin sections of 44 pulmonary small cell carcinomas, 15 pulmonary atypical carcinoids, 12 bronchial carcinoids, 28 medullary thyroid carcinomas, 27 phaeochromocytomas, and 17 insulinomas were immunostained with antibodies to c-erbB-2, c-myc, L-myc, and N-myc. Diffuse immunoreactivity was detectable for c-erbB-2 in 8 out of 44 (18 per cent) pulmonary small cell carcinomas, 3 out of 15 (20 per cent) pulmonary atypical carcinoids, and 6 out of 27 (22 per cent) phaeochromocytomas; for c-myc in 18 out of 44 (41 per cent) pulmonary small cell carcinomas and 5 out of 15 (33 per cent) pulmonary atypical carcinoids; for N-myc in 6 out of 28 (21 per cent) medullary thyroid carcinomas; and for L-myc in 4 out of 27 (15 per cent) phaeochromocytomas. There was considerable intratumoral and intertumoral heterogeneity and, in each tumour group, no relationship was found between tumour pattern, mitotic index, and oncoprotein immunoreactivity. These results suggest that oncogene products are present in a proportion of endocrine tumours, and that specific oncoproteins seem to be related to tumour type but not to other histopathological findings. Thus, oncoprotein detection may be a useful tool for identifying subsets of endocrine tumours that are not otherwise recognizable morphologically.

Histochemical and immunohistochemical investigations were performed on tissue obtained from the right heart side in three patients subjected to valve, replacement operations because of severe carcinoid heart disease. Extensive fibrotic changes were present on the endocardium of the right atrium, the papillary muscles of the tricuspid valve and the leaflets of the tricuspid and pulmonic valves of all patients. The main constituent of the lesions was a stroma with abundant acid mucopolysaccharides and collagen but devoid of stainable elastic components. The lesions were in some areas sharply delineated from the normal endocardium, but often also extended into the endocardium and myocardium. Small to medium sized vessels were demonstrated histochemically in the lesions and confirmed by positive immunoreaction against endothelial and smooth muscle cells. The moderate number of mesenchymal cells within the lesions had immunoreactivity consistent with muscle cells which seemed to have a very low proliferating activity. The histochemical and immunohistochemical techniques used confirmed some earlier observations in carcinoid heart disease but also rendered new information contradicting previous findings. The infiltrative nature of the carcinoid plaque gives a new dimension to the carcinoid heart disease. The etiology still remains obscure and well known growth factors for connective tissue such as platelet derived growth factor (PDGF) do not seem to be directly involved in the process.

The distribution of the mRNAs for chromogranin A and B was analyzed by in situ hybridization with 35S-labeled oligonucleotide probes in formalin-fixed paraffin-embedded carcinoid tumor tissues. All the 15 mid-gut carcinoid tumors examined contained both mRNAs for chromogranin A and B at high level in tumor cells. Sixteen of 18 bronchial carcinoid tumors but only 2 of 5 rectal carcinoid tumors expressed one or both species of chromogranin mRNAs. The same tendency was seen with the argyrophil reaction according to Grimelius where most of the mid-gut tumor cells were uniformly stained, while considerable variation in reactivity was seen in some of the bronchial and rectal carcinoid tumor cells. The sequential sections were stained with a monoclonal antibody against chromogranin A and a polyclonal antiserum which reacts with both chromogranins. The expression of the mRNA for chromogranin A on the carcinoid tumors was almost concordant with that of chromogranin B as well as with the chromogranin A protein, whereas almost all tumors stained positively with the polyclonal antibodies. Analyses of mRNA expression of chromogranin A before and after interferon therapy on 4 patients with mid-gut carcinoids indicated an inhibition at pre-translational level. In conclusion, the mRNAs for chromogranin A and B are good markers for the carcinoid tumors, especially of mid-gut origin. Fore-gut, mid-gut and rectal carcinoid tumors are different in their endocrine properties regarding the expression of the chromogranins.

The classical (mid-gut) carcinoids of the intestinal tract display a characteristic light microscopic morphology. However, sometimes intestinal tumours are seen resembling carcinoids and differential diagnostic difficulties arise. In the present study silver stains and immunoreactivities to chromogranin A + B, cytokeratins and epithelial membrane antigen (EMA) were evaluated as diagnostic adjuncts in six classical carcinoids and six intestinal carcinomas with carcinoid-like features. All classical carcinoids were argentaffin and argyrophil and contained a majority cell population with chromogranin immunoreactivity while only one carcinoid-like carcinoma was chromogranin-immunoreactive and the stained cells in that case represented a minority of the tumour cell population. The cytokeratins were shown to be non-discriminatory. However, EMA expression occurred in five intestinal carcinomas and in the majority of the tumour cells of four of these cases, while only one classical carcinoid displayed a few EMA positive cells. Thus, silver stains in combination with chromogranin A + B and EMA appears to be of value to discriminate between classical carcinoids and carcinoid-like intestinal carcinomas. Further when intestinal carcinoids and carcinoid-like carcinomas are diagnosed with the aid of various tumour markers both qualitative and quantitative considerations must be made.

The chemical background of silver stains used for visualization and characterization of peripheral neuroendocrine cells in the gastrointestinal tract and pancreas, and of their corresponding tumours, was studied in tissue sections and by a dot-blot technique. Sequential staining of pancreatic islets with an immunohistochemical procedure and silver staining of the same tissue section revealed that chromogranin A immunostained cells also displayed an argyrophil reaction with the Grimelius method, but no argentaffin reaction with the Masson technique. Accordingly, purified chromogranin A (15 micrograms or less) treated in formalin and applied to nitrocellulose did not show any argentaffin reaction but displayed a dose-related argyrophil reaction. Equal quantities of other polypeptide components did not give rise to any silver reaction. Further dot-blot studies showed that the tryptophan and tyrosine metabolites, dopamine, norepinephrine, 5-hydroxytryptamine and 5-hydroxinodole caused strongly argentaffin and argyrophil reactions while epinephrine, 5-hydroxyindole-3-acetic acid and 5-hydroxytryptophan gave only the former reaction. Among other chemical components studied, only guanine displayed weak silver staining. The results indicate that the reaction products between aldehydes and the granular content of biogenic amines synthesized from tryptophan and tyrosine display an argentaffin reaction and that the granular chromogranin A caused an argyrophil but no argentaffin reaction.

In 21 patients ultrasound-guided cutting biopsies, from carcinoid metastases of the liver, were taken before and after therapy with alpha-interferon. Each biopsy was examined under light microscopy and the amount of tumor tissue and connective tissue was quantified and then correlated to objective response to interferon therapy. A significant reduction of the amount of tumor tissue, in spite of unaltered metastatic size and a corresponding increase in connective tissue, was seen after interferon therapy. A more pronounced reduction of tumor tissue occurred after long-term interferon therapy. A positive correlation between objective therapy response and tumor tissue reduction was also present. Patients responding poorly, or not at all, to therapy did not show any significant decrease in tumor tissue. Since treatment with immune response modifiers is expected to increase in the near future, it is important to choose the right investigations for therapy monitoring, and since all patients in this investigation had unchanged tumor size on repeated radiological examinations, it is obvious that microscopic examination of core biopsies is a better method for evaluating effects of long-term therapy than tumor size measurement with radiological techniques. Further, the results may indicate that interferon exerts a cytotoxic effect on carcinoid tumor cells in vivo.

Three monoclonal IgG 2a antibodies were produced after immunization of mice with dispersed cells from a human mid-gut carcinoid tumor. Acetone-fixed cryosections of 57 primary and metastatic mid-gut carcinoid tumors as well as 2 hind-gut (rectal) carcinoids showed a conspicuous immunoreaction while a thymic carcinoid was essentially unstained with the antibodies. The 3 antibodies yielded a similar pattern of immunostaining. The immunoreaction comprised more than 95% of the carcinoid tumor cells, and it was more uniform and intense in primary tumors than in mesenteric, hepatic, and ovarian metastases of the mid-gut carcinoid tumors. Immunofluorescence studies on suspended carcinoid tumor cells showed that the antibodies bound to the surface membrane of the cells. The antibodies immunostained enterocytes of the small and large bowel, intestinal metaplasia of the stomach mucosa as well as colorectal adenocarcinomas. Endocrine pancreatic tumors producing vasoactive intestinal polypeptide, gastrin, somatostatin, and/or pancreatic polypeptide as well as the epithelium of pancreatic ducts were also stained with the antibodies, whereas a large number of other normal and abnormal human tissues, including benign and malignant insulinomas, were unreactive. The findings indicate that the antibodies recognize differentiation antigens on the carcinoid tumor cell surface preserved also on endocrine and nonendocrine cells of the normal bowel mucosa. The restricted tissue reactivity of the antibodies suggests that they may constitute useful tools in the histological characterization of carcinoid tumors. Further studies may reveal if they are applicable for immunolocalization and perhaps even immunotherapy of these neoplasms.

Effects of 5 weeks streptozotocin treatment on the mouse gastrointestinal tract were studied with special emphasis on enterochromaffin (EC) cells. In the streptozotocin-treated animals the frequency of EC cells was reduced in the antral area and in the small intestine, but increased in the colon. The length of the intestinal tract and the mucosal thickness were increased in these animals. Possible mechanisms underlying these abnormalities of the EC cell system are discussed.

Fourty-six bronchial carcinoids, twelve tumourlets and twenty areas of neuroendocrine cell dysplasia (NED) were immunohistochemically evaluated for various neuroendocrine markers, S-100 protein (S-100), myelin basic protein, intermediate filaments, actin, Leu-7 and several neurohormonal polypeptides. Eighteen of the bronchial carcinoids (39.1%) showed a biphasic cell pattern, with abundant stellate-shaped S-100 positive cells (SC). SC were not reactive for chromogranin A, myelin basic protein, cytokeratins, neurofilaments, glial fibrillary acidic protein or actin, and were only occasionally weakly positive for vimentin. SC were not detected in the tumourlets nor in the NED observed. For comparison a group of other neuroendocrine tumours (11 gastrointestinal carcinoids, 4 pheochromocytomas and 4 paragangliomas) were immunostained for S-100, chromogranin A and actin. SC similar to the ones detected in the bronchial carcinoids could be detected in appendiceal carcinoids, paragangliomas and in two out of four pheochromocytomas. Our present data are in keeping with a Schwannian/sustentacular nature of SC rather than that of a histiocytic or myoepithelial nature. We suggest that SC-rich bronchial carcinoids are biphasic tumours, which could be designed "paraganglioid" bronchial carcinoids. The relationship between SC-rich bronchial carcinoids and tumourlets/NED is a matter of further investigation: SC-rich bronchial carcinoids may either differentiate in a biphasic pattern during tumoural growth or may not be histogenetically related to tumourlets.

Twenty-four patients with gastric carcinoids were examined retrospectively with regard to the presence of achylia and pernicious anaemia (PA). Six patients (25%) had achylia preoperatively and half of these had an associated PA. The gastric carcinoids occurring in an achylic stomach were almost exclusively located to the corpus area and tended to be multiple. Further, the majority of cells in most of them were argyrophil (Sevier-Munger) positive, indicating that they were of the enterochromaffin-like cell type. At present, achylia with or without concomitant PA appears to be the most distinct condition associated with gastric carcinoids. The pathogenesis of these tumours which develop in an achylic stomach is discussed.

The prevalence of gastric carcinoid in fundic atrophic gastritis is probably greater than previously recognized. To help elucidate the clinicopathology of this syndrome, we report a series of 11 patients with solitary or multicentric carcinoid tumors. In these patients, basal gastrin levels and density of fundic mucosal endocrine cells were greater than that for patients with uncomplicated fundic atrophic gastritis (p = 0.02 and p = 0.002, respectively). The polypoid tumors, of which the largest measured 30 mm, frequently showed characteristic endoscopic features. They were all situated in the fundic mucosa, which showed micronodular endocrine cell hyperplasia. Small, endoscopically evident tumorlets, or "early carcinoids," limited to the lamina propria were observed in some patients. These lesions may represent intermediate stages between micronodules and invasive carcinoids, all of which infiltrated at least into the muscularis mucosae of the gastric wall. Although some consistent characteristics features were noted, there were structural variations. The cells were argyrophil but nonargentaffin and did not stain with conventional mucus stains. They did not stain significantly for carcinoembryonic antigen (CEA). The secretory product of these tumors remains to be identified. Ultrastructurally, some tumors were mainly composed of enterochromaffinlike (ECL) cells, but in other tumors most of the cells could not be classified.

The usefulness of argentaffin (Masson) and argyrophil (Davenport, Sevier-Munger and Grimelius) staining methods for identification of endocrine cell types in the gastrointestinal tract and pancreas is discussed and comments are made on the techniques themselves. The applicability of silver impregnation methods in the histopathological investigations of endocrine tumours in the above-mentioned organs is outlined, and the chemical background of the silver reactions is briefly reviewed.

A case of thymic carcinoid appearing in a 56-year-old man is presented. No endocrine disturbances were apparent. Mediastinal lymph node metastases were found at operation. The tumour was positive with Grimelius argyrophil silver stain. Ultrastructurally neurosecretory granules were seen.

The accuracy of various techniques for preoperative location of adrenal lesions was studied in 55 consecutive cases, and the perioperative course and outcome of surgery were analyzed. CT correctly located 94% of all adrenal tumours, was less accurate (42%) in hyperplasia and gave no false-positive results. Selective angiography revealed 75% of the tumours, but was likewise diagnostically inadequate in adrenal hyperplasia. Adrenal venous sampling correctly located 80% of tumours and of hyperplasia. In hyperplasia it was the single most successful procedure for detecting the site of adrenal hyperfunction. Phlebography gave misleading information in almost 45% of cases. The perioperative complications included splenic rupture necessitating splenectomy (3 cases), superficial wound infection (2), pneumonia (4) and subphrenic abscess (1). There were no perioperative deaths. The study indicated CT to be the principal procedure for preoperative location of adrenal disorders. In some cases with adrenal hyperfunction, venous sampling should be added. Adrenal surgery can be performed with low morbidity and no mortality and with favourable long-term outlook in cases of benign lesion.

The immunohistochemical expression of blood-group substances and carcinoma-associated antigens were compared in rectal carcinoids and adenocarcinomas. Rectal carcinoid tumors, in contrast to rectal carcinomas, were consistently negative for CEA, gastrointestinal cancer antigen GICA (or CA 19-9), and carcinoma-associated antigen CA-50 (except in one case where less than 10 percent of the cells expressed CA-50). The carcinoids and rectal carcinomas extensively expressed blood-group substance A, B, and H, Lewis B antigen, and difucosylated carbohydrate antigens (DFCA). Thus, rectal carcinoids and adenocarcinomas possess both similar and different tumor antigen profiles. The occurrence of discrepant antigen determinants may be used in the differential diagnosis of these two types of tumors. The coexpression of blood-group substance A, B, and H, Lewis B antigen, and DFCA is consistent with the opinion that both rectal carcinomas and carcinoids have a common entoderm origin, but carcinoids are considered to rise from the endocrine-differentiated and the adenocarcinomas from the nonendocrine-differentiated enterocytes.

In a prospective study of 103 patients with carcinoid tumors consecutively referred for medical treatment, the most common sites of the primary tumors were the ileum (73%), bronchi (7%), and jejunum (4%). All patients had local metastases, and 96 (93%) also had liver metastases. The most common initial symptoms were diarrhea (32%), ileus (25%), and flush (23%). The overall frequency of diarrhea was 84% and of flush was 75%. Heart insufficiency caused by cardiac valve disease was seen in 33% of the patients. The carcinoid syndrome, including flush, diarrhea, and elevated urinary 5-hydroxyindole acetic acid (5-HIAA) concentrations, was manifested by 69 patients (67%), 64 of whom (93%) had carcinoid tumors of mid-gut origin. Elevated urinary 5-HIAA was found in 91 patients (88%), of which 89 displayed liver metastases. The plasma concentration of the tachykinin neuropeptide K (NPK) was elevated in 67 patients (66%), 63 of whom had tumors of the mid-gut region. Serum pancreatic polypeptide (PP) and human chorionic gonadotrophin alpha levels were elevated in 43% and 28% of the patients, respectively, and the highest levels were found in patients with metastatic bronchial carcinoid tumors. Thirty-nine of the 103 patients are now dead; 18 died of tumor progression, whereas 14 patients died of heart failure secondary to a carcinoid tricuspidal valve insufficiency. The estimated median survival from the time of histologic diagnosis was 14 years, and from the time of carcinoid syndrome was 8 years.

Forty-five carcinoids from the gastrointestinal tract, classified on morphological grounds, were stained with an argentaffin technique (Masson-Hamperl, Virchows Arch [Pathol Anat] 266:509-548, 1927) and three argyrophil techniques (Grimelius, Acta Soc Med Upsal 73:243-276, 1968; Sevier-Munger, J Neuropathol Exp Neurol 24: 130-135, 1965; and Hellerström-Hellman, Acta Endocrinol (Kbh) 35:518-532, 1960). Most but not all carcinoids from the embryonal midgut were argentaffin, whereas most fore- and hindgut carcinoids were nonargentaffin. The argyrophil stain of Sevier-Munger stained all argentaffin tumors and in addition a few nonargentaffin foregut carcinoids. The Grimelius stain stained all and the Hellerström-Hellman stained none of the tumors, irrespective of their location. In the argentaffin carcinoids the frequency of argyrophil cells exceeded that of argentaffin cells.

The parafollicular cells of the thyroid gland were visualized by means of the Sevier-Munger silver technique in normal mice and in mice receiving 131I in amounts sufficient to completely destroy the thyroid tissue. The destruction of the C-cells was observed in all 131I injected mice, and no histologic signs of recovery were seen during a period of 40 days following the treatment.

The clinical picture in 30 patients operated on for gastric carcinoids was studied retrospectively. There were 12 men and 18 women, with an age range of 32-79 years (mean 57 years). The tumours were located in the corpus area of the stomach in more than half of the patients and in one-third were relatively small (less than or equal to 1 cm). Four patients had multiple tumours. Metastases were found in eight patients, mostly those with larger primary tumours. In no case was the carcinoid syndrome present. The patients presented symptoms simulating those of more common affections of the stomach, such as polyps, ulcer and carcinoma. Barium contrast study and gastroscopy did not reveal the true nature of the gastric disease and even biopsy of the stomach failed to give a correct preoperative diagnosis in four of five patients. Since six patients had achylia preoperatively, it is emphasized that the possibility of a gastric carcinoid, especially in the corpus area, is more likely in association with this condition.

The hormone content of 72 endocrine tumors was determined by immunofluorescence and their amyloid content was investigated. Seventeen of the 72 tumors contained amyloid. Amyloid was frequently found in tumors producing calcitonin, insulin, or growth hormone, but was rarely found in other tumors. Thus, there is a relationship between the occurrence of amyloid in an endocrine tumor and the type of hormone it produces. The reason for this is not known, but there is evidence that the amyloid fibrils contain proteins related to the hormone produced by the tumors.

The argyrophil reaction occurs in the parathyroid cells with the Grimelius silver nitrate stain. In the present study, Bouin's fluid was found to be superior to other tested fixatives (formalin, Karnovsky's fluid, GPA etc) in this respect. A study of the argyrophil reaction on the ultrastructural level showed an accumulation of silver particles in the secretory granules, which means that the Grimelius silver stain can be assumed to be an indicator of the secretory granules stored in the parathyroid cells. Whether the argyrophil reaction can be of help in differential diagnosis between different abnormal parathyroid conditions is not yet clear.

A case of lymphosarcoma of the heart in a 10-year-old boy is presented. The clinical picture simulated a pericarditis. Prednisolone had a remarkably good temporary effect. Post-mortem examination showed a highly differentiated lymphosarcoma invading the pericardium and myocardium from a primary mediastinal localisation. Cytologic examination for malignant cells in the pericardial fluid must not be forgotten in a case of unexplained pericardial effusion.

Bouin-fixed tissues from non-diabetic adult human pancreata display an argyrophil reaction mainly in the periphery of the islets with the silver technique of Sevier-Munger. The nature of these argyrophil cells was examined after restaining by an indirect immunocytochemical method using antibodies against insulin, glucagon, somatostatin and pancreatic polypeptide. After this procedure the argyrophil cells were identified as glucagon (A-) cells and pancreatic polypeptide (PP-) cells, although the latter exhibited a weaker reaction. The insulin (B-) cells and somatostatin (D-) cells were unreactive. The results show that the Seiver-Munger stain is of equal value to the Grimelius silver nitrate stain in adult human pancreatic islets after fixation in Bouin's fluid.

Deposition of amyloid is the most constantly present alteration in the islets of Langerhans in type 2 diabetes mellitus and is also quite common in insulin-producing tumors of the pancreas and it is very likely that these two amyloids are identical. We have isolated amyloid fibrils from an insulin-secreting human tumour and purified the fibrillar protein. N-terminal amino acid sequence of the protein is unique and does not resemble insulin or its precursors. Instead it has about 50% homology with the neuropeptide CGRP (calcitonin gene related peptide).

The activity and localization of alkaline phosphatase activity (AP) in aorta and heart, and the incidence of calcifications in aorta, heart and kidney as well as cardial fibrosis were studied in uraemic rats treated with 1,25-dihydroxycholecalciferol (1,25-DCHH) and Nifedipine. 1,25-DHCC treatment elevated the serum Ca x P product and aggravated the development of renal and aortic calcifications and cardial fibrosis. Nifedipine did not protect against calcifications, but decreased the incidence of cardial fibrosis. The activity of AP was increased in the thoracic aorta in uraemia independent of 1,25-DHCC or Nifedipine treatment or presence of calcification. No changes of the AP activity were found in the heart.

The increased knowledge of the pathobiology of gastrointestinal carcinoid (neuroendocrine) tumours and the improved therapeutic possibilities have brought a demand for more precise diagnosis. Although the carcinoid tumours can often be tentatively recognized in routinely processed microscopic slides, their more accurate identification requires additional diagnostic procedures. General neuroendocrine markers such as the argyrophil reaction of Grimelius and immunohistochemistry with application of antibodies against chromogranin A and of neuron-specific enolase are discriminatory staining methods which are used to reveal the neuroendocrine origin of almost all highly differentiated carcinoid tumours of the gastrointestinal tract. Mid-gut carcinoids, which predominate among these tumours almost unexceptionally contain serotonin. This biogenic amine can be demonstrated by the argentaffin reaction of Masson, serotonin immunoreactively or by formalin-induced fluorescence. The characteristic staining pattern of mid-gut carcinoids is almost invariably preserved in the metastatic deposits and consequently the staining methods for identifying serotonin can also be used on metastases to reveal a primary mid-gut carcinoid. The enterochromaffin-like (ECL) cell carcinoids of the body and fundic area of the stomach often seen in association with pernicious anaemia are argyrophil with the Sevier-Munger silver stain. Other neuroendocrine tumours, viz. antral, duodenal and rectal carcinoids should be studied by a battery of relevant peptide hormone antisera for adequate diagnosis. During the last decade new peptide hormones have been found in circulation in patients with carcinoid tumours, but serotonin and urinary 5-HIAA are still the most important markers for carcinoids of the mid-gut origin. Other clinically useful tumour markers are chromogranin A + B, pancreatic polypeptide, human chorionic gonadotropin alpha and beta subunits. For localizing procedures, angiography is the most reliable investigative method for primary tumours in the gut, whereas CT-scan and ultrasound investigations are good for detection of liver metastases. During the last five years, the therapy for malignant carcinoid tumours has been considerably improved. Chemotherapy has only revealed objective response rates in about 10-30% of the patients giving median survivals from start of therapy of about 10 months. Recently treatment with alpha interferons and the new somatostatin analogue octreotide have given objective responses in 50-75% of patients with malignant mid-gut carcinoid tumours. These patients have now a median survival from start of therapy of 70 months when treated with alpha interferons. In the future new therapies will come into use such as monoclonal antibodies and perhaps also agents blocking different growth factors.

Cytologic specimens of neuroendocrine tumors metastatic to the liver were examined with regard to their silver staining properties after the application of argentaffin and argyrophil staining techniques (Masson, Grimelius and Sevier-Munger). In tumors with a content of serotonin (small intestine carcinoids), the presence of this substance was demonstrated cytologically as an argentaffin reaction in individual tumor cells; however, formalin fixation was a prerequisite for positive staining. Melanin in malignant melanoma cells displayed a positive argentaffin reaction, irrespective of the fixation used (air drying, formalin, Bouin's fluid or acetone-alcohol). Thus, serotonin and melanin can be distinguished in cytologic samples of neuroendocrine tumors by the use of the Masson argentaffin reaction with different fixatives. The nonargentaffin-positive neuroendocrine tumor cells were weakly stained or unreactive with the Grimelius argyrophil technique. The Sevier-Munger argyrophil technique was negative or gave a disturbing nonspecific background staining reaction that was difficult to interpret in the cytologic samples. Thus, the Grimelius method appears to be the most useful silver stain for identifying neuroendocrine tumor cells in cytologic material, irrespective of their hormone content, since both argentaffin-positive and argentaffin-negative cell samples were stained at least to some degree.

125I-labelled alpha rat atrial natriuretic peptide (28 amino acids: Ser 99-Tyr 126) ([125I]alpha-rANP) was given i.v. to Sprague-Dawley rats and the distribution of radioactivity in the tissues was examined by whole-body and microautoradiography at intervals from 2 min to 4 h after the administration. Inhibition of uptake of the [125I]alpha-rANP by simultaneous injection of an excess of non-labelled alpha-rANP was taken as an indication that highly labelled structures in rats injected with [125I]alpha-rANP alone are due to an abundance of specific receptors for the peptide. In the rats given only the [125I]alpha-rANP a rapid and high radioactivity occurred in the renal glomeruli, the endocardium of the heart ventricles, the endothelium of the processus ciliares of the eyes, the portal vessels and a few larger vessels of the liver, the subcapsular vessels of the adrenal glands and the parenchyma of the lungs. Other tissues showing a distinct, but less prominent, radioactivity were the endocardium of the heart atria, the walls of the great afferent and efferent vessels in the thoracic cavity, the choroid plexuses of the brain ventricles, the pia mater, brown fat, the muscularis layer of the stomach and the intestines, the lamina propria of the villi in the small intestine and the walls of a few small blood vessels in the kidney medulla. The specific labelling was highest at 2 min after injection and then diminished at later intervals.(ABSTRACT TRUNCATED AT 250 WORDS)

A 64-year-old woman died after 12 years of progressive pulmonary disease which was initially diagnosed as sarcoidosis but later correctly identified as mineral oil pneumonia due to insufflation of paraffin instilled as nasal drops. In view of the potential damaging effect of liquid paraffin on the lungs, the current indications for its use must be strongly questioned.

A medullary carcinoma of the thyroid was preoperatively diagnosed on ultrasonically guided fine needle aspiration biopsies. After cytocentrifugation, the tumor cells displayed a dense cytoplasmic silver granulation with the Sevier-Munger technique when applied to air-dried or acetone-ethanol-fixed samples and an obvious calcitonin immunoreactivity after fixation in Bouin's fluid. These methods may prove useful in the identification of nonpalpable metastases and recurrences of medullary carcinomas of the thyroid, especially since the cytologic typing of medullary thyroid carcinoma cells may be difficult with routine stainings.

A case of splenic-gonadal fusion is described. In this case, as in most cases reported previously, the malformation was discovered accidentally. An ectopic spleen attached to the left testis is a rare differential diagnosis to left-sided testicular tumour.

The uptake of the nicotinamide adenine dinucleotide (NAD)-precursors nicotinamide, nicotinic acid and tryptophan in the pancreatic islets of mice was studied by use of autoradiographic methods. The ability of these substances to prevent streptoxotocin diabetes was studied in the same species. It was found that only nicotinamide was strongly accumulated in the pancreatic islets and nicotinamide was also the only NAD-precursor which protected against the streptoxotocin diabetes. Apparently there is a relationship between the ability of the NAD-precursors to be taken up in the pancreatic islets and their ability to prevent streptoxotocin diabetes.

Plasma atrial natriuretic peptide (ANP) concentration was determined and cardiac ultrasound studies were performed in 50 patients with malignant mid-gut carcinoid tumors. The extent of carcinoid-related heart disease varied among the patients. The patients with the most severe right-sided heart disease, who often had signs of right ventricular failure, had significantly (p less than 0.001) higher plasma ANP concentrations than either patients with less or no abnormal ultrasound findings or age- and sex-matched healthy control subjects. ANP levels were serially determined for 0.5 to 4 years (median 2.1) in 12 patients. The levels increased above the reference range in patients with clinical findings of right ventricular failure. In patients without cardiac decompensation the levels remained within the reference range. In 3 patients who had successful tricuspid and pulmonary valve replacements, signs and symptoms of right ventricular failure disappeared and plasma ANP levels declined and normalized. Five patients with progressive right ventricular failure and increasing plasma ANP levels during follow-up eventually died from cardiac decompensation. This study demonstrates the predictive value of serial determinations of plasma ANP in carcinoid heart disease. Such measurements can be an additional guide in the clinical management of these patients.

A comparative immunocytochemical study was performed of subepithelial neuroendocrine cells of the human small intestine and appendix and carcinoid tumours of these sites, using a monoclonal antibody to serotonin and polyclonal antisera against neuron-specific enolase (NSE) and S-100 protein. Subepithelial neuroendocrine cells were easily identified in the lamina propria of the appendix. These cells, which sometimes occurred in aggregates, displayed serotonin and NSE immunoreactivity and were surrounded by S-100 protein immunoreactive cells, presumably of Schwann cell origin. In the appendix scattered cells with corresponding morphological features and immunoreactivity were also observed deep in the submucosa. In addition, subepithelial neuroendocrine cells were sparsely present in the lamina propria of the small intestine, occurring only as single cells in the deeper part of the mucosa below or between the epithelial crypts. Most appendiceal carcinoid tumours (11 of 12 examined cases) were biphasic and consisted of neuroendocrine tumour cells with intermingled S-100 protein immunoreactive cells (Schwann cells) with long cytoplasmic extensions. However, small intestinal (11 cases) and caecal (10 cases) carcinoids lacked S-100 protein immunoreactive cells as an integral component. The results indicate that the appendiceal carcinoids are mostly closely related structurally to the subepithelial neuroendocrine and Schwann cell aggregates of the lamina propria and are thus presumed to be histogenetically related to this cell system, while the histogenesis of small-intestinal and caecal carcinoids remains less clear.

The Grimelius argyrophil reaction was studied in 27 rectal carcinoids. A majority of the tumour cells were silver-reactive in 16 tumours and a minority in 11 tumours. Both weakly and strongly stained tumours were found, as well as tumour cells with variable staining properties within the same tumour. Re-staining resulted in potentiation of the reaction, but this was most obvious in tumours with an initially easily identified argyrophilia. It is suggested that evaluation of the Grimelius argyrophil reaction in combination with routine histology increases the accuracy of the diagnosis of rectal carcinoids.

In a histopathological and immunocytochemical study of biopsy and/or operation specimens from 27 patients with endocrine tumors of the colon and rectum ("hind-gut carcinoids") enkephalin-immunoreactive tumor cells were observed in two cases. Both patients were obese women, about 50 years of age, with a history of constipation. The tumors were situated near the anus in the dorsal wall of the rectum. One tumor had metastasized to a lymph node, and the other showed vascular invasion. The tumor cells were non-argentaffin; some were argyrophil. One tumor contained only few enkephalin-immunoreactive cells but had numerous beta-endorphin-immunoreactive cells, which were distinct from the former. The other contained large numbers of enkephalin-immunoreactive cells but no beta-endorphin cells. Both tumors also harboured glucagon-immunoreactive cells; in one there were also cells containing immunoreactive pancreatic polypeptide. These cells were distinct from the enkephalin-storing ones. No 5-hydroxytryptamine could be detected in the two tumors.

Twenty patients with malignant carcinoid tumors were treated for 6 months with recombinant interferon alfa-2b (IFN alpha-2b; Intron-A; Schering Corp., Bloomfield, NJ) at a mean dose of 5.9 megaunits three times per week. Eleven of the 20 patients (55%) had a greater than 50% reduction of tumor markers (urinary 5-hydroxyindoleacetic acid or plasma neuropeptide K), showing objective tumor response. Six patients (30%) had stable disease with no significant change in tumor markers or tumor size, and three (15%) had progressive disease with an increase in tumor markers and size. These results are similar to those reported earlier for treatment with natural leukocyte IFN in patients with carcinoid tumors. Only two patients (35%) had a slight reduction of tumor size after 6 months of treatment. Three patients developed neutralizing antibodies to IFN alpha-2b. Two of these patients initially showed an objective response, which lasted until IFN antibodies developed. In one of these patients, a change to human leukocyte IFN resulted in normalization of antibody titers within 3 months, and the patient had a second objective clinical response. There was no correlation between development of IFN antibodies and development of autoimmune phenomena such as increased titers of antinuclear antibodies or thyroid autoantibodies. IFN alpha-2b seems to be as potent as human leukocyte IFN in the treatment of patients with malignant carcinoid tumors, but it is important to recognize that antibodies neutralizing IFN may develop in some patients, with concomitant loss of antitumor effects. A change to natural leukocyte IFN might be beneficial in these patients.