Human eosinophil cationic proteins (ECP and EPX) and their suppressive effects on lymphocyte proliferation.

ECP (eosinophil cationic protein) and EPX (eosinophil protein X) are two highly basic proteins contained in the granules of human eosinophils. In this study, the effect of ECP and EPX upon lymphocyte proliferation in vitro has been investigated. Peripheral blood mononuclear cells from normal donors were cultured in medium containing ECP or EPX at concentrations from 10(-10) to 10(-7) M. 3H-Thymidine incorporation in PHA-blasts or MLR-blasts was dose-dependently inhibited by both ECP and EPX. The effect was irreversible and was not due to cytotoxic damage. The suppressive effect of EPX may involve suppressor cells. The effect of ECP and EPX on lymphocyte proliferation at relevant in vivo concentrations suggests a regulatory role for the eosinophil in immunological reactions.

Polyclonal antibodies against human melanocortin MC1 receptor: preliminary immunohistochemical localisation of melanocortin MC1 receptor to malignant melanoma cells.

Peptides of 11 and 15 residue lengths were synthesised according to the sequence of the N-terminal region of the human MC1 melanocyte stimulating hormone receptor. The peptides were conjugated to thyroglobulin and used for preparation of antisera in the rabbit. Each of the conjugates raised antisera which showed high titre and specificity for its respective peptide antigen when evaluated in an ELISA test. Both types of antisera immunostained MC1 receptor expressing COS-7 cells. By contrast, the sera did not stain control COS-7 cells not expressing the MC1 receptor. Moreover, preimmune sera or antiserum preadsorbed with its respective peptide did not stain the MC1 receptor expressing cells. The antisera were used to immunostain sections of normal human skin, as well as samples of cutaneous malignant melanoma tumours obtained from a patient. The cells of the melanoma tumours were very strongly immunostained with the MC1 receptor antisera. By contrast, melanocytes which were present in the normal skin could not be visualised with our antisera.

Biliary tract carcinoma.

A series of 127 patients treated for cancer of the biliary tract during the period 1965-1974 was studied with a follow-up period of at least five years. The mean age was 66 years and the female:male ratio was 3:1. An early diagnosis of biliary tract carcinoma is difficult and the prognosis unfavourable although not hopeless. The surgical treatment given in the present series included a radical approach whenever possible. In the total series 56 patients died within a month, 24 lived for more than one year, 9 longer than three years and 5 longer than five years and 3 are still alive. A mean survival time of 25.6 months was reached when a radical operation was attempted while a palliative procedure gave a mean survival time of 5.5 months. These figures indicated that a careful selection of patients for an attempt of radical surgery should be done since removal of the tumour gives a more effective palliation, often a longer survival time, an improvement of quality of live and in a few patients cure.

Identification of children at high anaesthetic risk at the time of primary palatoplasty.

Closure of the palate is an operation with a higher risk of postoperative hypoxaemia than other plastic surgical procedures. An increase in anaesthetic complications was noticed among our children with a cleft palate who were operated on with a palatoplasty, and this was further investigated. The records of 154 patients operated on between 1979 and 1996 were eligible for the study. A total of 128 had isolated cleft palate (ICP), 19 had Pierre Robin syndrome (PRS), and seven had other identified syndromes (IS). The results showed that the risk of anaesthetic complications was four times greater when the operation was done when the child was less than 1 year old, and there was a sixfold increase when a more elaborate velopharyngoplasty technique was used. Children with Pierre Robin syndrome were further classified according to the degree of neonatal difficulties. The 12 children with most severe problems were at increased risk. When the diagnostic criteria as well as the type of operation were taken into account the incidence of anaesthetic complications was 7/11, which was the highest risk encountered in the present study. When the first operation on the palate in children with Pierre Robin syndrome is planned, it should be based on a classification of neonatal problems and an investigation of their clinical condition. In children at risk the palatoplasty procedure should be postponed until the age of 12-18 months.

The effect of growth factors and synovial fluid on chondrogenesis in perichondrium.

Reconstruction of cartilage with perichondrium depends on the chondrogenic property of the perichondrial fibrocytes. The present investigation concerns the conditions for the differentiation of fibrocytes into chondrocytes both in vivo and in vitro. For the in vivo studies specimens of rib and auricular perichondrium from adult rabbits were wrapped round silicon rods which were enclosed in dialysis bags. One was placed in the suprapatellar pouch of the knee joint and one was placed intraperitoneally in each rabbit. After two months the bags were extracted, the perichondrium prepared for microscopic examination, and the chondrogenesis evaluated. In vitro the perichondrium was divided into small pieces and incubated with tissue culture medium. The medium was supplemented with fetal calf serum, together with epidermal growth factor, platelet derived growth factor, synovial fluid, or with human serum albumin (control group). After three weeks the explants were prepared for microscopy. Chondrogenesis was judged by the degree of cellular enlargement, capsule formation, deposition of matrix, and activation of the outer fibrocytic layer. In vivo, good cartilage development was found in all specimens placed in the knee joint but, in those placed intraperitoneally, little if any chondrogenesis was seen. In vitro profound differentiation occurred in all cultures supplemented with epidermal growth factor and platelet derived growth factor. An equivalent differentiation was found in perichondrium that had been incubated with synovial fluid. We conclude that the differentiation of perichondrial fibrocytes is initiated in vitro by growth factors. In addition, we have shown that synovial fluid contains factors that promote and enhance the development of cartilage from perichondrium.

Expression of platelet-derived growth factor beta receptor in chondrogenesis of perichondrial transplants.

An experimental study was done in two rabbits to establish whether the early stages of cartilage generation involves stimulation via the platelet-derived growth factor beta receptor (PDGFR-beta). Rib perichondrium was dissected off and transplanted to the knee joint. The perichondrial grafts were removed for microscopic investigation at day 0 and day 6 and kept frozen until analysed by immunohistochemistry and in situ hybridisation for the presence of PDGF beta receptor protein and mRNA, respectively. Samples from day 0 showed sparse cells with a positive immunohistochemical reaction. In samples from day 6 there were signs of perichondrial proliferation and chondroid maturation and a more widespread immunohistochemical staining reaction could be seen mainly on proliferating perichondrial cells, but also on some chondrocytes. In situ hybridisation confirmed the expression of PDGF beta receptor mRNA in the same areas of serial sections. These findings may indicate that PDGF beta receptor activity is part of the early response after autotransplantation of perichondrium.

Substance P and enteroglucagon-like immunoreactivity in argentaffin and argyrophil midgut carcinoid tumours.

The presence of serotonin, substance P and enteroglucagon was investigated in 8 argentaffin and argyrophil midgut carcinoid tumours. All tumours were argentaffin and displayed formalin-induced fluorescence indicating a content of serotonin. In addition, all 8 tumours showed substance P immunoreactivity and 7 of them enteroglucagon immunoreactivity. The results indicate that the midgut carcinoid tumours are as a rule multihormonal.

Carcinoma of the junction of the main hepatic ducts.

Patients with carcinoma of the hepatic duct junction often present the surgeon with a difficult problem. If at all possible radical surgery may be performed either as a resection of the tumour and an anastomosis according to Roux-en-Y or a resection of the left or right hepatic lobe together with the obliterated hepatic duct or ducts. The present series consists of 17 patients treated for cancer of the hepatic duct junction during the period 1965-1974 with a follow-up period of at least five years. The surgical treatment given included a radical procedure in 8 cases, a palliative procedure in 7 cases and in 2 cases surgery of the hepatic ducts was not possible. Radical procedures included 4 patients with a resection of the left hepatic lobe and 4 patients with resection of the left and right hepatic ducts and anastomosis according to Roux. A mean survival time of 22.8 months was reached when a radical operation was attempted with a longest survival time of 8 years. A palliative procedure gave a mean survival time of 2.0 months. These figures indicate that a careful selection of patients for an attempt of radical surgery could be made since removal of the tumour gives a more effective palliation, often a longer survival time and an improvement of quality of life.

On the significance of nipple discharge in the diagnosis of breast disease.

Nipple discharge, especially the blood-stained type, is regarded as an important symptom in breast disease. In many reports the high incidence of malignancy is stressed and an active surgical approach has often been recommended. In this series of 80 women with nipple discharge the type of secretion and the result of exfoliative cytology were evaluated. Three women had cancer but the nipple discharge was important for the diagnosis in only one case. Six women had papilloma and in all the nipple discharge was essential for the diagnosis. Fibroadenosis and duct ectasia were the most common diagnoses. The blood-stained secretion was due to fibroadenosis in about half of the cases. No cancer was found in 46 women with a serous secretion. When a tumour is present nipple discharge is of little importance for the diagnosis and treatment. In the absence of a tumour and when exfoliative cytology gives no suspicion of cancer and no atypical cells or papillomatous clusters are present, an expectant attitude towards surgery seems satisfactory. With such an approach many surgical biopsies prove unnecessary, but a prerequisite is an organized follow-up.

Polypeptide hormones in argentaffin and argyrophil gastroduodenal endocrine tumors.

The morphology and histochemistry of gastroduodenal endocrine tumors from 16 patients were studied. All patients underwent operation, in most cases with a preoperative diagnosis of nonendocrine tumor, ulcer, o polyp(s). The argentaffin reaction was positive in three tumors, and the Hellerström--Hellman argyrophil reaction was positive in four tumors. All tumors reacted positively to the Grimelius argyrophil stain, and 13 were positive with the Sevier--Munger argyrophil stain. Gastrin immunoreactivity was found in eight tumors, and substance-P immunoreactivity in seven tumors. No enteroglucagon, adrenal cortex hormone, or pancreatic polypeptide was observed in any of the tumors. Three patients with Sevier--Munger-positive gastric tumors had concurrent pernicious anemia, and 2 patients with gastrin-immunoreactive tumors had acute or chronic gastroduodenal ulceration. The results indicate that the gastroduodenal endocrine tumor as a rule gives no endocrine symptoms and that the tumor type is an unexpected finding at operation. The tumors may contain gastrin, substance P, somatostatin, and serotonin and tend to be multihormonal.

Characterization of human lymphocyte surface receptors for mitogenic and non-mitogenic substances.

To compare the receptor patterns for mitogenic and non-mitogenic substances, surface glycoproteins of human lymphocytes were labelled with the lactoperioxidase-catalysed iodination technique and with a galactose oxidase-tritiated sodium borohydride technique. Labelled cells were detergent-solubilized, and the lysates were allowed to react with insolubilized purified mitogenic lectins, phytohaemagglutinin, leucoagglutinin and an insolubilized non-mitogenic lectin, oxidized leucoagglutinin. Lectin-reactive proteins were eluted with sodium dodecyl sulphate (SDS) buffer. Cell membrane components reactive with anti-lymphocyte globulin (ALG) were retrieved by indirect immunoprecipitation with protein-A-bearing staphylococcus Cowan I strain (SaCI). Lectin- and ALG-reactive proteins were analysed by SDS polyacrylamide gel electrophoresis. Iodinated glycoproteins regularly showed four major components with molecular weights of 120,000, 70,000, 60,000 and 43,000 daltons, respectively, on 7% gels. An additional broad peak in the molecular weight range 20,000--35,000 daltons was found on 10% gels. Tritiated glycoproteins also showed four major components with MW 120,000, 70,000, 60,000 AND 42,000, RESPECTIVely, which reacted with lectin and ALG. In addition, ALG reacted with some glycoproteins with MW between 150,000 and 230,000 daltons. On 10% gels additional lectin- and ALG-binding glycoproteins with MW around 30,000 daltons were found. The similarity in structures bound by mitogenic and non-mitogenic substances indicates that lymphocyte activation may depend on some property conferred by the mitogen.

The cartilaginous potential of the perichondrium in rabbit ear and rib. A comparative study in vivo and in vitro.

In an experimental study the cartilaginous protential of the rabbit ear perichondrium has been compared with that of the rib in vivo and in vitro. Perichondrium was transferred as free autologous grafts to the subcutaneous tissue on the scalp and as loose bodies into the knee joint. The presence of cartilage in the grafts was examined after six weeks. In vitro explants of rabbit perichondrium from the ear and the rib were maintained in an organ culture system. The presence of cartilage was analyzed after one to three weeks. Rabbit perichondrium from the rib appeared to have a greater cartilaginous potential than that from the ear both in vivo and in vitro. Chondrogenesis in perichondrium was demonstrated in vitro.

Enteroglucagon and substance P-like immunoreactivity in argentaffin and argyrophil rectal carcinoids.

A specific immunofluorescence for enteroglucagon or substance P or for both hormones was demonstrated in nine out of 12 examined rectal carcinoids. One tumor was argentaffin, contained ultrastructurally pleomorphic granules of the entero-chromaffin cell type, and showed immunofluorescence for substance P. The rest were non-argentaffin but were argyrophil with the Grimelius technique and contained round granules. The argyrophil carcinoids were immunoreactive to one or both hormones in eight cases and not fluorescent in three cases. In two of the non-argentaffin carcinoids a small number of argyrophil cells was found with the method of Sevier-Munger.