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Biomedical subjects

J Koudstaal

Publications and source records attributed to J Koudstaal.

At least 73 records · Page 4Linked to original sources

Determination of tissue iron and ferritin in liver pathology comparison of histochemical and biochemical results.

Results are presented from the determination of tissue iron and ferritin in 15 pathological livers (6 with steatosis, 8 with fibrosis/cirrhosis and 1 with haemochromatosis). The histological assessment according to the Rowe system, after Perls' staining, was compared with the measurement of the iron content in liver homogenate by flameless atomic absorption spectrophotometry. The results of the flameless atomic absorption spectrophotometry were less accurate than in normal livers, but the method can still be considered reliable with satisfactory precision. As in normal livers, the range of chemically determined values in the histological staining grades was considerable and there was quite an overlap between consecutive grades. The chemical determination of liver iron content is to be preferred. The levels of ferritin protein and ferritin iron follow the same pattern as the total iron content.

Ferritins↗

Accumulation of iron and iron compounds in liver tissue. A comparative study of the histological and chemical estimation of liver iron.

Unsatisfactory results obtained by histological evaluation of liver tissue in iron loading diseases prompted us to study the distribution of the total liver iron, haem iron and ferritin iron in post mortem human liver tissue from two different sites of the same liver. The total liver iron content was measured by flameless atomic absorption spectroscopy in native liver homogenates and in acid digested liver tissue from 60 consecutive autopsies, and the results from the two methods were compared. From the standard deviation of the duplicate analyses, it was deduced that the liver iron is possibly inhomogeneously distributed. The CVduplo (22%) of total iron, measured in acid digested tissue was higher than the CVduplo (14%) of total iron in homogenates from liver tissue from which non-homogenized tissue e.g. vessel walls, fibrotic tissue, had been removed. The CVduplo of ferritin iron and haem iron in liver homogenate was 14% and 30% respectively. The ferritin iron increased with an increasing total iron content until saturation of ferritin iron appeared to be reached at 2.5 micrograms ferritin iron per mg liver protein. When the results of total non-heam liver iron measurements are expressed properly (amount of iron per amount of homogenized liver protein), the distribution of iron is found to be homogeneous in both normal and pathological liver tissues. It was concluded that the estimation of liver iron content by visual microscopic evaluation is unsatisfactory, and that more reliable results are obtained by atomic absorption spectrophotometry.

Ferric Compounds↗

Measurement of liver iron content in paraffin-embedded biopsies.

A reliable method for the determination of total liver iron in formalin-fixed, paraffin-embedded tissue is presented. The correlation with total liver iron in fresh tissue is good (r = 0.92). Material, which is processed routinely in the pathological anatomical department and stored in the archives, can be used for a quantitative iron determination for clinical or research purposes.

Biopsy↗

Double immunohistochemical demonstration of antigen expression and DNA-incorporated 5-bromodeoxyuridine in frozen and plastic embedded sections.

In this study a double immunohistochemical staining procedure is described for the simultaneous visualization of antigen expressing cells and replicating cells. Cell surface antigen expression was marked with a monoclonal antibody against I a (His 19) or a monoclonal antibody against a membrane component of the cells of the monocyte-macrophage lineage (ED2). Replicating cells were detected by the incorporation of 5-bromodeoxyuridine. The method was applied sequentially. On frozen sections two peroxidase labeled reagents were used with two different substrates yielding a red and a dark-blue black reaction product. On plastic-embedded sections a peroxidase and an alkaline phosphatase labeled reagent were applied resulting in a brown and a blue reaction product.

Animals↗

A patient with methotrexate lung and granulomatous changes in the liver--or just sarcoidosis?

A 57 year old ex-miner, who had been treated for his psoriasis with methotrexate 5 mg 3 times a week, taken orally (in total +/- 12 g) developed in the spring 1985 the symptoms of an allergic pneumonia. In biopsies of lung and liver non-caseating granulomata were found, compatible with the diagnosis sarcoidosis. The question whether one clinical entity is concerned or two superponed diseases, remains unanswered.

Blood Gas Analysis↗

Simultaneous immunohistochemical demonstration of antigen expression and 5-bromodeoxyuridine incorporation in plastic embedded sections.

In this study a double immunohistochemical staining procedure is described for the simultaneous demonstration of antigen expressing cells and replicating cells in rat thymus. As markers for cell surface antigen expression a monoclonal antibody against Ia-expressing cells (His 19) and a monoclonal antibody against cells of the monocyte-macrophage lineage (ED2) were used. Replicating cells were demonstrated by the incorporation of 5-bromodeoxyuridine (BrdUrd). Tissue pieces were fixed in a periodate-lysine-paraformaldehyde fixative and embedded in glycol methacrylate. To demonstrate Ia-expressing cells or ED2-positive macrophages in plastic embedded sections a digestion with trypsin is necessary. The staining procedure was applied sequentially and was performed with a peroxidase and an alkaline phosphatase labeled reagent yielding respectively a brown and a blue reaction product. Results with this staining procedure on plastic embedded sections of rat thymus, an organ with a high DNA synthesizing capacity, showed incorporation of BrdUrd predominantly in the cortex. ED2-positive macrophages were only found in the cortex. The Ia-positive epithelial reticular cells demonstrated extremely well their stellate form.

Animals↗

An analysis of histology and DNA-ploidy in primary wilms tumors and their metastases and a study of the morphological effects of therapy.

In children with Wilms' tumours the length of survival is greatly influenced by success in preventing or controlling metastatic disease. The current study focuses on the morphological aspects of metastases when compared with the primary tumour. In 8 patients it appeared that blastema is the most likely component to metastasize, whereas epithelial and stromal components were hardly, if at all, represented in metastases. Furthermore, flow cytometric DNA ploidy determinations on 4 cases showed that both the primary tumours and the metastases had stemlines in the diploid and low aneuploid (hyperdiploid) range. Finally, in four cases the influence of therapy on morphology of the primary tumours was analyzed. In these cases blastema seemed to be the component most sensitive to therapy. Thus, blastema seems to play a central role in prognosis of Wilms' tumours; either reacting to therapy or, if insensitive, by metastasizing.

Antineoplastic Combined Chemotherapy Protocols↗

Gastrointestinal toxicity of chemotherapy and the influence of hyperalimentation.

The effect of combination chemotherapy on human small intestinal morphology and disaccharidase activities and their relation with clinical and chemical (fecal wet weight and K-excretion) parameters for gastrointestinal toxicity were evaluated in patients with disseminated malignant melanoma receiving enteral normoalimentation (NA). Also evaluated were the supposed protective effects on gastrointestinal toxicity of enteral hyperalimentation (HA) with an elemental diet. After chemotherapy, a comparable decrease in villus height, total mucosal height, and mitotic index was found in jejunal biopsy specimens of both groups. However, in the NA group, the crypt depth decreased (in contrast to the HA group), whereas the disaccharidase activities in the HA group deteriorated to lower values than in the NA group. The authors found no correlation between disaccharidase levels and mucosal morphology, nor was there a correlation between these variables, fecal parameters and clinical diarrhea, suggesting that diarrhea occurring after chemotherapy was not due to loss of mucosal tissue or decrease in enzyme activities. A protective effect of HA with an elemental diet on gastrointestinal toxicity could not be established.

Adult↗

Immunohistochemical demonstration of DNA-incorporated 5-bromodeoxyuridine in frozen and plastic embedded sections.

The application of an immunohistochemical method in the detection of replicating cells, that have incorporated 5-bromodeoxyuridine (BrdUrd), was studied on frozen and plastic embedded sections of different rat tissues. Hydrolysis conditions employed in the Feulgen procedure are essential in making the incorporated BrdUrd accessible to the monoclonal anti-BrdUrd antibodies. To demonstrate the incorporated BrdUrd in plastic embedded sections a subsequent etching with xylene and digestion with protease is necessary. Data obtained with this method are completely comparable with those found by the tritiated thymidine method. In comparison with the thymidine method, the BrdUrd method is much less time consuming and does not require precautions in working with radioactivity. The BrdUrd-method enables a more precise localization as is especially shown in the plastic embedded sections.

Animals↗

Evidence for a migratory capability of rat Kupffer cells to portal tracts and hepatic lymph nodes.

The present study concerns the migratory ability of Kupffer cells in the rat. Phagocytic cells were labeled with colloidal carbon or gold, these markers being administered intravenously either into a tail vein, which resulted in generalized reticuloendothelial uptake, or in low dose into the portal vein, which produced uptake by Kupffer cells alone. Cells containing marker were observed in the portal tracts and in hepatic lymph nodes from 1 to 3 days after injection into the portal vein. The direct movement of single marker particles to the portal tracts could be excluded. Since injection of marker into the portal vein labeled Kupffer cells exclusively, whereas blood cells, splenic and bone marrow macrophages remained unlabeled, the labeled cells in the portal tracts and hepatic lymph nodes appeared to be former Kupffer cells migrating which had migrated to these sites.

Animals↗

Nesidioblastosis and endocrine hyperplasia of the pancreas: a secondary phenomenon.

Diffuse endocrine cell proliferation (nesidioblastosis) and islet cell hyperplasia are considered causes of organic hyperinsulinism but have not been distinguished (by histometric or immunohistologic methods) from the normally variable pancreatic islet cell population during development and in adults. Therefore, in this study morphologic, immunohistologic (to detect insulin, glucagon, somatostatin, and pancreatic polypeptide), and morphometric features were evaluated in 1) normal pancreases (from fetal to adult; n = 49); 2) pancreases from patients with nesidioblastosis (n = 5); and 3) tumor-associated pancreases (TAP) from patients with insulin-producing islet cell tumors (n = 8). The study of normal postnatal development revealed that all features of fetal development remain present after birth and that the diagnosis of any diffuse endocrine disorder should therefore be based essentially on quantitative histometric parameters (total endocrine area, islet size distribution, distribution of each endocrine cell type). With these parameters endocrine cell hyperplasia was demonstrated in TAP from adults due to increased numbers of A and D cells. However, in the cases previously diagnosed as pathologic nesidioblastosis, all parameters were within the normal range. Thus, nesidioblastosis does not appear to be a pathologic entity. Careful re-examination of the pancreases, prompted by these data, revealed small islet cell tumors in three of these five cases. It is concluded that the endocrine pancreas can react rapidly, both morphologically and functionally, to changes in hormonal feedback, e.g., islet cell tumors. Therefore, the observation of a diffuse islet cell disorder in a patient with hyperinsulinism should not be considered an indication that an islet cell tumor is not present.

Adult↗

Graft-versus-host disease in the rat: cellular changes and major histocompatibility complex antigen expression in the liver.

Cellular changes in the liver were studied during an acute lethal graft-versus-host (GVH) disease in relation to the expression of major histocompatibility complex (MHC) antigens on different liver cells. Screening for MHC antigen expression revealed that control livers contained very few Ia+ cells: mainly cells in the portal tract interstitium and a small percentage of the Kupffer cells. The changes during an ongoing GVH reaction could be separated into those related to the sinusoid-associated cells, including the liver parenchyma, and those related to the portal-tract-associated cells, including periportal hepatocytes. In the sinusoids an increase in the number of Kupffer cells was seen, now all expressing Ia antigens. No damage to hepatocytes or other sinusoid-associated cells was observed. It is postulated that the increase in both number and Ia expression of the Kupffer cells is most probably due to an increased phagocytic uptake of blood-borne cellular debris and is not a result of extensive damage to hepatocytes. In the portal tracts expanding infiltrates were found composed of Ia+ T cells and macrophages (ratio 2:1). These infiltrates are probably due to a local accumulation of lymphocytes and macrophages as a result of an interaction of migrating donor-type alloreactive T cells with recipient type Ia+ cells present in the portal tract interstitium, which also interfered with normal recipient lymphocyte and macrophage traffic. Damage to portal-tract-associated cells was slight and confined to bile duct epithelial cells, which now expressed Ia antigens, and to periportal hepatocytes. In conclusion, these data do not indicate that damage to liver parenchyma plays a major role in the pathogenesis of an acute GVH reaction.

Acute Disease↗

Zonal heterogeneity of rat hepatocytes in the in vivo uptake of 17 nm colloidal gold granules.

The in vivo uptake in hepatocytes of intravenously injected colloidal gold granules with a diameter of 17 nm or 79 nm and coated with bovine serum albumin or with polyvinyl-pyrrolidone was studied. Irrespective of coating only the 17 nm granules were taken up in hepatocytes. Perivenous hepatocytes did take up much more gold granules than periportal hepatocytes. The gold granules were found in lysosomes around bile canaliculi. Two hours after injection hepatocytes contained the maximal amount of granules. At least a portion of the granules was discharged into the bile. The observed zonal gradient in the uptake of 17 nm gold granules might be caused by the greater supply of granules to the perivenous hepatocytes as a combined result of the higher porosity of the endothelial lining and the smaller number of Kupffer cells with a low endocytic activity in this zone.

Animals↗

The significance of the number of centroblasts in centroblastic/centrocytic lymphomas. A long term study in a large group of patients.

A group of 385 centroblastic/centrocytic lymphomas with long follow-up periods was subdivided on the basis of the relative number of centroblasts in the initial biopsies. Only minor differences in overall survival were found although a different survival pattern was observed between the groups. The transition into a high-grade malignant secondary centroblastic lymphoma correlated, at a statistically significant level, with the relative number of centroblasts in the initial biopsy.

Actuarial Analysis↗

Mesangial function and glomerular sclerosis in rats with aminonucleoside nephrosis.

The possible relationship between mesangial dysfunction and development of glomerular sclerosis was studied in the puromycin aminonucleoside (PAN) model. Five male Wistar rats received repeated subcutaneous PAN injections; five controls received saline only. After 4 weeks the PAN rats were severely proteinuric (190 +/- 80 mg/24 hr), and all rats were given colloidal carbon (CC) intravenously. At 5 months glomerular sclerosis was found in 7.6 +/- 3.4% of the glomeruli of PAN rats; glomeruli of the controls were normal. Glomeruli of PAN rats contained significantly more CC than glomeruli of controls. Glomeruli with sclerosis contained significantly more CC than non-sclerotic glomeruli in the same kidneys. CC was preferentially localized within the sclerotic areas of the affected glomeruli. Since mesangial CC clearance from the mesangium did not change during chronic PAN treatment, we conclude that this preferential CC localization within the lesions is caused by an increased CC uptake shortly after injection in apparent vulnerable areas where sclerosis will develop subsequently. Cluster analysis showed a random distribution of lesions in the PAN glomeruli in concordance with the random localization of mesangial areas with dysfunction in this model. Similar to the remnant kidney model in PAN nephrosis the development of glomerular sclerosis may be related to "mesangial overloading."

Animals↗