p53 mutations in defined structural and functional domains are related to poor clinical outcome in non-small cell lung cancer patients.

The prognostic value of p53 status in non-small cell lung cancer has been investigated in 148 patients with clinical stage I-IIIB disease. Tumor tissues were examined for mutations in exons 4-9, with emphasis on defined structural and functional domains. Eighty-four mutations were detected in 83 (54%) of the patients. Eighty-eight percent of the mutations were within exons 5-8, and 12% of the mutations were within exons 4 and 9. Missense mutations occurred in 67% of the tumors, and 30% were null mutations (10% stop mutations, 15% frameshift mutations, and 5% splice site mutations). Patients with mutations in p53 had a significantly higher risk for lung cancer-related death and for death from all causes than those with wild-type p53 [hazard ratio (HR) = 2.09 and 95% confidence interval (CI) = 1.20-3.64 and HR = 1.69 and 95% CI = 1.06-2.70, respectively]. Mutations in p53 related to even still poorer lung cancer-related prognosis were found at the following locations: (a) exon 8 (HR = 3.5; 95% CI, 1.59-7.71)]; (b) the structural domains L2 + L3 (HR = 2.36; 95% CI, 1.18-4.74), and (c) codons involved in zinc binding (HR = 11.7; 95% CI, 3.56-38.69). Together, the biologically functional group of severe flexible mutants (codons 172, 173, 175, 176, 179, 181, 238, 245, and 267) and severe contact mutants (248, 282) were significantly related to shorter lung cancer-related survival (HR = 4.16; 95% CI, 1.93-8.97). Squamous cell carcinoma was the dominant histological type in tumors involved in poor prognosis in exon 8 (HR = 3.19; 95% CI, 1.07-9.45). These results indicate that mutations in defined structural and functional domains of p53 may be useful molecular biological markers for prognosis and treatment strategy in non-small cell lung cancer patients.

Prognostic importance of various clinicopathological features in papillary thyroid carcinoma.

The influence of various pathological features on tumour recurrences and cancer deaths has been studied in 173 consecutive cases of surgically treated papillary thyroid carcinoma recorded in 1971-1985. During the follow-up (median 7.3 years), 18.6% of the 161 radically treated patients had recurrent disease, and 8.7% died of thyroid cancer. In the univariate life-table analysis, recurrence-free survival was significantly related to age, pTNM category, tumour size, presence of certain growth patterns, tumour necrosis, tumour infiltration in surrounding thyroid tissue and thyroid gland capsule, lymph node metastases, presence of extra-nodal tumour growth and number of positive lymph nodes, whereas only tumour diameter, thyroid gland capsular infiltration and presence of extra-nodal tumour growth remained as significant prognostic factors in the multivariate analysis. Regarding thyroid cancer deaths, sex, age, pTNM category, radicality of surgical treatment, tumour diameter, macroscopic appearance, cellular atypia, tumour necrosis, thyroid gland capsular infiltration, vascular invasion, extra-thyroidal extension and lymph node metastases were all significant variables in the univariate analysis. However, only sex, age, radicality of surgical treatment and vascular invasion were found to be significant predictors of thyroid cancer deaths in the final multivariate Cox model, whereas cellular atypia and necrosis showed a borderline significance. Our study thus documents the independent importance of certain histological features for morbidity and mortality in surgically treated cases of papillary thyroid cancer.

Chronic cold agglutinin disease of the "idiopathic" type is a premalignant or low-grade malignant lymphoproliferative disease.

We investigated the clinical, pathological, and immunological features of "idiopathic" cold agglutinin disease (CAD) in a population-based study. Fourteen patients were studied, giving a prevalence of about 14 per million with a mean age of 75 years. Haemolysis was present in all cases, but only eight patients had clinical symptoms of peripheral haemagglutination. Serum electrophoresis, immunofixation, morphological bone marrow evaluation, and flow cytometric immunophenotyping were used to detect any monoclonal lymphoproliferative disorder. Flow cytometry seemed to be a sensitive way to demonstrate a clonal B-cell proliferation. Some evidence of clonality was found in 13 patients, and a clonal lymphoproliferative disease was documented by flow cytometry or biopsy in 10 out of 11 patients. We conclude that CAD is a symptom-producing monoclonal lymphoproliferative disorder in nearly all patients.

Actual volume estimation of the paracortex in lymph nodes by morphometry. Significance of centrally placed sections.

Variation of paracortical size in lymph nodes has been demonstrated to be an important indicator of paracortical activation under chronic antigenic stimulation. This introduces a stereologic problem that can be solved by the analysis of serial sections. However in practical applications, a such procedure is too time consuming. Evidence is presented here that data from centrally placed, single sections in mice and guinea pigs may serve as a fairly good parameter for paracortical volume fractions when groups of lymph nodes are analyzed. However in predicting individual volume fractions, there is considerable risk of serious deviation from actual values.

Balloon cell melanoma mimicking clear cell carcinoma.

Balloon cells may occur in both benign nevi and malignant melanomas. Sometimes they dominate the histological appearance and cause difficulties in biopsy interpretation. There are no specific clinical characteristics. We report a metastatic balloon cell melanoma where the primary tumour was not identified and the histological appearance mimicked that of a clear cell renal carcinoma.

Increased content of chromium and nickel in lung tissues from patients with bronchial carcinoma.

Epidemiological studies have shown that occupational exposure to certain chromium and nickel compounds is followed by an increased lung cancer incidence. However, few data exist on the content of these metals in lung cancer patients in general. In the present study, central and peripheral lung tissue, bronchial tissue and hilar lymph nodes were collected from 20 patients with bronchial carcinoma and 21 control individuals, and the tissue concentration of chromium and nickel was measured by use of atomic absorption analysis. Increased levels of both metals were found in cancer patients as compared to controls. Lung tissue concentration of chromium was two-fold increased, while the bronchial wall content of nickel was three times the level in control individuals. Smokers showed a dose-related increase in the deposition of both chromium and nickel. Furthermore, in cancer patients an inverse relationship between smoking and the tissue level of chromium in regional lymph nodes was found, possibly indicating a depressive effect on pulmonary clearance mechanisms. Our results emphasize the possible role of small amounts of chromium and nickel as agents in bronchial carcinogenesis, unrelated to occupation and probably related to tobacco smoking.

Dynamic and morphological responses of draining lymph nodes to single and repeated applications of oxazolone to guinea-pig skin.

Auricular lymph nodes in guinea-pigs were studied for 3 weeks under continuous stimulation with oxazolone applied to the ear skin. Quantitation of 3H-Tdr labelled paracortical lymphocytes following pulse labeling, demonstrated a marked, but only transitory rise in the proportion of cells in DNA synthesis. In spite of this, the total number of cells in S-phase continued to rise during the remaining part of the observation time, as a result of a steady increase of the paracortical cell mass. The variation in the proportion of large, pyroninophilic blast cells, revealed a pattern similar to that of the labelling index. A high proportion of blast cells was found only in the initial phase of the reaction, while the chronic response was characterized by a low proportion, no different from the starting level. Paracortical enlargement appeared to be the most reliable morphological criterion on which a chronically stimulated paracortex could be distinguished from an "unstimulated". The initial paracortical response, similar to a primary reaction, was followed by an equally pronounced development of germinal centres and plasma cells. These were also persistent features under continued stimulation. The reported changes are most likely specific responses to oxazolone stimulation.

Morphological changes in paracortical high endothelial venules to single and repeated application of oxazolone to mouse skin.

Morphological changes in paracortical high endothelial venules (HEV) have been studied after single and repeated applications of oxazolone to mouse skin. The primary response was characterized by a rapid and marked vascular dilatation, by marked increase in the cross sectional area of the vessel wall and in the size of the individual endothelial cells, as well as increased accumulation of lymphocytes in HEV walls. These changes regressed (in the more long lasting response). The transitory structural alterations may be related to increase of blood flow, increased flow of cells from the thymus to the stimulated lymph nodes as well as increased transport capacity of endothelial cells based on augmentation of their surface area. These features are closely related to other cellular events such as a prominent blastoid reaction in the paracortex as well as an associated cellular depletion of the thymus.

Responses of the thymus and the paracortex of draining lymph nodes to repeated applications of oxazolone to mouse skin.

The oxazolone-induced response in the paracortex of draining lymph nodes is characterized by an early increase in the proliferative activity that decreases to control levels when stimulation is continued. The possibility that this may be a toxic side effect of the concentrated oxazolone solution used was investigated by simultaneous registration of the changes taking place in the thymus. These were found to be different from toxin-induced changes and compatible with cell loss due to massive emigration of cells. Repopulation of the thymus took place over the last 1 1/2 week of stimulation. It was concluded that the changes in the thymus as well as the decline of the proliferative activity in the paracortex, are most likely physiological responses. The most important factor in maintaining a high production of paracortical lymphocytes under chronic stimulation is the increase in the lymphocyte mass in the paracortex.

Dose dependency of the responses in draining lymph nodes and skin to repeated applications of oxazolone. A quantitative and histological study in mice.

The skin reactivity and lymph node responses to a large scale of different doses of oxazolone, were followed under continuous stimulation for a period of 12 days. It was found that the lack of a continuous high blast cell activity in the paracortex was a physiological response that could be observed with high dosage as well as with very low dosage. Low doses gave distinct stimulation of the paracortex without any detectable reaction in the cortex and in the medulla. Manifest stimulation of these compartments required considerably higher doses. A marked paracortical response was the only lymph node change required for development of a typical delayed type skin response. When germinal centres and plasma cells had developed, the skin reactivity was also found to change characteristically.

Repreoducibility of a method for a graded stenosis in tubes and vessels of small calibres. An in vitro and in vivo experiment.

The reproducibility of a previously described method for graded stenosis has been tested in a model experiment and by repeated stenosis of the portal vein in rats. In the model experiment, the method was tested in terms of flow variations; in the animal experiment, in terms of prestenotic pressure variations. In the model experiment, the coefficient of variation in repeated stenosis was less than 3%. In the animal experiment, repeated stenosis constantly produced the same high level of pressure. The present method is discussed in relation to a theoretical optimal standard and found to be satisfactory.

Prehepatic portal hypertension in the rat. Immediate and long-term effects on portal vein and aortic pressure of a graded portal vein stenosis, followed by occlusion of the portal vein and spleno-renal collaterals.

The creation of a portal vein stenosis with a diameter of 1.2 mm in Wistar rats resulted in a sustained elevation of the portal vein pressure to approximately twice the control value lasting for at least 8 weeks. The alterations in portal vein pressure following occlusion of the stenosed portal vein and splenorenal collaterals, either separately or in combination, indicate that a considerable quantity of portal blood is diverted into the inferior caval vein through the splenorenal collaterals, and that the development of these collaterals occurs during the first 3-4 weeks after the establishment of the stenosis. The concomitant reduction in aortic pressure and increase in heart rate following total occlusion of the portal vein were most pronounced during the first weeks after stenosis, and were probably due to diminished venous return to the heart.

Starch microsphere induced small intestinal ischaemia. Blood flow and morphologic investigations of late effects.

Severe temporary small intestinal ischaemia was induced in 15 cats by mesenteric embolization of degradable starch microspheres. After an observation period of 14 days, small intestinal histopathology and tissue blood flow were investigated. Intracellular starch fragments were found in tissue macrophages in ileal Peyer's patches in all animals, but no late adverse effects of starch microsphere embolization were observed.

Two-stage occulsion of the protal vein in the rat:survival related to weight variation and the interval between partial and total occulusion.

A portal vein stenosis with a diameter of 1.1 mm was performed in female Wistar rats of four different weight groups. The mortality rate was found to increase with increasing weight of the animals, a near linear correlation between mortality rate and body weight being found. Severe circulatory disturbances of the small intestine with haemorrhage and necrosis of the bowel wall were a prominent finding at death. In the survivors total occulsion was carried out at varying times, from 48 h after the first operation, and the mortality rates were recorded. A major contributing cause of death appeared to be ischaemic infarction of the liver. Only a slight congestion of the duodenum was found and no circulatory disturbance of the small intestine. Thus by 48 h the porto-systemic collaterals are sifficiently developed to prevent subsequent venous congestion. The survival rates showed a significant increase when the time interval between partial and total occlusion was extended from 48 to 72 h. This may be due to an increased blood flow through the hepatic artery and collateral arteries to the liver.

Differentiated thyroid carcinomas: the relevance of various pathological features for tumour classification and prediction of tumour progress.

In this series of 263 surgically treated cases of thyroid cancer, 12% were finally classified as benign lesions after histopathological review. Difficulty in the assessment of tumour capsule invasion in follicular neoplasms was the most frequent cause of diagnostic error. Squamous metaplasia, clusters of ground glass nuclei and psammoma bodies were found to be the most specific discriminators between papillary and follicular carcinomas. Among papillary carcinomas, tumour diameter above 30 mm, thyroid capsular invasion and regional lymph node metastases were found to be significant prognostic factors according to survival analyses. Of additional practical importance, our results indicate that tumour infiltration in the thyroid capsule should be reported as a marker of early extra-thyroidal extension.

[Pulmonary changes as primary manifestations in Waldenström macroglobulinemia]. / Lungeforandringer som primaer manifestasjon av Waldenströms makroglobulinemi.

We describe a patient with a 19-year history of lymphocytic lung infiltrations. A diagnosis of lymphocytic interstitial pneumonitis was made in 1982 after an open lung biopsy, but in 1995 a comprehensive re-evaluation led to the diagnosis of Waldenström's macroglobulinaemia with primary bronchopulmonary involvement. It could also be demonstrated by polymerase chain reaction and immunological techniques that this disease had been present since before 1982. In 1995 it was still difficult to demonstrate bone marrow involvement, even with new and sensitive methods. We discuss some diagnostic problems of organ manifestations of uncommon systemic diseases. Pulmonary manifestations of Waldenström's macroglobulinaemia or other diseases of the immune system should be considered in patients with atypical lung disorders.

Porto-hepatic bypass by organohepatopexy in rats with prehepatic portal hypertension. Formation of new venous porto-hepatic collaterals following omento-, jejuno-, and splenohepatopexy, and their effect on portal vein pressure.

Transposition to the liver of the greater omentum; the jejunum, and the spleen in 3 groups of rats with prehepatic portal hypertension due to a calibrated portal vein stenosis with a diameter of 1.2 mm resulted in the formation of new venous collaterals from the transposed organs into the liver parenchyma. The collaterals were visualized macro- and microscopically, as well as angiographically, and were found to drain into intrahepatic branches of the portal vein, sinusoids, and hepatic veins. The portal vein pressure 8 weeks after stenosis and transposition was found to be significantly lower in rats with transposed organs than in controls. The collaterals were sufficiently well developed to significantly reduce the increase in portal vein pressure that follows acute occlusion of the portal vein and the natural splenorenal collaterals.

A rapid method for estimating volume ratios.

A rapid stereological method is proposed for estimating the volume ratio V(K2)/V(K1) of two bodies K1, K2 such that K2 is embedded in K1. If K1, K2 fit approximately into a 'star specimen' model, then a single section, taken at a certain level through K1, provides a rather accurate estimate of V(K2,/V(K1). For a population of moderately similar star specimens, a constant sectioning level can be estimated from a sample using a least squares criterion. A pilot experiment involving mouse lymph nodes, aimed at estimating the mean and the variance of the individual fractions V (paracortex)/V(node), indicates a fair robustness against deviations from the model.

Stereological estimation of volume ratios by systematic sections.

Two related problems are explored. Firstly, a single opaque solid omega 2, is considered. The problem is to estimate the minimum number of systematic sections m, necessary to estimate the volume ratio v = V (omega 2)/V (omega 1) with a coefficient of error or at most gamma 0 with a probability 1 - alpha. Secondly, we consider a population of such specimens. The second problem is to estimate the optimum number of n of specimens to be sampled and the number n of systematic sections per specimen in order to estimate the mean volume ratio v of the population with a relative error of at most epsilon 0 with a probability 1 - alpha. General guidelines for solving the two problems are presented. Practical results applicable to two populations of mouse and guinea-pid lymph nodes, exhibiting a wide variation in size and shape, are obtained.

Portohepatic bypass by splenohepatopexy in rats with prehepatic portal hypertension. A long-term (12-month) study of the development of splenohepatic collaterals and their effect on portal vein pressure.

Transposition to the liver of the spleen in rats with prehepatic portal hypertension due to a calibrated portal vein stenosis with a diameter of 1.2 mm resulted in the development of new venous collaterals from the spleen into the liver parenchyma. The collaterals were visualized macro- and microscopically as well as angiographically and were found to drain into intrahepatic branches of the portal vein, sinusoids, and hepatic veins. The portal pressure 12 months after stenosis and transposition was found to be significantly lower in rats with transposed spleens than in rats with portal vein stenosis alone. The collaterals were sufficiently well developed to significantly reduce the increase in portal vein pressure that follows acute occlusion of the portal vein and the natural splenorenal collaterals.