Surfactant protein gene expression in metastatic and micrometastatic pulmonary adenocarcinomas and other non-small cell lung carcinomas: detection by reverse transcriptase-polymerase chain reaction.

Reverse transcriptase (RT)-PCR with primers specific for surfactant protein A (SP-A), B (SP-B), C (SP-C), and D (SP-D) genes was applied to detect metastatic non-small cell lung carcinomas. Forty-one paratracheal and subcarinal lymph nodes obtained from 41 patients with non-small cell lung carcinomas, 11 lymph nodes from 11 patients with extrapulmonary adenocarcinomas, and eight control lymph nodes from patients without cancer were analyzed using RT-PCR. PCR products corresponding to SP-B gene products were found in all eight control lymph nodes, offering evidence of SP-B gene expression in cells of lymphatic tissue. SP-A, SP-C, and/or SP-D transcripts were detected in 11 (84.6%) of 13 lymph nodes with histologically identifiable metastases of pulmonary adenocarcinomas and in 10 (55.5%) of 18 lymph nodes that were tumor free on routine histological examination. These findings provide evidence of micrometastatic nodal involvement which remains undetectable by conventional light microscopy but can be evaluated by surfactant RT-PCR. Gene expression of SP-A and SP-C was restricted to metastatic pulmonary adenocarcinomas but SP-D gene activity has been also detected in two of four metastases of pulmonary large cell carcinomas, one adenosquamous carcinoma, and nine extrapulmonary adenocarcinomas as well.

Differential expression of CD44 splice variants in intestinal- and diffuse-type human gastric carcinomas and normal gastric mucosa.

Immunohistochemical screening of gastric adenocarcinomas from 42 different patients revealed variant CD44 expression in all specimens tested. Adenocarcinomas of the intestinal type were strongly positive for epitopes encoded by variant exons v5 and v6, whereas diffuse-type adenocarcinomas predominantly expressed only exon v5. Normal stomach mucosa was stained by an exon v5-specific monoclonal antibody within the foveolar proliferation zone and on mucoid surface epithelium. Areas of intestinal metaplasia reacted positively with monoclonal antibodies specific for exons v5 and v6. Analysis of RNA expression revealed dramatic differences between normal mucosa and adenocarcinomas. Whereas in normal epithelium only two CD44 variant RNAs containing exons v5 and/or v6 could be detected, intestinal-type tumors yielded a much more complex pattern of amplification products which hybridized to exons v5 and v6. A similar complex expression pattern of CD44 variants was observed in three cell lines established from intestinal-type tumors. In a sample of a diffuse-type tumor, expression of exon v5, but not v6, could be detected, confirming the data obtained with immunohistochemistry. These differences in variant exon v6 expression observed between diffuse-type and intestinal-type stomach adenocarcinomas establish variant CD44-specific antibodies as a tool in gastric cancer diagnosis and also support the theory of different origins for these tumor types.

Importance of different CD44v6 expression in human gastric intestinal and diffuse type cancers for metastatic lymphogenic spreading.

In 42 human gastric adenocarcinomas of intestinal (n = 25) and diffuse types (n = 17) the expression of CD44v6 splice variants was investigated immunohistochemically and compared with the pattern of lymphogenic tumor spreading. Distinct differences were observed between the two cancer types: 92% of intestinal-type tumors expressed CD44v6 as in the intestinal metaplasia in chronic atrophic gastritis, while v6 expression occurred in only 17% of diffuse-type cancers. The analysis of RNA expression confirmed the immunohistochemical data. Intestinal-type cancers yielded a much more complex pattern of amplification products hybridizing to exon v6 than did normal mucosa, whereas diffuse-type tumors did not express exon v6. Also the pattern of lymphogenic spreading was quite different between the two cancer types: in diffuse-type tumors only a sinus carcinosis without CD44v6 expression was observed in a significantly higher number of lymph nodes than in intestinal-type cancers, which showed in particular infiltrative lymph node metastases always with CD44v6 expression as in the primary tumors. When infiltrative lymph node invasion occurred in v6-negative diffuse-type cancers, v6 neoexpression was also demonstrable in the lymph node metastases. Additionally, the number of infiltrative lymphogenic metastases increased with more extensive v6 expression in primary gastric cancers of both types. These data suggest that the expression of CD44v6 isoform is important for the infiltrative spreading of tumor cells into lymph nodes. Additionally, the phenotypic similarities in v6 expression between intestinal metaplasia and intestinal-type cancers, but not of tumors of diffuse type, may support the Correa hypothesis.

Early interstitial changes in hypertension-induced renal injury.

To elucidate the mechanisms of hypertensive renal injury, we investigated the time course and extent of changes in matrix composition, as well as cell proliferation and infiltration in two-kidney, one clip rats. The nonclipped kidneys from hypertensive and sham-operated control rats (n = 5 to 10 in each group) were studied at 7, 14, 21, and 28 days after clipping. Systolic blood pressure was elevated by day 7 (154 +/- 3 versus 111 +/- 4 mm Hg in sham group, P < .001, n = 10 each). Hypertension resulted in an early expansion of the interstitial volume by 37%, whereas hypertensive vascular changes and glomerular injury did not become evident until day 21. Immunofluorescence studies revealed an early interstitial accumulation of collagens I, III, IV, V, VI, and fibronectin by day 7. In contrast, the glomeruli showed a mild to moderate increase in collagens I, III, IV, V, laminin, and fibronectin but not collagen VI later in the established phase of hypertension. Staining for proliferating cell nuclear antigen as a marker of cell replication was increased in tubular epithelial but not interstitial or glomerular cells. A progressive infiltration of macrophages (16 +/- 2 versus 9 +/- 1 ED1+ cells/mm2, P < .05, n = 6) and T lymphocytes (93 +/- 15 versus 74 +/- 7 CD4+ cells/mm2, n = 8) in the cortical interstitium had already occurred by day 7. On the other hand, only macrophages increased in number within the glomeruli. Thus, renovascular hypertension leads to an early tubular cell proliferation, mononuclear cell recruitment, and deposition of matrix proteins primarily within the interstitium. We conclude that the injury producing nephrosclerosis in this model extends far beyond the glomeruli. Both the tubules and the interstitium are actively involved and may be the more important initial sites of injury.

Postoperative prognosis of pulmonary adenocarcinoma subtypes correlates with HLA-DR expression and the number of tumor-infiltrating cells.

Human pulmonary adenocarcinomas (AC) can be divided into two types with special morphologic and immunohistologic properties and a different number of tumor-infiltrating cells as shown by previous investigations. In the present study the relevance of this subdivision for patients' survival was investigated. 42 surgically resected pulmonary AC of stage I and II were subclassified using light and electron microscope. For immunohistologic phenotypization, reactions with monoclonal antibodies against HLA-DR, CD1 and CD3 were studied on fresh tumor specimens. Postoperative survival was evaluated after at least 24 months. AC of type I (N=23) with mucin production and ultrastructural properties of goblet cells showed almost no HLA-DR expression. Infiltration by CD1-positive dendritic cells Langerhans cells and CD3-positive T lymphocytes was significantly lower than in AC of type II (N=19), which expressed HLA-DR homogeneously and showed, ultrastructurally, Clara cell and/or type II pneumocyte properties. Patients' outcome was similar in stage I AC of both types: about 70% of patients were still alive after 24 months. However, significant differences were found between the two types in stage II AC with regional lymph node metastases: survival of patients with AC of type II corresponded roughly with stage I tumors (67%) but only 20% of patients with type I AC were still alive after 24 months. These results indicate that postoperative prognosis for patients with pulmonary AC of type II is more favourable than for mucinous AC of type I. This may be due to the homogeneous HLA-DR expression and higher number of immunologically competent tumor-infiltrating cells which possibly results in better tumor surveillance.

Tumor-specific apoptosis induced by the human monoclonal antibody SC-1: a new therapeutical approach for stomach cancer.

Stomach cancer is one of the most frequently occurring cancers worldwide, with a very poor prognosis, even after complete gastrectomy. We describe here an alternative therapeutical approach using a human monoclonal antibody (SC-1), which was isolated from a patient with diffuse-type gastric adenocarcinoma. We demonstrate that the antibody significantly reduces stomach cancer growth in vivo, by inducing tumor-specific apoptosis and that the antibody, even delivered in high doses, shows no toxic crossreactivity to other organs or tissues. The data presented here show that tumor-specific apoptosis can be induced and they give rise to the hope that human monoclonal antibodies with biological activity might present a completely new type of adjuvant cancer therapy.

Expression of p53 and bcl-2 in primary locally confined renal cell carcinomas: no evidence for prognostic significance.

p53 tumor suppressor gene mutations are present in a wide variety of human cancers, and the bcl-2 gene product is considered to prevent apoptosis. However, the significance of these gene products for the aggressiveness of the tumor and correspondingly for the prognosis of patients with renal cell carcinoma (RCC) is unclear. The expression of p53 and bcl-2 gene products was studied immunohistochemically using formalin-fixed paraffinembedded tumor samples of 31 locally confined RCC of patients treated with radical nephrectomy. The significance of these 2 parameters, in addition to tumor stage and malignancy grade, was tested with regard to survival and time of no recurrence using Kaplan-Meier-plots by the log rank test or Tarone's test and the Cox multiple hazard regression analysis (mean follow-up 5.4 years). Only 5 of the 31 RCCs stained positively for p53 and only 2 showed positive bcl-2 staining of tumor cells. Tumor stage (P < 0.002) and malignancy grade (P < 0.007) were statistically significant prognostic parameters for both survival and disease-free period by univariate analysis. In contrast, the detection of either p53 (P > 0.67) or bcl-2 gene product (P > 0.28) had no prognostic impact. Also in the multivariate statistical analysis, neither of the 2 parameters i.e. p53 and bcl-2 expression significantly improved the prognostic impact of the conventional prognosticators stage and grade, if applied in addition. The expression of p53 and bcl-2 seems unimportant as a prognostic factor in locally confined RCCs.

Multiple peptide hormone producing adenocarcinoma of lung with neurotensin and CRF-like immunoreactivity.

A pulmonary cancer of a 43 year-old-man was classified according to WHO-criteria as a poorly differentiated adenocarcinoma because of demonstration of mucinous substances in tumor cells and in rarely occurring acinar lumina. Immunohistochemical reactivity for several peptide hormones was detected in the predominant solid parts of the carcinoma. In many tumor cells an immunoreactivity to neurotensin, bombesin and - according to the elevated serum levels - to ACTH, CRF and calcitonin was found. Therefore the present lung cancer must be defined as a combined carcinoma composed of mucous producing and endocrine cell types. Because peptide hormone production is not restricted to the classical "neuro"-endocrine lung tumors, the histogenesis of this heterogeneous lung cancer is discussed.

Involvement of autophagic degradation in ACTH-induced skeletal muscle atrophy.

Intracellular autophagic breakdown was investigated in skeletal myocytes undergoing atrophy under the influence of ACTH. Adult male Sprague-Dawley rats received s.c. injections of ACTH (12.5 U/kg body wt) or physiological saline as control twice daily. 5 experimental and 5 control animals were sacrificed by perfusion fixation after 6, 9, and 12 days of treatment. While the weight of the left pectoralis minor muscle decreased significantly in the ACTH-treated animals compared with controls, the volume fraction of autophagic vacuoles, as determined by quantitative electron microscopy, increased significantly after 9 days (2.6-fold) and 12 days (2.9-fold) in the atrophying skeletal myocytes of experimental animals compared with controls. Only non-myofibrillar components were detected as contents of autophagic vacuoles. Since, however, the ratio of myofibrils to other cytoplasmic components remained constant, the degradation of myofibrils seems to occur via a non-lysosomal pathway. The data suggest that the breakdown of cytoplasmic components by cellular autophagy is important for the development of skeletal muscle atrophy as observed following long-term ACTH treatment or in Cushing's syndrome.

Ultrastructural investigations for reducing endothelial cell damage of vein grafts during CABG-operation and practical consequences.

BACKGROUND: In the present study the influence of different storage solutions on endothelial integrity or damage was investigated with direct methods particularly with transmission electron microscopy (TEM), scanning electron microscopy (SEM) and immunohistochemistry. METHODS: Saphenous vein segments of 10 cm in length were taken surgically from 6 male CABG-patients (aged 60-70) under standardized conditions. Each vein segment was cut into rings, which were incubated at room temperature for 45 minutes in different storage solutions, particularly in 0.9% sodium chloride solution and in buffered solution (M 199) with 5% human serum albumin respectively. Then, the vein segments were fixed in 3.5% glutaraldehyde and prepared for scanning and transmission electron microscopy to evaluate the endothelial damage. In addition, immunohistochemical staining (CD34, PECAM and Factor VIII) was performed. RESULTS: When using 0.9% sodium chloride solution, the SEM-examination revealed that 55% of the cell population was destroyed. In comparison to these findings only 26% of the endothelial cell population was damaged when the venous segment was stored in buffered solution with 5% albumin (p<0.01). In immunohistochemistry (CD34, PECAM, Factor VIII) these findings were supported. CONCLUSIONS: This study demonstrates the importance of storage solutions in regard to endothelial integrity. For best preservation of endothelium it is necessary to modify conventional storage methods. So, storage in buffered solution with albumin has shown much better endothelial cell preservation compared with physiological saline which might reduce the obliteration rate of CABG in future.

Electro-immortalization of B lymphocytes isolated from stomach carcinoma biopsy material.

The advantages of electrofusion were used to immortalize the small number of B-cells from fresh biopsy material taken from a gastric carcinoma of a patient. Two stable human antibody secreting clones could be produced which exhibited functional activity against the autologous tumour cells (inhibition of cell adhesion and immunofluorescence staining of the membranes). This shows that a variety of hitherto inaccessible B lymphocyte populations from other human organ biopsies can be immortalised by the improved electrofusion technique.

[Histologic studies in the course of programmed peritoneal lavage]. / Histologische Untersuchungen während des Verlaufs der programmierten Peritoneal-Lavage.

A programmed peritoneal lavage was done in 20 patients. During this lavage, tissue samples were withdrawn for histological control. During the therapeutical lavage, the morphological signs of inflammation decreased. For the sanitation of peritonitis, the cause of the disease must be eliminated. The morphological signs of healing start during the first three days postoperatively. Peritoneal repair prevails after the second or third postoperative day. Peritonitis will be lethal without focus sanitation and with microorganisms resisting any therapy, although lavage having been done.

[Fungal peritonitis]. / Die Pilzperitonitis.

Mycotic peritonitis can be demonstrated by microbiological, histological and serological tests. The disease can be proved histologically by a deep invasion of fungi. Initially, the mycotic peritonitis can be caused by polymicrobial infections and also by genuine mycotic invasion after perforation of the gastrointestinal tract. In the final phase of the disease only these fungi are of relevance. In most of the cases candida albicans can be verified. In the procedure of programmed peritoneal lavage the mycotic peritonitis provides a severe complication. Untreated, it would cause death by dissemination, fungemia and candida sepsis. 8 out of 12 patients with candida peritonitis died. Most of the patients had been severely ill previously and had shown several risk factors promoting mycotic disease. Antimycotic treatment has to be initiated as soon as possible, in order to diminish the high lethality.

[Synchronous growth of a mixed mesodermal tumor and an adenocarcinoma of the urinary bladder]. / Synchrones Wachstum eines mesodermalen Mischtumors und eines Adenokarzinoms in der Harnblase.

The case of synchronous growth of a mesodermal mixed tumour and an adenocarcinoma in the bladder is presented. This is a rare phenomenon. Mesodermal mixed tumours are malignancies with both epithelial (carcinoma) and mesenchymal (sarcoma) differentiation. The term carcinosarcoma is frequently used in the literature. In most cases the prognosis of mesodermal mixed tumours, which depends mainly on the high rate of local recurrences, is poor. Therefore, cystectomy with suprapubic urinary diversion is the treatment of choice. However, results reported in the literature show that transurethral resection of the tumour may be adequate when there is only superficial growth of mesodermal mixed tumours. Short follow-up intervals are absolutely essential for patients treated this way.

[Lipoma of the adrenal gland. Case report and review of the literature]. / Lipom der Nebenniere. Fallbericht und Literaturübersicht.

Adrenal lipomas are extremely rarely occurring benign tumors being hormonally inactive. The patient described in this report underwent surgical excision of an adrenal lipoma because of the associated hypertension. The blood pressure decreased postoperatively to normal levels. This case report and the review of the literature consider diagnosis and therapy of the adrenal lipomas.

Intracellular turnover and cardiac hypertrophy.

Ultrastructural evidence is presented that intracellular autophagic degradation of cytoplasmic constituents is reduced during pressure induced hypertrophy of left ventricular myocardium after supravalvular aortic constriction in rats. This anti-catabolic reaction has to be considered as an important mechanism for shifting the balance between synthesis and degradation to the positive side. Short term studies after administration of isoproterenol suggest a close functional relationship between work load on the one hand and the anti-catabolic reaction on the other.