PubMed Health⌕ Search

Biomedical subjects

F T Bosman

Publications and source records attributed to F T Bosman.

At least 145 records · Page 8Linked to original sources

DNA synthesis in the non-infarcted cardiac interstitium after left coronary artery ligation in the rat: effects of captopril.

Effects of Captopril. Journal of Molecular and Cellular Cardiology (1991) 23, 1245-1253. This study was undertaken to investigate the alterations in interstitial DNA synthesis and collagen content in the non-infarcted left and right ventricle after induction of a myocardial infarction (MI) in the rat. MI was induced by ligation of the left anterior descending coronary artery. All animals received 5-Bromo-2'-deoxyUridine (BrdU), via a subcutaneous osmotic minipump, one day before sacrifice, to quantitate DNA synthesis. A transient rise in BrdU incorporation was found in both ventricles. Peak levels were found at day 7 and 14 after infarct induction. BrdU incorporation had returned to control levels 3 weeks after infarct induction. By using anti BRDU--anti-laminin immuno- double staining DNA synthesis was localized mainly in the cardiac interstitium. Concomitantly, a sustained increase in collagen content, measured as the Sirius red positive area on cross sections, was found from day 7 after infarct induction. No changes were found in sham animals. In the second part of the study the effects of the angiotensin I converting enzyme inhibitor captopril and the arteriolar vasodilator hydralazine on MI induced interstitial DNA synthesis and collagen content were investigated. Captopril reduced both the increase in DNA synthesis and collagen content. Hydralazine did not affect interstitial DNA synthesis, but reduced the MI induced collagen content. Both drugs had no effects in sham animals. We conclude that induction of a myocardial infarction stimulates interstitial DNA synthesis and increases the collagen content in the non-infarcted areas of the heart. Interstitial DNA synthesis is dependent on the angiotensin I converting enzyme in a direct manner independent from afterload changes.

Animals↗

In vivo DNA synthesis is not uniformly increased in arterial smooth muscle of young spontaneously hypertensive rats.

We compared the distribution of DNA synthesis over the arterial tree of young normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR) with marginally elevated blood pressure. Six-week-old male SHR and WKY rats were therefore infused with 5-bromo-2'-deoxyuridine (BrdUrd) for 2 days and the nuclear incorporation of the thymidine analogue in the media of various arteries was determined by immunohistochemistry. In WKY rats, 2.5% of the arterial smooth muscle nuclei in elastic, muscular and resistance arteries incorporated BrdUrd. In SHR, DNA synthesis was more marked in large arteries than in resistance arteries. It was in addition significantly larger in the aorta, superior mesenteric, renal and femoral arteries of the SHR than in those of the WKY rats. However, nuclear incorporation of BrdUrd in vivo did not differ between SHR and WKY rats in aortic endothelium, carotid arterial smooth muscle, nor in mesenteric or renal resistance arteries. Between 6 and 20 weeks of age, the number of nuclear profiles per media cross-section did not increase in large arteries of WKY rats and SHR. During this period of time, however, carotid artery and thoracic aorta weight and DNA content increased. SHR large arteries gained more DNA than those of WKY rats. These data indicate that DNA synthesis is uniformly distributed over the arterial system in young WKY rats and that DNA synthesis is elevated in the smooth muscle of large arteries of 6-week-old SHR but not in their resistance arteries.

Animals↗

Angiotensin II induces smooth muscle cell proliferation in the normal and injured rat arterial wall.

The present study was undertaken to explore the possibility that neointimal smooth muscle cells, the characteristic cells of restenosis and atherosclerosis, are selectively stimulated to replicate by a hypertensive stimulus. Angiotensin II (AII) was infused by osmotic minipumps for 2 weeks in 4.5-month-old rats. Group A received AII (200 ng/min) 2 weeks after a balloon catheter-induced injury of the thoracic aorta and left common carotid artery. Group B received only AII, group C only balloon denudation, and group D neither balloon injury nor AII. During the AII or Ringer's solution infusion, all animals received [3H]thymidine via a second minipump to measure DNA synthesis. AII increased the systolic pressure by more than 40 mm Hg. AII significantly increased DNA synthesis in the media of the carotid artery from 0.2 +/- 0.2% in group C to 2.5 +/- 1.5% in group A (mean +/- SD, n = 5 or 6). DNA synthesis in the neointima of the carotid artery significantly increased with AII from 4.8 +/- 4.2% in group C to 19.8 +/- 13.9% in group A. Cross-sectional area of the neointima almost doubled during AII infusion, and it increased approximately 25% in the media. Comparable results were obtained in the aorta. In a second experiment, AII was infused (125 ng/min) for 2 weeks in 11-week-old rats. Concomitantly, [3H]thymidine was given. Control rats received Ringer's solution and [3H]thymidine in their pumps. Blood pressures were elevated to the same extent as in the older animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Angiotensin I converting enzyme inhibitors and cardiac remodeling.

Angiotensin I converting enzyme (ACE) inhibitors are widely used in the treatment of heart failure. It is not known whether the beneficial effects of ACE inhibition are only due to a reduction in pre- and afterload or whether ACE inhibition also has direct effects on cardiac remodeling processes after myocardial infarction. In addition, the effects of differential timing of the treatment are not known. The left coronary artery was ligated in rats to induce a myocardial infarction. Control rats received no treatment. Captopril was given via s.c. placed minipumps (500 micrograms/kg.h) in the first 3 weeks or in the third and fourth week after induction of the myocardial infarction. The structural changes of the heart were investigated. Early after ligation the left ventricle was dilated in association with a marked, but transient DNA synthesis in the remaining left ventricle; the amount of collagen also increased. Early captopril treatment blocked this response, while late treatment had no effects. We suggest that the negative effects of early captopril treatment are due to an interference with the normal adaptive responses of the heart to the loss of muscle, probably through interactions at the cellular level.

Adaptation, Physiological↗

Prognostic significance of type IV collagen and laminin immunoreactivity in urothelial carcinomas of the bladder.

Invasion of a carcinoma involves the degradation and penetration of the subepithelial basement membrane (BM). This phenomenon might be used for histopathologic evaluation of neoplasms of the bladder. The authors studied the clinicopathologic data and tissue specimens of 125 cases of urothelial carcinomas collected prospectively. Penetration of the BM was evaluated by immunohistochemical staining of the BM components laminin and type IV collagen. The use of this parameter as a prognostic indicator in bladder cancer was assessed. The 5-year survival rate of patients having tumors with an interrupted or absent BM was significantly lower than that of patients having tumors with an intact BM. The rate of progression was greater in tumors with an interrupted or absent BM than in tumors with an intact BM. No association was found between BM status and recurrence. However, a significant correlation between tumor stage and BM staining was found. A correlation was also found between ploidy and BM staining as well as between histologic grade and BM staining pattern. When evaluating histologic grade, stage, ploidy, age, and BM score as prognostic parameters, the stage of bladder carcinomas turned out to be the most important factor in predicting the survival rate and the progression-free survival. However, BM staining was found to be of value for early identification of microinvasion and is helpful for correct staging of urothelial carcinomas.

Basement Membrane↗

Solid-phase adsorption of antigens for efficient production of antibodies reactive with native and fixed tissue antigens.

A study has been made of the efficacy of different immunization protocols using low antigen levels for the generation of monoclonal antibodies capable of detecting antigens (ADCP) in processed tissues. Protocols using unprocessed native antigen immobilized on nitrocellulose were more efficient than soluble antigen in generating serum antibodies reactive with both native antigen and processed tissues. The derived monoclonal antibodies reacted with native but not processed antigen. The use of antigen immobilized on polyvinylidene (PVDF) and subsequently processed as for histochemistry was successful in generating monoclonal antibodies reactive with processed antigen.

Adenosine Deaminase↗

Adenosine deaminase complexing protein (ADCP) expression and metastatic potential in prostatic adenocarcinomas.

The expression of the adenosine deaminase complexing protein (ADCP) was investigated by immunohistochemistry in the normal and hyperplastic human prostate, in 30 prostatic adenocarcinomas, and in seven human prostatic adenocarcinoma cell lines grown as xenografts in athymic nude mice. In the normal and hyperplastic prostate, ADCP was localized exclusively in the apical membrane and the apical cytoplasm of the glandular epithelial cells. In prostatic adenocarcinomas, four distinct ADCP expression patterns were observed: diffuse cytoplasmic, membranous, both cytoplasmic and membranous, and no ADCP expression. The expression patterns were compared with the presence of metastases. We found an inverse correlation between membranous ADCP immunoreactivity and metastatic propensity. Exclusively membranous ADCP immunoreactivity occurred only in non-metastatic tumours. In contrast, the metastatic tumours showed no or diffuse cytoplasmic ADCP immunoreactivity. This suggests that immunohistochemical detection of ADCP might predict the biological behaviour of prostatic cancer. However, the occurrence of membranous ADCP immunoreactivity in the xenograft of a cell line (PC-EW), derived from a prostatic carcinoma metastasis, indicates that not only the tendency to metastasize modulates ADCP expression.

Adenocarcinoma↗

Is immunohistochemical analysis of oestrogen and progesterone receptors in endometrial carcinoma superior to the radioligand binding assay?

The aim of this study was to evaluate tissue and steroid receptor heterogeneity in endometrial carcinoma specimens as a possible source of discordance between biochemically assayed receptor status and response to endocrine treatment. For this purpose the oestrogen receptor (OR) and progesterone receptor (PR) levels in specimens from 16 endometrial carcinoma patients were analysed on adjacent tissue sections using both a radiochemical and an immunohistochemical assay. With immunohistochemical receptor analysis extensive tissue and tumour cell receptor heterogeneity was observed. Many tumour samples revealed up to 75 per cent contamination with benign tissue. In the majority of cases, evaluation of immunoreactivity in normal tissue elements of the specimen could explain the apparent discordance between semiquantitative immunohistochemical receptor scoring of tumour cells and radiochemical receptor assay. Immunohistochemical analysis of OR and PR in endometrial carcinoma specimens allows a more specific determination of tumour cell receptor content and hence may yield a more accurate prediction of response to endocrine therapy than the biochemical assay.

Adult↗

Muscle fiber typing in routinely processed skeletal muscle with monoclonal antibodies.

Muscle fiber typing is conventionally performed using mATPase enzyme histochemistry on cryostat sections. After pre-incubation of sections at pH 4.3, 4.6 and 10.3, based on the pattern of enzyme reactivity, the fibers can be classified in types I, II (subtypes A, AB and B) and the intermediate C (I and II) fibers. We have attempted to perform fiber typing of human psoas muscle by immunohistochemistry, using monoclonal antibodies R11D10 (specific for cardiac and type I skeletal myosin) and MY-32 (specific for fast muscle fibers) on cryostat as well as on paraffin sections. Staining of consecutive cryostat sections showed that type I fibers are R11D10 reactive whereas type II fibers are MY-32 reactive. Subtyping of type II fibers could not be performed by immunohistochemistry. Quantitative analysis of type I and II fibers showed that enzyme histochemical and immunohistochemical analysis are in close agreement.

Antibodies, Monoclonal↗

Absence of type IV collagen in the centre of the corneal epithelial basement membrane.

Type IV collagen is the basic structural component of all basement membranes (BM), and forms the backbone to which other BM components attach. We have found that in the centre of the adult human cornea the epithelium does not display a type IV collagen immunoreactive BM. In fetal corneas (14 and 22 weeks of gestation), however, the epithelial BM shows uninterrupted type IV collagen immunoreactivity. In similar experiments laminin immunoreactivity was observed in the entire corneal epithelial BM, in fetal as well as adult corneas. Ultrastructurally, a normal BM with a lamina lucida and a lamina densa can be observed in the conjunctiva. The adult corneal centre, however, shows epithelium without a lamina densa. Focal deposits of electron-dense material are observed in conjunction with hemidesmosomes and anchoring fibres. These observations indicate that in the development of the eye, the cornea is initially covered with an epithelium which attaches to a normal BM. Later on, however, the BM type IV collagen disappears from the corneal centre. Assuming that highly differentiated epithelium cannot produce a BM, this could be due to the high level of differentiation of central corneal epithelium, which is generated in the limbal proliferation zone. Alternatively, the acellular Bowman's layer might lack triggers to induce type IV collagen production by the epithelial cells.

Adult↗

Colonic mucosal changes in nude mice associated with orthotopic xenografts of human colon cancer cells.

We used the nude mouse tumour xenograft model to study the pathogenesis of mucosa alterations in the large bowel surrounding a carcinoma. In mouse colonic mucosa overlying HT-29 colonic carcinoma xenografts in the caecum, the crypts were elongated in comparison with those in distant mucosa and also frequently showed a shift towards sialomucin production. These features, which are comparable with socalled transitional mucosa (TM) in man, were absent in control animals inoculated with Indian Ink instead of HT-29 cells. Although the localization of the proliferative cell compartment in mouse colonic mucosa overlying HT-29 xenografts appeared to be confined to the lower half of the crypt as in normal mucosa, the relative length of the DNA synthesizing cell compartment along the crypts was slightly elongated. These data strongly suggest that TM should be regarded as a secondary phenomenon rather than a premalignant change in large intestinal epithelium and that higher proliferative activity of epithelial cells contributes little to the elongation of crypts in TM.

Animals↗

Southern and dot blot analysis of DNA from formalin-fixed, paraffin-embedded tissue samples from colonic carcinomas.

Two extraction methods for the isolation of DNA from formalin-fixed, paraffin-embedded tissue samples from colonic carcinomas were compared. The processed DNAs were compared with DNAs from fresh specimens of the same tumors. The two extraction methods gave similar results. Formalin-fixation and paraffin-embedding irreversibly denatured DNA and consequently decreased the extraction yield and interfered with the quantitative measurement of DNA. Southern blot and dot blot analysis of processed and native DNA was performed using a c-myc and an actin probe. The results show that for Southern analysis processed DNA can be used but, due to the generation of random breaks, the restriction fragments have to be small. Furthermore, the fixation-induced crosslinking of DNA appears to hamper hybridization. For these reasons processed DNA can be analyzed better by dot blot rather than Southern blot hybridization.

Blotting, Southern↗

Examination of bone marrow biopsy specimens and staging of small cell lung cancer.

Bone marrow biopsy specimens were evaluated retrospectively in 63 of 88 (72%) patients with small cell lung cancer (SCLC). Significant differences were not found between extensive disease (ED) patients with or without bone marrow metastases in survival nor in nadirs of leucocytes or platelets subsequent to chemotherapy. A panel of antibodies was used to investigate whether immunohistochemical analysis on routinely processed bone marrow biopsy specimens could detect marrow metastases more effectively than conventional microscopy. In histologically proven marrow metastases and in control SCLC sections a combination of an antibody against cytokeratin 8, 18 and 19 (NCL5D3) and an antibody against neurone specific enolase was validated for detection of metastases. In histologically negative marrow biopsy samples, however, this combination did not yield any additional tumour positive cases. Therefore, histological evaluation of a bone marrow biopsy specimen, even when analysed by immunohistochemistry, does not contribute information relevant for staging, therapy evaluation or prognosis in SCLC.

Aged↗

Dose-dependent effects of short-term dietary administration of the food additive butylated hydroxyanisole on cell kinetic parameters in rat gastro-intestinal tract.

Groups of ten 5-week old male Wistar rats were fed a diet containing 0, 0.25, 0.50, 0.75, 1.0 or 2.0% butylated hydroxyanisole (BHA) ad libitum for 2 weeks; another group of rats served as a pair-fed control (PFC) group for the 2% BHA-fed animals. Subsequently, rats were injected i.p. with the thymidine-analogue bromodeoxyuridine (BrdU) which was incorporated into the DNA of cells during DNA synthesis. Cell kinetic parameters in gastro-intestinal tract tissues were determined by means of bivariate BrdU/DNA analysis applying flow-cytometry to randomized tissue samples or by applying immunohistology to randomized tissue sections. In the forestomach, glandular stomach, small intestine and colon/rectum, mean tissue labelling index (LI) was significantly increased in rats on a diet supplemented with 2% BHA, in comparison with rats fed the basal diet (0% BHA) ad libitum or restricted to the mean daily intake of 2% BHA-fed rats (PFC). In the oesophagus of rats fed 2% BHA, the LI was significantly higher in comparison with their PFC group, but not with the group of rats fed 0% BHA ad libitum. In rat forestomach, an apparent no observed effect level for ad libitum fed rats was found at 0.5% BHA (LI) and at 0.75% BHA (potential doubling time). Thus, the oesophagus, glandular stomach, small intestine and large bowel, in addition to the forestomach, are possible target tissues in rats for the proliferation enhancing effects of BHA. At the time of termination of the experiment, plasma BHA concentrations were dose dependently increased and were in the range that is easily attained in man after ingestion of a dose equal to the acceptable daily intake for BHA (0.5 mg/kg).

Animals↗

Solitary and synchronous adenomas of the colon and rectum: comparison of malignancy parameters.

The histological, mucin histochemical and immunohistochemical features were evaluated of 74 solitary and 73 synchronous colorectal adenomas which were endoscopically removed from 124 patients. Of the patients, 60% had a single adenoma, whereas 40% had at least two adenomas in their colorectum. Comparing the incidence of synchronous adenomas in both sexes revealed a statistically significant higher incidence (P less than 0.005) in males. The localization of the solitary and synchronous adenomas in the large bowel was similar. Moreover, parameters of malignant change within the adenomas (size, predominant type of mucosal growth and degree of dysplasia-with the exception of severe dysplasia) as well as signs of dedifferentiation (relative proportions of goblet and columnar cells) were also similar. Mucin staining intensities (periodic acid-Schiff, high iron diamine and alcian blue) and the immunoreactivity patterns of secretory component and carcinoembryonic antigen, both cytoplasmic and on the surface of the epithelial cells, were also identical in both groups of adenomas. Thus, neither the routine histological nor the mucin- and immunohistochemical features differed between the groups, except for severe dysplasia. It is concluded that there is no inherent difference in malignant potential between solitary and synchronous adenomas, with the possible exception of the degree of dysplasia.

Adenoma↗