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Biomedical subjects

R H Beelen

Publications and source records attributed to R H Beelen.

At least 37 records · Page 2Linked to original sources
PubMed1999

Laparoscopic versus conventional Nissen fundoplication: the influence on plasma insulin-like growth factor-I and insulin-like growth factor binding protein-3.

BACKGROUND: Conventional surgery is known to influence postoperative insulin-like growth factor-I (IGF-I) levels and to increase protein breakdown. Recent animal data described that laparoscopic surgery, in contrast to conventional surgery, maintains postoperative IGF-I levels, suggesting a reduced postoperative catabolic response. AIM: The aim of this study was to demonstrate differences in the postoperative catabolic response between laparoscopic and conventional surgery. METHODS: Seventeen patients scheduled for Nissen fundoplication were randomly assigned to undergo either a laparoscopic or a conventional procedure. The catabolic response was assessed by measuring total IGF-I and insulin-like growth factor binding protein-3 (IGFBP-3). RESULTS: IGF-I levels decreased significantly 4 days after both types of surgery. IGFBP-3 decreased more rapidly after conventional surgery than after the laparoscopic technique; however, the differences between the groups were not significant. CONCLUSION: In contrast to animal data, no differences in plasma IGF-I or IGFBP-3 were seen in patients undergoing laparoscopic or conventional Nissen fundoplication.

Adult↗
PubMed1998

Milky spots in the greater omentum are predominant sites of local tumour cell proliferation and accumulation in the peritoneal cavity.

The role that milky spots in the greater omentum play in tumour cell spread in the peritoneal cavity is presently not fully understood. To study whether intraperitoneally injected tumour cells appear preferentially in milky spots of the greater omentum and to study the changes in the greater omentum, and especially in the cell population of milky spots after tumour cell infiltration, the following study was performed. A detailed temporal sequences of changes in morphology and cellular composition in milky spots of the greater omentum of Wag/Rij rats 5, 15, 30, 60 min, 2, 4, 8, 16, 24 h, 2, 4, 8 days and 2 and 4 weeks after intraperitoneal administration of 2.0 x 10(6) CC 531 tumour cells was investigated by light microscopy and electron microscopy (pre-embedding labelling). Our data showed that the milky spots in the greater omentum were the sites to which tumour cells migrated preferentially from the peritoneal cavity. The tumour cells infiltrated the milky spots and formed clusters within. The cellular population in milky spots reacted by a very rapid influx of young macrophages during the first hour and an increase of the total number of cells (P < 0.01). After 4 h tumour cells were also located on the greater omentum outside the area of the milky spots. Around these tumour cell deposits, new milky spots are formed, which increased the total number of milky spots. The cells present in milky spots are not capable of reversing the growth of tumours and finally a solid omental cake of tumour cells is formed.

Animals↗
PubMed1998

Functional and phenotypic differences of monocyte-derived dendritic cells from allergic and nonallergic patients.

BACKGROUND: Several studies suggest a role for dendritic cell in the pathogenesis of allergic disease. OBJECTIVE: The purpose of this study was to compare function and phenotype of monocyte-derived dendritic cells (MoDC) from allergic asthmatic patients and healthy control subjects. METHODS: MoDCs were developed by incubating adherent monocytes for 5 days with IL-4 and granulocyte-macrophage colony-stimulating factor. Phenotype was assessed with flow cytometry, and the antigen-presenting function was assessed with the allogeneic mixed leukocyte reaction and an autologous specific antigen presentation. RESULTS: The morphology of the MoDCs was characteristic for immature dendritic cells. MoDCs from allergic asthmatic patients showed phenotypic differences in the expression of HLA-DR, CD11b, and the high-affinity receptor for IgE. A clearly enhanced accessory potential of MoDCs from atopic asthmatic patients in the mixed leukocyte reaction was also shown. Moreover, house dust mite-specific T-cell proliferation was increased. CONCLUSION: This study suggests the involvement of dendritic cells in the pathogenesis of atopic asthma by an increased immunostimulatory capacity of MoDCs.

Antigen Presentation↗
PubMed1998

Chemokines produced by mesothelial cells: huGRO-alpha, IP-10, MCP-1 and RANTES.

Recently we showed the in vivo relevance of chemokines in cases of bacterial peritonitis in continuous ambulatory peritoneal dialysis (CAPD) patients. Mesothelial cells, the most numerous cells in the peritoneal cavity, are hypothesized to function as a main source of chemokine production. We investigated the time- and dose-dependent expression patterns of four chemokines by mesothelial cells at the mRNA and protein level in response to stimulation with physiological doses of proinflammatory mediators that are present at the site of bacterial inflammation. Besides the chemokines huGRO-alpha (attractant for neutrophils), MCP-1 and RANTES (monocyte attractants), the expression and production of IP-10 was analysed. Mesothelial cells were cultured and stimulated with either IL-1beta, tumour necrosis factor-alpha (TNF-alpha) or IFN-gamma or combinations of these. The time- and dose-dependent mRNA expression of the chemokines was determined by Northern blot analysis and the protein production by ELISA. It was concluded that mesothelial cells could indeed be triggered by the mentioned stimuli to induce mRNA and protein production (huGRO-alpha and IP-10) or to augment constitutive protein production (MCP-1). However, RANTES mRNA and protein production could only be induced in some cases and only in small amounts. The chemokine response of mesothelial cells was regulated differentially, depending on the stimulus and the chemokine measured. In distinct cases, combination of the stimuli led to synergy in mRNA expression and protein production. The presented in vitro data support our hypothesis that mesothelial cells in vivo are the main source of relevant chemokines in response to proinflammatory mediators, suggesting an important role for mesothelial cells in host defence.

Cells, Cultured↗
PubMed1998

Experimental models in peritoneal dialysis: a European experience.

BACKGROUND: The development of adequate animal models is important for the in vivo study of selected aspects of peritoneal dialysis (PD) that cannot be evaluated by an in vitro model, such as peritoneal membrane transport, the influence of local defense mechanisms, and for testing new osmotic agents and their biocompatibilities. METHODS: Our experience with animal models for PD, including the acute Stockholm model in non-uremic rats, the acute and chronic Amsterdam model in non-uremic rats, and the chronic Gent model in uremic rats, is described. RESULTS: The Stockholm model proved to be useful in understanding the normal physiology of peritoneal transport, and for testing new dialysis solutions and their biocompatibilities. It is a rather simple and inexpensive model, and thus is suitable for screening new solutions and additives. The Amsterdam model permits the study of chemokines and mesothelial cell regeneration in vivo, and is applied in a model of chronic peritonitis. The results of the Gent model suggest that chronic peritoneal dialysis in uremic rats is feasible for at least eight weeks. This model is, however, very laborious, time consuming, and expensive. CONCLUSION: Further improvement of the technique and increase of the dialysis dose should result in a better and more realistic model for peritoneal dialysis. It is hoped that in the future these models will be useful to test the effects of long-term intraperitoneal application of different dialysis solutions and additives in uremic animals.

Animals↗
PubMed1998

Talc-induced inflammation in the pleural cavity.

Talc administration into the pleural cavity induces pleurodesis. To obtain further insight into the inflammatory process that causes pleurodesis, the cellular kinetics in the pleural space after the administration of talc was studied, along with its relation to chemokine concentrations in the pleural fluid. Thirteen consecutive patients with idiopathic spontaneous pneumothorax and eight patients with malignant pleural effusions received talc pleurodesis. The first group was treated with talc poudrage, whereas the second group was treated with talc slurry. Pleural fluids were isolated before talc administration as well as 3, 6, 24, 48 and 72 h afterwards. The talc induced a rapid polymorphonuclear neutrophil (PMN) influx followed by an accumulation of macrophages. In addition, increased production of interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 was observed. The talc-induced PMN influx reached its maximum after 3-24 h, and was related to the IL-8 concentration. In contrast, the MCP-1 was not related to the macrophage accumulation. Talc-induced inflammation in patients with idiopathic spontaneous pneumothorax and malignant pleural effusion is characterized by an influx of polymorphonuclear neutrophils related to interleukin-8, followed by an accumulation of monocytes.

Adult↗
PubMed1998

Effect of peritoneal dialysis fluid measured in vivo in a rat-model of continuous peritoneal dialysis.

To study the long-term effects of dialysis fluids on the peritoneal cavity, an in vivo model for continuous peritoneal dialysis in rats was developed. Mini vascular access ports were implanted subcutaneously in the neck of the rats and an attached catheter was instilled into the peritoneal cavity. Rats were injected daily with 10 mL of standard 3.86% Dianeal or saline for a period up to 12 weeks. In the peritoneal cavity an initial increase in total cells was observed after 4 weeks of fluid instillation. This had declined after 12 weeks. A similar trend was also seen for macrophage and neutrophil numbers, whereas the percentage of lymphocytes kept increasing in time. An effect of fluid instillation was observed on the density and the morphology of the mesothelial monolayer of the rats. A higher density of cells was observed after 12 weeks, and foci of young mesothelial cells within activated mesothelium were found. The results show that the rat model presented can be compared with the situation in the peritoneal cavity of continuous ambulatory peritoneal dialysis (CAPD) patients, and therefore is suitable for intervention studies.

Animals↗
PubMed1997

Ontogeny of milky spots in the human greater omentum: an immunochemical study.

BACKGROUND: Milky spots in the human greater omentum are preformed specific accumulations of primarily macrophages within the stroma of the greater omentum. To obtain a better understanding of milky spots in the human greater omentum, the development and the earliest forms of milky spots in the human greater omentum were studied, with special attention to the macrophage population. METHODS: Specimens of human greater omentum were obtained from fetuses of 20 to 40 weeks gestation and one newborn three days old (n = 6). Using mature macrophages (RFD 7), activated macrophages (RFD 1), B-lymphocytes (CD 22), and T-lymphocytes (CD 2), and immunoperoxydase labeling, the percentage of these cells in developing milky spots and the development of milky spots were studied by light microscopy. A time-dependent increase in the percentage of positive staining cells and the size of clusters was analyzed using the non-parametric Spearman rank correlation test. RESULTS: Small accumulations of cells with about 50% monocytes/macrophages were present at 20 weeks of gestation. With increasing gestational age the number of clusters of cells increased significantly (P < 0.01) as well as their size (P < 0.01). Starting at 29 weeks, vascularized clusters of cells were seen; true milky spots were present at 35 weeks. A significant (P < 0.05) increase in the percentage of mature macrophages was found in developing milky spots, whereas no activated macrophages were seen. The percentage of B-lymphocytes and T-lymphocytes found in the clusters of cells and milky spots increased significantly (P < 0.05) but did not exceed 10% of the total number of cells. CONCLUSIONS: From our data it can be concluded that milky spots are specific structures in the greater omentum formed between the 20th and 35th week of gestation. Further, we concluded that immature cells (promonocytes) mature locally in developing milky spots.

Cell Aggregation↗
PubMed1997

A functional study on the migration of human monocytes to human leukemic cell lines and the role of monocyte chemoattractant protein-1.

In the present study the migration of human monocytes towards the supernatants of five different human myeloid leukemic cell lines, four different human lymphatic leukemic cell lines and blasts derived from three different patients with acute myeloid leukemia (AML) was studied and the role of monocyte chemoattractant protein (MCP)-1 was established with an ELISA assay. Large differences in migration of monocytes towards the leukemic cell supernatants were shown (variation of approximately 10 to 150% compared to positive control), but high amounts of monocyte migration was always restricted to myeloid leukemic cells (cell lines or patient blasts). MCP-1 turned out to play a major role in the migration, firstly since there was a direct correlation between the amount of migration and the concentration of MCP-1 in the supernatants, and secondly since the addition of anti-hMCP-1 was able to inhibit migration to background level in all cases. Cytotoxicity experiments with a MTT test using MCP-1-stimulated monocytes against two human myeloid leukemic cell lines showed no increase in cell death compared to unstimulated monocytes. It is concluded that monocyte migration towards leukemic cells is restricted to the myeloid lineage and is regulated by MCP-1, which is produced in different amounts by the leukemic cells. Besides, MCP-1 does not increase the direct toxic effects of monocytes on leukemic cells.

Cell Culture Techniques↗
PubMed1997

Granulocyte macrophage colony stimulating factor pretreatment induces an increase of rat Kupffer cells with enhanced cytotoxicity in vitro and prevention of tumor outgrowth in vivo.

Male Wag rats were pretreated for 7 days with 1000 U/ml recombinant murine granulocyte macrophage colony stimulating factor (rmGM-CSF). Rat Kupffer cells (KC) were isolated by a enzymatic method. We injected decreasing numbers of CC531 tumor cells in the portal system. Mean KC yield increased from 1.5 +/- 0.2 to 2.2 +/- 0.2 (p < 0.05). Mean percentage of KC-mediated cytotoxicity against CC531 increased from 20.0 +/- 0.5 to 42 +/- 1.0 after rmGM-CSF (p < 0.05). At 1 x 10(5) CC531 tumor cells we demonstrated prevention of formation of small foci of CC52+ tumor cells. We demonstrate increased isolated KC with enhanced cytotoxic capacity after rmGM-CSF. rmGM-CSF induced prevention of minimal residual disease in the rat liver.

Animals↗
PubMed1997

Production of IL-1 beta and TNF-alpha by peritoneal macrophages depends on the bacterial species and the inoculum.

Peritoneal macrophages (PMs) are very potent producers of proinflammatory stimuli, such as interleukin (IL)-1 beta and tumor necrosis factor (TNF)-alpha. After contact with invading micro-organisms, PMs produce different cytokines, both pro- and anti-inflammatory. Therefore, they are crucial in the regulation of inflammatory events. The aim of this study was to investigate whether the production of IL-1 beta and TNF-alpha is dependent on the bacterial species used. PMs were harvested from spent peritoneal dialysis effluent and subsequently stimulated with five strains of bacteria in two different concentrations. After 24 hours of stimulation, supernatants were harvested and analyzed for both IL-1 beta and TNF-alpha content. IL-1 beta was measured with a commercial ELISA, and TNF-alpha was determined with a bioassay. Both the IL-1 beta and the TNF-alpha production were species-dependent. One strain of Staphylococcus aureus and one strain of Staphylococcus epidermidis induced a markedly higher response in both IL-1 beta and TNF-alpha than the other species. This response was also dose-dependent, and this holds true for all species. In conclusion, the IL-1 beta and TNF-alpha response by PMs is both species- and dose-dependent.

Dialysis Solutions↗
PubMed1996

Omental milky spots in the local immune response in the peritoneal cavity of rats.

BACKGROUND: Milky spots have been described as reactive structures, their classification varying from inflamed or haematopoietic tissue to lymphoid organs. In this study we investigated the reactivity of the milky spots in the omentum of rats upon induction of a chronic immune response in the peritoneal cavity. METHODS: At different time points after intraperitoneal administration of Bacillus Calmette-Guérin (BCG), a peritoneal lavage was made, and the omentum and the draining parathymic lymph nodes were taken out. The cellular composition of these tissues was examined on the light microscopic level, using a panel of monoclonal antibodies, and also by electron microscopy. RESULTS: During the first 4 months after administering BCG, the number and size of the milky spots increased enormously. Separate macrophage, T, and B cell areas were formed, but interdigitating cells and follicular dendritic cells were not observed. The number of cells in the peritoneal cavity also increased, and the cellular composition showed a strong similarity with that of the milky spots. Especially during the onset of the experiment, most bacteria were observed in the macrophages in the milky spots rather than in the draining lymph nodes. A cellular immune response was observed in the parathymic lymph nodes but not in the milky spots. CONCLUSIONS: Milky spots, either unstimulated or stimulated, should be classified as perivascular infiltrates. They play a role in the initial clearance of bacteria from the peritoneal cavity. Although the large increase in cell number is predominantly caused by immigration of cells, the results do support the role of milky spots as a site for local proliferation and maturation of especially macrophages and also B cells. The obtained data, however, do not support the earlier made assumption that milky spots function as a secondary lymphoid organ in the peritoneal cavity.

Animals↗
PubMed1996

Role of Kupffer cells in arresting circulating tumor cells and controlling metastatic growth in the liver.

Metastasis to the liver is a common event in clinical oncology. Blood-borne tumor cells (TCs) arriving to the liver sinusoids run into a special vascular bed. The lining of liver sinusoids is shared by Kupffer cells (KCs) and endothelial cells. KCs, liver-fixed macrophages, are responsible for detection and removal of "non-self" particles. To investigate their role in arresting blood-borne TCs and controlling tumor growth, we injected a syngeneic colon carcinoma cell line into a mesenteric vein of two groups of rats; one group was without Kupffer cells and the other normal controls. We removed the liver of these animals at different time intervals and performed immunohistochemical analysis with monoclonal antibodies (MoAbs) against our tumor cell line, three macrophage subpopulations, natural killer cells, and B and T lymphocytes. Additionally, we showed in vitro spontaneous cytotoxicity of KCs against our tumor cell line. Results suggest that KCs play a relevant role in arresting circulating TCs at the liver sinusoid, although it is limited to a small number of malignant cells. They also seem to play a major role in clearing neoplastic cells from the liver parenchyma, in controlling tumor growth in the very early stages of metastatic development, and in modulating the host immune response to cancer cells.

Animals↗
PubMed1996

Effect of intraperitoneal administration of granulocyte/macrophage-colony-stimulating factor in rats on omental milky-spot composition and tumoricidal activity in vivo and in vitro.

Milky spots in the greater omentum are small accumulations of leucocytes that consist mainly of macrophages and have recently shown to be a selective dissemination site of intraperitoneal (i.p.) inoculated tumour cells. However, milky-spot macrophages show tumoricidal activity and may, therefore, be an excellent source of effector cells suited for local immunotherapy. In the present study we first examined whether granulocyte/macrophage-colony-stimulating factor (GM-CSF) treatment of isolated milky-spot macrophages affects the cytotoxicity against syngeneic colon carcinoma cells (CC531) in vitro. Secondly, we studied the influence of intraperitoneal GM-CSF administration on the number and antitumour activity of milky-spot and peritoneal macrophages. All studies were performed in Wag/Rij rats in which a syngeneic colon carcinoma cell line (CC531) is available. The results of the in vitro study showed that GM-CSF treatment of the omental macrophages led to an increased cytotoxicity against the tumour cell line. Intraperitoneal administration of 1000 U GM-CSF daily for 7 consecutive days demonstrated both an enhanced antitumour activity of the milky-spot macrophages and an increase in the milky-spot macrophage population. An increase in the proliferative capacity, according to bromodeoxyuridine incorporation, was shown in the milky-spot macrophages. Taking into account both the enhanced macrophage number and their enhanced activity upon i.p. GM-CSF treatment, the milky-spot macrophages may provide a rationale for local intraperitoneal immunotherapy in the prevention of intra-abdominal tumour growth.

Animals↗
PubMed1996

Detection of CD 14 on migrated monocytes by specific antibody: a possible quantification for blood monocyte chemotaxis.

In the present study we investigated the possibility to use antigen-antibody recognition for detection of monocyte chemotaxis in the 48-well microchamber assay. The described method is based on recognition of cell-specific antigenic determinants present on the migrated monocytes. After conventional 48-well chemotaxis, the migrated cells were incubated with an antibody against the monocyte surface marker CD14 (3C10 hybridoma). Subsequent incubation with enzyme-coupled antibodies and their substrate allowed the antigen and hence the migrated cells carrying this antigen, to be detected and measured in a microplate reader. Our results show that chemotaxis of normal blood monocytes towards the monocyte chemoattractants FMLP and MCP-1 could be detected with the anti-CD14 antibody 3C10 in combination with a horse-radish peroxidase coupled antibody, and that the optical density is a measure for cell number per well (positive correlation, r = 0.95). Incubation of monocytes with the applied chemoattractants FMLP and MCP-1 did not change the CD14 expression as was determined by FACScan analysis. Therefore we conclude that it is possible to use antibodies directed against antigenic determinants like CD14 to detect blood monocyte migration in a more objective way compared to subjective counting of cells on a filter. Eventually, this method can be valuable, especially for chemokine research since chemokines exert their effects on specific target cell populations. By varying the detection antibody, other cell populations besides monocytes may be quantified.

Antibodies, Monoclonal↗
PubMed1996

Ingestion of Staphylococcus aureus, Staphylococcus epidermidis, and Escherichia coli by human peritoneal mesothelial cells.

In the present study we examined whether mesothelial cells can ingest and digest bacteria. The results showed that all strains were ingested. Ingested staphylococci proliferated abundantly, and only a few were digested. Escherichia coli, however, was digested during the first 8 h, whereafter the mesothelial cells disintegrated and proliferation of bacteria could be observed. The clinical implications of these findings are discussed.

Cells, Cultured↗