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Biomedical subjects

A J Suurmeijer

Publications and source records attributed to A J Suurmeijer.

At least 19 recordsLinked to original sources
PubMed2001

Early expression of natriuretic peptides and SERCA in mild heart failure: association with severity of the disease.

BACKGROUND: We investigated changes in genetic expression of atrial and brain natriuretic peptides (ANP and BNP) and sarcoplasmic reticulum Ca(2+)-ATPase (SERCA) in patients with stable mild to moderate chronic heart failure (CHF), since data on this topic were primarily obtained in end-stage CHF. METHODS: We studied tissue from 25 patients with idiopathic dilated cardiomyopathy (IDC) in New York Heart Association (NYHA) class II (n=12) and III-IV (n=13). Myocardial tissue from normal hearts (n=10) served as controls. Messenger RNA (mRNA) expression of ANP, BNP, and SERCA was isolated, and correlated with severity of CHF, left ventricular function (LVEF), peak oxygen uptake (peak VO(2)), and wedge pressure. RESULTS: A significant trend for gradual changes in mRNA expression according to increasing NYHA class was found for ANP, BNP (P<0.0001) and SERCA (P=0.04), with a marked increase in patients with more advanced CHF (ANP and BNP: P<0.01 vs. controls; SERCA: NS) and less pronounced changes in patients with mild CHF. mRNA of ANP and BNP correlated strongly with LVEF (-0.621 and -0.621, respectively, both P<0.01) and peak VO(2) (-0.625 and -0.555, respectively, both P<0.01) and, to a lesser extent, with wedge pressure (0.440 and 0.488, respectively, both P<0.05). SERCA correlated most strongly with wedge pressure (-0.623, P<0.01), and weak, non-significant correlations with LVEF and peak VO(2) were found. CONCLUSIONS: Genetic expression of ANP, BNP, and SERCA is progressively altered in proportion to the severity of CHF, although this is more marked for ANP and to a lesser extent BNP, than for SERCA. These changes support the concept that already early in CHF, genetic expression is affected, which has implications for the understanding of the pathophysiology of CHF.

Adult↗
PubMed2001

Distribution of cytoskeletal proteins, integrins, leukocyte adhesion molecules and extracellular matrix proteins in plastic-embedded human and rat kidneys.

OBJECTIVE: To study the distribution of cytoskeletal proteins (actin, alpha-actinin, vinculin, beta-tubulin, keratin, vimentin, desmin), adhesion molecules for cell-matrix interations (very later antigens [VLA1-61, beta1, beta2 [CD18], vitronectin receptor [alphavbeta3], CD 11b), leukocyte adhesion molecules (ICAM-1) and extracellular matrix proteins (collagen IV, fibronectin, laminin, vitronectin) in human and rat kidneys by using a superior processing and immunohistochemical staining technique. STUDY DESIGN: Human and rat kidneys were fixed in 2% paraformaldehyde, dehydrated in acetone and processed in a new, low-toxic glycol, methacrylate mixture, especially developed for immunohistochemistry. Both the glomeruli and the interstitial areas were carefully examined and scored semiquantitatively. RESULTS: Immunostained plastic sections showed excellent morphology combined with remarkably well preserved antigenicity. CONCLUSION: The above mentioned provides an excellent tool for the accurate localization of a wide variety of antigens at the light microscopic level.

Animals↗
PubMed2000

Additional use of immunostaining for active caspase 3 and cleaved actin and PARP fragments to detect apoptosis in patients with chronic heart failure.

BACKGROUND: Apoptosis might play a role in the pathophysiology of chronic heart failure (CHF). Detection of apoptosis in tissue mostly relies on the terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) method. Recent studies showed, however, that TUNEL-positive cardiomyocytes were not in apoptosis. Therefore, additional methods are warranted. In this study, we tested the specificity of the TUNEL method by additional immunostaining for enzymes and cleaved proteins that are considered specific for apoptosis. METHODS AND RESULTS: We tested 25 myocardial tissue sections from 16 patients with severe CHF (caused by either idiopathic or ischemic cardiomyopathy) for the existence of apoptosis. We compared TUNEL with immunostaining for caspase 3, active caspase 3, cleaved actin, and poly (ADP-ribose) polymerase (PARP). TUNEL-positive myocytes were seen in 68% of the specimens. Immunostaining in myocytes for active caspase 3 and caspase 3 was positive in only a few patients, whereas staining for cleaved actin and PARP was negative in all patients. CONCLUSION: We found variable myocyte labeling with TUNEL in the failing myocardium of patients with CHF and only obtained scarce additional immunohistochemical evidence for myocyte apoptosis with immunostaining for active caspase 3 and caspase 3 and no positive myocytes with immunostaining for cleaved actin or PARP fragments. These findings support the notion that TUNEL-positive cardiomyocytes are not conclusive for the existence of apoptosis.

Actins↗
PubMed1999

Pretreatment in a high-pressure microwave processor for MIB-1 immunostaining of cytological smears and paraffin tissue sections to visualize the various phases of the mitotic cycle.

In many pathology laboratories, both microwave ovens and pressure cookers are used for pretreatment of cytologic smears and paraffin sections to allow MIB-1 staining. For both methods there are two problems. First, the results cannot be used for quantitation because standardization is impossible. Second, the staining results are often suboptimal, resulting in negative staining of cells in the G(1)- and S-phases. When pretreatment is performed in a microwave processor, allowing microwave heating under pressure, precise temperature monitoring becomes possible. In addition, the importance of the pH of the buffer was studied using a test battery series. Optimal staining is achieved at a temperature of 115C, 10 min, pH 6. This method proved to be highly reproducible. Because the immunostaining results are optimal, the various phases of the cell cycle can be defined in the sections and smears. In addition, the perinucleolar staining of the late G(1)-phase is optimally visualized and nuclei of the stable pKi-67 pathway can be identified. Under suboptimal conditions, in particular, the number of cells in the late G(1)-phase are underestimated in the MIB-1 counts.

Antigens, Nuclear↗
PubMed1999

Drug resistance-associated markers P-glycoprotein, multidrug resistance-associated protein 1, multidrug resistance-associated protein 2, and lung resistance protein as prognostic factors in ovarian carcinoma.

Intrinsic and/or acquired resistance to chemotherapy is the major obstacle to overcome in the treatment of patients with ovarian carcinoma. The aim of the present study was to investigate the prognostic value of drug resistance-associated proteins P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1), canalicular multispecific organic anion transporter (c-MOAT/MRP2), and lung resistance protein (LRP) in ovarian carcinoma. Expression of P-gp, MRP1, MRP2, and LRP was determined by immunohistochemistry of frozen tissue sections of 115 ovarian carcinoma patients and related to clinicopathological factors, response to chemotherapy, and progression-free survival. P-gp expression was observed in 20 of 115 (17%), MRP1 in 51 (44%), MRP2 in 19 (16%), and LRP in 85 (74%) tumors. Expression of MRP1 was related to MRP2 (P<0.0001) and P-gp (P<0.001) expression, whereas LRP expression was more frequently observed in patients with early stage (P<0.01), lower grade (P<0.05), and smaller residual tumor (P<0.05). Early stage (P<0.001), smaller residual tumor (P<0.001), and lower differentiation grade (P<0.05) were related to longer (progression-free) survival. P-gp, MRP1, MRP2, and LRP expression were neither related to response to first-line chemotherapy in 59 evaluable patients nor to progression-free survival in all patients. On multivariate analysis, only stage and residual tumor were independent prognostic factors for survival. In conclusion, in ovarian carcinoma, MRP1 expression is associated with MRP2 and P-gp expression, whereas LRP expression is associated with favorable clinicopathological characteristics. Assessment of P-gp, MRP1, MRP2, or LRP does not allow prediction of response to chemotherapy or survival in ovarian carcinoma.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1999

[Expression and prognostic value of the drug resistance markers P-gp, Mrp1, Mrp2, and Lrp in ovarian carcinoma].

BACKGROUND: Intrinsic and/or acquired chemoresistance is the major obstacle to overcome in the treatment of patients with ovarian carcinoma. The aim of the present study was to investigate the prognostic value of drug resistance associated proteins P-glycoprotein (P-gp), multidrug resistance related protein (Mrp1), canalicular multispecific organic anion trans-porter (c-MOAT or Mrp2) and lung resistance protein (Lrp) in ovarian carcinoma. METHODS: Expression of P-gp, Mrp1, Mrp2 and Lrp was determined by immunohistochemistry of frozen tissue sections of 115 ovarian carcinoma patients and associated to clinico-pathological factors, response to chemotherapy and (progression free) survival. RESULTS: Expression of P-gp was observed in 20 out of 115 (17%), Mrp1 in 51 out of 115 (44%), Mrp2 in 19 out of 115 (16%) and Lrp in 85 out 115 (74%) tumors. Expression of Mrp1 was related to Mrp2 (p < 0.0001) and P-gp (p < 0.001) expression, while Lrp expression was more frequently observed in patients with stage I/II versus stage III/IV tumors (p < 0.01), grade I/II versus III tumors (p < 0.05) and residual tumor < 2 cm versus > 2 cm after laparotomy (p < 0.05). Lower stage (p < 0.001), small residual tumor after first laparotomy (p < 0.001) and lower differentiation grade (p < 0.05) were related to longer (progression free) survival. P-gp, Mrp1, Mrp2, and Lrp expression was neither related to response to first line chemotherapy (59 evaluable patients) nor to (progression free) survival (all patients). On multivariate analysis only stage and residual tumor after first laparotomy were independent prognostic factors for (progression free) survival. CONCLUSIONS: In ovarian carcinoma Mrp1 expression is associated with Mrp2 and P-gp expression, while Lrp expression is associated with favorable clinicopathological characteristics. Assessment of P-gp, Mrp1, Mrp2 or Lrp does not allow prediction of response to chemotherapy or (progression free) survival in ovarian carcinoma.

ATP Binding Cassette Transporter, Subfamily B↗
PubMed1998

Antigen retrieval in formaldehyde-fixed human brain tissue.

Microwave-stimulated antigen retrieval has become a widely accepted method in both pathology and research laboratories. Since the introduction of the method in 1991, many groups have tried to optimize and standardize it. This review describes the present state of the art. A standard method for microwave-stimulated antigen retrieval in formaldehyde-fixed paraffin-embedded and nonembedded tissue is presented that results, in general, in very good staining for antibodies used in neuroscience. However, there are still a few antigens that are retrieved not at all or not in an optimal manner. Factors of importance for microwave antigen retrieval are the pH of the retrieval solution and, related to the pH, the temperature and duration of heating. These factors are discussed.

Antigens↗
PubMed1995

Confocal microscopy of false-negative breast aspirates.

Even when using a novel optimized FNA-21 for breast fine-needle aspiration biopsy, our series of 163 breast lesions with consecutive histology, including 110 carcinomas, contained eight cancer cases with a negative cytologic diagnosis. Two consisted exclusively of thick, undiagnosable epithelial fragments and were devoid of detached malignant cells on which the proper diagnosis could have been made. Optical sectioning of the original smears by the confocal microscope allowed us to reclaim from these epithelial fragments the diagnostic abnormal chromatin patterns, even in the air-dried smears. In the thin optical sections the cancerous tubular or cribriform growing pattern was visualized. The high spatial resolution of the confocal microscope can be further exploited with the application visualization system (AVS). The AVS three-dimensional reconstructions allow us to see the surface of these malignant cells in a detail approaching that of scanning electron microscopy. Furthermore, the internal aspects of the malignant nuclei can be visualized with these techniques.

Biopsy, Needle↗
PubMed1995

Value of P-glycoprotein, glutathione S-transferase pi, c-erbB-2, and p53 as prognostic factors in ovarian carcinomas.

PURPOSE: To determine the prognostic value of immunostaining of P-glycoprotein (P-gp), glutathione S-transferase (GST) pi, c-erbB-2, and p53 in patients with advanced-stage ovarian carcinoma. PATIENTS AND METHODS: Immunostaining of P-gp, GST pi, c-erbB-2, and p53 was performed on 89 primary tumors and 38 residual tumors after chemotherapy (P-gp and GST pi) in patients with advanced ovarian carcinoma treated with platinum- and doxorubicin-containing chemotherapy. The results of immunostaining were related to clinicopathologic prognostic factors, response to chemotherapy, and progression-free survival (PFS) and overall survival. RESULTS: P-gp and GST pi immunoreactivity were present in 13 (15%) and 79 cases (89%), respectively, and were not associated with any other prognostic factor or PFS or overall survival. C-erbB-2 immunoreactivity was present in 18 cases (20%) and was associated with undifferentiated histiotype (P < .05), but not with PFS or overall survival. p53 immunoreactivity was present in the nuclei of 31 cases (35%) and cytoplasm of nine cases (10%). Nuclear p53 staining was associated with grade III tumors, presence of more than 1-L ascites, and residual tumor after first laparotomy more than 2 cm. Nuclear p53 staining was associated with shorter PFS (relative risk [RR], 3.3; 95% confidence interval [CI], 2.0 to 5.6) and overall survival (RR, 2.6; 95% CI, 1.7 to 3.8). After adjustment for presence of more than 1-L ascites or age more than 50 years, nuclear p53 staining did not retain independent prognostic significance in stage III/IV tumors. The frequency of P-gp staining in residual tumors after chemotherapy (18 of 38 cases) was higher in comparison to untreated tumors (13 of 89 cases) (P < .001). No combination of prognostic parameters was able to predict response to chemotherapy adequately. CONCLUSION: Nuclear immunoreactivity of p53 in ovarian carcinomas is associated with shorter PFS and overall survival and determinants of more aggressive tumor growth. The higher frequency of P-gp immunoreactivity in residual tumors after chemotherapy points to induction of P-gp in ovarian carcinomas by doxorubicin-containing combination chemotherapy. The determination of P-gp, GST pi, c-erbB-2, and p53 does not permit more adequate prediction of response to chemotherapy.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1991

Glycol methacrylate embedding in diagnostic pathology. A standardized method for processing and embedding human tissue biopsy specimens.

A standardized "low-cost" processing and embedding procedure is described for the preparation of semithin glycol methacrylate (GMA) sections of human tissue biopsy specimens for use in routine diagnostic pathology. The method is highly reliable and reproducible and is based on many years of experience in fundamental GMA research. The procedure combines the application of new, less toxic, plasticizers and the preparation of high-quality GMA mixtures. Microwave irradiation was used to improve staining qualities and to reduce staining times. An improved reticulin staining procedure is presented as well.

Biopsy↗
PubMed1989

Accelerated immunogold silver and immunoperoxidase staining of paraffin sections with the use of microwave irradiation. Factors influencing results.

With the use of microwave irradiation, incubation times for the immunotechniques used in immunomicroscopy can be shortened significantly. The authors found that factors influencing results are irradiation time, power output, and presence of gold label. Precise temperature control is essential, and techniques to achieve this are discussed. For beta-human chorionic gonadotropin (beta-HCG), the results with the immunoperoxidase technique were inferior to those of the conventional technique, and with the immunogold silver technique an enhancement of labeling results was achieved. Immunogold silver staining for beta-HCG on paraffin sections can be performed with the use of microwave irradiation within one hour.

Histological Techniques↗
PubMed1987

Major improvement in microscopic-image quality of cryostat sections. Combining freezing and microwave-stimulated fixation.

In this article the authors describe a procedure to obtain cryostat sections of superior quality using microwave stimulation of a fixation reagent with suitable microwavability properties. In fact, the light microscopic images are of a quality that is comparable to that of the best conventional paraffin technique. The new procedure requires less time than the existing cryostat technique. The only additional piece of equipment needed in this procedure is a relatively inexpensive commercially available microwave oven.

Fixatives↗
PubMed1987

Microwave-stimulated staining of plastic embedded bone marrow sections with the Romanowsky-Giemsa stain: improved staining patterns.

Staining plastic sections with the Romanowsky-Giemsa method is both time-consuming and difficult. This paper reports how the staining time can be reduced to 25 min using microwave irradiation of the staining solution. It is shown that staining results depend on the fixative used, staining temperature, dye concentration and pH of the staining solution as well as on several parameters of the microwave irradiation technique. The staining patterns are improved when compared with those obtained by conventional staining of plastic sections. The colors are more brilliant and greater contrasts are observed. Basophilia, polychromasia, and orthochromasia accompanying red cell maturation are more pronounced. For white cell maturation the initial appearance of specific granules (neutrophil, basophil, and eosinophil) is more evident. Thus, cell classification is easily accomplished using the described technique. It is suggested that microwave-stimulated staining be considered for routine use.

Animals↗
PubMed1985

Protein composition of cyst fluids from mature teratoma in patients with nonseminomatous germ cell tumors of the testis.

Protein analyses were performed in 15 cyst fluids (CF) from mature teratoma (TD) and in 15 corresponding sera from 9 nonseminomatous germ cell tumor patients. Qualitatively, many similarities between the protein compositions of CF and corresponding sera were seen. Quantitative comparisons suggested free diffusion of plasma proteins into the cyst lumen in nine cases, whereas in five CF a decreased size selectivity of the blood-TD barrier was observed. From the quantitative data it was concluded that the significantly increased CCF/Cserum concentration ratios for the tumor markers alpha-fetoprotein (8/14), human chorionic gonadotropin (3/14), and carcinoembryonic antigen (13/13) as well as for lysozyme (12/13), ferritin (12/13), and fibronectin (3/6) were either due to local synthesis or to concentrating properties of the TD cells. The results of the current study encourage further research for new tumor-associated proteins in cyst fluids.

Blood Proteins↗
PubMed1985

Significance of human chorionic gonadotropin, alpha-fetoprotein, and pregnancy-specific beta-1-glycoprotein in the detection of tumor relapse and partial remission in 126 patients with nonseminomatous testicular germ cell tumors.

After surgery and initial treatment, 126 patients with nonseminomatous germ cell tumors of the testis were followed by estimation of serum human chorionic gonadotropin (HCG), alpha-fetoprotein (AFP), and pregnancy-specific beta-1-glycoprotein (SP-1). The median follow-up time was 39 months (range, 12-69 months). An evaluation was made of the sensitivity, specificity, and predictive value of these markers to detect tumor relapse (TR) and partial remission (PR) rates. Five of the 35 (14%) Stage I patients had a TR, and in 4 the TR was recognized early by increasing serum levels of HCG (four times) and AFP (one time). In Stage II and III disease 17 of the 91 (18%) patients had a TR or PR. In 15 of the 17 patients TR and PR were associated with elevated serum levels of HCG (nine times), AFP (seven times), and SP-1 (five times). Rising levels of HCG and/or AFP preceded a clinical confirmation of TR and PR by 4 to 8 weeks. Serum SP-1 did not add useful information. The combined use of serum HCG and AFP gives a sensitive indicator for TR and PR: 19 of 22 (86%) of such patients were recognized. The specificity (98% and 100%, respectively) and the predictive value of positive tests (87% and 100%, respectively) were high for both HCG and AFP.

Chorionic Gonadotropin↗