Biomedical subjects

H Rogatsch

Publications and source records attributed to H Rogatsch.

35 records · Page 2Linked to original sources

PubMed1999

Mature dendritic cells induce T-helper type-1-dominant immune responses in patients with metastatic renal cell carcinoma.

We performed a pilot study on a dendritic cell (DC)-based vaccine in 4 patients with advanced renal cell carcinoma. The vaccine consisted of cultured blood DCs loaded with autologous tumor cell lysate plus keyhole limpet hemocyanin (KLH) and matured with a combination of tumor necrosis factor alpha and prostaglandin E(2). We describe the immune response against KLH induced by DC-based immunization in a patient undergoing an objective partial response and compare it with the responses observed in patients with either stable or progressive disease. The patient with the clinical response developed strong delayed-type hypersensitivity (DTH) against KLH after a single vaccination with antigen-loaded DCs, whereas the other patients failed to develop DTH reactivity even after repeated vaccinations. Antigenic stimulation of mononuclear cells (MNCs) induced proliferation and IFN-gamma but not IL-4 production as well as expression of the chemokine receptor CXCR3 consistent with a T-helper (Th) type-1 bias. Exogenous IL-12 enhanced and exogenous IL-4 diminished IFN-gamma production. In the 2 patients with stable disease two or more vaccinations were required to induce maximal MNC responses. In the patient with progressive disease MNC responses were hardly detectable. Anti-KLH antibodies appeared with different kinetics but could be detected in the serum of all patients. Isotype analysis revealed the presence of IgM, IgG(1), IgG(2) and IgG(3) as well as IgA and complete absence of IgE. The patient with progressive disease also developed IgG(4) antibodies indicative of a deviation towards Th2. Cultured blood DCs can be a potent vaccine for the antigen-specific immunization of patients with advanced kidney cancer. KLH serves as a tracer molecule which allows determination of the magnitude, kinetics and Th bias of the cellular and humoral immune response induced by DC-based immunization. The data also suggest that Th type-1-dominant immune responses involving DTH reaction are required for the induction of tumor regression.

Adjuvants, Immunologic↗

PubMed1999

Malignant large cell calcifying Sertoli cell tumor of the testis (LCCSCTT). Report of a case in an elderly man and review of the literature.

Malignant Large Cell Calcifying Sertoli Cell Tumor of the Testis (LCCSCTT) is a rare histological variant of sex cordstromal tumors. It usually arises in young males, sometimes is associated with endocrine abnormalities and has a benign course. It is exceptional in elderly men and the outcome is rarely fatal. We report a case of LCCSCTT in a 73 year-old man with fatal outcome. The tumor involved the right testis and several areas of the tunica albuginea were grossly invaded. Serum levels of HCG, LH and testosterone were normal. Lymphoangiography performed after orchiectomy showed an involvement of the iliac and preaortic lymph nodes. X-ray of the chest showed no lung metastases. A thorough study of the light microscopic, immunohistochemical and ultrastructural appearances was performed. Immunohistochemistry revealed positivity to vimentin, S-100 and NSE. Our observations confirm the previous findings concerning malignant LCCSCTT and point out the histogenesis of the tumor from Sertoli cells.

Age of Onset↗

PubMed1998

Improvement of specificity in PSA-based screening by using PSA-transition zone density and percent free PSA in addition to total PSA levels.

BACKGROUND: The clinical value of prostate-specific antigen (PSA) density in differentiating between prostate cancer and benign prostatic hyperplasia has been the subject of several studies. In this context the question has been raised about the diagnostic benefit of PSA transition-zone density (PSA-TZ density = total PSA/transition-zone volume) in the detection of prostate cancer. In the following study the value of PSA-TZ density alone and in combination with free PSA was investigated. METHODS: Between August 1995-May 1996, 308 first-line screening volunteers with elevated total PSA levels ranging from 2.5-10.0 ng/ml were evaluated. All patients underwent digital rectal examination, transrectal ultrasound, and transrectal ultrasound-guided biopsy of the prostate. Prior to these investigations, serum was obtained and total as well as free PSA levels were obtained. PSA transition-zone density (PSA-TZ density) was defined as follows: PSA-TZ density = total PSA/transition-zone volume. RESULTS: ROC curve analyses for PSA-TZ density showed that by using a PSA-TZ density of more than 0.22 ng/ml/cc as a biopsy criterion, 24.4% of negative biopsies could be avoided; ROC curve analyses for free PSA showed that by using percent free PSA <20% as a biopsy criterion, 45.5% of negative biopsies could be eliminated. When combining these two diagnostic tests, 54.2% of negative biopsies could be avoided. CONCLUSIONS: We conclude that PSA-TZ density, in addition to total and free PSA, is a new opportunity which renders it possible to calculate the likelihood of detecting prostate cancer on repeat biopsies in an individual patient.

Biopsy↗

PubMed1998

[Urothelial tumors and preneoplasias. Diagnostic problems and their clinical consequences].

The surgical pathology of biopsies or electroresection specimens taken from clinically suspicious or overt tumors in the urinary bladder encompasses the evaluation of a few parameters. This should allow the segregation of bladder tumor patients into subgroups with distinct clinical features and biological behavior, thus providing a rationale for choosing the best available therapy. In essence, the pathologist's role entails a careful morphologic assessment of the primary tumor, including evaluation of the histologic type, the growth pattern, the tumor grade, the tumor stage, and finally the presence and type of primary or tumor-associated flat intraurothelial lesions. Whereas the growth pattern of a lesion can be readily recognized, the correct grading and staging of papillary tumors are often more dependent on the complexity of the individual case and the experience of the pathologist due to the inherent subjectivity of the field and a lack of standardized criteria. These problems of intra- and interobserver variability are intimately coupled with the characteristics of the material, that is, bad orientation and tangential sectioning, thermal injury, crush and fixation artifacts, and limitations of the size of the samples. The correct evaluation and interpretation of flat intrauorothelial lesions suffer from similar difficulties and are further complicated by a confusing categorization and terminology. Although new modalities and molecular approaches have been introduced in recent years in an effort to overcome some of these obstacles, morphology still remains the most effective means to assess the biological behavior and prognosis of urothelial bladder cancer. The present article therefore addresses some of the diagnostically and clinically most relevant controversies and aims to give some useful hints for the evaluation of the above-mentioned morphological parameters. In addition, it adds some remarks on the morphological basis and diagnostic validity of urinary cytology in primary diagnosis and, more importantly, monitoring of bladder cancer patients.

Adenocarcinoma↗

PubMed1997

Microvessel density in core biopsies of prostatic adenocarcinoma: a stage predictor?

Microvessel density was recently reported to be an independent correlate of tumour stage in whole mount prostatectomy specimens. This prompted an investigation of whether the quantitation of tumour microvessels could also be reliably applied to prostatic core biopsies, as a presurgical determinant of local tumour extension. The study was performed on a series of 46 unselected patients with prostatic adenocarcinomas undergoing radical prostatectomy. Intratumoural microvasculature was highlighted immunohistochemically using an antibody against CD31 and subsequently evaluated at x 400 magnification in both biopsies and corresponding prostatectomies. The highest microvessel count was reported for each case. Ten cases (22 per cent) had to be excluded because of insufficient measurable tumour areas in core biopsies. The remaining 36 cases (16 pT2; 20 pT3) showed a high degree of correlation between microvessel density in biopsies and prostatectomies (P < 0.0001). Similarly, pre- and post-operatively determined microvascular counts correlated well with tumour stage (P < 0.0001). Furthermore, the median microvessel density in core biopsies and tumours, i.e., 34, distinguished well between organ-confined and organ-extending tumours (positive predictive value for pT3 tumours 94.4 per cent; sensitivity 85 per cent). These data indicate that the evaluation of microvessels in core biopsies, eventually combined with other parameters, could be a reliable method for the individual prediction of the post-surgical tumour stage of prostatic adenocarcinoma.

Adenocarcinoma↗

PubMed1996

Expression of the gene for tear lipocalin/von Ebner's gland protein in human prostate.

Northern analysis of human multiple tissue blots containing poly A+ RNA from spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes revealed that a prostate specific transcript hybridizes to a tear lipocalin/von Ebner's gland protein (TL/VEGP) gene probe. To characterize this transcript, the corresponding cDNA was amplified by reverse transcription (RT)-PCR. Cloning and sequence analysis showed that it was identical to the tear lipocalin cDNA isolated from human lachrymal glands. Immunohistochemical analysis on thin layer sections of human prostate using a tear lipocalin specific antiserum confirmed the expression of this cDNA in prostate. Thus, our results clearly argue against a unique function of TL/VEGP in human tear fluid or saliva. The human cDNA was expressed in E. coli using the pQE system yielding a recombinant protein which shows biochemical properties identical to the native TL/VEGP.

Amino Acid Sequence↗

PubMed1996

Expression of vimentin, cytokeratin, and desmin in Sertoli cells of human fetal, cryptorchid, and tumour-adjacent testicular tissue.

The intermediate filament of mature human Sertoli cells is vimentin. A co-expression of vimentin together with cytokeratin has been demonstrated in Sertoli cells during embryonal development and under pathologic conditions in adult testes. We analysed the presence of vimentin, cytokeratin, and desmin in Sertoli cells of fetal testes (n=20), in seminiferous tubules of cryptorchid testes (n=10) and adjacent to testicular germ cell tumours (n=47) using specific monoclonal antibodies and single and double-labelling immunohistochemistry. During embryonal development prominent cytokeratin expression disappears after the 20th week of gestation. Interestingly, we also found desmin in immature intratubular Sertoli cells between weeks 11 and 14. In adult cryptorchid testes and in peritumour tubules, desmin was also prominently present in Sertoli cells in the vast majority of the cases investigated, as well as vimentin and cytokeratin co-expression. This first description of desmin immunoreactivity may shed some light on the ontogeny of human Sertoli cells and demonstrates that this cell type is able to express three types of intermediate filaments in a complex manner.

Adult↗

PubMed1996

Lamina propria of sex cords in human fetal testis: an immunohistological and stereological study.

Testicular peritubular cells are located in the lamina propria of seminiferous tubules. These cells, significantly contributing to the basal membrane of seminiferous epithelium, have been studied in a number of species. However, there is a lack of data on the development of the lamina propria in the human testis. The aim of our survey was to investigate the characteristics of the lamina propria and, in particular, peritubular cells in the fetal human testes by immunohistological and stereological methods. Therefore, testes (14-39 weeks of gestation, n = 45) were dissected and fixed in a 4% buffered paraformaldehyde solution. Several pieces of each testis were embedded in paraffin and processed for immunohistochemical and stereological analysis. All investigated testes have shown sex cords in the process of development and differentiation. Morphologically, peritubular cells in the lamina propria can be divided into two types: fibroblast-like (FL) and myoid-like (ML) type (cells which much resemble mature myoid cells). By immunohistochemistry, both FL and ML cells are found to be strongly positive for the intermediate filament desmin, but negative for alpha-smooth actin. While FL cells intensively express Ki-67 demonstrating proliferative activity, ML cells are found to be negative. The basement membrane of sex cords as well as the blood vessels of the interstitium show strong positivity to collagen IV and laminin. Concerning the correlation between the appearance of the investigated antigens with the gestational age, all antigens have been expressed (in the manner described above) already in the 14th week of gestation. The stereological analysis of the number (Nv) and volume (Vv) of peritubular cells indicates a pulsatile development of these cells in the lamina propria of the human fetal testis. While the stereological variables determined for FL cells show a gradual decrease, the same variables determined for ML cells demonstrate a successive increase. It appears that the lamina propria of the fetal human testes shares many of the properties previously discovered in rodents.

Cell Count↗

PubMed1996

CD30 expression in seminoma.

In testicular germ cell tumors the CD30 antigen has been shown to be regularly expressed in embryonal carcinoma and was thus suggested as a marker for this particular neoplasm. Very recently, it has been proven that the monoclonal antibody Ber-H2 is suitable for the detection of this membrane antigen in paraffin sections. We conducted an immunohistochemical study to investigate the CD30 expression in a large series of different presentations of seminoma (ie, pure, mixed, and spermatocytic) because there is evidence from several sources that embryonal carcinoma is histogenetically closely related to, and probably derives from, seminoma. Sections from formalin-fixed, paraffin-embedded tissue from 38 cases of testicular seminomas were immunostained for the demonstration of the CD30 antigen using the monoclonal antibody Ber-H2, cytokeratins, and placental alkaline phosphatase following an indirect streptavidin-peroxidase regimen. In selected cases, immunostainings were performed on consecutive sections to investigate a possible colocalization of CD30 and cytokeratins in seminoma. Specific immunostaining for CD30 in seminoma cells could be detected in single minute foci in 4 of 21 cases of pure classic seminoma. Seminomatous components of mixed tumors showed CD30 positivity in single, but also multiple, foci in 7 of 14 cases. CD30 immunoreactivity in seminoma cells occurred with and without colocalized expression of cytokeratin. Spermatocytic seminoma (n = 3) as well as intratubular germ cell neoplasia in tumor adjacent parenchyma (n = 36) were negative in all cases investigated. We conclude that in testicular germ cell tumors, the expression of CD30 is not restricted to embryonal carcinoma but can also be found focally in seminoma, adding further evidence for a close relationship between these two tumors. The prevalence of CD30 expression in seminomatous components of mixed tumors, as well as the coexpression with cytokeratins, suggest that CD30 expression in seminomas might indicate their upcoming transformation to embryonal carcinoma. This conclusion coincides with a model featuring seminoma in a central role of germ cell tumor development.

Adolescent↗

PubMed1996

Structure of small blood vessels in the testes of infertile men.

In the testis, 'hyalinization' of the lamina propria of seminiferous tubules is often accompanied by similar changes within the walls of testicular blood vessels. The aim of our study was to investigate the structure of small blood vessels in hyalinized human testes by means of immunohistochemistry, electron microscopy and image analysis methods. Results of immunohistochemical analysis indicated that, despite hyalinization, testicular small blood vessels retained positive immunostaining for desmin and actin. Their basement membranes remained immunopositive for collagen IV and laminin. No proliferative (Ki-67) activity was observed in the blood vessel walls in testes from both control and infertile men. P-170 glycoprotein was found to be expressed only in primary spermatocytes. No difference in expression and localization of this antigen was observed between control and affected testes. Electron microscopy revealed a number of testicular arterioles with a notably narrow lumen due to enlarged endothelial cells in infertile men. Such arterioles also had a thickened subendothelial layer and an abundant tunica adventitia rich in connective tissue fibres and ground substance. Some venules in hyalinized testes displayed increased connective fibres and ground substance in the subendothelial layer, between the smooth muscle cells of the tunica media and the tunica adventitia. However, no changes were found in the capillary network, when compared to controls. Image analysis data showed a statistically significant increase in the surface of tunica intima and adventitia of arterioles and tunica media of venules. It is concluded that hyalinization mostly affects testicular arterioles and venules, but not capillaries. Our immunohistochemical data indicate that the 'nature' and/or extent of hyalinization in testicular small blood vessels differs from that described previously for the lamina propria of seminiferous tubules.

Adult↗

PubMed1995

Testicular seminomas are aneuploid tumors.

BACKGROUND: Distinct ploidy patterns of germ cell tumors and aneuploidy of classic seminoma are of central importance in a current concept designating seminoma as a stage in tumor evolution preceding all nonseminomatous germ cell tumors. Whereas cytogenetic studies almost constantly revealed numerical chromosomal aberrations in seminomas in line with this pathogenetic model, ploidy determination by DNA cytometry yielded highly controversial results on the frequency of diploid seminomas. EXPERIMENTAL DESIGN: Twenty-two classic seminomas were investigated in a comparative manner by DNA flow and DNA image cytometry on nuclei isolated from fresh as well as formalin-fixed tumors. The cellular composition of the tumors, i.e., the number of neoplastic, interstitial, and lymphoid cells was quantified in each case. RESULTS: Irrespective of the tissue preparation and fixation method, all of the tumors that were analyzed were aneuploid by image cytometry. Diploid cases were only found by flow cytometry. Thus, 1/22 cases from paraffin-embedded and 4/11 cases of native tumors showed a single diploid stemline by flow cytometry, however, they exhibited aneuploidy by image cytometry. The quantity of tumor infiltrating lymphocytes was clearly correlated with false diploidy detected by flow cytometry. CONCLUSIONS: Classic testicular seminomas are aneuploid tumors. Previous reports on diploid cases detected by flow cytometry have most likely been caused by extensive lymphocytic infiltration.

Aneuploidy↗

PubMed1995

Seminomas positive for Epstein-Barr virus by the polymerase chain reaction: viral RNA transcripts (Epstein-Barr-encoded small RNAs) are present in intratumoral lymphocytes but absent from the neoplastic cells.

A possible role of Epstein-Barr virus (EBV) in the pathogenesis of testicular germ cell neoplasms has been suggested repeatedly, but direct evidence for an association of testicular cancer with EBV is lacking. We examined 26 cases of classical seminoma, two spermatocytic seminomas, and 12 cases of nonseminomatous or combined germ cell tumors for the presence and cellular location of EBV with a combined approach using the polymerase chain reaction and nonradioactive in situ hybridization for EBV-encoded small RNAs (EBER1/2). After exclusion of cases without amplifiable DNA, 4/21 (19%) seminomas, but none of the other tumors, were positive for EBV by polymerase chain reaction. In situ hybridization for EBER1/2 showed rare positive lymphocytes, probably latently infected B-cells, in two of these four EBV-positive cases. No EBER-positive tumor cells were found in any of the analyzed tumors. The occurrence of EBV-positive lymphoid cells was not correlated to the frequency of intratumoral lymphocytes, including B-cells, which were present in seminomas in significant numbers. Our study demonstrates the absence of EBV from the neoplastic cells of testicular germ cell tumors and makes a direct role of EBV in the development of these malignancies improbable. Whether the presence of EBER-positive lymphocytes in some seminomas simply reflects the normal occurrence of latently infected cells in lymphoid tissue of seropositive individuals or is influenced by local factors remains to be determined.

Antibodies, Monoclonal↗

PubMed1994

Carcinoma in situ of the testis detected by DNA flow cytometry of testicular fine-needle aspirates.

Testicular carcinoma in situ (CIS) is usually diagnosed histologically on surgical biopsies. The present study was performed to test an alternative approach--DNA flow cytometry (FCM) of testicular fine-needle aspirates (FNA)--for the detection of this lesion. FNAs from 18 cases of testicular germ cell tumors (TGCT) and tumor adjacent parenchyma were analysed by DNA FCM. DNA histograms of all cell nuclei and histograms representing selectively the hyperdiploid compartment were analysed. The presence and extention of CIS were determined by histology and immunohistochemistry. In 16 of 18 cases, CIS was histologically present, whereas aneuploid peaks were detected in only 11 cases in DNA histograms of all cell nuclei. In the analysis of the histograms of the hyperdiploid region, 4 additional cases of CIS could be identified increasing the sensitivity to 15/16 cases (93.8%). In all but one case, the DNA indices of CIS and invasive tumors were identical. The study demonstrates that DNA FCM of testicular FNAs using the described data acquisition and analysis could also be applicable for detection of CIS in a clinical situation.

Aneuploidy↗

PubMed1994

In vitro investigations of interphase and metaphase argyrophilic nucleolar organizer regions and cellular proliferation in the human urothelial cancer cell line HOK-1.

Detailed investigation of cell growth and nucleolar organizer region associated argyrophilic proteins (Ag-NORs) is necessary to assess a possible impact of Ag-NOR quantification on the diagnosis and prognosis of tumours. In this study, cellular proliferation of the transitional-cell carcinoma cell line HOK-1 was modulated over a period of 11 days by starvation and subsequent medium addition. Proliferation was determined daily by DNA flow cytometric estimation of S-phase fraction (SPF) and mitotic index (MI) calculation. The number and area of interphase Ag-NORs were quantified by automated image analysis daily and the number of Ag-NOR bearing chromosomes in metaphase was counted. In interphase nuclei, Ag-NOR area showed a highly significant correlation with SPF (p < 0.0001) whereas interphase Ag-NOR number showed significant correlation with MI (p < 0.05). A positive relationship between the number of Ag-NOR bearing chromosomes in metaphases and cellular proliferation was also observed. There is variability in Ag-NOR quantity during interphase and metaphase depending on growth conditions in vitro. Correlations of the number of interphase Ag-NORs with the MI on one hand and Ag-NOR area with SPF on the other provide further evidence that distribution and quantity of Ag-NORs are strongly influenced by the cell cycle phase within the structural-functional unit of the nucleolus.

Carcinoma, Transitional Cell↗

PubMed1994

Quantification of spermatogenesis by dual-parameter flow cytometry.

OBJECTIVE: To investigate the usefulness of dual-parameter flow cytometry of testicular fine-needle aspirates for the quantification of spermatogenesis. Immunofluorescence staining for vimentin was introduced to discriminate vimentin-negative germ cells from vimentin-positive stromal cells. The results of flow cytometry were compared with testicular morphology and immunohistochemistry in cases of regular and disturbed germ cell maturation. DESIGN: Testicular fine-needle aspiration and surgical biopsy were performed in 50 autopsy cases. The fine-needle aspirates were double stained for the intermediate filament vimentin by indirect immunofluorescence and DNA propidium iodide and analyzed by flow cytometry. Surgical biopsies were examined by light microscopy. The distribution of vimentin-positive cells was evaluated by immunohistochemistry. RESULTS: A comparison of morphology and flow cytometric analysis yielded characteristic quantitative distribution patterns of the different germ cell and somatic cell populations in regular and disturbed spermatogenesis. All three ploidy compartments of the germ epithelium correlated with high statistical significance with the respective histologic diagnoses. Moreover, a marked quantitative increase of stromal cells could be demonstrated in spermatogenetic disorders. CONCLUSIONS: The simultaneous analysis of the cellular DNA content and the intermediate filament vimentin by flow cytometry enables a detailed investigation of spermatogenetic disorders. Quantitative changes of the relationships between germ cells and somatic cells can be selectively investigated.

Biopsy, Needle↗

PubMed1992

Deoxyribonucleic acid flow cytometry and semiquantitative histology of spermatogenesis: a comparative study.

OBJECTIVE: To compare deoxyribonucleic acid (DNA) flow cytometry with a semiquantitative histologic method for the assessment of spermatogenesis and to investigate the influence of various preparative techniques on the results of DNA measurements. DESIGN: Testicular tissue from 52 autopsies was examined for spermatogenesis by DNA flow cytometry and by conventional semiquantitative histology. Flow cytometry was performed on single nuclei suspensions from fine needle aspiration biopsies, resuspended imprints, as well as on specimens obtained by mechanical disaggregation of surgical biopsies of the same testis. RESULTS: In all cases, flow cytometry revealed a characteristic distribution pattern of haploid (n), diploid (2n), and tetraploid (4n) cells independent of the preparation technique used. The percentage of haploid cells of the testes showed statistically highly significant correlations (P = 0.0002) with the results of semiquantitative histology. Fine needle aspiration biopsy always contained more haploid cells compared with resuspended imprints and mechanically disaggregated samples. CONCLUSION: Our results confirm the suitability of DNA flow cytometry as a fast and reliable quantitative method for the assessment of spermatogenesis and demonstrate its optimal feasibility in combination with fine needle aspiration biopsy.

Adult↗

PubMedDate not supplied

Improved technique for investigations on archival formalin-fixed, paraffin-embedded tumors by interphase in-situ hybridisation.

In-situ hybridisation techniques are powerful tools for the analysis of chromosomes in a variety of specimens. Due to hybridisation problems caused by extensive formalin-fixation in tissue samples mainly isolated nuclei were used for chromosomal analysis. In archival tissue, for sections to retain both an optimum of well preserved morphology and hybridisation efficiency a large-scale adjustment of protease digestion steps for each tissue block was required. We sought to develop an easy, reproducible technique by pretreatment in a 90 degrees C glycerol solution and subsequent denaturation in an autoclave 1 bar for 10 minutes. Our experiments were performed on eight up to ten years old human bladder cancer tissue blocks. Comparison with established pretreatment techniques did not reveal hybridisation differences while the morphology of tissue sections kept intact. This technique facilitates retrospective interphase cytogenetic analyses and is a reliable method for detection of chromosomal anomalies in formalin-fixed archival tumors.

Centromere↗