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Biomedical subjects

D W Visscher

Publications and source records attributed to D W Visscher.

At least 37 records · Page 2Linked to original sources
PubMed1996

Cell cycle analysis of normal, atrophic, and hyperplastic breast epithelium using two-color multiparametric flow cytometry.

We performed two-color flow cytometric synthesis phase fraction (SPF) determinations on cytokeratin-labeled benign epithelial populations from 142 breast specimens (41 mastectomy, 70 diagnostic biopsy, 31 reduction mammoplasty). There was wide variability of SPF, ranging from 0.1 to 3.5%, with a frequency distribution skewed to higher values (mean 0.75%, median 0.5%). The mean SPE for women less than 29 years was 0.91%, vs. 0.89% for 30-42 years, 0.66% for 43-49 years, and 0.56% for > or = 50 years (P = 0.05). Histologically atrophic tissue samples exhibited a mean SPF approximately half that of morphologically normal tissue from premenopausal age women (0.79% vs. 0.36%, P = 0.02). Tissues showing histologically proliferative fibrocystic features had a greater mean SPF than non-proliferative fibrocystic tissues (0.59% vs. 0.92%); however, due to the wide spread of values within each of these categories, this difference was not statistically significant and neither group was significantly different from 'normal' tissue samples. Patients with histologically normal breast tissue, though, were significantly younger (mean = 34.6 years) than those with fibrocystic changes (non-proliferative mean = 53.4 years vs. proliferative mean = 42.8 years, P = 0.005). Synchronous right- and left-sided specimens obtained from reduction mammoplasty demonstrated significantly correlated SPF determinations (R = 0.77). We conclude that selective analysis of epithelial populations using two-color flow cytometry provides cell cycle information in benign breast tissue which is analogous to that obtained by labor-intensive nucleotide labeling studies. This study also confirms the biologic variability and age-dependence of breast epithelial proliferation. Finally, the data imply that derangements of cell proliferation in fibrocystic conditions are heterogeneous, complex and incompletely correlated with histologic parameters such as hyperplasia.

Adult↗
PubMed1995

Detection of chromosome aneuploidy in breast lesions with fluorescence in situ hybridization: comparison of whole nuclei to thin tissue sections and correlation with flow cytometric DNA analysis.

We compared flow-cytometric DNA histogram pattern to counts of 4 fluorescent-labelled centromeric probes (chromosomes 1, 7, 8, and 17) in whole nuclei (WN) and in nuclei from the corresponding formalin-fixed deparaffinized thin tissue section (TS) in 25 breast lesions (9 invasive carcinomas, 1 duct carcinoma-in-situ, 5 fibroadenomas, 10 fibrocystic change). In benign lesions, signal gains (i.e., trisomic nuclei) were never observed in greater than 10% of nuclei from either WN or TS preparations. Loss of signal in benign breast lesions, however, varied considerably (0-43%) between individual case and between chromosome probes. The mean incidence of signal loss in WN of benign lesions ranged from 8.9% (chromosome 7) to 14.4% (chromosome 1) of nuclei. These signal loss frequencies exceeded those of benign lymphoid control cells. In three benign lesions, signal loss in WN (with one probe) was observed in at least 25% of nuclei. Signal losses in benign TS, on average, were 50-150% greater than in matched WN preparations (chromosome 1-21.7%, chromosome 7-21.5%). Malignant lesions generally, but not always, displayed fewer monosomic nuclei and more trisomic nuclei in WN compared to TS, compatible with a slicing (i.e., nuclear truncation) artifact. Signal counts in carcinomas correlated well with flow cytometric DNA index; however, they were also characterized by evidence of genetic instability, manifest as signal gains in a subset of nuclei (10-25%) with individual probes in diploid range cases, as well as intratumoral heterogeneity, reflected as discrepancies in probe counts between WN and TS samples. We conclude that signal losses with centromeric probes are largely, but not entirely, explained by nuclear slicing.(ABSTRACT TRUNCATED AT 250 WORDS)

Aneuploidy↗
PubMed1995

Clinicopathologic analysis of macrophage infiltrates in breast carcinoma.

We compared macrophage density, assessed by enumeration of peritumoral mononuclear cell immunoreactivity for HAM 56, to clinicopathologic features and to immunostaining for two "invasion-associated" proteases (Cathepsin D and Urokinase plasminogen activator) in 80 breast carcinomas. Diffuse (2+) infiltrates of HAM 56- positive mononuclear cells were present in 27 cases (34%) and 43 (54%) exhibited focal (1+) infiltrates. Presence of 2+ macrophage infiltrates correlated significantly with poor differentiation. None of the seven well-differentiated cases exhibited 2+ infiltrates, whereas 9/43 (21%) moderately differentiated and 18/30 (60%) poorly differentiated tumors were diffusely infiltrated (p = .001). Wide-spread macrophage infiltrates were also more frequent in cases with advanced stage (23% of node negative vs 40% of node positive cases, p = NS). Forty-four percent of the cases with diffuse macrophage infiltrates were cathepsin D positive (i.e. in host derived cells) vs only 18% with focal macrophage infiltrates (p = .002). A similar relationship was observed between staining for HAM 56 and urokinase-type plasminogen activator (p = .02). Disease recurrences (50 months median follow-up) were more frequent in patients with 2+ (17/27, 63%) as opposed to 0+ (1/10, 10%) macrophage infiltrates (p = .01). We conclude that the density of stromal macrophage infiltrates is associated with clinical aggressiveness in breast carcinomas. Further, this relationship may reflect contribution of host derived macrophages to invasion and metastasis through elaboration of proteases which putatively mediate degradation and remodeling of extracellular matrix.

Breast Neoplasms↗
PubMed1995

Coordinate expression of the PRM1, PRM2, and TNP2 multigene locus in human testis.

Maintenance of the transcriptionally inert state of the mature human spermatozoon requires the expression of the various members of the human protamine gene cluster prior to the final stages of spermatogenesis. During this process, known as spermiogenesis, round spermatids morphologically differentiate into mature spermatozoa. The expression of the PRM1, PRM2, and TNP2 genes facilitates the compaction and condensation of the genetic material within the developing spermatid. To understand better the coordinate control governing this transformation, we have examined the localization and distribution of the human protamines PRM1 and PRM2 and transition protein TNP2 transcripts during human spermatogenesis. The stage-specific expression of these transcripts was determined by in situ hybridization analysis using [alpha-35S]-labeled cRNA probes. PRM1, PRM2, and TNP2 transcripts were abundant in association with round and elongating spermatids, located in the adluminal region of the seminiferous epithelium. They were not observed in association with spermatogonia, spermatocytes, Sertoli cells, or interstitial cells. These data indicate that the human PRM1, PRM2, and TNP2 transcripts are expressed postmeiotically in round and elongating spermatids. The quantitative evaluation of each transcript was determined as a function of the relative optical density per unit area. In all cases examined, the relative level of each transcript was consistent with the following pattern, PRM2 > PRM1 congruent to TNP2.

Chromosomal Proteins, Non-Histone↗
PubMed1995

Evaluation of chromosome 12 copy number in ovarian granulosa cell tumors using interphase cytogenetics.

Trisomy 12 is frequently observed in karyotypes of ovarian sex cord-stromal tumors, including adult and juvenile granulosa cell tumors (AGCTs and JGCTs). We assessed the ability to detect this abnormality in deparaffinized tissue sections of 19 ovarian GCTs (17 AGCTs, two JGCTs) and in one fibrothecoma by simultaneous in situ hybridization with fluorescent-labeled centromeric probes to chromosomes 12 (fluorescein isothiocyanate conjugated) and 17 (rhodamine conjugated). In order to quantitate the artifact introduced by nuclear slicing, such analyses were performed both on intact tissue sections and on cytospins of nuclei prepared by enzymatic dissociation from the corresponding tissue block. The series was also evaluated for numerical abnormalities of chromosome X, a less common cytogenetic finding in GCT. Twelve of 19 cases (63%) displayed evidence of trisomy 12 (defined as signal gain in > or = 10% of nuclei) in the intact section, the cytospin, or both. Trisomy for chromosome 17 was present in one case, and trisomy X was present in two cases. In tissue sections the incidence of signal gain for the chromosome 12 probe varied from 0-45% of nuclei (mean 19%). In cytospin preparations, the percentage signal gain for chromosome 12 ranged from 0 to 84% (mean 33%). This study supports the presence of trisomy 12 as a common, but not defining, cytogenetic anomaly in ovarian GCTs. Its presence, however, within only a minority of tumor cells may be partly explained by slicing artifact associated with intact tissue sections, although partial involvement of intact nuclei suggests that trisomy 12 may also be encountered as a heterogeneous abnormality within neoplastic populations of GCTs.

Adult↗
PubMed1995

A universal method for the mutational analysis of K-ras and p53 gene in non-small-cell lung cancer using formalin-fixed paraffin-embedded tissue.

The p53 tumor suppressor gene has been found to be altered in almost all human solid tumors, whereas K-ras gene mutations have been observed in a limited number of human cancers (adenocarcinoma of colon, pancreas, and lung). Studies of mutational inactivation for both genes in the same patient's sample on non-small-cell lung cancer have been limited. In an effort to perform such an analysis, we developed and compared methods (for the mutational detection of p53 and K-ras gene) that represent a modified and universal protocol, in terms of DNA extraction, polymerase chain reaction (PCR) amplification, and nonradioisotopic PCR-single-strand conformation polymorphism (PCR-SSCP) analysis, which is readily applicable to either formalin-fixed, paraffin-embedded tissues or frozen tumor specimens. We applied this method to the evaluation of p53 (exons 5-8) and K-ras (codon 12 and 13) gene mutations in 55 cases of non-small-cell lung cancer. The mutational status in the p53 gene was evaluated by radioisotopic PCR-SSCP and compared with PCR-SSCP utilizing our standardized nonradioisotopic detection system using a single 6-microns tissue section. The mutational patterns observed by PCR-SSCP were subsequently confirmed by PCR-DNA sequencing. The mutational status in the K-ras gene was similarly evaluated by PCR-SSCP, and the specific mutation was confirmed by Southern slot-blot hybridization using 32P-labeled sequence-specific oligonucleotide probes for codons 12 and 13. Mutational changes in K-ras (codon 12) were found in 10 of 55 (18%) of non-small-cell lung cancers. Whereas adenocarcinoma showed K-ras mutation in 33% of the cases at codon 12, only one mutation was found at codon 13. As expected, squamous cell carcinoma samples (25 cases) did not show K-ras mutations. Mutations at exons 5-8 of the p53 gene were documented in 19 of 55 (34.5%) cases. Ten of the 19 mutations were single nucleotide point mutations, leading to amino acid substitution. Six showed insertional mutation, and three showed deletion mutations. Only three samples showed mutations of both K-ras and p53 genes. We conclude that although K-ras and p53 gene mutations are frequent in non-small-cell lung cancer, mutations of both genes in the same patient's samples are not common. We also conclude that this universal nonradioisotopic method is superior to other similar methods and is readily applicable to the rapid screening of large numbers of formalin-fixed, paraffin-embedded or frozen samples for the mutational analysis of multiple genes.

Base Sequence↗
PubMed1995

Idiopathic bronchiolitis obliterans with organizing pneumonia. An acute and life-threatening syndrome.

Idiopathic bronchiolitis obliterans with organizing pneumonia (BOOP) is a clinicopathologic syndrome characterized by an indolent course and favorable prognosis. This report describes five patients with a fulminating and life-threatening variant of this syndrome. Four patients presented with respiratory failure requiring respiratory assistance and positive pressure ventilation. Early recognition of the entity and prompt initiation of corticosteroid therapy in three patients was instrumental in preventing mortality. Our findings suggest that idiopathic BOOP may be the underlying pathology in a number of patients presenting with ARDS. Since corticosteroid therapy may improve survival in these patients, clinicians should heighten their index of suspicion for this entity. Early histologic diagnosis and initiation of corticosteroid therapy should be considered in patients with unexplained ARDS.

Adrenal Cortex Hormones↗
PubMed1995

Assessment and significance of diploid-range epithelial populations in DNA aneuploid breast carcinomas using multi-parametric flow cytometry.

A 2-color (PI, cytokeratin--FITC) multi-parametric analysis of intact cells was used to reveal diploid-range epithelial populations by flow cytometry in 108 consecutive DNA aneuploid breast carcinomas. Thirty-eight tumors (35%) contained a significant diploid range epithelial population, defined as cytokeratin-positive cells having a DNA content indistinguishable from that of endogenous lymphocytes and comprising at least 20% of all cytokeratin-positive cells. An additional 23 cases (21%) contained a minor diploid range epithelial population having a normal DNA content and comprising only 5-20% of all cytokeratin-positive cells. Multiple DNA aneuploid stemlines were present in 24 cases (22%). Diploid-range populations were more frequent (91%) in tetraploid cases than in hyperdiploid (32%), hypodiploid (17%) or hypertetraploid cases (20%). The presence of diploid epithelial populations and/or multiple aneuploid stemlines correlated with histologic parameters, including an extensive intraductal component (unimodal--4% vs. multi-modal--57%, P = 0.001), heterogeneous differentiation (unimodal--0% vs. multi-modal--52%, P = 0.001), and multi-focal growth with residual interspersed benign tissue (unimodal--8% vs. multimodal--57%, P = 0.01). These data show that diploid-range epithelial cells are frequent in aneuploid breast carcinomas analyzed by flow cytometry. In some tumors, these populations undoubtedly reflect the presence of residual benign epithelium. The numerical dominance of other histograms by near-diploid measurements suggests the presence of diploid-range neoplastic stemlines which would be 'hidden' by contaminating host-derived cells in single parameter DNA histograms. Finally, the correlation of DNA content heterogeneity with distinctive histologic patterns of breast neoplasia implies that co-existing stemlines may have biological significance in the progression of some tumors.

Aneuploidy↗
PubMed1995

Biologic and clinical significance of basic fibroblast growth factor immunostaining in breast carcinoma.

Acetone-fixed cryostat sections of 79 breast carcinomas were immunostained with antibodies to basic fibroblast growth factor (bFGF) and urokinase-type plasminogen activator (uPA). Staining intensity was then compared with microvessel density assessed by manually counting vascular spaces highlighted by immunostaining vascular basal lamina (Type IV) collagen. Extensive (2+) bFGF immunoreactivity was present in neoplastic cells of 30 tumors (38%) and in host-derived stromal cells of 29 cases (37%). Disease recurrence correlated with bFGF staining: 0 to 1+ stromal staining, 30% recurred versus 2+ stromal staining, 73% recurred (P = 0.001) (54-mo median follow-up). Neither stromal nor epithelial bFGF staining correlated significantly with microvessel count; however, there was a statistically significant association between stromal cell bFGF staining and uPA staining of peritumor host cells: absent bFGF--0% 2+ uPA versus weak bFGF--9% 2+ uPA versus 2+ bFGF--29% uPA (P = 0.01). We conclude that elevated expression of bFGF in breast carcinomas is associated with aggressive clinical behavior. Its biologic significance, however, appears more closely related to extracellular matrix remodeling than to induction of prominent neovascularization per se.

Breast Neoplasms↗
PubMed1994

Enhanced expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) in the stroma of breast carcinomas correlates with tumor recurrence.

The 72-kDa (MMP-2, gelatinase A) and the 92-kDa (MMP-9, gelatinase B) matrix metalloproteinases have been associated with tumor cell invasion and metastasis. Immunohistological staining of MMP-2 and MMP-9, basal lamina collagen IV and TIMP-2 were performed on frozen sections of 83 invasive breast carcinomas. MMP-2 and MMP-9 were associated with neoplastic cell plasma membrane in 72% of cases and exhibited inter-tumoral variability of staining intensity. MMP-2 and MMP-9 staining was not correlated with presence of metastases at time of diagnosis or with disease outcome. TIMP-2 was detected in the peri-tumoral stroma and was present in 87% of cases. Residual benign breast tissue was negative for TIMP-2 staining. Neoplasms with diffuse TIMP-2 staining (24%) recurred significantly more frequently (75% recurred) than cases with focal (42% recurred) or absent (27% recurred) TIMP-2. Presence of collagen IV was negatively correlated with gelatinase staining. We conclude that up-regulation of MMP-2 and MMP-9 expression in breast tumor cells is reciprocally correlated to collagen IV staining. Clinical outcome, however, is more closely related to the presence of TIMP-2 than the corresponding MMPs. Enhanced TIMP-2 expression, therefore, may denote a stromal response to tumor invasion, indicative of aggressive behavior in a subset of breast carcinomas.

Antibodies, Monoclonal↗
PubMed1994

c-erbB-2 promoter-specific DNA-binding protein isolated from human breast cancer tissues displays mitogenic activity.

Amplification and overexpression of the c-erbB-2 gene appears to play a role in the pathogenesis of human breast and other cancers. Frequent amplification (20-30%) of the c-erbB-2 gene was observed in human adenocarcinomas of kidney, pancreas, lung, ovarian, and breast cancer. The gene product is a 185-kDa glycoprotein that has intrinsic tyrosine kinase activity and is believed to be a receptor. Several candidate ligands have been described. In the present study we have identified and purified a novel DNA-binding protein from malignant human breast tissues. The protein binds to a core element (-22 to +9, +1 being the transcription start site) of the c-erbB-2 promoter region in a sequence-specific manner. The affinity-purified protein has the ability to induce DNA synthesis in quiescent NIH/3T3 cells, suggesting that the factor has mitogenic activity. The purified protein induces c-erbB-2 expression on the surface of microinjected NIH/3T3 cells. This DNA-binding protein is a sequence-specific cellular factor that is associated with high level expression of the c-erbB-2 gene and appears to play a role in cell transformation. Understanding the control and expression of this DNA-binding protein may shed light on the mechanism(s) of c-erbB-2 gene regulation and its potential role in the pathogenesis of human adenocarcinomas.

3T3 Cells↗
PubMed1994

Interaction of transforming growth factor-alpha and epidermal growth factor receptor in breast carcinoma. An immunohistologic study.

BACKGROUND: Interaction of transforming growth factor-alpha (TGF-alpha) with its receptor, epidermal growth factor receptor (EGFR), has been implicated as an autoregulatory autocrine mechanism of breast epithelial proliferation. METHODS: To examine the interrelationship and clinical relevance of TGF-alpha and EGFR in breast carcinoma, methanol-fixed cryostat sections from 73 patients were immunostained with monoclonal antibodies to epidermal growth factor (EGF), EGFR, and TGF-alpha. RESULTS: Neither EGFR nor TGF-alpha staining was diagnostic or specific for the detection of malignant neoplastic cells. Both exhibited staining along the basal lamina of most benign ducts and lobules. TGF-alpha staining was observed in neoplastic cells in 41% and in non-neoplastic cells (peritumoral stroma and benign duct/lobular epithelium) in 36% of patients. Staining for EGF and TGF-alpha failed to correlate with node status or grade; however, TGF-alpha negative tumors were more frequently positive for estrogen receptor (ER) (70% versus 14%; P = 0.03). The presence of EGFR correlated with positive lymph node status (P = 0.004), poor differentiation (P = 0.001), and negative ER status (P = 0.0001). EGFR staining was more common in neoplasms which recurred, but this approached significance only in the group with node-negative disease (mean follow-up, 52 months; P = 0.06), and neoplastic cell TGF-alpha correlated with disease recurrence in patients with node-positive disease (no recurrence, -13% positive versus recurrence, -52% positive; P = 0.01). Concurrent TGF-alpha/EGFR staining, present in 18% of tumors, also was predictive of disease recurrence (no recurrence, 3% positive for both versus recurrence, 31% positive for both, P = 0.03). CONCLUSIONS: TGF-alpha is heterogeneously expressed in neoplastic and host-derived components of breast tumors. Concurrent EGFR/TGF-alpha immunostaining may characterize a clinically aggressive subset of breast carcinomas, possibly reflecting autocrine interaction, and conferring growth advantage or metastatic phenotype.

Breast Neoplasms↗
PubMed1994

Chromosomal aneuploidy in proliferative breast disease.

Although some forms of proliferative breast disease have been associated with increased risk of breast cancer, substantial confirmatory evidence that the lesions are biologically premalignant has not been presented. Our intent was to identify cytogenetic aberrations in proliferative breast disease using fluorescence in situ hybridization probes selected for their relationship to aberrations previously reported in breast cancer. Application of fluorescence in situ hybridization techniques to paraffin tissue sections using pericentromeric probes for chromosomes 1, 16, 17, 18, and X revealed chromosome aneuploidy in proliferative and malignant lesions of the breast. Sectioning artifact that may result in nuclear truncation was controlled by establishing expected baseline frequencies for gain and loss in normal tissues from the same breast. Localization of chromosomal aberrations to proliferative breast disease lesions with concomitant retention of a normal chromosome complement in corresponding normal breast tissues indicates biologic significance of the results. The similarities of losses involving chromosomes 16, 17, and 18 in hyperplastic lesions and in malignant breast lesions suggest that some hyperplasias may be part of a sequence of progression to malignancy in breast cancer. Gains of chromosome 1 in both in situ and invasive carcinoma are consistent with reports of polysomy 1q as a common cytogenetic change in breast cancer. Its localization to advanced lesions suggests that this trisomy is probably not the initial cytogenetic change in breast cancer tumorigenesis.

Aneuploidy↗
PubMed1994

Angiogenesis in breast carcinoma--clinicopathologic relevance and potential use as a quantifiable surrogate endpoint biomarker.

Although stochastic acquisition of structural genetic aberrations has become the dominant paradigm for progression of solid tumors, the process of tumor cell invasion and metastasis in large part represents a cooperative interaction with host tissues. Such interactions occur at multiple levels and include the exchange of cytokines/growth factors, the induction of neovascularization (angiogenesis), proteolytic degradation and synthetic alterations of the native extracellular matrix, and fibroblast/inflammatory cell chemotaxis. Many classical histologic and mammographic features of breast carcinoma, such as desmoplasia, are a direct reflection of these tumor-host relationships. Further, sustained growth is not possible without a coordinated interaction between diverse neoplastic and native cell populations. In recent years, quantifiable parameters of tumor-host interaction, such as protease elaboration, have been shown to correlate with intrinsic properties of neoplastic cells and to predict clinical outcome. It is thus reasonable to propose that functional cellular alterations imposed by chemopreventive agents may be reflected in tumor-host interaction parameters. The objective of this review is to briefly summarize the literature addressing one parameter of tumor-host interaction--angiogenesis--emphasizing its applicability as a quantifiable biomarker in breast neoplasia.

Biomarkers, Tumor↗
PubMed1994

Clinicopathologic significance of cathepsin B immunostaining in transitional neoplasia.

Fifty Stage heterogeneous urinary bladder carcinomas were immunostained for cathepsin B, a lysosomal endoproteinase putatively associated with tumor invasion. Neoplastic cell CB immunoreactivity was strongly correlated to both grade (I/II--42% positive versus III--68% positive, P = 0.01) and invasion beyond the lamina propria (Ta/T1--42% positive versus T2/T3--68% positive, P = 0.02). Most low grade, papillary tumors displayed a granular cytoplasmic staining pattern, compatible with lysosomal distribution, in contrast to high grade tumors, in which diffuse staining was present in the cytoplasm. Staining was also accentuated at the advancing front of invading tumors and in angiolymphatic tumor emboli. Non-neoplastic mononuclear inflammatory cells, particularly those at the host-tumor interface, displayed variable, sometimes intense staining. Strong tumor-cell CB was more frequent among recurrent TCC than in patients who remain free of disease (55% versus 29%, n = 18, T2-3, cystectomy, 5-yr min. follow-up). We conclude these observed staining patterns and grade/stage associations are compatible with an important biological role for CB in facilitating host invasion in some bladder tumors. Levels and/or distribution of CB may also be of potential value in defining clinically aggressive tumor subsets.

Carcinoma, Transitional Cell↗