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Biomedical subjects

R E Hurst

Publications and source records attributed to R E Hurst.

At least 37 records · Page 2Linked to original sources
PubMed1995

Advances in cancer research: report from the eighty-sixth annual meeting of the American Association for Cancer Research.

A meeting of the American Association for Cancer Research was held in Toronto, Canada on March 18-22, 1995 to discuss advances in cancer research. Growing interest was shown in the molecular mechanisms of prostate cancer. Both breast cancer and colorectal cancer continued to be major scientific topics of interest. Research into the genetics of cancer was dominated by the issue of genomic instability due to failures in the mismatch DNA repair gene system of cells. Instability appeared to represent the key mechanism for the generation of mutations. Another common theme was the identification of putative tumor suppressor and metastasis loci using loss of heterozygosity and cell chromosome fusion approaches. A few novel sequences with a potential suppressive function were reported. Also, relationships between growth factor receptors, tumor suppressor genes, oncogenes, and proteins controlling cell cycle were analyzed. Progress in gene therapy and chemoprevention of malignant tumors was frequently discussed.

American Cancer Society↗
PubMed1994

Structure, function, and pathology of proteoglycans and glycosaminoglycans in the urinary tract.

The roles of glycosaminoglycans and proteoglycans in the physiology of the urinary tract are reviewed. The structures of proteoglycans and glycosaminoglycans are reviewed together with their role in control of epithelial differentiation through stromal-epithelial interactions and as modulators of cytokines. Heparan sulfate proteoglycans appear to be important in maintaining selectivity of the kidney tubular basement membrane, and the majority of the glycosaminoglycan found in the urine appears to come from the upper tract. Evidence suggesting that a dense layer of glycosaminoglycans on the urothelial surface is important to maintaining urothelial impermeability is reviewed and new data showing a high density of proteoglycans on the lumenal surface of the urothelium is presented. The role of this layer in maintaining antibacterial adherence and impermeability was discussed together with data suggesting that failure of this layer is an etiologic factor in interstitial cystitis. A model of the bladder surface is also presented to illustrate the role of proteoglycans and exogenous glycosaminoglycans in the defenses of normal bladder lumen and the failure of these defenses in the interstitial cystitis bladder.

Animals↗
PubMed1994

Identification of proteoglycans present at high density on bovine and human bladder luminal surface.

The proteoglycans on the bladder luminal surface that have been implicated in producing impermeability of the urothelium were investigated. Large-scale isolation was effected with bovine bladder, and smaller scale identification was achieved with human bladder. Little difference was noted between bovine and human bladders. About 80 to 90% of the total surface glycosaminoglycan was bound as integral membrane proteins released with 4 M. guanidinium chloride. About 55% of the protein-bound glycosaminoglycan (proteoglycan) was heparan sulfate, 29% was chondroitin sulfate, and the remainder was either not identified or was dermatan sulfate. Four main proteoglycan species (molecular weight between 85 and 240 kD) were seen, with two minor components present. The proteoglycans were present at very high densities on bladder surface, being stacked 5 to 60 deep. This density of charge will produce a bound water layer that may explain the bladder impermeability.

Animals↗
PubMed1993

Bladder cancer risk assessment with quantitative fluorescence image analysis of tumor markers in exfoliated bladder cells.

BACKGROUND: The detection of potentially highly curable low-grade bladder cancers by noninvasive techniques remains an unsolved problem. Conventional cytology detects such tumors with 50% sensitivity, and addition of DNA measurements to cytology only improves sensitivity incrementally. Tumor-associated antigens potentially offer an additional diagnostic marker. METHODS: In this study, the M344 antibody against a tumor-associated antigen expressed mainly by low-grade tumor cells was tested for its sensitivity and specificity, alone and in combination with DNA ploidy and cytology. Voided urine samples from 69 asymptomatic control subjects, urines and bladder washings from 59 patients with cancer, and 195 symptomatic control patients were collected. Cells were double-labeled with M344 monoclonal antibody and Hoechst. Each case was blinded, and the number of positive cells was scored by two independent observers. RESULTS: High-grade and low-grade transitional cell carcinomas (TCC) were detected with equal efficiency (78%, P < 0.001 versus symptomatic control patients). Urine samples proved higher specificity in detecting cancers. Patients being monitored for recurrence, but without current detectable cancer, were intermediates between control subjects and patients with cancer, suggesting that this marker also responds to dysplasia or field disease. Patients with outlet obstruction did not significantly differ from patients with previous TCC (P = 0.95). When combined with DNA ploidy measurements and cytology, the sensitivity for low-grade and high-grade tumors was 88% and 95%, respectively. CONCLUSIONS: The M344 antibody potentially could improve the specificity and sensitivity of detection of low-grade bladder tumors in symptomatic and asymptomatic patients as well as monitoring for recurrence, therapeutic response, and assessment of individual risk.

Antibodies, Monoclonal↗
PubMed1993

Alterations in phenotypic biochemical markers in bladder epithelium during tumorigenesis.

Phenotypic biochemical markers of oncogenesis and differentiation were mapped in bladder biopsies to investigate changes that occur in bladder tumorigenesis and to identify markers for increased bladder cancer risk. Touch preparations from biopsy specimens from 30 patients were obtained from tumors, the adjacent bladder epithelium, and random distant bladder epithelium. Markers, including DNA ploidy, epidermal growth factor receptor (EGFR), and oncoproteins, were quantified in individual cells by using quantitative fluorescence image analysis. Cluster analysis revealed the markers fell into three independent groups: (i) G-actin and EGFR; (ii) ploidy, cytology, and p185 (HER-2/neu oncoprotein) (ERBB2); and (iii) p300, a low-grade tumor antigen. Each marker displayed a gradient of abnormality from distant field to adjacent field to tumor. Different patterns for each marker suggested a developmental sequence of bladder cancer oncogenesis; G-actin was altered in 58% of distant biopsies (vs. 0/6 normals, P < 0.001), ploidy and cytology were altered in < 20% of distant fields and approximately 80% of tumors, and the other markers were intermediate. Patterns of EGFR and p185 suggest low-and high-grade tracks diverge early (P < 0.05 by Mann-Whitney U test for EGFR and ANOVA for p185). In conclusion, this study shows that a sequence of phenotypic changes accompanies development and progression of bladder cancers. Biochemical alterations in cells of the bladder field are often detectable before abnormal pathology, and markers previously thought to be limited to tumors were found in the field. The hierarchy of expression may be useful in identifying high-risk patients, assessing completeness of response to therapy, and monitoring and predicting recurrence.

Analysis of Variance↗
PubMed1993

Effect of exogenous heparin on anchorage-independent growth of fibroblasts induced by transforming cytokines.

Heparin at concentrations below 100 micrograms/ml stimulated anchorage-independent growth of NRK 49F (normal rat kidney fibroblasts, American type culture collection) rat fibroblasts at suboptimal cytokine concentrations but inhibited it at higher heparin concentrations regardless of the cytokine concentration. Heparin did not stimulate growth above that seen at optimal cytokine concentrations, suggesting that it alters the cellular response to the cytokines. These data suggest natural protein-glycosaminoglycan interactions play a role in modulating or mediating the actions of transforming cytokines and suggest they may play a role in acquisition of the transformed phenotype.

Agar↗
PubMed1993

Urinary glycosaminoglycan excretion as a laboratory marker in the diagnosis of interstitial cystitis.

Urinary macromolecular uronate and glycosaminoglycan uronate concentrations were determined in 209 urine specimens obtained from 192 interstitial cystitis patients, 47 asymptomatic normal individuals and 32 spinal cord injury patients. As a group the concentration of macromolecular uronate or glycosaminoglycan uronate was significantly less in the interstitial cystitis patients than in the normal controls. Some interstitial cystitis patients showed low values of macromolecular uronate and glycosaminoglycan uronate. Analysis of the population distributions suggested that macromolecular uronate may have significant value in diagnosis of interstitial cystitis.

Adult↗
PubMed1992

Intermediate endpoint biomarkers for chemoprevention.

The understanding of intermediate endpoint biomarker expression in relation to the sequential events in bladder tumorigenesis establishes a useful approach for evaluating chemopreventive agents. Biomarkers may be genotypic or phenotypic and function as biomarkers of susceptibility, exposure, effect, or disease. This paper reviews several years of research on biomarkers and their use in monitoring chemoprevention therapy. In initial animal experiments, mice were dosed with N-butyl-N-(4-hydroxybutyl)nitrosamine (OH-BBN) while co-administering N-(4-hydroxyphenyl)retinamide (4-HPR). 4-HPR did not statistically reduce tumor incidence, but did affect tumor differentiation and, consequently, nuclear size and DNA ploidy. These results suggest that nuclear size and ploidy may function as intermediate endpoint biomarkers of effect for oncogenesis and that epigenetic as well as genetic mechanisms may be primary in the oncogenic process. Early biomarkers of effect which occur prior to genetic effects or chromosome aberration may portend a higher probability of being modulated by differentiating agents such as retinoids. In vitro studies demonstrated that RPMI-7666 cells cultured with a phorbol ester tumor promoter (12-O-tetradecanoyl-phorbol-13-acetate) could be redifferentiated with 13-cis-retinoic acid and dimethyl sulfoxide (DMSO). F-actin, a cytoskeletal biomarker with a presumed function in the epigenetic mechanisms of carcinogenesis, could also be normalized in HL-60 cells treated with 4-HPR or DMSO. A clinical evaluation of F-actin in patients with varying degrees of risk confirmed the value of F-actin as a differentiating biomarker useful for bladder cancer risk assessment. The clarification of when the phenotypic changes of F-actin occur in the oncogenic process was achieved when a variety of biochemical changes were mapped in the patients with bladder cancer. These studies confirmed that G-actin, a reciprocal form of F-actin, is increased relatively early in bladder cancer oncogenesis when multiple biomarkers are quantitated in the field, adjacent area, and the tumor. Comparison of each individual biomarker's expression from field, adjacent to tumor, and tumor, and subsequent cluster analysis of these biomarkers, indicated that the possible sequence of phenotypic expression of biomarkers in bladder cancer oncogenesis is from G-actin, to p300 antigen, to epidermal growth factor receptor (EGFR), to p185 (neu oncogene product), to DNA aneuploidy and, finally, to visual morphology.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1991

Cellular F-actin levels as a marker for cellular transformation: correlation with bladder cancer risk.

Previous findings in cultured cells that differentiated cells had markedly higher F-actin levels than undifferentiated cells (Cancer Res., 50: 2215-2220, 1990) suggested that quantitative F-actin measurements in urinary cells might provide diagnostic or prognostic information by identifying those individuals with cells tending towards a lower degree of differentiation. The feasibility of such an approach was investigated using a risk stratification schema. Bladder wash samples were obtained from 163 symptomatic patients being evaluated for bladder cancer and 41 asymptomatic controls without hematuria or other symptoms consistent with bladder cancer. F-actin levels were evaluated by flow cytometry using a fluorescent phalloidin probe. The risk of bladder cancer was stratified according to biopsy, either DNA ploidy by flow cytometry or quantitative fluorescence image analysis cytology, previous bladder cancer history, and hematuria. A strong correlation between the presence of cells with abnormally low F-actin content in cells obtained by bladder wash from 38 patients and biopsy-proved bladder transitional cell carcinoma (P less than 0.001) was observed. A strong correlation was also observed between the presence of cells with low F-actin content and risk of bladder cancer assessed by either stratification schema (P less than 0.0001). The correlation was more consistent with the stratification by quantitative fluorescence image analysis cytology because of the 37% false-positive rate of ploidy analysis by flow cytometry among the control patients. Further evidence that low F-actin was correlated with cellular abnormality was obtained from simultaneously labeling cells for F-actin and with M344 antibody, a monoclonal antibody against a low-grade bladder tumor-related antigen. These studies showed that the F-actin content of the M344-positive cells was lower than that of the M344-negative cells. These results suggest that F-actin could be an early and sensitive marker for bladder cancer detection and risk prognostication.

Actins↗
PubMed1991

Identification of a high risk subgroup of grade 1 transitional cell carcinoma using image analysis based deoxyribonucleic acid ploidy analysis of tumor tissue.

The use of deoxyribonucleic acid (DNA) cytometry to identify a subset of patients with grade 1, stage Ta or T1 transitional cell carcinoma at high risk for death or recurrence was investigated in a retrospective study using paraffin blocks from 88 low grade transitional cell carcinomas of the bladder with an absorptiometric video-based image analysis system. Tumors were evaluated for ploidy (70 diploid, 16 aneuploid and 2 tetraploid) and the presence of cells with greater than 5C DNA. Survival analysis of 62 patients with adequate followup (15 to 20 years) showed that 43 of 62 (69%) suffered recurrences and 13 (21%) died of bladder cancer. The single most important predictors of death and recurrence were stem line aneuploidy and the presence of cells with greater than 5C DNA, respectively.

Carcinoma, Transitional Cell↗
PubMed1991

Quantitative fluorescence image analysis of deoxyribonucleic acid ploidy in urine from normal children.

Quantitative fluorescence image analysis incorporates the 2 diagnostic techniques of cytological analysis with quantitation of deoxyribonucleic acid (DNA). Exfoliated urinary cells are ideal for analysis by this method, which allows the identification of "rare event" abnormal cells. We evaluated the urine from 50 children who had undergone cystoscopy or were catheterized for other reasons. The urine was free of infection by urinalysis. Cytological analysis demonstrated normal or atypical cells in all patients. Of the patients 1 (2%) had greater than 2 of 500 cells analyzed with greater than 5C DNA and 4 (8%) had greater than 2 of 500 cells with greater than 5C double stranded nucleic acid. These data suggest that it may be "normal" for urine to contain "rare event" abnormal cells. The significance of this finding is unclear at present.

Adolescent↗
PubMed1990

Cellular F-actin levels as a marker for cellular transformation: relationship to cell division and differentiation.

Transformation is associated with profound structural and quantitative changes in the cytoskeleton. Herein we report studies using F-actin, a major cytoskeletal protein, as a quantitative marker for transformation cells, focusing on separating the effects of the cell cycle, cell differentiation, and transformation. The model system for these studies consisted of three lymphoblastic cell lines, one untransformed line (RPMI) and two transformed lines, one (HL-60) of which can be induced to differentiate and the other (Daudi) which cannot. The relation of F-actin levels to cell cycle was studied by flow cytometry with the use of fluorescein-phalloidin to label F-actin and propidium iodide to label DNA. F-actin levels in transformed Daudi and HL-60 lines were only two-thirds that of the untransformed RPMI cells. Histograms of the distribution of F-actin showed that the transformed lines consisted of two cell populations, one having an F-actin content near that of untransformed cells and the other having much less. Cell cycle analysis showed that F-actin in untransformed cells increased 10-15% as cells entered the S compartment, remaining approximately constant through G2 + M phases of the cell cycle, but in transformed cells the major increase in F-actin occurred during G2 + M phase. Double-label studies with rhodamine-phalloidin for F-actin and KI-67 monoclonal antibody for dividing cells (cells at late G1, S, G2, and M) measured with quantitative fluorescence image analysis showed that the mean F-actin content of dividing cells was twice that of nondividing cells. These results suggested that most of the cell division-related F-actin increase occurred during late G1 phase in untransformed cells. Differentiation of HL-60 cells with dimethyl sulfoxide or retinoic acid normalized the F-actin content of the nondividing cell population, but dimethyl sulfoxide and retinoic acid produced no detectable change in F-actin in the undifferentiable Daudi cells. A tumor promoter (12-O-tetradecanoylphorphol-13-acetate) inhibits differentiation of hematopoietic cells, resulted in a 32% decrease in the mean F-actin content of RPMI cells due to the appearance of a new subpopulation of low F-actin content. The 12-O-tetradecanoylphorbol-13-acetate-induced changes reversed slowly after removal of 12-O-tetradecanolyphorbol-13-acetate but more rapidly in the presence of retinoic acid. These results indicate that F-actin quantification can serve as a marker for cellular transformation and provides a tool for studying the mechanisms of cellular differentiation that may lead to a better understanding of the oncogenic process.

Actins↗
PubMed1990

Quantitative immunofluorescence, anti-ras p21 antibody specificity, and cellular oncoprotein levels.

A general approach to investigating specificity and saturation of antibodies by quantitative immunofluorescence is applied to monoclonal antibodies generated against p21 or ras oligopeptides to quantify ras p21 oncoprotein in cultured cells. Ras 10, a panreactive mouse monoclonal antibody, appears to be a superior probe for detection of p21 in cell extracts or fixed cells because it binds a 21 kD protein on SDS-PAGE/western blots and labels the cytoplasmic membrane in a saturable and competitive manner. RAP-5, a widely used mouse monoclonal antibody generated against an oligopeptide of ras p21, does not recognize p21 in denaturing immunoblots or in immunofluorescence of cultured cells.

Animals↗
PubMed1990

Early detection of colorectal cancer by quantitative fluorescence image analysis of exfoliated cells.

Early-stage colorectal cancer is potentially curable. In the present study, we applied quantitative fluorescence image analysis (QFIA) cytology to the detection of experimental colorectal cancer in a rodent model. QFIA cytology combines visual cytologic examination with quantitation of DNA content in single exfoliated cells. Cancer was induced by treating 110 rats with subcutaneous 1,2-dimethylhydrazine. Sequential colon washes were obtained weekly from each animal for 20 weeks. Control animals were treated identically except for the administration of carcinogen. Cells that were cytologically abnormal or had increased DNA content were found starting in the second week. By the eighth week, roughly 50 percent of animals had positive results, and this level remained approximately constant for the duration of the study. Tissue pathologic results were normal during weeks 1 to 7. Dysplasias became common during weeks 8 to 15 whereas most cancers appeared during weeks 16 to 21. These results indicate that QFIA cytology is a highly sensitive method for detecting even preneoplastic changes resulting from carcinogen administration and may prove useful in detecting human colorectal cancer.

Animals↗
PubMed1990

Decreased urinary uronic acid levels in individuals with interstitial cystitis.

The pathogenesis of interstitial cystitis currently is unknown. A possible etiology is that the transitional epithelium is defective, leading to molecular leaks that initiate the disease complex. An important surface defense mechanism is the glycosaminoglycans or polysaccharides that line the bladder epithelium and act as a nonspecific antiadherence factor blocking access of bacteria, microcrystals, proteins and ions to the underlying transitional cells. We examined the excretion of urinary macromolecular uronic acid and glycosaminoglycans in normal individuals and those with interstitial cystitis. A total of 37 controls had a mean macromolecular uronic acid level of 56 nmol. per mg. creatinine, compared to 40.2 nmol. per mg. creatinine in 43 patients with active disease (differences were significant, p equals 0.03). The median excretions of glycosaminoglycan uronate for controls and patients were 15.1 and 11.1 nmol. per mg. creatinine, respectively. (There was an over-all tendency to decrease excretion in patients with a p value of 0.06.) Specimens obtained at cystoscopy from patients with active interstitial cystitis had ureteral macromolecular uronic acid levels of 40.5 nmol. uronate per mg. creatinine compared to 43.6 nmol. uronate per mg. creatinine from the bladder. Interstitial cystitis patients had 16.0 nmol. glycosaminoglycan uronate per mg. creatinine compared to 14.6 nmol. per mg. creatinine in normal controls. Neither of these differences was statistically significant. It would appear that there is a tendency to lower macromolecular uronic acid and polysaccharide excretion in individuals afflicted with this syndrome.

Creatinine↗
PubMed1990

Quantitative fluorescence image analysis in bladder cancer screening.

Quantitative fluorescence image analysis (QFIA) cytology combines image analysis to measure DNA with visual cytology for bladder cancer detection. QFIA sensitivity is 76% to 81% and 95% to 100% for low- and high-grade tumors respectively, with 94% specificity in asymptomatic controls. QFIA screening of 504 persons within a beta-naphthylamine exposed cohort found DNA hyperploidy correlated with the duration of carcinogenic exposure and smoking history; marker prevalence was 23% for exposed workers who smoked and 2% for nonexposed nonsmokers. In prospective animal studies, QFIA was useful in monitoring carcinogenesis and chemoprevention with retinoids. Current QFIA research focuses on measurements of oncogenes, growth factors and their receptors, cytoskeleton, and tumor-associated antigens to improve sensitivity and specificity to low-grade tumors and to identify premalignant conditions. Profiles of biochemical and immunological markers on single cells may further assist in the study of high-risk cohorts and individual risk assessment.

Biomarkers, Tumor↗
PubMed1990

Molecular and cellular biological approaches and techniques in the detection of bladder cancer and enhanced risk for bladder cancer in high-risk groups.

Recent advances in molecular cell biology, immunology, and toxicology enhance detection of actual and incipient disease and the definition of risk. Tumor-associated antigens may greatly improve detection of low-grade tumor cells. Several new strategies depend upon detecting molecular lesions of the genes controlling cell growth (oncogenes) at either the gene or protein levels. Highly sensitive techniques detecting DNA adducts are refining the ability to detect specific DNA damage shortly after exposure. The recognition of controls that actively stop proliferation (tumor suppressor genes), regulate differentiation or of relationships between metabolism and carcinogenesis may lead to new ways to identify persons genetically at increased risk from carcinogenic exposure. Current technologies, those on the near horizon, and long-term research needs are analyzed critically in terms of screening objectives.

Biomarkers, Tumor↗