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A D Deitch

Publications and source records attributed to A D Deitch.

At least 37 records · Page 2Linked to original sources
PubMed1989

Measurement variability in DNA flow cytometry of replicate samples.

A Bladder Cancer Flow Cytometry Network study has been carried out aimed at identification of the sources of inter- and intralaboratory variability. Replicate "cocktail" samples containing a mixture of peripheral blood lymphocytes and an aneuploid cell line and samples of peripheral blood lymphocytes serving as a DNA reference standard were distributed to five network laboratories. The samples were stained for DNA using propidium iodide, with each laboratory using its own staining protocol. Sets of these samples were analyzed by flow cytometry to obtain cellular DNA distributions. DNA index and hyperdiploid fraction were calculated for each histogram using an automated technique. Results were evaluated by analysis of variance to identify sources of variability. Three important sources of variation were found that affect flow cytometry in general and- the transportability of flow cytometry results to routine clinical use in particular. The significant variation among laboratories that is constant across time most probably represents stable differences in instrumentation, instrument set-up, and laboratory techniques. This variation can be compensated for, if it is known and stable, to develop transportable classification criteria. The second type of variation, termed the interaction component, represents differences among laboratories that are not constant across time. Sources of this variation include inconsistency in sample preparation, staining, and analysis. The elimination of this type of variation is required for meaningful comparison of data within and among laboratories and the creation of interlaboratory data-bases. The third type of variation represents pure measurement variability and affects the sensitivity of the technique.

Cell Line↗
PubMed1989

The clinical application of aspiration deoxyribonucleic acid flow cytometry to neurologically impaired men entering an electroejaculation program.

For the neurologically impaired patient electroejaculation with a rectal probe can furnish sperm for artificial insemination. However, documentation of pregnancies in this patient group has been limited. Deoxyribonucleic acid flow cytometry has been shown to be a reproducible quantitative method with which to evaluate spermatogenesis. We assessed the ability of deoxyribonucleic acid flow cytometry on testicular aspirates to predict the quality of sperm production in 13 men undergoing electroejaculation. Semen analysis was performed on antegrade and retrograde specimens. Deoxyribonucleic acid histograms from testicular aspirates were evaluated for the relative proportions of haploid, diploid and tetraploid cells. These ratios were compared to the means reported for our own series of 25 patients who at vasovasostomy had sperm intraoperatively and to the similar figures from a group of 10 accident victims. Five patients had normal testicular ploidy compartments defined as within 2 standard deviations from control means. Of the 5 patients 4 had normal sperm counts and motilities (30 per cent or greater). Two patients have contributed sperm for artificial insemination; 1 has resulted in pregnancy. Three patients had moderately abnormal (2 to 4 standard deviations) testicular ploidy classes. These patients had adequate sperm counts but low motilities (5 per cent or less). Of the 5 patients with markedly abnormal (greater than 4 standard deviations) histograms 4 remained azoospermic despite repeated attempts at electroejaculation. Our data support the use of deoxyribonucleic acid flow cytometry on testicular aspirates as a predictor of which patients might succeed and those who are unlikely to succeed in an electroejaculation program.

Adult↗
PubMed1989

Deoxyribonucleic acid flow cytometry of benign prostatic disease.

Deoxyribonucleic acid flow cytometry was performed on aspirated prostatic cells from 198 patients who had benign cytological or histological findings. Unsatisfactory acellular histograms were obtained from 10.6 per cent of the cases. Three-fourths of the satisfactory samples (more than 5,000 cells after subtracting debris) showed the expected single peak deoxyribonucleic acid diploid to near diploid histograms. Unexpectedly, the remaining samples were deoxyribonucleic acid aneuploid, most having 2 peridiploid peaks (deoxyribonucleic acid index 0.82 to 1.31). Usually, proliferation was low with less than 20 per cent hyperdiploid cells and with 2.5 +/- 1.5 per cent G2 cells. In 10 per cent of the single peak histograms there was evidence of inflammation, identified as an increase in hyperdiploid cells without an increased percentage of G2 cells but with a tail of high channel values. The aforementioned histogram features were considered to be benign findings. Seven per cent of the samples had deoxyribonucleic acid histograms suggestive of prostate cancer. Of these samples 7 had diploid or peridiploid aneuploid histograms with high proliferation (more than 20 per cent hyperdiploid cells with 8.5 +/- 3.8 per cent G2 cells), while 5 had histograms with deoxyribonucleic acid aneuploidy other than peridiploidy.

DNA↗
PubMed1989

Canine 563 cells: an established canine salivary gland cell line.

Canine mandibular salivary glandular epithelial cells were established in culture and passaged more than 250 times. The cultivated cells show epithelial morphology, and ultrastructurally, show desmosomes, desmosome-like specializations (punctae adhaerens), cytoplasmic tonofilaments, and neoluminae suggestive of glandular epithelium. The cells showed modest PAS reactivity and were negative for alcian blue. Using immunohistochemistry, the 563 cells were shown to express cytokeratins, vimentin and epithelial membrane antigen. Chromosome analysis of the cells revealed a modal number of 76 plus 2 sex chromosomes, and the DNA histogram showed major peaks at 2 C (diploid) and 4 C (tetraploid). Attempts to demonstrate a retrovirus by particle labelling with 3H-uridine or by isolation of 70S RNA gave negative results. A nucleic acid hybridization test for mycoplasma contamination gave negative results. It is anticipated that this established cell line derived from normal canine glandular epithelium will prove useful for canine virologic studies, or as a substrate for the growth of type C retroviruses from heterospecies.

Animals↗
PubMed1988

Interinstitutional variability in DNA flow cytometric analysis of tumors. The National Cancer Institute's Flow Cytometry Network Experience.

Flow cytometric DNA analysis of human urinary bladder specimens may be clinically useful for prognosis in transitional cell (urothelial) carcinoma and for detecting recurrence after treatment. However, many important methodological differences exist among institutions which have described this technique, and it has not previously been shown that data from different institutions are comparable. The National Cancer Institute has created a Flow Cytometry Network to address the need for technology assessment of flow cytometry. This report describes the independent flow cytometric analysis and interpretation of "unknown" paraffin-embedded bladder tumor specimens by the five Network institutions. Although important differences in method existed among the institutions, substantial agreement was achieved in actual data generated and their interpretation. This suggests that a consensus regarding acceptable laboratory performance of this technique could be reached, which should facilitate its more widespread clinical implementation.

Carcinoma, Transitional Cell↗
PubMed1988

Comparison of automated and manual techniques for analysis of DNA frequency distributions in bladder washings.

Quantitative methods for interpretation of flow cytometry DNA histograms are required for the widespread clinical use of this technology. The usefulness of a histogram analysis technique in this setting requires that it be operator independent, easy to implement in a clinical laboratory, and provide high sensitivity to the desired information. Additionally, the technique must be tolerant of the relatively low signal-to-noise ratios often found in DNA distributions obtained from clinical samples. Among the factors that have been used to assess the malignant potential of tumors are the presence of an aneuploid population, the proportion of hyperdiploid cells, the width of the G1 peak, the DNA index, and the fraction of cells in S. A computer-based method has been developed for extraction of the above-mentioned features from DNA histograms. The program detects peaks in the histogram and uses straight-line fits to the cumulative frequency distribution to define cell population bounds. A test set of 44 histograms compiled from bladder irrigation specimens obtained from patients with a present or past history of transitional cell carcinoma (TCC) was analyzed by five collaborating laboratories forming a Network sponsored by the National Cancer Institute (NCI). This test set was used to evaluate the performance of the computer-based method by comparing results with those of four expert observers. In this preliminary analysis, perfect agreement was found in the detection of aneuploid cell populations by all observers and the computer-based method. Correlation of percent hyperdiploid cell fraction was also excellent.(ABSTRACT TRUNCATED AT 250 WORDS)

Algorithms↗
PubMed1988

Characterization of urinary keratin number 18 using a new assay.

The levels of the low-molecular weight keratin 18 appearing in the voided urine of 130 healthy volunteers was studied using a new immunoradiometric assay for keratin 18 (IRMAK-18). Keratin 18 was detected in 79 urines with an average level of 2.5 +/- 4.7 ng./ml. The assay does not detect complex epidermal keratins or non-keratin intermediate filaments. Only 50% of the purified keratin added to urine could be recovered. The recovered keratin had the same molecular weight as the input molecules, implying that substances in the urine physically inhibited the assay rather than destroyed the keratin. The independent variables of cell count, dead cells, and protein concentration were not found to be important in evaluating the results. However, the urine concentration, as measured by creatinine, might be an important consideration for assessing the levels of keratin detected.

Adult↗
PubMed1988

The predictive value of flow cytometric information in the clinical management of stage O (Ta) bladder cancer.

Two blind retrospective flow cytometric studies were performed on archival stage O (Ta) papillary grades 1 to 2 transitional cell carcinomas of the bladder to learn whether deoxyribonucleic acid histogram patterns predict disease recurrence and progression. All patients with aneuploid histograms as predicted experienced recurrent disease. In 20 of 28 cases of recurrent disease the ploidy pattern changed during recurrence. Because of this finding we predicted disease progression if 2 or more histograms in the sequence were aneuploid. Two-thirds (8 of 12) of those having more than 1 aneuploid tumor incident experienced invasion, while only approximately a third (6 of 16) lacking repetitive aneuploidy had invasion. Furthermore, of the patients with 2 aneuploid lesions half had progression within 1 year and two-thirds within 2 years after the second aneuploid lesion. Therefore, it appears that in this disease deoxyribonucleic acid histograms can provide prognostic information beyond that obtained from tumor grade and stage.

Carcinoma, Transitional Cell↗
PubMed1988

Urine: a suitable sample for deoxyribonucleic acid flow cytometry studies in patients with bladder cancer.

Deoxyribonucleic acid flow cytometry of bladder washings has proved to be a valuable procedure for the diagnosis and followup of patients with transitional cell carcinoma. However, for this procedure to gain maximal acceptance, it should be possible to use voided urine specimens instead of bladder washings. To evaluate this possibility we compared histogram findings for 114 bladder washings and 122 concomitantly obtained urine samples (voided and catheterized) from 89 consecutive patients who had active or a history of transitional cell carcinoma. Unsatisfactory histograms were obtained in 4 per cent of the urine samples and in 2.6 per cent of the bladder washing samples. The satisfactory rate for voided or catheterized urine samples was the same. We conclude that in this patient population satisfactory deoxyribonucleic acid histograms can be obtained from samples of voided urine.

Carcinoma, Transitional Cell↗
PubMed1986

Flow cytometry: role in monitoring transitional cell carcinoma of bladder.

The ability to detect and monitor the course of transitional cell carcinoma (TCC) of the bladder using DNA histograms obtained from flow cytometry was studied. Voided urine and barbotage specimens were collected from patients with active TCC or a past history of TCC. These specimens were submitted to routine cytologic and flow cytometric analyses. Samples were considered to be positive if they met one or more of three criteria: if they had aneuploid or tetraploid peaks, if the DNA index of the major G1 peak was shifted more than 10 per cent from that of diploid cells, or if 15 per cent or more of the cells fell to the right of the major diploid G1 cell population thereby constituting a significant hyperdiploid cell population. Using these methods for patients with active disease, the detection rate was 91 per cent. In patients with a past history of TCC, positive histograms preceded the appearance of visible tumor in one third of the cases. Flow cytometry proved to be an excellent way of following patients with a past history of TCC or of screening patients suspected of having active disease. Following this protocol, few biologically active tumors go undetected. However, in 112 patients without a history of bladder cancer, the false positive or suspicious rate was 38 per cent. Before flow cytometry can be recommended as a widespread screening method for patients thought to be at risk of TCC of the bladder developing, this suspicious group will have to be eliminated.

Carcinoma, Transitional Cell↗
PubMed1985

The influence of cytoreductive surgery on the response to chemotherapy of a rat renal cancer.

The potential ability of cytoreductive surgery to increase the effectiveness of chemotherapy (vindesine) was tested utilizing male Wistar Lewis rats transplanted simultaneously with intraperitoneal and flank implants of a spontaneously arising renal adenocarcinoma. Cytoreduction was accomplished in some animals by removing the flank tumor 5-7 weeks following implantation; all animals received vindesine (IP injection of 0.5 mg/kg on two successive weeks). While vindesine reduced tumor growth, in no case did the addition of cytoreductive surgery enhance the effect of chemotherapy. The addition of cytoreductive surgery to marginally effective chemotherapy was found to be ineffective or even detrimental.

Animals↗
PubMed1984

Comparison of histologic and flow cytometric evaluation of cyclophosphamide-induced testicular damage.

This study evaluates the ability of DNA histograms obtained by flow cytometry to detect and quantify reversible alterations in spermatogenesis induced by cyclophosphamide, a known inhibitor of spermatogenesis. Evaluation of per cent of cells in each of the haploid (lc), diploid (2c), and tetraploid peaks (4c) as determined by flow cytometry in treated and control Balb/C mice over a six-week period, and comparison with routine histologic evaluation have led us to conclude that DNA histogram evaluation is a rapid and accurate means of identifying testicular damage and recovery. This technique may be useful in sequential monitoring of the effects of malignancy and/or treatments applied on spermatogenesis in young men.

Animals↗
PubMed1984

Testicular torsion: temporal considerations.

Unilateral testicular torsion may result in contralateral testicular alterations which appear immunologically mediated and avoidable by immunosuppression or orchiectomy of the twisted testicle within 24 hours. This study was instituted to assess three temporal aspects of these observations: (1) The effect of prepubertal torsion was studied. It was found that after prepubertal torsion, the contralateral testicle underwent normal development. (2) The duration of torsion necessary to result in contralateral testicular alterations in adult rats was studied. Deoxyribonucleic acid (DNA) histograms were utilized to assess disturbed spermatogenesis. After greater than 8 hours of torsion, detorsion offered no protection to the contralateral testicle. Marked alterations occurred in DNA histograms of 60% to 80% of the animals. (3) The duration and significance of these alterations were assessed. The alterations persisted in the contralateral testis 6 months, and the fertility rates were significantly lower than for control animals.

Animals↗
PubMed1984

Alterations of physical and biochemical parameters of the R3327-CP rat prostate adenocarcinoma following hormonal manipulation of the host.

Several physical and biochemical parameters of a rapidly growing, hormonally responsive, poorly differentiated strain of Dunning R3327 rat prostatic adenocarcinoma (the CP strain) were monitored for 1 month during growth in control and hormonally manipulated male Fischer X Copenhagen rats. The tumor was implanted into control rats and into rats 1 month following orchiectomy. Twenty-nine days following tumor implantation, 1 group of unoperated rats was orchiectomized while the rats implanted subsequent to orchiectomy were repleted with pharmacological doses of testosterone. At 2 and 4 weeks following treatment, half the original number of rats from each group were sacrificed and the growth rate (doubling time), per cent of aneuploid cells and androgen receptor levels (total, cytoplasmic and nuclear) were determined for each tumor. Orchiectomy increased tumor doubling time, while testosterone repletion decreased it, demonstrating the hormonal dependence of this tumor strain. Orchiectomy also decreased the levels of aneuploid cells in the tumor; however, repletion of testosterone to rats orchiectomized prior to implantation did not restore the aneuploid cell number to control levels. A sensitive indicator of the hormonal status of the tumor was the per cent of androgen receptors in the nucleus. Tumors grown in rats orchiectomized after implantation had the lowest percentage of androgen receptor in the nucleus while orchiectomized rats repleted with testosterone had the highest percentage. Comparison of the levels of androgen receptors in the tumors from the various groups (androgen receptor per gram of tissue) unexpectedly revealed that tumors grown in the orchiectomized rats had slightly higher total receptor levels than did control tumors, while the tumors of orchiectomized rats repleted with testosterone had lower amounts than did the control tumors. In contrast to these findings, the prostates of orchiectomized rats replenished with testosterone had higher levels of total androgen receptor than did the prostates of control rats.

Adenocarcinoma↗