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Biomedical subjects

D Haag

Publications and source records attributed to D Haag.

At least 19 recordsLinked to original sources

Limitations of controlled experimental systems as models for natural systems: a conceptual assessment of experimental practices in biogeochemistry and soil science.

Experimental systems in which phenomena are studied under controlled conditions allow scientists to infer causal relationships from observable effects. When investigating ecosystems, however, scientists face complex systems. The conventional approach is to divide the system into conceptual units and to prepare experimental systems accordingly. Experimental systems are used as models for ecosystems: initially, scientists assume an analogy between the experimental system and ecosystem, then encode the experimental system into a formal system by measuring variables, and decode statements from the formal system to the ecosystem. We distinguish three types of experimental systems, i.e. laboratory, container and field set-ups, further divided into seven subtypes. Starting from the premises of experimental systems, we comment on the possibilities and limitations of experimentally derived causal relationships and on their significance for ecosystem understanding and prediction, illustrated by examples from soil science and the environmental sciences. Experimental set-ups have a characteristic duration, degree of structural integrity, internal variability and boundaries, which relate to conceptual closure and experimental control: control tends to be maximum on short time scales, in homogeneous set-ups with analytical boundaries, and in systems with few parameters to be observed. Complexity is increased at the expense of control. The higher the degree of manipulation, however, the better is reproducibility, but the larger is the deviation from unique ecosystems with their infinite number of factors. The material realization of closed systems is preceded by a conceptual closure of the system. Closure is relative to the domain of phenomena of interest, the theory and the list of variables selected by the scientist. Successful decoding from experimental systems to ecosystems largely depends on the validity of the chosen analogy. Laboratory systems are idealized systems which contain a limited number of a priori defined variables, and which are shielded from environmental influences. In contrast, ecosystems are materially and conceptually open, non-stationary, historical systems, in which system-level properties can emerge, and in which variables are produced internally. We conclude that when conducting experiments, causal factors can be identified, but that causal knowledge derived from insufficiently closed systems is invalid. In ecosystems, innumerous factors interact, which may enhance, reduce or neutralize the effect of an experimentally determined factor. Thus, experimental model systems need to be evaluated for concrete, well-defined ecosystems with a concrete history. Increasingly detailed studies of isolated phenomena in the laboratory will probably not contribute much to ecosystem-level understanding. When conducting experiments, scientists should aim at the maximum degree of complexity they can actually handle and they should justify the chosen analogy.

Ecosystem↗

IMDA/aldol strategy for transforming carbohydrates into functionalized trans-decalins.

L-Rhamnal is readily converted into an allyl 2, 3-unsaturated-C-glycopyranoside. The (S) configuration of the alphaL-anomer defines the stereochemical outcome of the future IMDA reaction, leading to the absolute stereochemistry for the trans-decalin moiety in naturally occurring terpenoids. Selective cleavage of the terminal double bond of the allyl group provides an aldehydo function which serves for an aldol/Claisen addition with ethyl sorbate. Of the four possible diastereomers, one is obtained in pure form and processed to give the IMDA precursor. Cyclocondensation is achieved by heating in xylene to give a tricyclic trans-decalin whose structure is established by NMR and X-ray analysis.

Carbohydrates↗

A model for enhancing worldwide personal health and wellness.

Individuals must increasingly take control of managing their own health affairs. This requires access to quality information that is not easily obtained from traditional social institutions. NetWellness is an electronic consumer health information service that provides a model for reaching the goal of enhancing personal health and quality of life. [1] We present a vision of consumer health information delivery in the 21st century, and a model for reaching that vision. Our experience to date, progressing through a five-phase model, is aimed at providing the best health information possible to the widest population possible.

Community Participation↗

[DNA cytometry in breast carcinoma. Review of method and value in assessing prognosis].

DNA flow cytometry (FCM) has become a routine method in breast cancer diagnosis for evaluation of ploidy and proliferation kinetics (cell cycle analysis). Image cytometry is less practicable and provides less information than flow cytometry. An optimized technique with a low coefficient of variation is required for optimal results in flow cytometry. The S-phase fraction and the proliferation index (sum of S-phase fraction and G2M fraction) provide prognostic and therapeutically relevant information. A profound knowledge of the technique and its limitations is indispensable for the interpretation of FCM results. It remains to be established whether immunohistological evaluation of cell proliferation has the same prognostic value. Future developments are to be expected from multiparametric analysis and the improvement of mathematical analysis of FCM measurements.

Breast↗

An electronic consumer health library: NetWellness.

NetWellness is a community-based, consumer-defined grant program supporting the delivery of electronic health information to rural residents of southern Ohio and urban and suburban communities in the Greater Cincinnati tri-state region. NetWellness is a collaboratively developed and publicly and privately funded demonstration project. Information is delivered via ISDN, standard dial, dedicated network connections, and the Internet. TriState Online (Greater Cincinnati's Free-Net) and other southern Ohio Free-Nets are key access points in the larger project communities. The other access points are more than forty workstations distributed at public sites throughout the project's primary geographical area. Design strengths and limitations, training initiatives, technical issues, and the project's impact on medical librarianship are examined in this paper. Also discussed are ways of determining community needs and interest, building political alliances, finding and developing funding sources, and overcoming technical obstacles. NetWellness's Internet address is: http:@www.netwellness.org.

Community Participation↗

FDG uptake, tumor proliferation and expression of glycolysis associated genes in animal tumor models.

To determine the influence of tumor cell proliferation and changes in the genetic program in malignant cells on the fluorodeoxyglucose (FDG) uptake we performed PET studies in several animal tumors: spontaneous mammary fibroadenoma, chemically-induced mammary adenocarcinoma and Dunning prostate adenocarcinoma. The expression of the glucose transporter (GLUT1) and of hexokinase (Hk) was measured using 32P-labeled cDNA probes and densitometry. Furthermore the proliferative activity was determined with one-dimensional flow cytometry. The FDG uptake and the proliferation parameters were not correlated. The normalized amounts of GLUT and Hk mRNA were lower in spontaneous fibroadenomas and prostate tumors than in chemically induced mammary. The FDG uptake was correlated to GLUT1 expression with r = 0.83 and to Hk expression with r = 0.77. Multiple regression analysis revealed a relation of FDG uptake to GLUT1 and HK with r = 0.87. Our results show that the FDG uptake in our study was related not to differences in proliferation, but rather to differences in the transcription of glycolysis associated genes.

Animals↗

Glucose uptake, perfusion, and cell proliferation in head and neck tumors: relation of positron emission tomography to flow cytometry.

The uptake of 18F-Deoxyglucose (FDG) was studied in vivo in relation to the proliferation rate of human head and neck tumors. Forty-two patients with histologically proven squamous-cell carcinoma of the head and neck and four patients with metastases of head and neck tumors were examined with PET and FDG prior to surgery. In 35 of these patients, a flow cytometric analysis of the DNA content and the proliferation rate was done using one-dimensional flow cytometry rate was done using one-dimensional flow cytometry (DAPI staining). In 17 cases, perfusion studies with 15O-labeled water were performed. Twenty-seven specimens were evaluable by flow cytometry. The analysis of the distribution of the FDG uptake revealed two groups, showing a high and a lower uptake pattern. In both groups the FDG uptake and the proliferation rate were correlated with an r-value of 0.64 and 0.8 respectively. However, the slope of the regression function was flat. No correlation was found between the perfusion and the proliferation rate. It is suggested that these differences in uptake in histologically identical tumor populations may correspond to differences at the molecular level, e.g., differences in the amount of the glucose carrier, perhaps caused by oncogenic transformation.

Carcinoma, Squamous Cell↗

DNA index and cell cycle analysis of primary breast cancer and synchronous axillary lymph node metastases.

The DNA Index (DI) and the percentage of cells in S-phase (S-phase fraction, SPF) were measured by flow cytometry in 80 primary breast carcinomas and in 80 accompanying axillary lymph node metastases. The DI in primary tumors and metastases agreed in 61 cases (76%). Cases with diploid primary tumors revealed more constancy of the DI in comparison to the metastases than the cases with aneuploid primary tumors (91% and 70% respectively). The mean values of the SPF were in close agreement in the primary tumors and in the lymph node metastases (6.1% and 6.0% respectively). Differences between the SPF of the two groups could be detected only by the consideration of case-related data pairs. In 50 cases (62%), the percentage of SPF agreed approximately in primary tumors and in the correspondent metastases. The cases with diploid primary tumors revealed more agreement of the SPF in the primary site and the metastases than did cases with aneuploid primary tumors (78% and 56% respectively). In conclusion, diploid carcinomas and their metastases revealed more constancy of the DI and the percentage of SPF than aneuploid carcinomas. These findings agree well with a better prognosis of diploid mammary carcinomas, as reported in the literature. Comparisons between the DI and the SPF in primary tumors and the corresponding metastases could be a source of valuable information on the biological behaviour and the aggressiveness of mammary carcinomas.

Aneuploidy↗

Correlation of DNA flow cytometric results and other prognostic factors in primary breast cancer.

The percentage of cells in S-phase and DNA-ploidy have been measured in 300 primary mammary carcinomas by means of DNA-flow cytometry (FCM). The data were compared with the age and menopausal status of the patients as well as with the size, regional lymph-node involvement, histologic type, grade and concentration of estrogen (ER) and progesterone (PR) receptors of the tumors. A DNA-diploid distribution of the G0/1-peak was found in 37.6% of the cases. The mean percentage of S-phase fractions was 4.83. DNA-aneuploid tumors had significantly higher amounts of S-phase fractions (6.12%) than DNA-diploid tumors (2.66%). There was also a significant correlation between the DNA measurement data (DNA-ploidy and S-phase fractions) and histologic grade, as well as the content of ER and PR, but not between DNA-ploidy, S-phase fractions, tumor size (T) and evidence of axillary lymph-node metastases. DNA-FCM gives a biological characterization of the tumor in addition to the histopathologic examination. The method can be used as a routine procedure because of the reliability and reproducibility of the results as well as the short time needed for the measurements.

Age Factors↗

Comparison of S-phase fractions measured by flow cytometry and autoradiography in human transplant tumors.

The 3H-thymidine labeling index (TLI) and the percentage of cells in the S-phase have been determined by autoradiography and by flow cytometry, (FCM), respectively, in six malignant tumors of human origin transplanted on athymic nude mice. The Dean and Jett model and the graphical model were used to determine the percent of S-phase cells by FCM. Cell cycle analysis was performed using 1) no correction for background; 2) an algebraic function for background correction; and 3) an exponential function for background subtraction. Each of these three data sets was evaluated using both the Dean and Jett model and a graphical model for the evaluation of DNA histograms. The S-phase fractions (SPF) were compared to the corresponding labeling index results. SPF without background correction were 1.54 times higher than the TLI. SPF, after correction using the algebraic model, were 1.29-fold higher than the TLI, whereas SPF obtained after background subtraction according to the exponential model were only 1.05-fold higher than the TLI. Student's t-test revealed significant differences between the mean TLI values (16.25 +/- 9.06) and the mean SPF obtained by FCM without background correction (mean 25.0 +/- 9.36, P less than 0.01), but not between the mean TLI values and the mean SPF percentages after algebraic (mean 21.0 +/- 10.29) and exponential background correction (mean 17.11 +/- 11.59), P greater than 0.05 each. There was no difference between the results obtained using the Dean and Jett model and those obtained using the graphic evaluation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of systematic errors on the evaluation of the S phase portions from DNA distributions of solid tumors as shown for 328 breast carcinomas.

The portion of cells in S phase has proved to be a valuable prognostic indicator of early relapse and life expectancy, particularly in breast carcinoma. Comparisons of published data on samples of primary breast carcinoma biopsies showed that the values obtained by analyses of flow cytometric DNA distributions were generally higher than those of determinations based on the tritiated thymidine (3H-ThdR) labeling index (LI). Flow cytometric DNA analyses of 328 biopsy samples of primary breast carcinomas revealed that these differences could be explained by varying contributions of debris background. Since this influence is inversely proportional to the cell counts in each channel, it may cause considerable errors, particularly in the S phase channels, which normally contain the lowest counts of the DNA distributions. Two different mathematical rationales were tested in order to separate DNA distributions from the debris superimposition. No appreciable differences were found with respect to the essential results. After appropriate subtraction of the background levels, the previously reported discrepancies between cytometrically determined S phase portions and 3H-ThdR LI values disappeared, and good agreement was achieved for the comparable tumor samples of the present study. In conclusion, debris background subtractions should generally precede the DNA histogram analyses, particularly of solid tumors, in order to obtain reliable S phase values.

Breast Neoplasms↗

Heterogeneity of cultured leukemic lymphoid progenitor cells from B cell precursor acute lymphoblastic leukemia (ALL) patients.

Colony assays were performed for 50 patients with B cell precursor acute lymphoblastic leukemia (ALL). Blast colony formation was observed for 33 patients, and the plating efficiency (PE) showed a marked interpatient variation, which indicates a pronounced biological heterogeneity at the level of leukemic progenitor cells. Notably, the mean PE of leukemic B cell precursors from patients with a pseudodiploid or near-diploid karyotype with structural chromosomal abnormalities (SCA) was significantly higher than the mean PE of normal diploid or hyperdiploid cases. All patients who had SCA involving 7p13, 11q23-24, or 12p11-13, and patients with a Philadelphia chromosome had high PE values. The S phase percentage, expression of CD19 antigen, and relapse status were also correlated with PE. Significantly, colony blasts had slightly different surface marker profiles in each case and were common ALL antigen negative in 33% of cases, which indicates the existence of a marked immunological heterogeneity at the level of leukemic progenitor cells.

Adolescent↗

Ex vivo elimination of lymphoblastic leukemia cells from human marrow by mafosfamid.

Studies were performed to evaluate the anti-tumor activity of mafosfamid, a new synthetic derivative of cyclophosphamide. We tested its ability to eliminate lymphoblastic leukemia cells from autologous bone marrow grafts following a 30 min preincubation in a highly sensitive clonogenic assay. Treatment with 50-100 micrograms mafosfamid/ml eliminated more than 4 logs of contaminating clonogenic tumor cells from a 200-fold excess of normal bone marrow. Flow cytometric studies showed differences in cell cycle kinetics between mafosfamid-resistant and mafosfamid-susceptible tumor cell clones. Compared to drug susceptible clonogenic tumor cells, clones that resisted treatment with 100 micrograms mafosfamid/ml exhibited a smaller percentage of cells in S-phase, indicating that mafosfamid is mostly cytotoxic to rapidly cycling tumor cells. The combination of mafosfamid and a target cell selective immunotoxin containing pokeweed anti-viral protein was superior to mafosfamid alone or immunotoxin alone for purging mafosfamid-resistant leukemic cells from human marrow.

Bone Marrow↗

DNA flow cytometry of ovarian tumors with correlation to histopathology.

Tissue samples from 49 women with seven benign and 42 malignant ovarian tumors were examined by means of DNA flow cytometry (FCM). The FCM data (DNA-ploidy and the number of S-phase fractions) were compared with the International Federation of Gynecology and Obstetrics (FIGO) stage, the histologic grade of differentiation and in some cases with the clinical outcome. The benign ovarian tumors were all diploid with a mean of 2.1% (+/- 1.66) S-phase fractions. Adenocarcinomas with the histologic grade 1 were diploid and had a mean of 2.1% (+/- 1.17) S-phase fractions. Grade 2 adenocarcinomas were aneuploid in eight of nine cases and revealed an increased proliferative activity (7.7% +/- 2.67 S-phase fractions). A high number of aneuploid cases (nine of 13) and an increased DNA synthesis were found in grade 3 adenocarcinomas (12.0% +/- 5.72 S-phase fractions). Four of six malignant nonepithelial tumors also had high numbers of S-phase fractions (9.7-14.5%). A significant correlation between the histologic grade and the DNA synthesis, FIGO stage, and DNA-ploidy was found. DNA-FCM may be used as an additional diagnostic tool supplementing routine histopathologic examination of ovarian tumors for better biological characterization, especially for those with uncertain grading, for grade 2 neoplasms, and for malignant nonepithelial tumors.

Adenocarcinoma↗

[DNA measurement of malignant tumors by impulse cytophotometry. The principles and importance for assessing growth behavior and the degree of abnormality].

Monometric DNA impulse cytophotometry of ploidism state (diploid or aneuploid tumors with stemmline shift, polyploidism ) and proportion of DNA-synthesized cells (S-phase proportion) were determined on over 500, mostly malignant, tumors of different sites. The results were compared with histopathological findings (TNM stage and degree of differentiation). Large diploid carcinomas of the oral cavity have lower S-phase activity than aneuploid tumors. Carcinomas of the breast, stomach and ovary are frequently diploid. Aneuploid carcinomas of these regions generally have high synthesizing activity. Mucoid signet-ring carcinomas of the stomach have lower S-phase activity than non-mucoid carcinomas. In the colorectal region aneuploid carcinoma predominates, in the colon more than in the rectum. Tumor metastases from colon and rectum into the liver predominantly are from aneuploid primary tumors. For tumors of the breast, corpus uteri and ovary with high and moderate differentiation there is a direct correlation between histological grade of differentiation and the S-phase proportions. Aneuploid tumors vary from low to high synthesizing activity. Diploid meningioma and glioma have a low S-phase proportion, aneuploidism correlates with an increase in growth. Supplementation of histopathological diagnosis by determining ploidism and S-phase activity makes an important contribution in the assessment of the degree of malignancy, as well as for therapeutic purposes.

Aneuploidy↗

The proliferative index (PI) of human breast cancer as obtained by flow cytometry.

As known from previous reports, the DNA synthesis fraction of mammary carcinoma cells is correlated with the course of the disease and response to adjuvant therapy. Quantitative parameters of the proliferative activity can be determined by the classic 3HTdR-labelling technique as well as by the more rapid flow cytometric (FCM) DNA analysis. The values of DNA-cytometric S-phase fractions in breast cancers reported up to now were consistently higher than those obtained by the 3HTdR labelling index (LI). This discrepancy was surmounted in the present study by corrections for systemic errors of the method. The fractions of cells in stages of the cell cycle as well as the DNA indices (DI) in 155 cases of primary resectable breast cancers were analyzed by DNA flow cytometry. The patients were aged between 24 and 88 years. As indicated by a bimodal age distribution with a decrease at age 60, the patient population was a representative of the generally known breast cancer incidence. 54% of the patients had aneuploid tumor cells with preferences of DNA indices 1.6 and 2.0. In the remaining 46%, no cell populations with deviating DNA content could be detected, in part possibly due to very small differences beyond the limits of detection. No correlations were found between age, menopausal status, histologic type of tumors, tumor size, fractions in stages of the cell cycle and proliferative index (PI = S + G2 + M in %), except a significant correlation between the S-phase fractions and the (G2 + M)-phase fractions in a ratio of approximately 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗