Search PubMed⌕ Search

Biomedical subjects

M A Davis

Publications and source records attributed to M A Davis.

At least 91 records · Page 5Linked to original sources
PubMed1995

Lack of dependence of 5-fluorodeoxyuridine-mediated radiosensitization on cytotoxicity.

It has been proposed that fluoropyrimidine-mediated cytotoxicity and radiosensitization are closely correlated. We have shown that HT29 human colon cancer cells transfected with the E. coli dUTPase gene are resistant to 5-fluorodeoxyuridine (FdUrd)-mediated cytotoxicity, presumably through more effective elimination of dUTP. We used these cells to assess the association between radiosensitization and cytotoxicity produced by FdUrd. The radiation sensitivities of the clones expressing elevated dUTPase activity (dutE clones) were similar to those of untransfected HT29 cells or HT29 cells which had been transfected with only the expression vector for the E. coli gene (con clones). We found that FdUrd produced similar increases in radiation sensitivity regardless of dUTPase activity. Levels of dUTPase in the dutE clones remained elevated during the entire period of FdUrd exposure, demonstrating that the lack of difference between dutE and Con clones was not a reflection of down-regulation of dUTPase activity by FdUrd. Flow cytometry showed that all clones progressed past the G1/S-phase boundary and into early S phase during FdUrd treatment. These data suggest that the mechanisms of FdUrd-mediated cytotoxicity and radiosensitization are not closely linked. These findings, combined with our previous investigations, are consistent with the hypothesis that radiosensitization occurs in cells which progress past the G1/S-phase boundary in the presence of FdUrd.

Cell Survival↗
PubMed1995

RP 73870, a gastrin/cholecystokinin-B receptor antagonist with potent anti-ulcer activity in the rat.

RP 73870, the racemic potassium salt of (([N-(methoxy-3-phenyl)-N-(N-methyl-N-phenyl-carbamoylmethyl)- carbamoylmethyl]-3-ureido)-3-phenyl)-2-ethylsulfonate-(RS) is a potent, reversible antagonist of both gastrin and cholecystokinin-B receptors in guinea pig and rat tissues. This compound is a potent inhibitor of pentagastrin-stimulated gastric acid secretion in the perfused rat stomach. RP 73870 also inhibits basal gastric acid secretion in the rat, although at doses higher than that required for inhibition of pentagastrin-stimulated gastric acid secretion. RP 73870 is a potent inhibitor of aspirin-induced gastric damage in the rat. In the prevention of aspirin-induced gastric damage, RP 73870, given p.o., was 10-fold less potent than when given i.v. RP 73870 was as potent as a H2 receptor antagonist or proton pump inhibitor in the prevention of cysteamine-induced duodenal ulcers in the rat. Relative to other gastrin/cholecystokinin-B antagonists, RP 73870 demonstrates greater affinity to gastrin binding sites, and possesses a unique spectrum of in vivo biological activities appropriate for an anti-ulcer indication.

Animals↗
PubMed1994

A multistate outbreak of Escherichia coli O157:H7-associated bloody diarrhea and hemolytic uremic syndrome from hamburgers. The Washington experience.

OBJECTIVE: To determine the source of and describe a large outbreak of Escherichia coli O157:H7 infections in Washington State. DESIGN: Case-control study; environmental investigation; provider-based surveillance for E coli O157:H7 infections. SETTING: Chain of fast-food restaurants, hospitals, physician offices, local laboratories, and local health departments. PARTICIPANTS: Patients with diarrhea and neighborhood controls. A case was defined as diarrhea with culture-confirmed E coli O157:H7 infection or postdiarrheal hemolytic uremic syndrome (HUS) occurring from December 1, 1992, through February 28, 1993, in a Washington State resident. Controls were age- and neighborhood-matched friends of the first 16 case patients. INTERVENTIONS: Announcement to the public; recall of implicated hamburger lots. MAIN OUTCOME MEASURE: Abatement of outbreak due to E coli O157:H7. RESULTS: Infection was associated with eating at a fast-food chain (chain A) in the 10 days before symptoms began. Twelve (75%) of 16 case patients but no controls had eaten at chain A (matched odds ratio undefined; lower 95% confidence interval, 3.5; P < .001). In total, 501 cases were reported, including 151 hospitalizations (31%), 45 cases of HUS (9%), and three deaths. Forty-eight patients (10%) had secondary infections. Of the remaining 453 patients (90%), 398 (86%) reported eating at a Washington chain A restaurant; 92% of them reported eating a regular hamburger. The pulsed-field gel electrophoresis pattern of the E coli O157:H7 strains isolated from all regular hamburger lots of a single production date shipped to Washington was identical to that of the strains isolated from patients. Ten (63%) of 16 regular hamburgers cooked according to chain A policy had internal temperatures below 60 degrees C. Public health action removed more than 250,000 potentially contaminated hamburgers, preventing an estimated 800 cases. CONCLUSIONS: This E coli O157:H7 outbreak, the largest reported, resulted from errors in meat processing and cooking. Public health surveillance through state-mandated reporting of E coli O157:H7 infection as is carried out in Washington State was critical for prompt outbreak recognition and control. Measures should be developed to reduce meat contamination. Consumers and food service workers should be educated about cooking hamburger meat thoroughly.

Adolescent↗
PubMed1994

Nitrate reductase of the ascomycetous fungus, Leptosphaeria maculans: gene sequence and chromosomal location.

The nitrate reductase (niaD) gene was isolated from the phytopathogenic loculoascomycete Leptosphaeria maculans by screening a genomic DNA library with the Aspergillus nidulans niaD gene. The L. maculans niaD gene is the first protein-encoding gene characterised from this fungus. It encodes a predicted protein of 893 amino acids and contains four putative introns at positions in the gene equivalent to those of four of the six introns in the A. nidulans niaD gene. Mutants defective in niaD and molybdenum cofactor gene(s) of L. maculans have been isolated. Transformation of a L. maculans niaD mutant with a 3.8 kb SacII fragment containing the L. maculans niaD gene restored wild-type growth on nitrate as a sole nitrogen source. The niaD gene is present as a single copy on a chromosome which ranges in size from 2.6 to 2.8 Mb between the different L. maculans isolates examined.

Amino Acid Sequence↗
PubMed1994

Dependence of 5-fluorouracil-mediated radiosensitization on DNA-directed effects.

PURPOSE: Although 5-fluorouracil (FUra) has been demonstrated to be a radiation sensitizer both in the laboratory and the clinic, it is not known whether radiosensitization results primarily from FUra's DNA or RNA-directed effects. METHODS AND MATERIALS: We studied the radiosensitizing effects of FUra +/- thymidine (dThd)) on HT29 human colon cancer cells, which are relatively sensitive to the DNA-directed action of FUra, in comparison to SW620 and HuTu80 human colon cancer cells, which are relatively resistant to FUra's DNA-directed effects. We hypothesized that if FUra were acting chiefly through DNA dependent mechanisms, HT29 cells would (a) show greater radiosensitization than SW620 and HuTu80 cells under the same conditions of exposure; and (b) demonstrate selective reversal of radiation sensitivity (compared to cytotoxicity) in the presence of FUra + dThd, compared to FUra alone. RESULTS: We found that the enhancement ratio produced by a 24 h exposure to 10 microM FUra was significantly greater in HT29 cells compared to SW620 and HuTu80 cells (enhancement ratios of 2.1 +/- 0.1; 1.1 +/- 0.1, and 1.3 +/- 0.1, respectively). Furthermore, in HT29 cells, dThd blocked FUra-mediated radiosensitization to a greater extent than FUra-mediated cytotoxicity. Thus, our hypotheses were confirmed. CONCLUSION: These findings support the concept that the manipulation of FUra's DNA-dependent actions, for example, through modulators of thymidylate synthase (TS) activity, may increase radiosensitization in clinical trials in the treatment of gastrointestinal cancers. However, since resistance to the DNA-directed effects of fluoropyrimidines can result from mechanisms unrelated to TS inhibition, additional strategies will be required to potentiate fluoropyrimidine-mediated radiosensitization.

Cell Cycle↗
PubMed1994

Kinetics of bromodeoxyuridine elimination from human colon cancer cells in vitro and in vivo.

We have previously shown that the thymidine analogue radiation sensitizer bromodeoxyuridine (BrdUrd) is incorporated into human tumors to a greater extent than into the livers of athymic mice bearing these tumors as xenografts. However, incorporation into the intestine and bone marrow exceeds that of the tumor (T. S. Lawrence, M. A. Davis, J. Maybaum, S. K. Mukhopadhyay, P. L. Stetson, D. P. Normolle, P. E. McKeever, and W. D. Ensminger, Cancer Res., 52: 3698-3704, 1992). We hypothesized that the ratio of tumor incorporation to intestinal or bone marrow incorporation might increase during a period of drug elimination following the termination of an infusion. To test this hypothesis, we infused athymic mice bearing HT29 human colon cancer xenografts with BrdUrd and measured incorporation in the tumor and normal tissues up to 7 days after the infusion was discontinued. In addition, we assessed the effect of exposure to BrdUrd on subsequent incorporation in vitro and in vivo through the use of a stable isotope of BrdUrd ("isotopic BrdUrd"), which could be differentiated from normotopic BrdUrd using the gas chromatographic-mass spectrometric assay. We found a significant increase in the ratio of BrdUrd in the tumor compared to bone marrow and intestine during the drug elimination period. We also found that BrdUrd incorporation slowed the kinetics of subsequent BrdUrd incorporation and elimination. These findings suggest that when the radiation dose-limiting organ is rapidly proliferative, such as the intestine or bone marrow, delivering radiation during a drug elimination period may improve the therapeutic index.

Animals↗
PubMed1994

Induction of resistance to fluorodeoxyuridine cytotoxicity and DNA damage in human tumor cells by expression of Escherichia coli deoxyuridinetriphosphatase.

Recent studies from our laboratory suggested that, in some human colorectal tumor cell lines, sensitivity to fluorodeoxyuridine may depend upon the extent of dUTP accumulation that occurs following drug treatment and that elevation of dUTPase activity might be the basis for some instances of resistance to fluoropyrimidines. To test this model, we expressed Escherichia coli dUTPase in an established human tumor cell line (HT29) and measured the effect of this manipulation on response to fluorodeoxyuridine. As predicted, HT29 derivatives containing dUTPase activity 4-5-fold higher than controls were protected from fluorodeoxyuridine-induced loss of clonogenicity and from formation of DNA double strand breaks. These data provide the first direct evidence that alteration in a component of the uracil misincorporation/misrepair pathway can confer resistance to fluoropyrimidines in human tumor cells.

Base Sequence↗
PubMed1994

Sensitization of radiolabeled monoclonal antibody therapy using bromodeoxyuridine.

BACKGROUND: Although treatment with radiolabeled monoclonal antibodies (MoAb) against tumor-associated antigens offers the potential for targeted therapy, the efficacy of this approach is limited by the low dose-rate delivered. This could be overcome by increasing tumor sensitivity through the use of radiation sensitizers. METHODS AND RESULTS: In vitro studies using LS174T human colon cancer cells showed that exposure to 1 microM bromodeoxyuridine (BrdUrd), a thymidine-analog radiation sensitizer, at a plasma concentration easily achievable through systemic administration in both animals and patients, increased the cytotoxicity of continuous low dose-rate irradiation delivered by a cesium-137 irradiator (at 12 cGy/h which resembles the dose-rate delivered by radiolabeled MoAb therapy). It was found that 1 microM BrdUrd produced marked radiosensitization (enhancement ratio of 1.42 +/- 0.03) but did not affect cell cycle distribution. Systemic administration of BrdUrd in athymic nude mice bearing LS174T xenografts was performed using osmotic pumps. Animals tolerated infusions of 200 mg/kg/day BrdUrd for 4 days, which resulted in 8.4 +/- 0.8% of thymidine replacement by BrdUrd in tumors. However, bone marrow incorporation was 15.8 +/- 1.1% under these conditions. To improve the ratio of incorporation in the tumor compared to that in the bone marrow, animals were given an infusion of BrdUrd and the pumps were removed. Incorporation of BrdUrd in the bone marrow and intestine decreased rapidly after the infusion was completed. In contrast, there was relatively little change in the incorporation into the tumor after an initial decrease. Based on these data, experiments were performed comparing the effects of 500 microCi iodine-131-labeled 17-1A MoAb alone to the same dose of iodine-131-labeled 17-1A administered 1 day after discontinuation of the infusion of BrdUrd (200 mg/kg/day for 4 days). BrdUrd tended to increase the delay in tumor growth produced by iodine-131-labeled 17-1A administration. CONCLUSIONS: Radiosensitization by BrdUrd in vitro appears to be caused not by cell cycle effects but by increased radiation sensitivity. The in vivo data suggests that BrdUrd improved the efficacy (tumor growth inhibition) of radiolabeled MoAb.

Animals↗
PubMed1994

Health behaviors and survival among middle-aged and older men and women in the NHANES I Epidemiologic Follow-up Study.

BACKGROUND: Since the 1960s there has been a decline in mortality rates for older U.S. adults, suggesting the importance of examining the role of prevention and health promotion in improving the health and survival of older adults. Epidemiologic studies of age and gender differences in the impact of health behaviors on survival for older U.S. adults are needed to provide information for intervention and health promotion efforts for older Americans. METHODS: We examined whether health behavior risk factors (smoking, drinking, physical activity, and body weight) for mortality vary by age and gender among 6,109 adults 45-74 years old in the National Health and Nutrition Examination Survey 1971-1975 (NHANES I) who were traced during the 1982-1984 NHANES I Follow-up Survey. RESULTS: For middle-aged men (45-54 years old) and for older men (65-74 years old), both smoking and nonrecreational physical activity were predictors of survival time. Additionally, for older men, drinking and low body mass index were associated with shorter survival time. Among women, there was less consistency of associations across age groups. As with men, nonrecreational physical activity and low body mass index were associated with shorter survival among older women. CONCLUSIONS: These findings suggest that health behaviors are associated with survival in older adults as well as in middle-aged adults, although the specific behavioral risk factors may vary by age and gender.

Adult↗
PubMed1994

Long-term physical functioning in persons with knee osteoarthritis from NHANES. I: Effects of comorbid medical conditions.

This study examined the effects of comorbid medical conditions (heart disease, pulmonary disease, hypertension and obesity) on the association of radiographic knee osteoarthritis (OA) with long-term difficulty in physical function. Data are from the National Health and Nutrition Examination Survey, 1971-1975 (NHANES I), a prospective epidemiologic cohort study, and the NHANES Epidemiologic Follow-up Study, 1982-1984 (NHEFS) and included 4059 persons who were 45-74 years old and participated in the detailed examination component of NHANES I. Knee OA was ascertained by anterior-posterior bilateral radiographs of the knee and self-report of knee pain, heart and pulmonary disease by self report of disease or symptoms, and hypertension and obesity by blood pressure and weight measurements. The presence of symptomatic knee OA at NHANES I was associated with reported difficulty at NHEFS 1982-84 in functions which used the lower extremity (ambulation and transfer). The presence of coexistent chronic conditions, particularly heart disease, pulmonary disease and obesity, increased the likelihood of subsequent disability. These findings suggest that knee OA is associated with long-term physical disability, and that the presence of coexistent chronic disease may increase the amount of long-term disability from knee OA.

Activities of Daily Living↗
PubMed1994

The homologous operons for P1 and P7 plasmid partition are autoregulated from dissimilar operator sites.

The plasmid-partition regions of the P1 and P7 plasmid prophages in Escherichia coli are homologues which each encode two partition proteins, ParA and ParB. The equivalent P1 and P7 proteins are closely related. In each case, the proteins are encoded by an operon that is autoregulated by the ParA and ParB proteins in concert. This regulation is species-specific, as the P1 proteins are unable to repress the P7 par operon and vice versa. The homologous ParA proteins are primarily responsible for repression and bind to regions that overlap the operon promoter in both cases. The DNA-binding domain of the P7 autorepressor lies in the amino-terminal end of the P7 ParA protein. This region includes a helix-turn-helix motif that has a clear counterpart in the P1 ParA sequence. However, despite the common regulatory mechanism and the similarity of the proteins involved in repression, the promoter-operator sequences of these two operons are very different in sequence and organization. The operator is located downstream of the promoter in P1 and upstream of it in P7, and the two regions show little, if any, homology. How these differences may have arisen from a common ancestral form is discussed.

Amino Acid Sequence↗
PubMed1994

Excess demand and cost relationships among Kentucky nursing homes.

This article examines the influence of excess demand on nursing home costs. Previous work indicates that excess demand, reflected in a pervasive shortage of nursing home beds, constrains market competition and patient care expenditures. According to this view, nursing homes located in underbedded markets can reduce costs and quality with impunity because there is no pressure to compete for residents. Predictions based on the excess demand argument were tested using 1989 data from a sample of 179 Kentucky nursing homes. Overall, the results provide partial support for the excess demand argument. Factors that may counteract the influence of excess demand are considered. Finally, the role of competition in nursing home markets and difficulties associated with making it operational are discussed.

Bed Occupancy↗
PubMed1994

Influence of cell cycle phase on radiation-induced cytotoxicity and DNA damage in human colon cancer (HT29) and Chinese hamster ovary cells.

We have previously shown that fluorodeoxyuridine (FdUrd) radiosensitizes HT29 human colon carcinoma cells. Since treatment with FdUrd arrests cells at the G1/S-phase interface, a condition associated with increased radiation sensitivity in some cells, it seemed possible that redistribution of cells in the phases of the cell cycle might account for FdUrd-mediated radiosensitization. To begin to test this, HT29 cells were separated by centrifugal elutriation according to cell cycle phase and assessed for radiosensitivity, using a clonogenic assay, and radiation-induced DNA damage, using pulsed-field gel electrophoresis. We found that all of the elutriated fractions (which contained cells enriched in G1, G1/early S, mid to late S or G2/M phase) had the same radiation sensitivity and expressed a similar extent of radiation-induced DNA damage. To determine if the techniques used in this study could detect differences between the radiation sensitivity of cells in different phases of the cell cycle, analogous experiments were carried out using Chinese hamster ovary (CHO) cells. In contrast with the results of experiments with HT29 cells, but in agreement with previous studies, CHO cells separated under the same conditions as were used for HT29 cells showed a marked dependence on cell age of both clonogenic survival and radiation-induced DNA damage. Thus, within the limitations of the purity of separation obtained using elutriation, the radiation sensitivity of HT29 cells does not vary substantially as a function of cell cycle phase. Therefore, it seems unlikely that cell cycle redistribution alone explains the radiation sensitivity produced by exposure to FdUrd.

Animals↗
PubMed1993

The use of biphasic linear ramped pulsed field gel electrophoresis to quantify DNA damage based on fragment size distribution.

PURPOSE: The development of biphasic linear pulse ramping gel electrophoresis has permitted resolution of DNA fragments from 200 Kbp to 6 Mbp in a single gel. We used this technique to measure radiation-induced DNA damage based on fragment size. METHODS AND MATERIALS: Human colon cancer cells (HT29 and LS174T) and Chinese hamster ovary cells were embedded in agarose, deproteinized, irradiated with 5-80 Gy, and assessed for DNA double strand breakage using pulsed field gel electrophoresis. The frequency of DNA double strand breakage determined using a previously published method was compared to the breakage frequency calculated using the fragment size distribution. RESULTS: Both methods produced similar estimates for breakage frequency of approximately 5 x 10(-9) breaks Gy-1 bp-1. CONCLUSIONS: These findings suggest that biphasic linear pulse ramping gel electrophoresis can yield a quantitative estimate of DNA fragment distribution resulting from irradiation. The ability to quantify the distribution of DNA fragment sizes produced by irradiation should yield important information concerning the mechanisms of both DNA double strand break induction and repair.

Animals↗
PubMed1993

Phosphonate-modified Gd-DTPA complexes. III: The detection of myocardial infarction by MRI.

The potential of a phosphonate-modified-Gd-DTPA for MR image enhancement of myocardial infarction has been demonstrated in imaging experiments on rats. The agent, 1-hydroxy-3-aminopropane-1,1-diphosphonate-modified-Gd-DTPA (Gd-DTPA-HPDP) accumulates in two models of myocardial infarction, (i.e., drug-induced diffusely infarcted whole hearts and in focal acute myocardial infarction from a left coronary artery ligation). The time course of the accumulation of the agent in the focal model of infarction and subsequent washout has also been followed in vitro. Results of this kinetics demonstrate that the agent first perfuses all normal fluid spaces and then slowly diffuses into the occluded zone where it is retained for a prolonged period, in sufficient quantities to be useful as an MRI contrast agent. Wash-out of the agent from normal myocardium is fast and complete with MR signal returning to background in minutes. The specificity of Gd-DTPA-HPDP for soft-tissue calcification and its retention within the infarcts permitted imaging at 1 to 2 h postinjection, (after unbound material has cleared the normal tissues). Infarcted tissue appeared as regions of increased signal intensity in T1-weighted images (> 200% enhancement), and correlated with histopathology. Unmodified Gd-DTPA was not retained under identical conditions. Gd-DTPA-HPDP permits a more accurate infarct delineation than is possible with the unmodified agent.

Animals↗
PubMed1993

Differential effect of cyanohydroxybutene on glutathione synthesis in liver and pancreas of male rats.

1-Cyano-2-hydroxy-3-butene (CHB), an aliphatic nitrile found in cruciferous vegetables, causes a two- and sevenfold elevation in reduced glutathione (GSH) in rat liver and pancreas, respectively, after oral administration of 200 mg/kg. While this dose is also associated with pancreatotoxicity, a single 100 mg/kg dose or multiple lesser doses show the same effect, although somewhat reduced in magnitude, with no concomitant toxicity. In an attempt to identify the mechanism of this increase, we investigated the effect of CHB on GSH synthesis by examining the effect of buthionine sulfoximine (BSO), an inhibitor of GSH synthesis, on CHB-induced GSH elevation. Male Fischer 344 rats received 3 mmol BSO/kg ip 24 and 34 hr following CHB or corn oil. The CHB-mediated elevation in hepatic and pancreatic GSH was eradicated by BSO, suggesting that increased synthesis was responsible. The rate-limiting step in synthesis is gamma-glutamyl cysteine synthetase (GCS); the limiting substrate is cysteine. Therefore, CHB effects on GCS activity and hepatic and pancreatic cysteine equivalents were investigated. When rats were treated by gavage with CHB (100 mg/kg), hepatic GCS mRNA concentrations were increased 24 hr after treatment and hepatic cysteine equivalents were significantly elevated 4 hr following CHB. No significant elevation in hepatic GCS activity was observed, however, even 24 hr following CHB. Pancreatic cysteine equivalents were elevated at both 4 and 8 hr after CHB treatment. However, there was no detectable GCS mRNA or activity in pancreas, in either control or treated animals. Furthermore, CHB had no direct effect on the activity of GCS purified from kidney, regardless of whether GSH was present or absent. These results suggest that the mechanism of CHB-mediated induction of GSH may involve early increases in GSH precursors as well as a later increase in GCS mRNA. The mechanism of GSH elevation identified in these studies may hold therapeutic or prophylactic implications.

Alkenes↗
PubMed1993

Fungal catabolic gene regulation: molecular genetic analysis of the amdS gene of Aspergillus nidulans.

Aspergillus nidulans is an excellent experimental organism for the study of gene regulation. Genetic and molecular analyses of trans-acting and cis-acting mutations have revealed a complex pattern of regulation involving multiple independent controls. Expression of the amdS gene is regulated by the facB and amdA genes which encode positively acting regulatory proteins mediating a major and a minor form of acetate induction respectively. The product of the amdR gene mediates omega amino acid induction of amdS. The binding sites for each of these proteins have been localised through amdS cis-acting mutations which specifically affect the interaction with the regulatory protein. The global controls of nitrogen metabolite repression and carbon catabolite repression regulate the expression of many catabolic genes, including amdS. Nitrogen control is exerted through the positively acting areA gene product and carbon control is dependent on the creA gene product. Each of the characterized regulatory genes encodes a DNA-binding protein which recognises particular sequences in the amdS promoter to activate or repress gene expression. In addition, there is evidence for other genetically uncharacterized proteins, including a CCAAT-binding complex, which interact with the 5' region of the amdS gene.

Amidohydrolases↗