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R W Hart

Publications and source records attributed to R W Hart.

180 records · Page 10Linked to original sources

DNA repair in late-passage human cells.

Monolayer cultures of WI-38 cells from passages 18 to 60 were exposed to ultraviolet radiation, or to N-acetoxy-AAF, or to gamma-rays. The cultures were incubated in 3H-dThd, and unscheduled DNA synthesis--a measure of DNA repair--was measured radioautographically. Late passage cultures showed less repair, and many cells showed none. Double-label radioautographs indicated that there is an excellent correlation between cells that do scheduled synthesis and cells that do unscheduled synthesis. We interpret our results as indicating that failure of repair is not a causal event in the failure of late passage cells to divide, but that as cells age the ability to do the many coordinated steps in repair declines.

Acetoxyacetylaminofluorene↗

Overview of cancer and aging: a mechanistic perspective.

Cancer and aging are approached from the standpoint of damaging agents affecting target cell, local cellular environment, and the organism as a whole. The effects of exogenous agents are represented in a model emphasizing absorption, organismic disposition, and cellular disposition. Endogenous agents are represented similarly. The extent of endogenous damage is illustrated. Factors in the expression of damage as a toxic endpoint are emphasized, with the example of caloric restriction used as an example of environmental modulation of response.

Aging↗

Cell-cycle defect of DNA repair in progeria skin fibroblasts.

We examined the temporal regulation of DNA repair during synchronous cell proliferation in normal and progeroid human fibroblasts. Ultraviolet light-induced (254 nm, 20 J/m2) unscheduled DNA synthesis was measured at 4-h intervals after serum stimulation, for up to 32 h. Normal cells regulated DNA repair in a defined temporal sequence, showing a peak in the induction of DNA repair just before DNA synthesis. Progeroid skin fibroblasts failed to show an increase in nucleotide excision repair before scheduled DNA synthesis, but the background level of DNA repair was not significantly different from that in controls. Regulation of repair in progeroid human fibroblasts appeared similar, but not identical to that previously reported by Gupta and Sirover (1984b) for xeroderma pigmentosum complementation group C. Our results suggest that patients with Hutchinson-Gilford progeria may have a defect in DNA repair; the results offer nominal evidence that the average level of UV-induced DNA is decreased, and that individuals with this disease lack both the normal enhancement of DNA repair before scheduled DNA synthesis and the temporal control of DNA repair.

Cell Cycle↗

Evolution and dietary restriction.

Placing lifespan in the context of the life history of an organism, Alex Comfort's work has stimulated the analysis of dietary restriction (DR) and its effects on lifespan in an evolutionary context. DR results in the curtailment of energy-intensive nonfood-gathering activities, increased efficiency of food utilization, an increase in food acquisition activity, an increase in the reproductive lifespan, and an increase in the protection of genomic integrity. These result in further refinement of the Adaptive-Longevity Related Process Theory of the effects of dietary modulation to include increased protection of the genomic integrity of cells that result from delayed reproduction, and increased ability to compete for available food. These effects are discussed in the context of the "spacecraft" metaphor for the evolution of senescence. Also, the apparent paradox that increased body weight seems to be directly correlated to survival across species and inversely correlated to survival within a species is discussed in light of the importance of the cephalization index (a modified ratio of brain weight to body weight) for longevity.

Aging↗

Survival, body weight, and spontaneous neoplasms in ad Libitum-fed and food-restricted Fischer-344 rats.

Ad libitum-fed (AL) and food-restricted (FR) Fischer-344 male and female rats were monitored for survival, body weight, and spontaneous neoplasms. Mean and maximal lifespans for each group were inversely related to mean body weights. AL males were the shortest lived (mean lifespan 101 wk) followed by AL females (118 wk), FR males (125 wk), and FR females (132 wk). Gross and microscopic examinations were performed on 851 rats from cross-sectional and longevity components of the study. In FR groups, the incidence of mammary gland fibroadenomas, testicular interstitial cell tumors, and pituitary neoplasms was decreased while the latency of these neoplasms was increased. In longevity components, most FR groups had a higher incidence of leukemia than AL cohorts, but all FR groups had a higher mean age at death for the rats with leukemia. Higher leukemia rate in the FR groups was thought to be a result of their extended mean lifespan.

Animals↗

Age-related neoplasia in a lifetime study of ad libitum-fed and food-restricted B6C3F1 mice.

Longevity, body weight, and age-specific neoplasia were determined in 1,064 B6C3F1 mice as part of a coordinated study of food restriction (FR). Restricted animals were offered 60% of the diet consumed by the ad libitum (AL) group. Longevity data were derived from a set of 56 animals of each sex from each diet group, which were examined whenever dead or moribund. For cross-sectional data, a parallel set of 210 animals were sacrificed in groups of 12-15 at 6-mo intervals. Lifetime body weight was reduced in the FR mice approximately proportional to restriction (i.e., 40%). Food restriction increased the age at 50% survival (median) by 36% in both sexes and increased the maximal lifespan (mean age of oldest 10%) by 21.5% in males and by 32.5% in females. In 56 males of the longevity groups, there were 89 neoplasms in the AL subgroup versus 53 in FR; 56 AL females had 100, versus 58 in 55 FR females. Increase in lifespan of the restricted animals was achieved primarily by decrease in incidence and delay of onset of fatal tumors, of which lymphoma was the most prominent. This report catalogs all of the neoplasms (1,103) observed in longevity and cross-sectional groups, by diet, sex, and age. These data add to the existing knowledge base needed for future studies of dietary restriction and aging as well to evaluate nutrition of animals used in bioassays.

Aging↗

Longevity, body weight, and neoplasia in ad libitum-fed and diet-restricted C57BL6 mice fed NIH-31 open formula diet.

Groups of C57BL6 mice of each sex were assigned to one of 2 dietary regimens, ad libitum (AL) or dietary restriction (DR), to study effects of food restriction on body weight, survival, and neoplasia. The AL and DR groups were subdivided into a scheduled sacrifice group for examination at 6-mo intervals, and a lifetime group to provide longevity data. Necropsies and microscopic examinations were conducted on 911 animals. In the lifetime group food consumption averaged 33.6 and 34.4 g per week by AL males and AL females, respectively; the DR counterparts were given 40% less. The diet contained 4.35 kcal/g. The average lifetime body weights were 34.8, 26.8, 22.6, and 21.6 g for AL males, AL females, DR males, and DR females, respectively, and their age at 50% survival was 27.5, 26.9, 31.7, and 33.5 mo. Maximal lifespan was increased 18% in DR males and females. Lifetime incidence of tumor-bearing mice was 89% and 86% for AL males and females, versus 64% for each sex of DR mice. Dramatic reduction occurred in female DR mice in lymphoma (9% vs 29%), pituitary neoplasms (1% vs 37%), and thyroid neoplasms (0.4% vs 8%). In males, hepatocellular tumors were reduced to 1% from 10% by DR. In contrast, the incidence of histiocytic sarcoma was increased in DR females and unaffected in DR males. Tumor onset was delayed in DR animals; 87% of all neoplasms in males and 95% in females had occurred in the AL mice by 24 mo, whereas the DR animals had only 52% and 39% of their lifetime incidence, respectively, by that age. This study provided comparative AL and DR data from C57BL6 mice examined randomly at 6-mo intervals (cross-sectional group) in parallel with data from animals in similar cohort that was unsampled and allowed to succumb naturally (longevity group). Dietary restriction reduced the lifetime percentage of tumor-bearing animals and the number of tumors per animal, and delayed the age at onset of most neoplasms.

Animals↗

FDA points-to-consider documents: the need for dietary control for the reduction of experimental variability within animal assays and the use of dietary restriction to achieve dietary control.

Standard protocols for conducting chronic toxicity and carcinogenicity studies have been refined over the years to carefully control for many variables. Nevertheless, over the last 2 decades, there has been a steady increase in variability, a decrease in survival, an increase in tumor incidence rates, and an increase in the average body weight of control animals among the various rodent species and strains used for toxicity testing. These observations have prompted an evaluation of chronic study designs to determine what factor(s) may be responsible for such confounding changes. Ad libitum feeding and the selection of successful breeders with rapid offspring growth is believed to be at least partially responsible for the heavier, obese rodents with which many laboratories are coping today. As a result of these changes, some studies used for the evaluation of safety have been deemed inconclusive or inadequate for regulatory purposes and either additional supportive studies have been requested and/or studies per se have been repeated. Research on the molecular mechanisms of caloric restriction and agent-induced toxicity at the Food and Drug Administration (FDA) National Center for Toxicological Research stimulated the first international conference on the biological effects of dietary restriction in 1989; this was followed in 1993 by an FDA workshop exploring the utility of dietary restriction in controlling reduced survival in chronic tests and an international conference in 1994 exploring the implications for the regulatory community of using dietary restriction in toxicity and carcinogenicity studies used in support of a sponsor's submission or in risk assessments. The outcome of that conference was the FDA's commitment to develop Points-to-Consider documents that address the issue of dietary control in chronic toxicity and carcinogenicity studies.

Animal Feed↗

Differential effects of dehydroepiandrosterone and clofibrate on the binding of 7,12-dimethyl benz(a)anthracene to hepatic DNA in vivo--a preliminary study.

The effects of feeding two compounds, dehydroepiandrosterone (DHEA), an adrenal steroid, and clofibrate (CLOF) to rats (which are both hypolipidemic, hepatomegaly inducing and hepatic peroxisome proliferating agents) on the binding of 7,12-dimethylbenz(a)anthracene (DMBA) to hepatic DNA in vivo is compared. Male Sprague Dawley rats (two-three months old) were fed either DHEA or CLOF for 14 days at a dietary level of 0.8%. Control rats were pair fed. An increase in liver weight followed by increases per whole liver in total protein, without much change in DNA content was observed. Subsequently, all the animals were given a single intraperitoneal dose of [3H]DMBA (133 mumol/kg body weight, 102 microCi/rat) in 250 microliters dimethyl sulfoxide. Forty-eight hours later, binding of DMBA to hepatic DNA was determined. The results indicate that DMBA binding to DNA was reduced by 67% in DHEA-fed rats whereas in clofibrate-fed rats it was not significantly different from that of the controls.

Animals↗

Modulation of biological processes by mineral fiber adsorption of macromolecules in vitro.

Three classes of macromolecules (i.e., DNA, RNA, and protein) were shown to be adsorbed to asbestiform minerals. The cytotoxicity exerted by the fibers on a normal human fibroblast cell line, which may be an indicator of the carcinogenic potential of mineral fibers, correlated positively with the degree of macromolecular adsorption of the fiber, namely: chrysotile greater than amosite greater than glass fiber. Asbestiform fibers also induce an alteration in in vitro DNA hydrolysis by bovine pancreatic deoxyribonuclease. This phenomenon suggests that adsorption by asbestiform minerals may modulate biological processes by inducing a conformational change in biological macromolecules as a result of coulombic interaction between the surface charge of the fiber and the hydrophilic groups on the macromolecule.

Adsorption↗

Effect of food restriction on the metabolism of dimethylnitrosamine (NDMA) in rats.

The influence of food restriction on the macromolecular interactions of the hepatocarcinogen dimethylnitrosamine (NDMA) in the livers of male Sprague Dawley rats was investigated. Two-three month old rats were food restricted (FR) (40% with respect to ad libitum fed rats) for three weeks. The liver weight, total protein, microsomal and cytoplasmic protein, cytochrome P-450 and DNA content per whole liver were all reduced significantly in food restricted rats. Five hours after a single dose of (14C) NDMA (28 mg/k.b.w., 21 microCi/rat) the levels of 7-methylguanine increased in restricted rats by 32%. Cytochrome P-450 mediated generation of HCHO from NDMA (1.8 fold) in restricted rats was greater. Binding of NDMA derived radioactivity to total hepatic proteins decreased by 46% in restricted rats. These results suggest that food restriction enhances the metabolic activation of dimethylnitrosamine in Sprague Dawley rats.

Animals↗

DNA damage and repair in vivo.

DNA damage has been implicated in carcinogenesis, mutagenesis, and aging. DNA excision repair has been implicated as an ameliorating factor for such damage. It has been proposed that there is an error-prone post-replication repair system which is both mutagenic and carcinogenic. Assay of DNA damage and repair may thus illuminate the mechanism of carcinogenesis and serve as an indicator of the carcinogenic potential of environmental agents. DNA damage induction and repair can differ in vitro and in vivo. In order to rationally evaluate environmental health effects, it is therefore important to examine DNA damage induction and repair in vivo. An in vivo method is defined here as one in which the DNA is damaged and repaired in cells in situ in the organism. In vivo methods for studying DNA adducts and excision repair, strand-breaks and strand-break repair, post-replication repair, and photoreactivation repair, and the current state of knowledge of DNA damage induction and repair in vivo, are reviewed and evaluated.

Animals↗