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Biomedical subjects

J Caldwell

Publications and source records attributed to J Caldwell.

At least 109 records · Page 6Linked to original sources
PubMed1992

Problems and opportunities in toxicity testing arising from species differences in xenobiotic metabolism.

The processes of metabolism and disposition have a major bearing upon the biological properties of xenobiotics, determining both the chemical natures and target concentrations of the compound-derived materials in the body. The occurrence of major quantitative and qualitative differences between animal species in the metabolism of xenobiotics is well documented. Interspecies differences in metabolism represent a major complication in toxicity testing, being responsible for important differences both in the nature and magnitude of toxic responses. As such, they may be both help and hindrance and this presentation will illustrate both these possibilities with appropriate examples. In particular, these differences represent probably the single greatest complicating factor in the use of animal toxicity data as an indication of potential human hazard. Although it is considered desirable to identify a species which metabolizes the test compound like man, this ideal is generally not attainable. While metabolic and toxicokinetic data should be used in the selection of animal species, in reality the choices are constrained by other major factors such as availability, background pathological knowledge and regulatory acceptability. On the other hand, species differences in metabolism may present exploitable opportunities for insights into mechanisms of toxicity and with appropriate supporting data may thereby increase confidence in the animal-to-human extrapolation. The application of the new technologies of transgenesis to the creation of animals expressing the genes for human drug metabolizing enzymes offers the promise of increasing the opportunities and minimizing the problems presented by species differences in xenobiotic metabolism.

Animals↗
PubMed1991

Co-expression of full-length and truncated Ig mu-chains in human B lymphocytes results from alternative splicing of a single primary RNA transcript.

A survey of Ig synthesis among mu-heavy chain-producing human B cell lines indicated that roughly one-third co-express full length (mu+) and a particular type of truncated mu-chain (designated mu'). The relative molecular size of the intracellular form of this truncated mu-chain and its normal pattern of N-glycosylation suggested that mu'-chains were missing a single Ig domain at the protein level. Cell-free translation of polyA+ RNA from mu'-producing human B cell lines generated appropriate mu'-translation products, and Northern blot analysis demonstrated the presence of correspondingly truncated mu'-transcripts in these lines. These results pointed to a pretranslational basis for mu+/mu' co-expression. Sequencing of mu+- and mu'-cDNA clones from two human B cell lines showed that mu+- and mu'-transcripts derive from the same primary transcript, with mu'-mRNA formed by a direct leader-to-C mu 1 exon splice such that the heavy chain variable region exon is excluded via a cassette-type alternative splicing mechanism. Southern blot analysis of the rearranged Ig heavy chain genes in one B cell line confirmed that the co-expressed mu+- and mu'-mRNA derive from the same rearranged Ig heavy chain gene. mu'-cDNA clones were readily isolated from normal human bone marrow lymphocytes, whereas peripheral B cells do not appear to express mu'-transcripts. The frequent occurrence of mu'-mRNA in B cell lines, and its high relative expression in untransformed bone marrow lymphocytes attest to a mode of post-transcriptional control of Ig gene expression that may have implications for human B cell development.

B-Lymphocytes↗
PubMed1991

Culture perfusion schedules influence the metabolic activity and granulocyte-macrophage colony-stimulating factor production rates of human bone marrow stromal cells.

The metabolic function and GM-CSF production rates of adherent human bone marrow stromal cells were investigated as functions of medium and serum feeding rates. A range of medium exchange schedules was studied, ranging from a typical Dexter culture protocol of one weekly medium exchange to a full media exchange daily, which more closely approximates what bone marrow cells experience in situ. Glucose consumption was found to be significantly higher at full daily exchange rate than at any other exchange schedule examined. However, the lactate yield on glucose was a constant, at 1.8 mol/mol, under all conditions considered. Differential serum vs. medium exchange experiment showed that both serum supply and medium nutrients were responsible for the altered behavior at high exchange rates. Glutamine consumption was found to be insignificant under all culture conditions examined. A change in exchange schedule from 50% daily medium exchange to full daily medium exchange after 14 days of culture was found to result in a transient production of GM-CSF and a change in metabolic behavior to resemble that of cultures which had full daily exchange from day one. These results suggest that both stromal cell metabolism and GM-CSF production are sensitive to medium exchange schedules. Taken together, the data presented indicate that attempts to model the function of human bone marrow in vitro may be well served by beginning with medium exchange schedules that more closely mimic the in vivo physiologic state of bone marrow.

Adult↗
PubMed1991

UV spectroscopy of Titan's atmosphere, planetary organic chemistry and prebiological synthesis. II. Interpretation of new IUE observations in the 220-335 nm range.

We report on new observations of Titan with the International Ultraviolet Explorer in the mid-UV range (lambda approximately 220-335 nm). We use these data to determine upper limits for the abundances of simple organic compounds in the gas phase and to further constrain the properties of the high altitude haze on Titan. As a baseline, we adopted the parametrized microphysical model of McKay et al. (1989) which is successful at reproducing Titan's thermal structure while satisfying several other observational constraints in the visible and IR regions. However, we find that such a model--in which all particles at a given altitude are assumed to have the same size--cannot match simultaneously the IUE observations and the visible/IR data, even when allowance is made for a wide range of values in the adjustable parameters. On the other hand, a good overall agreement is obtained when considering a biomodal size distribution, with small haze particles or "polymers" (r < 0.02 micrometer) acting as strong Rayleigh absorbers below 300 nm and larger haze particles (r approximately 0.1-0.5 micrometer) being responsible for the characteristics of the albedo spectrum in the near-UV, visible, and IR regions. This approach is consistent with the results of several previous investigations of the properties of Titan's haze, although our preferred vertical structure for the haze + polymer material somewhat differs from earlier solutions. On the basis of simple dynamical considerations, we adopted a uniformly mixed layer between 150 and 600 km. The IUE data allow us to place fairly stringent constraints on the abundance of the Rayleigh absorbers, if we assume that their optical properties are similar to those of laboratory-synthesized "tholins": The column-mass density of this material--the essential observable that can be determined from our study--is of the order of 5 micrograms cm-2. This would correspond to number-densities between 10(3) and 10(7) cm-3 in the 150-600 km altitude range, if the average particle radius is between 0.001 and 0.02 micrometer. Such high number densities are a priori at odds with the estimated coagulation lifetime for particles of that size. Thus, our proposed bimodal size distribution is plausible only if inhibiting processes act to slow down considerably the coagulation of polymers in Titan's stratosphere.

Acetylene↗
PubMed1991

Congestive heart failure symptoms in patients with preserved left ventricular systolic function: analysis of the CASS registry.

The clinical characteristics and long-term survival of 284 patients from the Coronary Artery Surgery Study (CASS) registry data base who had moderate to severe congestive heart failure symptoms and a left ventricular ejection fraction greater than or equal to 0.45 were studied. A control group consisting of registry patients with an ejection fraction greater than or equal to 0.45 who did not have heart failure was used for comparison. Patients who had heart failure were older and more likely to be female and to have a higher incidence of hypertension, diabetes and chronic lung disease than registry patients who did not have heart failure. As a group, patients with heart failure had more severe angina and were more likely to have had a prior myocardial infarction than were registry patients without heart failure. At 6 year follow-up, 82% of patients in the heart failure group survived compared with 91% of patients in the control group (p less than 0.0001). Multivariate analysis using the Cox proportional hazards model identified the following independent predictors of mortality: regional ventricular systolic dysfunction, number of diseased coronary arteries, advanced age, hypertension, lung disease, diabetes, increased left ventricular end-diastolic pressure and heart failure symptoms. Among patients with heart failure, the 6-year survival rate of those who had three-vessel coronary artery disease was 68% compared with 92% for the group without coronary artery disease. However, the 6-year survival rate for patients with heart failure who underwent surgical revascularization of diseased coronary arteries was not significantly improved compared with that of patients treated medically.(ABSTRACT TRUNCATED AT 250 WORDS)

Coronary Artery Bypass↗
PubMed1990

The dispositional enantioselectivity of indobufen in rat and mouse.

The plasma pharmacokinetics and urinary elimination of the enantiomers of indobufen, a novel platelet aggregation inhibitor, have been studied in rats and mice given either the racemic compound or the individual enantiomers (rat 8 mg/kg racemate, 4 mg/kg enantiomers; mouse 25 mg/kg racemate, 12.5 mg/kg enantiomers). Enantiospecific analysis of indobufen in plasma and urine was achieved by HPLC of its L-leucinamide diastereoisomers. In rat, the two enantiomers have very different plasma elimination half lives (S, 3.9 hr; R, 12.2 hr), irrespective of the optical form administered. The plasma concentration-time curves of S-indobufen were identical after racemic or S-indobufen, but the plasma levels of R-indobufen were lower after the R-enantiomer than after the racemate. Urinary recovery of free and conjugated indobufen was less than 3% of the dose, independent of the optical form administered. In the mouse, R-indobufen was cleared from plasma more rapidly than its S-antipode (elimination T1/2 R, 2.5 hr; S, 3.8 hr) but differences were smaller than those seen in the rat. The plasma concentration-time curves of the S-enantiomer were the same after racemic or S-indobufen, but levels of its R-antipode were much lower when it was given alone than after administration of the racemate. The urinary recovery of free and conjugated indobufen also exhibited enantioselectivity, with preferential elimination of the S-enantiomer.

Animals↗
PubMed1990

Percutaneous absorption of benzyl acetate through rat skin in vitro. 1. Validation of an in vitro model against in vivo data.

The percutaneous absorption in vitro of the fragrance agent benzyl acetate has been evaluated in flow-through diffusion cells using shaved full-thickness skin from male Fischer 344 rats. After the application of neat [methylene-14C]benzyl acetate to the epidermal surface of the skin and occlusion with parafilm 1.3 cm above the skin surface, the absorption of the chemical across the skin and into the receptor fluid was rapid and extensive, commencing within 1 hr of application, and reaching 49.8 +/- 3.2% (mean +/- SD, n = 4) of the applied dose after 48 hr. The coefficient of variation for absorption at 48 hr between four identical experiments was 6.4%. The extent of absorption at 48 hr of benzyl acetate applied in 50% (v/v) ethanol was not significantly different from that after application neat, although absorption at earlier times was enhanced, with a maximum increase of 8.5% of the applied dose at 12 hr. Over the dose range studied there was a linear relationship (r = 0.996) between the amount of benzyl acetate applied to the skin (1.66-33.13 mg benzyl acetate/cm2) and the amount absorbed into the receptor fluid at 24 hr (0.66 +/- 0.04-10.27 +/- 0.51 mg/cm2). The extent of absorption of benzyl acetate through rat skin in vitro was compared with the extent of absorption in vivo at 24 hr and a correlation coefficient of 0.993 was obtained. These data support the use of this in vitro system as a model to predict in vivo absorption and indicate the suitability of the system to study factors influencing the disposition of topically applied benzyl acetate.

Animals↗
PubMed1990

Structure-specificity of the genotoxicity of some naturally occurring alkenylbenzenes determined by the unscheduled DNA synthesis assay in rat hepatocytes.

A number of alkenylbenzenes related to safrole and estragole are known to be hepatocarcinogenic in rats and/or mice, apparently by a genotoxic mechanism. However, they are not bacterial mutagens in the Ames test. We have studied the ability of a series of carcinogenic and non-carcinogenic congeners to induce unscheduled DNA synthesis (UDS) in freshly isolated rat hepatocytes in primary culture. The cytotoxicity of these compounds was assessed by lactate dehydrogenase leakage. There was an excellent correlation between UDS induction and known rodent hepatocarcinogenicity, with safrole, estragole and methyleugenol all inducing UDS. Anethole, isosafrole, eugenol and allylbenzene, for which evidence of carcinogenicity is equivocal or negative, did not induce UDS. All compounds were markedly cytotoxic at concentrations between 10(-3) and 10(-2) M, irrespective of their structural features. The data are discussed with reference to the known structure dependence of the disposition of the alkenylbenzenes, notably their metabolic activation, with which there are excellent correlations. The demonstration of the genotoxicity of rodent hepatocarcinogenic alkenylbenzenes in cells cultured from the in vivo target organ will allow the direct investigation of factors influencing these processes and facilitate the safety evaluation of these important natural flavours.

Animals↗
PubMed1990

The stereoselectivity of 1,2-phenylethanediol and mandelic acid metabolism and disposition in the rat.

1. The steps involved in determining the chirality of the mandelic acid excreted by rats after administration of ethylbenzene and styrene were investigated by studying the fate of racemic, (R)- and (s)1,2-phenylethanediol, a precursor of mandelic acid. These investigations indicate the occurrence of two alternative routes of metabolism for 1,2-phenylethanediol, one involving retention of configuration and the other resulting in the loss of the chiral centre. 2. The stereoselectivity of the disposition of mandelic acid was investigated; rats were dosed with mandelic acid either as the racemate or as the individual enantiomers, G.1.c.-mass spectrometry and h.p.l.c. were used to determine the enantiomers of mandelic acid. 3. There were at least two routes by which mandelic acid could be metabolized and/or excreted; there is a stereoselective pathway in rat for (s)-mandelic acid, which gives rise to phenylglyoxylic acid. 4. The chiral inversion of (s)-mandelic acid to (R)-mandelic acid is reported; although this has been observed in bacteria it has not previously been observed in mammals. 5. The extent to which mandelic acid is metabolized to phenylglyoxylic acid is dependent on the enantiomeric composition of the mandelic acid administered. There is no evidence to indicate significant ketone-alcohol conversion, that is phenylglyoxylic acid is not significantly reduced to mandelic acid in vivo.

Animals↗
PubMed1989

Characterization of immunoglobulin mRNA expression in Burkitt lymphoma cell lines.

Immunoglobulin heavy- and light-chain mRNA of 11 Burkitt lymphoma (BL) cell lines (9 African and 2 American) were analyzed for various structural characteristics. In agreement with previous results at the protein level, all the BL cell lines express heavy-chain mRNA transcripts of the mu class. Surprisingly, a high mu s/mu m mRNA ratio was found in 2 IgM-producing BL cell lines (Raji and CCL85), that do not secrete immunoglobulin. Variable region gene use was also assessed in the cell lines: while 4 out of 7 endemic BL cell lines use VH genes that belong to the VH3 gene family, no clear bias in the expression of particular VH or VL gene families among this sampling of BL lines was found. Northern blot analysis of immunoglobulin transcripts in endemic BL cell lines did show that 2 such lines (AG876 and HTB62) expressed truncated heavy-chain transcripts; RNA sequence analysis of the VH region demonstrated different abnormal 5'-localized RNA splicing events for the 2 shortened mu transcripts. The light-chain mRNA in these 2 cell lines also showed structural abnormalities and, in the case of HTB62, 3 different kappa light-chain transcripts are produced (of elongated, native and truncated sizes). In vitro translation of mRNA from HTB62 showed mu and kappa chain proteins corresponding with the relative size for each message.

Amino Acid Sequence↗
PubMed1989

Species differences in the chirality of the carbonyl reduction of [14C] fenofibrate in laboratory animals and humans.

The prochiral carbonyl group of fenofibrate (isopropyl 2-[4-(4-chlorobenzoyl)phenoxy]-2-methyl propionate) is reduced during its metabolism giving rise to a chiral secondary alcohol, "reduced fenofibric acid." Chiral and diastereomeric HPLC methods have been developed for the determination of its enantiomeric composition and these have been applied to the measurement of the "reduced fenofibric acid" enantiomers in urine of rats, guinea pigs, dogs, and human volunteers given [14C]fenofibrate. In the three animal species, the reduction is markedly enantioselective for the (-)-isomer, the enantiomeric ratios (-/+) being 95:5. This was not due to differences in the excretion of the enantiomers, since when racemic "reduced fenofibric acid" was given to rats it was recovered in the urine with the same enantiomeric composition as the dose form. In humans the ratio was 52:48 showing the lack of stereoselectivity of reduction in this species.

Animals↗
PubMed1989

Sensitization to illness and the risk of death: an explanation for Sri Lanka's approach to good health for all.

Recent research has shown clearly that levels of mortality are not determined solely by income levels and the provision of modern health services. There appears to be an important sociocultural component so that the same density of health services produces lower mortality in one country than in another. Sri Lanka has achieved an expectation of life at birth of 68 years with an annual per capita income of U.S. $330 and no greater provision of health services than most countries at that income level. This paper records the investigation of how such success has been attained. A collaborative program of the University of Colombo and the Australian National University, beginning in 1985, employed both demographic and anthropological methods to study almost 11,000 persons in seven localities of southwestern Sri Lanka. It was found that the major geographical mortality differentials are no longer urban-rural but between the richer and poor areas of Colombo. The low mortality was found to be based on a great sensitivity to illness and the risk of dying. This is ancient in origin but has been heightened by a nineteenth century religious reform movement and twentieth century mass education. It has been further increased by universal death registration, the carrying out of post-mortems wherever necessary with the informing of the police if the death was unnecessary, and by the erection of a system for examining the circumstances of sudden death. This sensitivity was not sufficient to produce low death rates in conjunction with traditional medicine but only with modern medicine. Traditional medicine is still widely practiced, and exorcism is a major cause of expenditure in many families. Nevertheless, the key findings were two. First, where modern medicine can save a life, the sick person nowadays is nearly always exposed to it while there is still sufficient time for a cure. Modern medicine is now usually obtained at a shorter distance and more cheaply than traditional medicine. Second, and of central importance, is the continuing evaluation of sickness from its first appearance and the quick resort to new treatments if the present treatment does not appear to be efficacious. Because of an apprehension of an unnecessary death occurring during their treatment, healers frequently refer cases, from traditional to modern medicine and from general practitioner to hospital. Nevertheless, most changes of treatment are decided by the sick themselves and their relatives so that unsuccessful treatment is changed on average every 5 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1989

Chronic toxicity/carcinogenicity study of trans-anethole in rats.

A chronic feeding study was carried out in rats with trans-anethole. The test substance was administered in the diet to groups (n = 26-78) of 312 male and 312 female Sprague-Dawley rats at concentrations of 0, 0.25, 0.5 and 1% for 117-121 wk. The average intakes of trans-anethole varied from 105-550 mg/kg body weight/day. No apparent treatment-related reactions were noted. The only effect was a transient retardation of body-weight gain. No excess mortality was caused by the treatment. No abnormalities related to treatment were seen on necropsy except for reduced adiposity in the highest dose groups. Haematological assessments did not reveal any changes related to treatment. Histological examination revealed certain non-neoplastic and neoplastic lesions common in older rats. The incidence of some hepatic lesions was significantly higher in some treated groups than in controls: altered cell foci (females of the 1% group), nodular hyperplasia (males of the 0.5% group and males and females of the 1% groups), benign tumours (females of the 1% group) and malignant tumours (females of the 1% group). The results are compared with those of previous investigations. The authors of this study stress that the low incidence of hepatocarcinomas is restricted to a single species and sex and to the highest dose tested. This pattern of species, sex and dose dependency strongly suggests that metabolic and pharmacokinetic studies will be helpful in interpreting the significance of the rat tumours with regard to the safe consumption of trans-anethole by man. The changes observed in this chronic feeding study are not thought to be of genetic origin and consequently trans-anethole does not constitute a significant carcinogenic risk to man. Further studies are in progress to substantiate this conclusion.

Allylbenzene Derivatives↗
PubMed1989

The biochemical pharmacology of fenofibrate.

Fenofibrate is metabolized in several stages. First, the carboxyl ester moiety is cleaved by hydrolysis, resulting in fenofibric acid, the main pharmacologically active compound. Fenofibric acid, in turn, undergoes carbonyl reduction, resulting in a pharmacologically active metabolite referred to as reduced fenofibric acid. Both fenofibric acid and reduced fenofibric acid may be conjugated to form glucuronides. There are important species differences in the metabolism and elimination patterns of fenofibrate. In the rat and dog, fenofibric acid and reduced fenofibric acid are the principal metabolites. In humans, the glucuronide of fenofibric acid is predominant. In the rat and dog, approximately 70-80% of fenofibrate and its metabolites are recovered in the feces, whereas in humans approximately 65% of the dose is excreted in the urine. Several mechanisms contribute to fenofibrate's hypolipidemic action, including inhibition of fatty acid synthesis, stimulation of fatty acid beta-oxidation, inhibition of triglyceride synthesis, and enhancement of lipoprotein lipase activity. Fenofibrate's hypocholesterolemic action is a result of both decreased biosynthesis of cholesterol through inhibition of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity and increased low-density lipoprotein (LDL) clearance via modulation of hepatic LDL receptors. Fenofibrate also has three other actions that may result in the prevention or at least slowing of atherogenesis, namely inhibition of cholesterol esterification, platelet aggregation, and platelet-derived growth factor. The native acyl glucuronide of fenofibric acid is very stable, and is unlikely to have any toxic potential. Although the elimination half-life of fenofibrate is prolonged in the elderly and in patients with impaired hepatic function, the area under the curve and its clearance are not altered because of compensatory changes in the volume of distribution.

Cholesterol↗
PubMed1989

The metabolism of ethylbenzene and styrene to mandelic acid: stereochemical considerations.

1. The stereochemistry of mandelic acid, produced as a major urinary metabolite of ethylbenzene and styrene in rat and man has been investigated. Although these solvents are both achiral they are metabolized to chiral metabolites, via a series of chiral intermediates. 2. Analytical methods (g.l.c.-mass spectrometry, h.p.l.c. and 19F-n.m.r.) have been developed for the determination of the enantiomeric composition of mandelic acid in urine. 3. These methods have been applied to the study of the metabolic stereochemistry of ethylbenzene and styrene in rats dosed orally (100 mg/kg body weight) and in human volunteers exposed to atmospheres containing these solvents at the upper limits prescribed for workplaces by the UK Health and Safety Executive (100 ppm in air). 4. Results show that whereas only the R-enantiomer of mandelic acid was excreted after ethylbenzene exposure, the mandelic acid formed from styrene was essentially racemic. In three workers occupationally exposed to styrene, ratios of R to S isomers of 1.16, 1.27 and 1.14 were found. A synthetic R/S mixture of mandelic acid had an R/S ratio of 1.03. 5. The implications of these findings for the biological monitoring of workers occupationally exposed to stryrene and/or ethylbenzene are discussed.

Animals↗
PubMed1988

The role of intestinal microflora in the formation of the methylthio adduct metabolites of paracetamol. Studies in neomycin-pretreated and germ-free mice.

The contribution of the gastrointestinal microflora to the formation of methylthio adducts from paracetamol has been studied by comparing the fate of this drug in conventional mice with that in germ-free and neomycin-treated animals. In both germ-free and neomycin-treated mice there was a highly significant reduction in the urinary excretion of 3-methylthioparacetamol, its glucuronic acid and sulfate conjugates and its sulfoxide, with no other systemic alteration to the overall fate of the drug. These data are consistent with the gut flora playing a major role in the C-S cleavage of paracetamol-3-cysteine, thereby reducing the excretion of the array of methylthio adducts subsequently formed by tissue enzymes from 3-thioparacetamol, the putative product of the C-S cleavage.

Acetaminophen↗