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Biomedical subjects

J F Young

Publications and source records attributed to J F Young.

At least 37 records · Page 2Linked to original sources

Retinal pigment epithelial cell function on substrates with chemically micropatterned surfaces.

Model substrates with desired chemical micropatterns were fabricated using a microcontact printing technique. The substrate surfaces contained organized arrays of circular glass domains with a diameter of either 10 or 50 microm surrounded and separated by regions modified with octadecyltrichlorosilane (OTS) self-assembled monolayers (SAMs). The effects of surface patterning on in vitro cell attachment, proliferation, morphology, and cytoskeletal organization were evaluated using a human retinal pigment epithelium (RPE) cell line. Both micropatterns affected initial RPE cell attachment, limited cell spreading, and promoted the characteristic cuboidal cell morphology throughout the culture period. In contrast, RPE cells on plain glass control were elongated and appeared fibroblast-like prior to confluence. In addition, cells seeded at 30,000 cell/cm2 on the patterned surfaces maintained a normal pattern of actin and cytokeratin expression, and formed confluent monolayers within 4 days of culture. The cell density increased about 30-fold on both micropatterns by day 7. These results show that it is feasible to control RPE cell shape and expression of differentiated phenotype using micropatterned surfaces.

Actins↗

Effect of parsley (Petroselinum crispum) intake on urinary apigenin excretion, blood antioxidant enzymes and biomarkers for oxidative stress in human subjects.

Seven men and seven women participated in a randomized crossover trial to study the effect of intake of parsley (Petroselinum crispum), containing high levels of the flavone apigenin, on the urinary excretion of flavones and on biomarkers for oxidative stress. The subjects received a strictly controlled diet low in flavones and other naturally occurring antioxidants during the 2 weeks of intervention. This basic diet was supplemented with parsley providing 3.73-4.49 mg apigenin/MJ in one of the intervention weeks. Urinary excretion of apigenin was 1.59-409.09 micrograms/MJ per 24 h during intervention with parsley and 0-112.27 micrograms/MJ per 24 h on the basic diet (P < 0.05). The fraction of apigenin intake excreted in the urine was 0.58 (SE 0.16)% during parsley intervention. Erythrocyte glutathione reductase (EC 1.6.4.1; GR) and superoxide dismutase (EC 1.15.1.1; SOD) activities increased during intervention with parsley (P < 0.005) as compared with the levels on the basic diet, whereas erythrocyte catalase (EC 1.11.1.6) and glutathione peroxidase (EC 1.11.1.9) activities did not change. No significant changes were observed in plasma protein 2-adipic semialdehyde residues, a biomarker of plasma protein oxidation. In this short-term investigation, an overall decreasing trend in the activity of antioxidant enzymes was observed during the 2-week study. The decreased activity of SOD was strongly correlated at the individual level with an increased oxidative damage to plasma proteins. However, the intervention with parsley seemed, partly, to overcome this decrease and resulted in increased levels of GR and SOD.

Adult↗

A direct comparison of the activities of two humanized respiratory syncytial virus monoclonal antibodies: MEDI-493 and RSHZl9.

Two humanized monoclonal antibodies, MEDI-493 and RSHZ19, were developed independently as potential improvements over RSV-IGIV for prevention of respiratory syncytial virus (RSV) infection. RSV-IGIV is a polyclonal human antibody preparation for intravenous infusion enriched for RSV neutralizing activity. A phase III clinical trial showed that MEDI-493 significantly reduced hospitalizations due to RSV infection. In a separate trial, RSHZ19 failed to show significant efficacy. In new studies, the in vitro and in vivo activities of MEDI-493 and RSHZ19 were compared to determine whether the different clinical results are related to differences in biologic activity. MEDI-493 was consistently 4- to 5-fold more potent than RSHZ19 in antigen binding, RSV neutralization, and fusion inhibition assays. Although both MEDI-493 and RSHZ19 were effective against A and B subtypes of RSV in the cotton rat model of RSV infection, 2- to 4-fold higher doses of RSHZ19 were required for similar protection. The enhanced activity of MEDI-493 compared with RSHZ19 may, in part, explain its better clinical effect.

Animals↗

Effect of fruit juice intake on urinary quercetin excretion and biomarkers of antioxidative status.

BACKGROUND: Epidemiologic studies suggest that foods rich in flavonoids might reduce the risk of cardiovascular disease. OBJECTIVE: Our objective was to investigate the effect of intake of flavonoid-containing black currant and apple juice on urinary excretion of quercetin and on markers of oxidative status. DESIGN: This was a crossover study with 3 doses of juice (750, 1000, and 1500 mL) consumed for 1 wk by 4 women and 1 man corresponding to an intake of 4.8, 6.4, and 9.6 mg quercetin/d. RESULTS: Urinary excretion of quercetin increased significantly with dose and with time. The fraction excreted in urine was 0.29-0.47%. Plasma quercetin did not change with juice intervention. Plasma ascorbate increased during intervention because of the ascorbate in the juice. Total plasma malondialdehyde decreased with time during the 1500-mL juice intervention, indicating reduced lipid oxidation in plasma. Plasma 2-amino-adipic semialdehyde residues increased with time and dose, indicating a prooxidant effect of the juice, whereas erythrocyte 2-aminoadipic semialdehyde and gamma-glutamyl semialdehyde concentrations, Trolox-equivalent antioxidant capacity, and ferric reducing ability of plasma did not change. Glutathione peroxidase activity increased significantly with juice dose. CONCLUSIONS: Urinary excretion of quercetin seemed to be a small but constant function of quercetin intake. Short-term, high intake of black currant and apple juices had a prooxidant effect on plasma proteins and increased glutathione peroxidase activity, whereas lipid oxidation in plasma seemed to decrease. These effects might be related to several components of the juice and cannot be attributed solely to its quercetin content.

Adult↗

Pharmacokinetic considerations of dexamethasone-induced developmental toxicity in rats.

Dexamethasone (DEX) has been shown to elicit growth stunting and cleft palate in rat fetuses. This investigation characterized DEX dosimetry as various pharmacokinetic parameters and evaluated their impact on developmental toxicity endpoints. DEX pharmacokinetics was evaluated as a single dose on either gestation day (GD) 9 or 14, as well as on GD 14 after multiple daily dosing from GD 9 to GD 14. An additional set of pregnant rats was dosed with DEX on GD 9 through GD 14, pharmacokinetic evaluation was conducted on GD 14 through GD 16, and teratological evaluation was conducted following sacrifice on GD 20. For all pharmacokinetic evaluations, a subcutaneous (sc) injection of 0.8 mg DEX/kg body weight together with 50 microCi 3H-DEX was administered to Sprague-Dawley rats. Blood, urine, and feces were collected for 24 or 48 h. At GD 20 sacrifice, maternal tissues as well as fetal brain and liver samples were collected as part of the laparotomy. All samples were assayed using scintillation spectrometry. DEX pharmacokinetic parameters remained similar whether dosing occurred early (GD 9) or late (GD 14) in organogenesis, or dosing occurred on multiple sequential days (GD 9-14). DEX produced maternal and fetal weight loss, fetal lethality, and cleft palate. DEX a-half-life was positively correlated with the percentage of implants affected [(number of non-live + number with cleft palate)/number of implants]/litter. Neither the area under the concentration-time curve (AUC), the maximum maternal plasma concentration (Cmax), nor the terminal phase beta-half-life correlated with any fetal outcome parameters. The correlation between the percentage of the litter that was affected and half-life was improved if AUC was added in a stepwise multiple regression. These data suggest that the length of time that DEX is present in the maternal plasma at a sufficiently high concentration (i.e., slower tissue distribution of DEX) appears to be important in determining the risk of an adverse outcome in the offspring.

Abnormalities, Drug-Induced↗

Haematology and serum chemistry parameters of the pregnant rat.

This study describes the baseline haematology and serum chemistry values found in non-pregnant, pregnant (gestational days [GD] 2-21) and lactating (postnatal days 1-9) Sprague Dawley rats (n = 3-10/day) from the NCTR breeding colony of Crl:COBS CD(SD)BR strain. Maternal body weights on GD0 ranged from 250 to 300 g. Multiple analytes were measured in both whole blood and serum of dams. Amniotic fluid, fetal serum, and postnatal pup serum analyte values were also acquired. Maternal blood was collected from the heart under subterminal carbon dioxide (CO2) anaesthesia. Most pregnant dam blood values were not appreciably different from values for non-pregnant dams until near term; near-term values for some analytes (red blood cells, haemoglobin, haematocrit, mean corpuscular haemoglobin concentration, alkaline phosphatase, albumin, total protein, glucose, total bilirubin, sodium, and chloride) decreased but returned to near-normal values soon after delivery. The most dramatic change was a three-fold elevation of serum triglyceride levels near term with a subsequent decrease at birth. Most serum chemistry analytes measured in progeny increased after birth except for alkaline phosphatase, calcium and potassium levels which decreased.

Amniotic Fluid↗

Metabolism of chloral hydrate in mice and rats after single and multiple doses.

Chloral hydrate is a hepatocarcinogen in mice but not rats. To examine this species-related difference, male and female B6C3F1 mice and Fischer (F344) rats were treated by gavage with 1 or 12 doses of chloral hydrate, and concentrations of the drug and its metabolites were determined in plasma at 0.25, 7, 3, 6, and 24 h and 2, 4, 8, and 16 d after the last treatment. Maximum levels of chloral hydrate were observed at the initial sampling time of 0.25 h. By 1 h, levels dropped substantially, and by 3 h, chloral hydrate could not be detected. Trichloroacetic acid was the major metabolite found in the plasma, with peak levels being observed 1-6 h after dosing. The concentrations then slowly decreased such that by 2 d this metabolite could no longer be detected. Trichloroethanol was assayed as both the free alcohol and its glucuronide. Maximum levels of trichoroethanol occurred at 0.25 h, and by 1-3 h approached the limits of detection. A pharmacokinetic model was constructed to describe the metabolic data. The plasma half-life values of chloral hydrate were similar in both species. In mice, the rate of elimination of trichloroacetic acid was significantly increased after multiple doses; this difference was not observed with rats. The half-life of trichloroethanol and its glucuronide was significantly greater in rats as compared to mice. None of the metabolic parameters appears to account for the hepatocarcinogenicity of chloral hydrate seen in mice but not rats.

Animals↗

Physiologically-based pharmacokinetic model for pregnancy as a tool for investigation of developmental mechanisms.

There is no known mechanism of teratogenesis! Although receptor occupancy has been implicated, associated and/or deemed necessary for some malformations in newborns (e.g. estrogen receptors, retinoic acid receptors), the upstream and downstream events from receptor occupancy that definitively tie a xenobiotic exposure with a resultant malformation are essentially unknown. One part of the puzzle that can be delineated is the xenobiotic target-tissue exposure curve. Physiologically-based pharmacokinetic (PBPK) models are designed to provide time-course exposure curves for organs, tissues and fluids of human or animal systems. In this context pregnancy requires special considerations in that the PBPK model must represent the dynamic growth of both the maternal and embryo/fetal systems.

Abnormalities, Drug-Induced↗

Physiological "constants" for PBPK models for pregnancy.

Physiologically based pharmacokinetic (PBPK) models for pregnancy are inherently more complex than conventional PBPK models due to the growth of the maternal and embryo/fetal tissues. Physiological parameters such as compartmental volumes or flow rates are relatively constant at any particular time during gestation when an acute experiment might be conducted, but vary greatly throughout the course of gestation (e.g., contrast relative fetal weight during the first month of gestation with the ninth month). Maternal physiological parameters change during gestation, depending upon the particular system; for example, cardiac output increases by approximately 50% during human gestation; plasma protein concentration decreases during pregnancy; overall metabolism remains fairly constant. Maternal compartmental volumes may change by 10-30%; embryo/fetal volume increases over a billionfold from conception to birth. Data describing these physiological changes in the human are available from the literature. Human embryo/fetal growth can be well described using the Gompertz equation. By contrast, very little of these same types of data is available for the laboratory animal. In the rodent there is a dearth of information during organogenesis as to embryo weights, and even less organ or tissue weight or volume data during embryonic or fetal periods. Allometric modeling offers a reasonable choice to extrapolate (approximately) from humans to animals; validation, however, is confined to comparisons with limited data during the late embryonic and fetal periods of development (after gestation d 11 in the rat and mouse). Embryonic weight measurements are limited by the small size of the embryo and the current state of technology. However, the application of the laser scanning confocal microscope (LSCM) to optically section intact embryos offers the capability of precise structural measurements and computer-generated three-dimensional reconstruction of early embryos. Application of these PBPK models of pregnancy in laboratory animal models at teratogenically sensitive periods of development provides exposure values at specific target tissues. These exposures provide fundamentally important data to help design and interpret molecular probe investigations into mechanisms of teratogenesis.

Animals↗

Mathematical analysis for teratogenic sensitivity.

A mathematical structure is described for determining teratogenic sensitivity or susceptibility from analysis of malformation incidence, dose-response, and pharmacokinetic data obtained during pregnancy as a result of exposure to a teratogenic agent. From the dosage or exposure of laboratory animals, embryonic and maternal concentrations of the xenobiotic are calculated using a physiologically based pharmacokinetic (PBPK) model. Malformations observed in the progeny are linked to the PBPK-derived target tissue concentrations with a model for the sensitivity calculated as a function of the embryonic age. The PBPK model for internal disposition of chemicals during pregnancy was developed previously. This report focuses on the development of the mathematical relations for the sensitivity of the embryo and effect functions on different organs. The concentrations of a xenobiotic calculated for the site of action or target tissue(s) in the embryo are weighted using both a nonlinear dose-response curve and a sensitivity distribution function that depends on the age or stage of development of the embryo. This weighted "exposure" of the target tissue is regressed with the number of observed malformations to quantify the parameters of the model. This approach lends itself to integration of diverse sources of experimental data, with hydroxyurea data taken from several sources in the literature as an example. This sensitivity function obtained from laboratory animal data serves as a vehicle for prediction and extrapolation to human pregnancy for the teratogenic potential of a substance.

Abnormalities, Drug-Induced↗

A computer model and program for xenobiotic disposition during pregnancy.

A physiologically based pharmacokinetic computer model and program have been developed that depict internal disposition of chemicals during pregnancy in the mother and embryo/fetus. The model is based on human physiology but has been extended to simulate laboratory animal data. The model represents the distribution, metabolism, and elimination of two chemicals in both the maternal and embryo/fetal systems; the program handles the two chemicals completely independently or interactively with the two chemicals sharing routes of metabolism and/or elimination. The FORTRAN program computes the concentration of the two chemicals in 26 organs/tissues in the pregnant mother and 15 organs/tissues in the embryo/fetus using a 486DX4 or Pentium PC. Adjustments for embryo/fetal organ and tissue volumes as a function of developmental age are made utilizing the Gompertz growth equation for the developing embryo/fetus and allometric relationships for the developing organs. Various changes in the maternal compartments which could affect the distribution of a xenobiotic during pregnancy are also included in the model. Input files require estimates of binding coefficients, first- and/or second-order metabolism constants, level of interaction between the two chemicals, and dosing information. Different possible routes of administration are included (e.g., i.v., infusion, oral, dermal, and inhalation, as well as repeated doses or exposures). Regression analysis can be conducted on any combination of these various parameters to fit actual data. Output concentration-time curves are available simultaneously from all 82 differential equations. An illustrative example compares observed data with simulations for imipramine and its demethylated metabolite, desipramine, in both the maternal rat and her fetuses. Methyl mercury data for the non-pregnant and pregnant rat also are compared with human data. Based on parameters determined from analysis of rat data, the model is readjusted for human physiology and predicts human maternal and fetal tissue concentrations as a function of time.

Animals↗

Development of a humanized monoclonal antibody (MEDI-493) with potent in vitro and in vivo activity against respiratory syncytial virus.

Neutralizing polyclonal antibody to respiratory syncytial virus (RSV) has been shown to be an effective prophylactic agent when administered intravenously in high-risk infants. This study describes the generation of a humanized monoclonal antibody, MEDI-493, that recognizes a conserved neutralizing epitope on the F glycoprotein of RSV. The affinity of MEDI-493 was found to be equal to or slightly better than an isotype-matched chimeric derivative of the parent antibody. In plaque reduction, microneutralization, and fusion-inhibition assays, MEDI-493 was significantly more potent than the polyclonal preparation. Broad neutralization of a panel of 57 clinical isolates of the RSV A and B subtypes was demonstrated. Pretreatment of cotton rats with MEDI-493 resulted in 99% reduction of lung RSV titers at a dose of 2.5 mg/kg, corresponding to a serum concentration of 25-30 microg/mL. Further, MEDI-493 did not induce increased RSV infection or pathology in either a primary or a secondary challenge.

Amino Acid Sequence↗

Developmental toxicity of sarin in rats and rabbits.

Sarin (Agent GB, isopropyl methylphosphonofluoridate) is an organophosphate cholinesterase inhibitor. Sarin (Type I or Type II) was administered by gavage to CD rats on d 6-15 of gestation at dose levels of 0, 100, 240, or 380 micrograms/kg/d and to New Zealand White (NZW) rabbits on d 6-19 of gestation at dose levels of 0, 5, 10, or 15 micrograms/kg/d. Females were weighed on gestational days (GD) 0, 6-16 for rats and 6-20 for rabbits, and immediately prior to termination (GD 20 for rats and GD 29 for rabbits). All animals were monitored daily for clinical signs of toxicity throughout dosing and until sacrifice. At necropsy, gravid uteri were weighed and examined for the number and status of implants (live, resorbed, or dead). Individual fetal body weight, malformations, and variations (external, visceral, and skeletal) were recorded. Rat and rabbit dams in the high-dose groups exhibited significant signs of maternal toxicity and increased maternal mortality. Examination of gravid uteri revealed no statistical differences among treatment groups in the incidence of resorptions or of dead or malformed fetuses, or in average body weight of live fetuses per litter. These results show no evidence or developmental toxicity in the CD rat or NZW rabbit following exposure to either Type I or Type II sarin during embryonic differentiation and major organogenesis, even at a dose that produced maternal toxicity.

Administration, Oral↗

Synthesis and analysis of the enantiomers of calmidazolium, and a 1H NMR demonstration of a chiral interaction with calmodulin.

Calmidazolium [R24571, 1-[bis(4-chlorophenyl)methyl]-3-[2-(2,4-dichlorophenyl)-2-[(2,4- dichlorophenyl)methoxy]ethyl]-1H-imidazolium chloride] is a potent calmodulin inhibitor. This paper describes the synthesis and properties of the enantiomers of calmidazolium from the enantiomers of miconazole [1(N)-(2-(2,4-dichlorobenzyloxy)-2-(2,4 dichlorophenyl))-ethyl imidazole], prepared from the racemate by chiral preparative scale high performance liquid chromatography. Overlap between ligand and protein resonances in the aromatic region of the 1H NMR spectrum of the calmidazolium-calmodulin complexes has been obviated by preparation of the protein with all of its nine phenylalanine rings deuterated (Phe-d5 calmodulin). This has been accomplished by the overexpression of calmodulin derived from Trypanosoma brucei rhodiesiense in E. coli in a medium supplemented with ring-deuterated phenylalanine. The kinetics of binding of each enantiomer are slow on the 1H NMR time scale as judged by the behaviour of the H2 resonance of Histidine-107, which is clearly visible under the sample conditions used. The aromatic spectral regions of the protein-bound (+) and (-) enantiomers contrast strikingly, reflecting differences in bound environment and/or conformation.

Calmodulin↗

Investigation of fecal incontinence with endoanal ultrasound.

PURPOSE: This study was undertaken to audit the results of endoanal ultrasound in patients with fecal incontinence. METHODS: Endoanal ultrasound was used to investigate 53 patients with fecal incontinence. Data for endoanal ultrasound were collected prospectively. Results were compared with clinical and obstetric history, obtained retrospectively from case notes, and were compared with manometric and operative findings. RESULTS: Sphincter abnormalities were identified in 42 of 53 patients. A total of 28 anterior defects were thought to be obstetric in origin. Fourteen other defects were secondary to anal pathology or surgery. Patients with anterior external sphincter defects either had complete defects (4 patients; mean age, 31 years) or proximal defects (24 patients; mean age, 55 years). For patients with a proximal defect, 38 percent gave a history of obstetric tear, episiotomy, or forceps delivery, and the rest declared having had an apparently normal delivery. Only 50 percent had a sphincter weakness that was evident on clinical examination. Of those studied with manometry, only 21 percent had low squeeze pressures consistent with an external sphincter defect. CONCLUSIONS: Sphincter defects seen on ultrasound may not have a history of obstetric trauma or abnormal clinical and manometric findings. Endoanal ultrasound is recommended in all patients with fecal incontinence to detect occult sphincter defects.

Adult↗

A modification of single stapled colorectal anastomosis using the transanal insertion device (TAIS).

A modification of single stapled colorectal anastomosis utilizing a reusable stapler head (transanal insertion device [TAIS]) is described. This modification minimizes the time that the proximal bowel is open by allowing early placement of the circular stapler head, and minimizes the risk of rectal injury that may be associated with the passage of a circular stapling device without the head in situ.

Anastomosis, Surgical↗

Mathematical representation of organ growth in the human embryo/fetus.

During human pregnancy, there is a huge increase in the total weight of the embryo/fetus from conception to term. The total growth, which is the summation of growth of the various organs and tissues that make up the organism, was analyzed in a previous paper and fitted to the Gompertz equation for growth. In the present study, allometry, the quantitative representation of the consequence of size, was utilized to describe the correlation of individual fetal organ/tissue weights with the total fetal weight. The organ/tissue weight and the total fetal weight data used in the analyses were pooled from various sources that provided data ranging from 25 days to 300 days post-conception. Allometric equations are presented for 16 embryo/fetal organs and tissues. The standard allometric equation gave adequate fits for embryo/fetal adrenal, bone, bone marrow, brain, heart, liver, pancreas, plasma, skeletal muscle, extracellular water, thymus and thyroid; but it was necessary to use a quadratic form of the allometric equation for embryo/fetal fat, kidney, lung and spleen. Parameters were also calculated for crown-to-rump and crown-to-heels for fetal lengths that occur during pregnancy.

Adipose Tissue↗