Biomedical subjects

J B Thomas

Publications and source records attributed to J B Thomas.

At least 19 recordsLinked to original sources

PubMed2006

The impact of social status on the erythrocyte beta-adrenergic response in rainbow trout, Oncorhynchus mykiss.

The objective of the present investigation was to determine whether chronic increases in circulating cortisol concentrations, resulting from the occupation of subordinate status in rainbow trout social hierarchies, resulted in an enhancement of the erythrocyte adrenergic response. Rainbow trout (Oncorhynchus mykiss) were confined in fork length matched pairs for 6 h, 18 h, 48 h or 5-7 days, and social status was assigned through observations of behaviour. Erythrocyte adrenergic responsiveness, determined in vitro as changes in water content following incubation with the beta-adrenoreceptor agonist isoproterenol, was significantly greater in subordinate than dominant fish at 48 h of social interactions but not after 5-7 days, nor when assessed as changes in extracellular pH (pHe). However, the activity of the Na+/H+ exchanger (beta-NHE), assessed in vitro as the pHe change following incubation with the permeable cyclic AMP analogue 8-bromo-cyclic AMP, was significantly lower in subordinate fish. The number of erythrocyte membrane-bound adrenergic receptors (Bmax) was significantly higher in subordinate than dominant fish at 48 h, but had decreased by 5-7 days to a value that was not significantly different from that for dominant fish. The apparent dissociation constant (KD) of these receptors was not significantly impacted by either social status or interaction time. Finally, the relative expressions of beta-3b adrenergic receptor (AR) and beta-NHE mRNA were determined using real-time PCR and were found to be minimally affected by social rank. Relative to a control group, beta-3b AR mRNA was significantly up-regulated in both dominant and subordinate trout at all time periods, whereas the expression of beta-NHE was in general significantly down-regulated. Unlike the situation in rainbow trout treated with exogenous cortisol, elevations in circulating cortisol resulting from low social status did not "pre-adapt" the erythrocyte adrenergic response, but rather may have served to offset the potentially adverse effects elicited by plasma catecholamines, which were elevated during social hierarchy formation.

8-Bromo Cyclic Adenosine Monophosphate↗

PubMed2006

Effects of ageing on the biomechanical properties of rat articular cartilage.

Previous studies have demonstrated that male Sprague Dawley (SD) rats experience age-related bone loss with the same characteristics as that in ageing men. As articular cartilage, like bone, is a critical component of the health and function of the musculoskeletal system, the authors hypothesized that articular cartilage in the untreated male SD rats could be a suitable model for studying the age-related deterioration of articular cartilage in men. To test this hypothesis, male SD rats were killed at between 6 and 27 months. The right femur of each rat was removed. The effects of ageing on the structural integrity of the distal femoral articular cartilage were studied by biomechanical testing with a creep indentation apparatus. The aggregate modulus, Poisson's ratio, permeability, thickness, and percentage recovery of articular cartilage were determined using finite element/non-linear optimization modelling. No significant differences were observed in these biomechanical properties of the distal femoral articular cartilage as a function of age. Therefore, untreated male SD rats appear to be unsuitable for studying the age-related changes of articular cartilage as they occur in men. However, and more intriguingly, it is also possible that ageing does not affect the biomechanical properties of articular cartilage in the absence of cartilage pathology.

Aging↗

PubMed2001

Expression and function of the LIM homeodomain protein Apterous during embryonic brain development of Drosophila.

We analyzed the expression and function of the LIM-homeodomain transcription factor Apterous (Ap ) in embryonic brain development of Drosophila. Expression of Ap in the embryonic brain begins at early stage 12 and is subsequently found in approximately 200 protocerebral neurons and in 4 deutocerebral neurons. Brain glia do not express Ap. Most of the Ap-expressing neurons are interneurons and project their axons across the midline to the contralateral hemisphere; a smaller subset projects their axons into the ventral nerve cord. A few Ap-expressing neurons project to the ring gland, suggesting that they are neurosecretory cells. In ap loss-of-function mutants, some of the protocerebral and deutocerebral interneurons that express Ap in the wild type show axon pathfinding errors and fasciculation defects in the brain, notably in the fascicles of the brain commissure. In contrast, the interneurons that project to the ring gland do not appear to be affected in ap mutants. Thus, in brain development, Ap is required for correct axon guidance and fasciculation of interneurons, and Ap-expressing cells may also be involved in the brain neuroendocrine system.

Animals↗

PubMed2001

Identification of the first trans-(3R,4R)- dimethyl-4-(3-hydroxyphenyl)piperidine derivative to possess highly potent and selective opioid kappa receptor antagonist activity.

A structurally novel opioid kappa receptor selective ligand has been identified. This compound, (3R)-7-hydroxy-N-((1S)-1-[[(3R,4R)-4-(3-hydroxyphenyl)-3,4-dimethyl-1-piperidinyl]methyl]-2-methylpropyl)-1,2,3,4-tetrahydro-3-isoquinolinecarboxamide (JDTic, 10) demonstrated high affinity for the kappa receptor in the binding assay (kappa K(i) = 0.3 nM) and highly potent and selective kappa antagonism in the [(35)S]GTP-gamma-S assay using cloned opioid receptors (kappa K(i) = 0.006 nM, mu/kappa ratio = 570, delta/kappa ratio > 16600).

Animals↗

PubMed2001

Opioid peptide receptor studies. 15. Relative efficacy of 4-[(N-allyl-3-methyl-4-piperidinyl)phenylamino]-N,N-diethylbenzamide and related compounds at the cloned human delta-opioid receptor.

Previous data obtained from both binding and functional assays demonstrated that (-)-4-[(N-allyl-3-methyl-4-piperidinyl)phenylamino]-N,N-diethylbenzamide [(-)-RTI5989-54] displays selective binding and full agonist activity relative to (+/-)-RTI5989-54 for the delta opioid receptor. The present study was conducted to evaluate the activities of structurally diverse opioid receptor delta ligands in the [(35)S]GTP-gamma-S binding assay, comparing the relationship between receptor binding, activation, efficacy, and intrinsic efficacy. The data, obtained with cloned human delta receptors, demonstrated that (-)-RTI5989-54 behaves like the highly selective delta agonist SNC80. Addition of the hydroxyl group to RTI5989-54 (RTI5989-61) or replacement of the allyl group with the trans-crotyl group on the piperidine nitrogen of RTI-5989-61 (RTI5989-62) increased binding affinity, produced full agonist activity, and decreased intrinsic efficacy at the delta opioid receptor. The order of potency for the EC(50) (GTP-gamma-S) was RTI5989-62 (0.20 nM) > RTI5989-61 (0.43 nM) > SNC80 (1.92 nM) > DPDPE (3.50 nM) > (-)-RTI5989-54 (17.6 nM) > (+/-)-RTI5989-54 (65.6 nM) > (+)-RTI5989-54 (483 nM). RTI5989-61 and RTI5989-62 were fully efficacious, but had intrinsic efficacy values that were 2.2-3.1 times lower than that of DPDPE and SNC80. Comparison of the binding K(i) in competitively inhibiting [(125)I]IOXY binding to the functional K(i) for delta antagonists [Ki (IOXY)/Ki (GTP-gamma-S)] shows that antagonists might antagonize agonist-evoked neurochemical effects with equal magnitude while occupying different proportions of target receptors.

Animals↗

PubMed2001

Factors influencing agonist potency and selectivity for the opioid delta receptor are revealed in structure-activity relationship studies of the 4-[(N-substituted-4-piperidinyl)arylamino]-N,N-diethylbenzamides.

A study of the effect of transposition of the internal nitrogen atom for the adjacent benzylic carbon atom in delta-selective agonists such as BW373U86 (1) and SNC-80 (2) has been undertaken. It was shown that high-affinity, fully efficacious, and delta opioid receptor-selective compounds can be obtained from this transposition. In addition to the N,N-diethylamido group needed as the delta address, the structural features identified to promote delta receptor affinity in the set of compounds studied included a cis relative stereochemistry between the 3- and 4-substituents in the piperidine ring, a trans-crotyl or allyl substituent on the basic nitrogen, the lack of a 2-methyl group in the piperidine ring, and either no substitution or hydroxyl substitution in the aryl ring not substituted with the N,N-diethylamido group. Structural features found to be important for mu affinity include hydroxyl substitution in the aryl ring, the presence of a 2-methyl group in a cis relative relationship to the 4-amino group as well as N-substituents such as cyclopropylmethyl. It was also determined that mu receptor affinity could be increased while maintaining delta receptor affinity, especially when hydroxyl-substituted compounds are considered. Additionally, it was discovered that the somewhat lower mu/delta selectivities observed for the piperidine compounds relative to the piperazine-based ligands appear to arise as a consequence of the carbon-nitrogen transposition which imparts an overall lower delta and higher mu affinity to the piperidine-based ligands. This higher affinity for the mu receptor, apparently intrinsic to the piperidine-based compounds, suggests that ligands of this class will more easily be converted to mu/delta combination agonists compared to the piperazine ligands such as 1. This is particularly important since analogues of 1, which show both mu- and delta-type activity, are now recognized as important for their strong analgesia and cross-canceling of many of the side effects found in agonists operating exclusively from either the delta or mu opioid receptor.

Animals↗

PubMed2001

The derailed guidance receptor does not require kinase activity in vivo.

The Drosophila Derailed (DRL) receptor tyrosine kinase (RTK) controls key guidance events in the developing nervous system and mesoderm. Like other members of the "related to tyrosine kinases" (RYK) subfamily of RTKs, DRL has several highly unusual amino acid substitutions within the catalytic domain, raising the possibility that members of this subfamily are catalytically inactive. To test the role of DRL kinase activity in vivo, we mutated the invariant lysine required for catalytic activity of known kinases and examined the ability of this mutant to function in two assays: a dominant gain-of-function axon switch assay in the nervous system and phenotypic rescue of muscle attachment in drl mutants. We show that this predicted kinase-deficient DRL mutant is capable of functioning in both assays. Our results indicate that DRL does not require kinase activity in vivo and suggest that members of the RYK subfamily of RTKs transduce signals unconventionally.

Animals↗

PubMed2001

Opioid peptide receptor studies. 14. Stereochemistry determines agonist efficacy and intrinsic efficacy in the [(35)S]GTP-gamma-S functional binding assay.

Previous data obtained with the cloned rat mu opioid receptor demonstrated that stereochemistry affects the four parameters of the ligand-receptor interaction: potency (ED(50)), efficacy (maximal stimulation), intrinsic efficacy (effect as a function of receptor occupation), and binding affinity. This study evaluated the activities of structurally diverse opioid receptor ligands in the [(35)S]GTP-gamma-S binding assay, comparing the relationship between receptor binding, activation, efficacy, and intrinsic efficacy. The data, obtained with cloned rat mu receptors, demonstrated that an analgetic, (-)-5-m-hydroxyphenyl-2-methylmorphan (NIH8508), and its (+)-isomer (NIH8509), behave as partial agonists, but had different intrinsic efficacy in the [(35)S]GTP-gamma-S binding assay. Replacement of the methyl group with the phenethyl group on the piperidine nitrogen of NIH8508 and NIH8509 [(1R,5S)-AH019 and (1S, 5R)-AH019] increased affinity for the mu receptor and eliminated any agonist effect, supporting the hypothesis that certain structural features make these compounds antagonists. These study also show that all of the fully efficacious mu agonists studied here had high levels of intrinsic efficacy, producing a 50% response at about 10% receptor occupancy. Comparison of the binding K(i) in competitively inhibiting [(125)I]IOXY binding to the functional K(i) for opioid antagonists [K(i)(IOXY)/K(i)(GTP-gamma-S)] provides more detailed evidence that the [(35)S]GTP-gamma-S binding assay can be used to reliably determine apparent functional antagonist K(i) values in addition to agonist ED(50), efficacy and intrinsic efficacy.

Analgesics, Opioid↗

PubMed2001

Effect of black blood MR image quality on vessel wall segmentation.

Black blood MRI has become a popular technique for measuring arterial wall area as an indicator of plaque size. Computer-assisted techniques for segmenting vessel boundaries have been developed to increase measurement precision. In this study, the carotid arteries of four normal subjects were imaged at seven different fields of view (FOVs), keeping all other imaging parameters fixed, to determine whether spatial resolution could be increased at the expense of image quality without sacrificing precision. Wall areas were measured via computer-assisted segmentation of the vessel boundaries performed repeatedly by two operators. Analysis of variance (ANOVA) demonstrated that the variability of wall area measurements was below 1.5 mm(2) for in-plane spatial resolutions between 0.22 mm and 0.37 mm. An inverse relationship between operator variability and the signal difference-to-noise ratio (SDNR) demonstrated that semi-automatic segmentation of the wall boundaries was robust for SDNR >3, defining a criterion above which subjective image quality can be degraded without an appreciable loss of information content. Our study also suggested that spatial resolutions higher than 0.3 mm may be required to quantify normal wall areas to within 10% accuracy, but that the reduced SNR associated with the higher resolution may be tolerated by semi-automated wall segmentation without an appreciable loss of precision.

Adult↗

PubMed2001

Effects of conjugated linoleic acids and docosahexaenoic acid on rat liver and reproductive tissue fatty acids, prostaglandins and matrix metalloproteinase production.

Long chain n-6 and n-3 fatty acids play important roles in labor and delivery. These effects may be mediated by prostaglandin (PG) synthesis and by regulation of matrix metalloproteinases (MMPs), both of which play roles in uterine contraction, cervical ripening and rupture of fetal membranes. The effects of altering dietary n-6:n-3 long chain fatty acid ratios, and the addition of dietary conjugated linoleic acids (CLA) and docosahexaenoic acid (DHA) on fatty acid composition of reproductive tissues, PG synthesis in liver and reproductive tissue and serum MMP levels were examined in pregnant rats. Modified AIN-96G diets with n-6:n-3 ratios of 7:1 and 34:1 with and without added 1.1% (by weight) conjugated linoleic acid (CLA) and/or 0.3% (by weight) DHA were fed through day 20 of gestation. Reproductive tissues readily incorporated both DHA and CLA. CLA significantly (P<0.05) depressed PGF(2 alpha)synthesis in placenta, uterus and liver by 50% when the n-6:n-3 ratio was 7:1 and by 66% at 34:1 ratio. Significant differences (P<0.05) in PGE(2)synthesis in uterus and liver were seen only between groups fed the high ratio of n-6:n-3 without CLA, and the low ratio with CLA. Addition of CLA to DHA containing diets depressed PGF(2alpha) by one-third in uterus and liver (P<0.05). Serum MMP-9 and active MMP-2 were suppressed (P<0.05) by addition of either CLA or DHA.

Animals↗

PubMed2001

Use of contrast echocardiography in the diagnosis of subacute myocardial rupture after myocardial infarction.

Myocardial rupture is an uncommon and catastrophic complication after acute myocardial infarction. It can present in an acute form or in a subacute form, with slower hemorrhage and thrombus formation at the site of rupture. These patients can survive several hours or days before the diagnosis is confirmed and the myocardial ruptured repaired. Two-dimensional Doppler echocardiography is very useful in the diagnosis of this complication, but the number of false-positive diagnoses is high, even in the presence of a large amount of pericardial effusion. In these patients, administration of a contrast agent can be useful to demonstrate active bleeding into the pericardium. We report a case of subacute myocardial rupture for which contrast echocardiography was useful in demonstrating the presence of persistent hemorrhage into the pericardium. To reduce the number of false-positive diagnoses, contrast echocardiography should be considered in patients with possible subacute myocardial rupture.

Aged↗

PubMed2001

A semi-automatic technique for measurement of arterial wall from black blood MRI.

Black blood magnetic resonance imaging (MRI) has become a popular technique for imaging the artery wall in vivo. Its noninvasiveness and high resolution make it ideal for studying the progression of early atherosclerosis in normal volunteers or asymptomatic patients with mild disease. However, the operator variability inherent in the manual measurement of vessel wall area from MR images hinders the reliable detection of relatively small changes in the artery wall over time. In this paper we present a semi-automatic method for segmenting the inner and outer boundary of the artery wall, and evaluate its operator variability using analysis of variance (ANOVA). In our approach, a discrete dynamic contour is approximately initialized by an operator, deformed to the inner boundary, dilated, and then deformed to the outer boundary. A group of four operators performed repeated measurements on 12 images from normal human subjects using both our semiautomatic technique and a manual approach. Results from the ANOVA indicate that the inter-operator standard error of measurement (SEM) of total wall area decreased from 3.254 mm2 (manual) to 1.293 mm2 (semi-automatic), and the intra-operator SEM decreased from 3.005 mm2 to 0.958 mm2. Operator reliability coefficients increased from less than 69% to more than 91% (inter-operator) and 95% (intra-operator). The minimum detectable change in wall area improved from more than 8.32 mm2 (intra-operator, manual) to less than 3.59 mm2 (inter-operator, semi-automatic), suggesting that it is better to have multiple operators measure wall area with our semi-automatic technique than to have a single operator make repeated measurements manually. Similar improvements in wall thickness and lumen radius measurements were also recorded. Since the semi-automatic technique has effectively ruled out the effect of the operator on these measurements, it may be possible to use such techniques to expand prospective studies of atherogenesis to multiple centers so as to increase access to real patient data without sacrificing reliability.

Adult↗

PubMed2001

Drosophila wing development in the absence of dorsal identity.

The developing wing disc of Drosophila is divided into distinct lineage-restricted compartments along both the anterior/posterior (A/P) and dorsal/ventral (D/V) axes. At compartment boundaries, morphogenic signals pattern the disc epithelium and direct appropriate outgrowth and differentiation of adult wing structures. The mechanisms by which affinity boundaries are established and maintained, however, are not completely understood. Compartment-specific adhesive differences and inter-compartment signaling have both been implicated in this process. The selector gene apterous (ap) is expressed in dorsal cells of the wing disc and is essential for D/V compartmentalization, wing margin formation, wing outgrowth and dorsal-specific wing structures. To better understand the mechanisms of Ap function and compartment formation, we have rescued aspects of the ap mutant phenotype with genes known to be downstream of Ap. We show that Fringe (Fng), a secreted protein involved in modulation of Notch signaling, is sufficient to rescue D/V compartmentalization, margin formation and wing outgrowth when appropriately expressed in an ap mutant background. When Fng and alphaPS1, a dorsally expressed integrin subunit, are co-expressed, a nearly normal-looking wing is generated. However, these wings are entirely of ventral identity. Our results demonstrate that a number of wing development features, including D/V compartmentalization and wing vein formation, can occur independently of dorsal identity and that inter-compartmental signaling, refined by Fng, plays the crucial role in maintaining the D/V affinity boundary. In addition, it is clear that key functions of the ap selector gene are mediated by only a small number of downstream effectors.

Animals↗

PubMed2001

Selenium status of cats in four regions of the world and comparison with reported incidence of hyperthyroidism in cats in those regions.

OBJECTIVE: To assess selenium (Se) status of cats in 4 regions of the world and to compare results for Se status with reported incidence of hyperthyroidism in cats in those regions. ANIMALS: 50 cats (30 from 2 regions with an allegedly high incidence of hyperthyroidism and 20 from 2 regions in which the disease is less commonly reported). PROCEDURE: Hematologic samples (heparinized whole blood, plasma, and RBC fractions) were obtained from 43 healthy euthyroid cats and 7 hyperthyroid cats. Plasma concentration of Se and activity of glutathione peroxidase (GPX) in whole blood and plasma were determined. RESULTS: Plasma concentration of Se and GPX activity in whole blood or plasma did not differ significantly among cats from the 4 regions. However, cats had a plasma concentration of Se that was approximately 5 times the concentration reported in rats and humans. The GPX activity in whole blood or plasma in cats generally was higher than values reported in rats or humans. CONCLUSIONS AND CLINICAL RELEVANCE: Cats have higher Se concentrations in plasma, compared with values for other species. However, Se status alone does not appear to affect the incidence of hyperthyroidism in cats. High Se concentrations may have implications for health of cats if such concentrations are influenced by the amount of that micronutrient included in diets.

Animals↗

PubMed2001

Agonist-, antagonist-, and inverse agonist-regulated trafficking of the delta-opioid receptor correlates with, but does not require, G protein activation.

In this study, we explored the relationship between ligand-induced regulation of surface delta opioid receptors and G protein activation. G protein activation was assessed with [(35)S]guanosine-5'-O-(3-thio)triphosphate (GTP gamma S) binding assays conducted at both 37 and 0 degrees C. Ligand-independent (constitutive) activity of the delta-receptor was readily observed when the [(35)S]GTP gamma S binding assay was performed at 37 degrees C. We identified a new class of alkaloid inverse agonists (RTI-5989-1, RTI-5989-23, RTI-5989-25), which are more potent than the previously described peptide inverse agonist ICI-174864 (N,N-diallyl-Tyr-Aib-Aib-Phe-Leu). Treatment with these inverse agonists for 18 h caused up-regulation of surface receptors. Eighteen-hour treatment with etorphine resulted in approximately 90% loss of surface receptor, whereas fentanyl, diprenorphine, and morphine caused between 20 and 50% loss. The abilities of ligands to modulate [(35)S]GTP gamma S binding at 37 degrees C showed a strong correlation with their abilities to regulate surface receptor number (r(2) = 0.86). Interestingly, the ability of fentanyl to activate G proteins was markedly temperature sensitive. Fentanyl showed no stimulation of [(35)S]GTP gamma S binding at 0 degrees C but was as efficacious as etorphine, morphine, and diprenorphine at 37 degrees C. Neither the ligand-induced receptor increases nor decreases were perturbed by pertussis toxin pretreatment, suggesting that functional G proteins are not required for ligand-regulated delta-opioid receptor trafficking.

Biotransformation↗

PubMed2000

NIST micronutrients measurement quality assurance program: characterizing the measurement community's performance over time.

The Micronutrients Measurement Quality Assurance Program (M2QAP) at the National Institute of Standards and Technology was created in 1984 with the goal of improving among-participant measurement comparability for fat-soluble vitamin-related compounds in human serum. We recently described improved tools for evaluating comparison exercise data; we here extend and apply these tools to the evaluation of the measurement community's performance over the entire 15-year history of the M2QAP. We here display measurement performance characteristics for the 14 measurands most commonly reported by the M2QAP community. We confirm that among-participant comparability for total beta-carotene cannot be much improved without improving average long-term within-participant measurement stability. We demonstrate that improved measurand definition and/or identification of interferences may help participants improve comparability for many of the M2QAP's other commonly reported measurands. The reported measurement performance characteristics may be of interest to clinical, nutritional, and epidemiological studies involving any of these measurands. The data analysis techniques utilized may be applicable to other programs.

Humans↗

PubMed2000

NIST micronutrients measurement quality assurance program: characterizing individual participant measurement performance over time.

The mission of the Micronutrients Measurement Quality Assurance Program (M2QAP) at the National Institute of Standards and Technology is enhanced interlaboratory measurement comparability for fat-soluble vitamin-related measurands in human serum. We recently described improved tools for evaluating individual participant measurement performance in single interlaboratory comparison exercises; we here apply and extend these tools to the evaluation of participant performance over the entire 15-year history of the M2QAP. We describe and illustrate a set of interconnected graphical reporting tools for identifying long-term trends and single-exercise events. We document and discuss recurrent patterns we observe in the measurement performance characteristics for M2QAP participants. The graphical analysis techniques utilized may be applicable to other interlaboratory comparison programs.

Humans↗

PubMed2000

Determination of vitamins in food-matrix Standard Reference Materials.

In recent years, the National Institute of Standards and Technology (NIST) has developed several food-matrix Standard Reference Materials (SRMs) characterized for vitamins and other organic nutrients. NIST uses several "modes" for assignment of analyte concentrations in SRMs, one of which includes the use of data provided by collaborating laboratories. Certification modes and liquid chromatographic methods that were used by NIST for value assignment of vitamin concentrations in recently introduced food-matrix SRMs are described in this paper. These materials and methods include vitamins D and E in coconut oil (SRM 1563) by gravimetry and multi-dimensional liquid chromatography (LC); vitamins A, E, and several B vitamins by reversed-phase LC and vitamin C by ion-exchange chromatography in infant formula (SRM 1846); and carotenoids and vitamins A and E by reversed-phase liquid chromatography in a baby food composite (SRM 2383).

Chromatography, Liquid↗