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Biomedical subjects

A Davis

Publications and source records attributed to A Davis.

At least 37 records · Page 2Linked to original sources

Metal distribution in Clark Fork River sediments.

Accurate sampling of pore waters and sediments in gravel river beds is problematic because cobbles preclude most coring techniques, while the angularity of sand grains destroys the membranes of standard pore water samplers. In the Clark Fork River, the recipient of over 100 years of mining activity, a modified single tube core-freezing device was used to profile bed sediment (approximately 65% cobbles and 35% gravel substrate), and a novel pore water sampler was developed to collect interstitial water. In the sediment, the <63 microm fraction comprised <4 wt % yet contained approximately an order of magnitude higher metal concentrations compared to the 63 microm-2 mm fraction (constituting approximately 20% of the mass). However, on a mass basis the sand fraction contained 60% of the metals, compared to 40% in the clay/silt fraction in the gravels, while in point bars the distribution was approximately 50:50. The metals occur predominantly in sulfides frequently armored with an oxide rim and other sparingly soluble phases that may explain the low pore water metal concentrations. These data demonstrate that consideration of multiple particle size cutoffs is necessary to accurately characterize fluvial bed sediment metal conditions and that the form of the metal is important in understanding metal solubility in the benthos.

Animals↗

Eukaryotic-like adenylyl cyclases in Mycobacterium tuberculosis H37Rv: cloning and characterization.

Screening the Mycobacterium tuberculosis H37Rv genomic library for complementation of catabolic defect for cAMP-dependent expression of maltose operon produced the adenylyl cyclase gene (Mtb cya, (1997)) annotated later as Rv1625c (Cole, S. T., Brosch, R., Parkhill, J., Garnier, T., Churcher, C., Harris, D., Gordon, S. V., Eiglmeier, K., Gas, S., Barry, C. E., III, et al. (1998) Nature 393, 537-544). The deduced amino acid (aa) sequence (443 aa) encoded by Mtb cya contains a single hydrophobic domain of six transmembrane helices (152 aa) in the amino-terminal half of the protein. Flanking this domain are an arginine-rich (17%) amino-terminal cytoplasmic tail (46 aa) and a carboxyl-terminal cytoplasmic domain (245 aa) with extensive homology to the catalytic core of eukaryotic adenylyl cyclases. Site-directed mutagenesis of Arg(43) and Arg(44) to alanine/glycine showed a loss of adenylyl cyclase activity, whereas mutagenesis to lysine restored the activity. Hence it is proposed that the formation of the catalytic site in Mtb adenylyl cyclase requires an interaction between Arg(43) and Arg(44) residues in the distal cytoplasmic tail and the carboxyl-terminal cytoplasmic domain. Mtb adenylyl cyclase activity at the physiological concentration of ATP (1 mm) was 475 nmol of cAMP/min/mg of membrane protein in the presence of Mn(2+) but only 10 nmol of cAMP/min/mg of membrane protein in the presence of Mg(2+). The physiological significance of the activation of Mtb adenylyl cyclase by Mn(2+) is discussed in view of the presence of manganese transporter protein in mycobacteria and macrophages wherein mycobacteria reside.

Adenosine Triphosphate↗

p38 stress-activated protein kinase inhibitor reverses bradykinin B(1) receptor-mediated component of inflammatory hyperalgesia.

The effects of a p38 stress-activated protein kinase inhibitor, 4-(4-fluorophenyl)-2-(-4-methylsulfonylphenyl)-5-(4-pyridynyl) imidazole (SB203580), were evaluated in a rat model of inflammatory hyperalgesia. Oral, but not intrathecal, administration of SB203580 significantly reversed inflammatory mechanical hyperalgesia induced by injection of complete Freund's adjuvant into the hindpaw. SB203580 did not, however, affect the increased levels of interleukin-1beta and cyclo-oxygenase 2 protein observed in the hindpaw following complete Freund's adjuvant injection. Intraplantar injection of interleukin-1beta into the hindpaw elicited mechanical hyperalgesia in the ipsilateral paw, as well as in the contralateral paw, following intraplantar injection of the bradykinin B(1) receptor agonist des-Arg(9)-bradykinin. Oral administration of SB203580 1 h prior to interleukin-1beta administration prevented the development of hyperalgesia in the ipslateral paw and the contralateral bradykinin B(1) receptor-mediated hyperalgesia. In addition, following interleukin-1beta injection into the ipsilateral paw, co-administration of SB203580 with des-Arg(9)-bradykinin into the contralateral paw inhibited the bradykinin B(1) receptor-mediated hyperalgesia. In human embryonic kidney 293 cells expressing the human bradykinin B(1) receptor, its agonist des-Arg(10)-kallidin produced a rapid phosphorylation of endogenous p38 stress-activated protein kinase. Our data suggest that p38 stress-activated protein kinase is involved in the development of inflammatory hyperalgesia in the rat, and that its pro-inflammatory effects involve the induction of the bradykinin B(1) receptor as well as functioning as its downstream effector.

Animals↗

An analysis of soil arsenic records of decision.

In 1986 the US EPA created the National Priority List (NPL) that now comprises in excess of 2,000 sites nationwide, with arsenic the second most common inorganic constituent. A survey of 69 Records of Decision (RODs) written between 1985 and 1998 for which arsenic was a major driver found that 84% of cleanup goals were risk-driven and 16% were background-driven, with a wide range of soil-arsenic cleanup standards for 10(-6) residential risk goals (2-305 mg/kg). In comparison, the range of background-based cleanup goals was much narrower (8-21 mg/kg). ROD soil arsenic concentrations exhibit no statistically significanttemporal trend, but on a geographic basis, EPA Regions 6, 8, 9, and 10 had some of the higher decisions. The risk assessment process is important in defining cleanup goals; however routine use of site-specific variables (i.e., bioavailability, realistic tenure in both residential and occupational settings, natural attenuation of arsenic in groundwater, etc.) is necessary to ensure an accurate assessment of potential site risks and to preclude over-remediation that may result from the use of default risk variables.

Arsenic↗

Are all wheezing disorders in very young (preschool) children increasing in prevalence?

BACKGROUND: Distinct wheezing disorders co-exist in young (preschool) children, some of which (early transient wheeze and viral wheeze) are thought to be unrelated to atopy. Investigation of changes in prevalence of wheezing disorders in preschool children could provide important clues about underlying mechanisms responsible for increasing prevalence of asthma in schoolchildren. METHODS: Repeated population surveys of the prevalence of respiratory symptoms were done by parent-completed postal questionnaires in random samples of 1650 (1990) and 2600 (1998) caucasian children aged 1-5 years living in the county of Leicestershire, UK. FINDINGS: The response rates were 86% (1422 of 1650) in 1990 and 84% (2127 of 2522) in 1998. Between 1990 and 1998, there was a significant increase in the prevalance of reported wheeze ever (16% to 29%, p<0.0001), current wheeze (12% to 26%, p<0.0001), diagnosis of asthma (11% to 19%, p<0.0001), treatment for wheeze (15% to 26%, p<0.0001), and admission for wheeze or other chest trouble (6% to 10%, p<0.0001). The increase occurred both in children with viral wheeze (9% to 19%) and in those with the classic asthma pattern of wheezing with multiple triggers (6% to 10%). There was also an increase in transient early wheezers (3% to 5%), persistent wheezers (5% to 13%), and late-onset wheezers (6% to 8%), and in all severity groups. The increase could not be accounted for by putative household risk factors because these declined between the 2 years. INTERPRETATION: The fact that all preschool wheezing disorders increased (including viral wheeze) makes it probable that factors unrelated to atopy are implicated in the changing epidemiology of wheeze in childhood.

Asthma↗

A peripheral mechanism for behavioral adaptation to specific "bitter" taste stimuli in an insect.

Animals have evolved several chemosensory systems for detecting potentially dangerous foods in the environment. Activation of specific sensory cells within these chemosensory systems usually elicits an aversive behavioral response, leading to avoidance of the noxious foods. Although this aversive behavioral response can be adaptive, there are many instances in which it generates "false alarms," causing animals to reject harmless foods. To minimize the number of false alarms, animals have evolved a variety of physiological mechanisms for selectively adapting their aversive behavioral response to harmless noxious compounds. We examined the mechanisms underlying exposure-induced adaptation to specific "bitter" compounds in Manduca sexta caterpillars. M. sexta exhibits an aversive behavioral response to many plant-derived compounds that taste bitter to humans, including caffeine and aristolochic acid. This aversive behavioral response is mediated by three pairs of bitter-sensitive taste cells: one responds vigorously to aristolochic acid alone, and the other two respond vigorously to both caffeine and aristolochic acid. We found that 24 hr of exposure to a caffeinated diet desensitized all of the caffeine-responsive taste cells to caffeine but not to aristolochic acid. In addition, we found that dietary exposure to caffeine adapted the aversive behavioral response of the caterpillar to caffeine, but not to aristolochic acid. We propose that the adapted aversive response to caffeine was mediated directly by the desensitized taste cells and that the adapted aversive response did not generalize to aristolochic acid because the signaling pathway for this compound was insulated from that for caffeine.

Adaptation, Physiological↗

Isothermal titration calorimetric analysis of the interaction between cationic lipids and plasmid DNA.

The effects of buffer and ionic strength upon the enthalpy of binding between plasmid DNA and a variety of cationic lipids used to enhance cellular transfection were studied using isothermal titration calorimetry at 25.0 degrees C and pH 7.4. The cationic lipids DOTAP (1,2-dioleoyl-3-trimethyl ammonium propane), DDAB (dimethyl dioctadecyl ammonium bromide), DOTAP:cholesterol (1:1), and DDAB:cholesterol (1:1) bound endothermally to plasmid DNA with a negligible proton exchange with buffer. In contrast, DOTAP: DOPE (L-alpha-dioleoyl phosphatidyl ethanolamine) (1:1) and DDAB:DOPE (1:1) liposomes displayed a negative enthalpy and a significant uptake of protons upon binding to plasmid DNA at neutral pH. These findings are most easily explained by a change in the apparent pKa of the amino group of DOPE upon binding. Complexes formed by reverse addition methods (DNA into lipid) produced different thermograms, sizes, zeta potentials, and aggregation behavior, suggesting that structurally different complexes were formed in each titration direction. Titrations performed in both directions in the presence of increasing ionic strength revealed a progressive decrease in the heat of binding and an increase in the lipid to DNA charge ratio at which aggregation occurred. The unfavorable binding enthalpy for the cationic lipids alone and with cholesterol implies an entropy-driven interaction, while the negative enthalpies observed with DOPE-containing lipid mixtures suggest an additional contribution from changes in protonation of DOPE.

Calorimetry↗

In vitro and in vivo anticancer activities of synthetic macrocyclic ketone analogues of halichondrin B.

Halichondrin B is a highly potent anticancer agent originally found in marine sponges. Although scarcity of the natural product has hampered efforts to develop halichondrin B as a new anticancer drug, the existence of a complete synthetic route has allowed synthesis of structurally simpler analogues that retain the remarkable potency of the parent compound. In this study, we show that two macrocyclic ketone analogues of halichondrir B, ER-076349 and ER-086526, have sub-nM growth inhibitory activities in vitro against numerous human cancer cell lines as well as marked in vivo activities at 0.1-1 mg/kg against four human xenografts: MDA-MB-435 breast cancer, COLO 205 colon cancer, LOX melanoma, and NIH: OVCAR-3 ovarian cancer. ER-076349 and ER-086526 induce G2-M cell cycle arrest and disruption of mitotic spindles, consistent with the tubulin-based antimitotic mechanism of halichondrin B. This is supported further by direct binding of the biotinylated analogue ER-040798 to tubulin and inhibition of tubulin polymerization in vitro by ER-076349 and ER-086526. Retention of the extraordinary in vitro and in vivo activity off halichondrin B in structurally simplified, fully synthetic analogues establishes the feasibility of developing halichondrin B-based agents as highly effective, novel anticancer drugs.

Animals↗

beta-Amyloid protein induces the formation of purine dimers in cellular DNA.

Experimental evidence implicates oxidative free radical reactions as central in the processes of neurodegenerative diseases. In particular, cellular interactions with the beta-amyloid protein have been linked to neuron cell death in Alzheimer's disease. Also, uncharacterized dimeric purine moieties have been detected in oxidized DNAs. It has been suggested that inadequate excision-repair of such products plays a functional role in the neurological degeneration observed in familial Alzheimer's disease, Down's syndrome, and xeroderma pigmentosum. Therefore, in order to obtain a reagent to monitor the presence of such products, the purine dimer 8-8-(2'-deoxyguanosyl)-2'-deoxyguanosine-5'-monophosphate was used as a hapten for elicitation of rabbit anti-purine dimer antiserum. This antiserum specifically recognizes various purified 8-8-bideoxyribonucleosides and 8-8-bideoxyribonucleotides. We found that DNA oxidized by the Fenton reaction is specifically recognized by this antiserum. This reagent can therefore be used to demonstrate formation and excision of DNA purine dimers. Moreover, incubation of cultured rat pheochromocytoma PC-12 cells with the beta-amyloid protein resulted in formation of these purine dimers in cellular DNA. These dimers were subsequently removed from cellular DNA. From these results we conclude that the free radicals generated by A beta cause oxidative DNA alterations including purine dimers. Deficient repair of this type of DNA damage might result in neural cell loss via apoptosis. Our findings suggest mechanisms for the roles of beta-amyloid and oxidative free radicals in neurodegenerative diseases and the role of DNA excision-repair in the prevention of lethal neurotoxicity.

Amyloid beta-Peptides↗

Metabotropic glutamate receptor subtype 5 (mGlu5) and nociceptive function. I. Selective blockade of mGlu5 receptors in models of acute, persistent and chronic pain.

The excitatory neurotransmitter, glutamate, is particularly important in the transmission of pain information in the nervous system through the activation of ionotropic and metabotropic glutamate receptors. A potent, subtype-selective antagonist of the metabotropic glutamate-5 (mGlu5) receptor, 2-methyl-6-(phenylethynyl)-pyridine (MPEP), has now been discovered that has effective anti-hyperalgesic effects in models of inflammatory pain. MPEP did not affect rotarod locomotor performance, or normal responses to noxious mechanical or thermal stimulation in naïve rats. However, in models of inflammatory pain, systemic administration of MPEP produced effective reversal of mechanical hyperalgesia without affecting inflammatory oedema. In contrast to the non-steroidal anti-inflammatory drugs, indomethacin and diclofenac, the maximal anti-hyperalgesic effects of orally administered MPEP were observed without acute erosion of the gastric mucosa. In contrast to its effects in models of inflammatory pain, MPEP did not produce significant reversal of mechanical hyperalgesia in a rat model of neuropathic pain.

Animals↗

mGlu5 receptors and nociceptive function II. mGlu5 receptors functionally expressed on peripheral sensory neurones mediate inflammatory hyperalgesia.

Previous studies have demonstrated that the metabotropic glutamate receptor subtype 5 (mGlu5 receptor) is expressed in the cell bodies of rat primary afferent neurones. We have further investigated the function and expression of mGlu5 receptors in primary afferent neurones, and their role in inflammatory nociception. Freund's complete adjuvant-induced inflammatory hyperalgesia of the rat hind paw was significantly reduced by intraplantar, but not by intracerebroventricular or intrathecal microinjection of the selective mGlu5 receptor antagonist, 2-methyl-6-(phenylethynyl)-pyridine (MPEP). Pharmacological comparison in vivo of the nociceptive effects of glutamate, and ionotropic and metabotropic glutamate (mGlu) receptor agonists applied to the rat hind paw, indicated that group I mGlu receptor agonists induce a dose-dependent decrease in paw withdrawal threshold (mechanical hyperalgesia). Group I mGlu agonist-induced hyperalgesia was inhibited by co-microinjection of MPEP, but not by the mGlu1 receptor antagonist (S)-4-carboxy-phenylglycine (4-CPG). Carrageenan-induced inflammatory hyperalgesia was inhibited by pre-treatment of the inflamed hind paw with MPEP, but not following MPEP injection into the contralateral hind paw. Dorsal horn neurones receiving peripheral nociceptive and non-nociceptive afferent input were recorded in anaesthetized rats following microinjection of CHPG into their peripheral receptive fields. CHPG significantly increased the frequency and duration of firing of dorsal horn wide dynamic range (WDR) neurones and this activity was prevented by co-administration of CHPG and MPEP into their receptive fields. Immunohistochemical experiments revealed the co-expression of mGlu5 receptor protein and betaIII tubulin in skin from naive rats, indicating the constitutive expression of mGlu5 receptors on peripheral neurones. Double-labelling of adult rat DRG cells with mGlu5 receptor and vanilloid receptor subtype 1 antisera also supports the expression of mGlu5 receptors on peripheral nociceptive afferents. These results suggest that mGlu5 receptors expressed on the peripheral terminals of sensory neurones are involved in nociceptive processes and contribute to the hyperalgesia associated with inflammation.

Analgesics, Opioid↗

Resident and faculty perceptions of a surgical residency program merger.

To evaluate resident and faculty perceptions of a residency merger process.Survey of faculty and residents of a recently merged general surgical residency. Nineteen separate program characteristics were evaluated via a numerical scoring system, and additional written commentary regarding dominant perceived benefits and detriments of the merger was solicited. Statistical significance was evaluated on numerically scored items by applying the Mann-Whitney U test to median values expressed with interquartile ranges, comparing resident and faculty responses.Scoring system responses from faculty and residents were generally similar. The merger was seen as neutral to positive in its impact on academic issues, but it had more negative effects on issues related to overall program atmosphere and morale. Statistically significant differences between resident and faculty responses were noted in 2 areas: teaching conference timing and overall program effectiveness in preparing for practice. Both of these areas were more favorably impacted by the merger from the residents' perspective, and more negatively as judged by the faculty (p < 0.05). Written commentary by both groups similarly emphasized areas of academic strengthening as a positive effect of the merger, and relationship and morale issues as being more negatively impacted.As reflected by resident and faculty perceptions, program mergers may provide opportunities to strengthen and enhance the academic and clinical foundation of residency. This may, however, occur at the expense of morale and relational issues, which may be negatively impacted by program administrative and geographic expansion.

Journal Article↗

Development, test reliability and validation of a classification for revision hip arthroplasty.

The objective of the study was to develop a valid and reliable classification system for failed hip arthroplasties. The study uses research principals derived from multi-attribute utility theory and consensus group techniques. The development of the severity measure was done in two phases. Phase I of the study included: (a) questionnaire development, (b) submission of the questionnaire to the respondents, (c) data synthesis of the responses and item reduction, and (d) classification development and inter-observer reliability testing. Phase II included: (a) resubmission of the instrument to the respondents for suggestions/feedback, (b) instrument revision by the co-investigators based on the respondents' second feedback, and (c) inter-observer reliability testing and intraoperative validity testing of the instrument. The questionnaires sought to capture expert opinion as to what clinical determinants obtained preoperatively (during patient interview, physical exam and review of plain radiographs - AP pelvis and hip lateral) that would in their clinical experience reveal intraoperative severity. There was an 80% (16/20) response rate from the outside experts invited to participate in the study. Based on item reduction and test retest analysis, a five-grade radiographic classification for the acetabulum as well as the femur was developed. This system was then reviewed by 13 of the initial outside experts (16, 80%) who participated in the first round. Inter-rater reliability testing of the final format of the classification revealed a weighted kappa statistic value of 0.88 between the two-blinded raters (inter-rater reliability) and 0.87 between the blinded raters and the reference standard (intraoperative validity). We conclude that the study developed a reliable and valid radiographic classification system for failed hip arthroplasty.

Arthroplasty, Replacement, Hip↗

Mood changes in adolescents using depot-medroxyprogesterone acetate for contraception: a prospective study.

STUDY OBJECTIVE: Depot-medroxyprogesterone acetate (DMPA) is thought to cause changes in mood among patients using it for contraception. The purpose of this study was to evaluate changes in negative and positive affect among adolescent females using DMPA as a contraceptive agent. DESIGN, SETTING, PARTICIPANTS: This prospective study was set in an urban hospital adolescent clinic. Thirty-nine adolescents choosing DMPA as a contraceptive agent and 24 adolescents not using any hormonal contraception were enrolled as subjects and controls, respectively. Two standardized questionnaires, the Beck Depression Inventory (BDI) and the Multiple Affect Adjective Checklist-Revised (MAACL-R), were administered at baseline to all participants and readministered at 3, 6, and 12 months. MAIN OUTCOME MEASURES: Changes in negative affect from baseline to 3, 6, and 12 months were evaluated by the BDI and by "dysphoria" subscale scores of the MAACL-R. Paired t-tests were used to measure these changes in subjects and controls separately. RESULTS: The mean change in BDI scores from baseline to one year for those who completed one year was -4.8 for subjects (P =.02) and +.3 (P =.84) for controls. The mean change in the dysphoria subscale scores was -5.7 (P =.21) for the subjects and -.1 (P =.98) for the controls while the change in the positive affect scores over a period of one year were -2.1 (P =.46) and +.1 (P =.98) for subjects and controls, respectively. CONCLUSIONS: Adolescents using DMPA do not show depressive symptoms when using DMPA as a contraceptive agent over a period of 12 months as measured by the BDI and show no significant changes in negative or positive affect as measured by the MAACL-R.

Adolescent↗

Efficient infection of primitive hematopoietic stem cells by modified adenovirus.

Almost all studies of adenoviral vector-mediated gene transfer have made use of the adenovirus type 5 (Ad5). Unfortunately, Ad5 has been ineffective at infecting hematopoietic progenitor cells (HPC). Chimeric Ad5/F35 vectors that have been engineered to substitute the shorter-shafted fiber protein from Ad35 can efficiently infect committed hematopoietic cells and we now show highly effective gene transfer to primitive progenitor subsets. An Ad5GFP and Ad5/F35GFP vector was added to CD34(+) and CD34(-)lineage(-) (lin(-)) HPC. Only 5-20% of CD34(+) and CD34(-)lin(-) cells expressed GFP after Ad5 exposure. In contrast, with the Ad5/F35 vector, 30-70% of the CD34(+), 50-70% of the CD34(-)lin(-) and up to 60% of the CD38(-) HPC expressed GFP and there was little evident cellular toxicity. Because of these improved results, we also analyzed the ability of Ad5/F35 virus to infect the hoechst negative 'side population' (SP) of marrow cells, which appear to be among the very earliest multipotent HPC. Between 51% and 80% of marrow SP cells expressed GFP. The infected populations retained their ability to form colonies in two short-term culture systems, with no loss of viability. We also studied the transfer and expression of immunomodulatory genes, CD40L (cell surface expression) and interleukin-2 (secreted). Both were expressed at immunomodulatory levels for >5 days. The ability of Ad5/F35 to deliver transgenes to primitive HPC with high efficiency and low toxicity in the absence of growth factors provides an improved means of studying the consequences of transient gene expression in these cells.

Adenoviridae↗

A prospective case-control study of 50 consecutive patients presenting with hyperlipidaemia.

The literature contains many references which refer to a causal relationship between hyperlipidaemia and hearing loss, but the majority of reports lack adequate controls, or are based on a series of cases which may represent incidental findings. This prospective case-control study compared a restricted population of 50 consecutive hyperlipidaemic patients attending a lipid clinic whose fasting lipid levels were > 2 SDs above the population mean with a control population recruited from patients undergoing nasal surgery for structural abnormalities (n = 159). The National Study of Hearing data was also compared with both groups in order to provide external validity to the control group. This study showed no consistent differences in the hearing thresholds of the hyperlipidaemic group compared with either control group. The Null hypothesis that sensorineural hearing loss is no greater in a population whose fasting blood lipids are raised > 2 SDs above the population mean level than in a control population cannot be refuted given the variability of the data.

Adult↗