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D M Smith

Publications and source records attributed to D M Smith.

At least 55 records · Page 3Linked to original sources
PubMed2001

Enzyme catalysis of 1,2-amino shifts: the cooperative action of B6, B12, and aminomutases.

Ab initio molecular orbital theory is used to investigate 1,2-amino shifts catalyzed by aminomutases, coenzyme B12, and vitamin B6 (in the form of pyridoxal 5'-phosphate or PLP). Our calculations suggest essential catalytic roles for each of B12, B6, and the enzyme in aminomutase-catalyzed reactions. In the first place, coenzyme B12 provides a source of abstracting radicals, allowing the rearrangement reaction to take place on the radical surface. The involvement of radicals is supported by comparison of experimental and theoretical electron paramagnetic resonance parameters. Next, B6 allows the enzyme to lower the barrier height by introducing a double bond (allowing a low-energy intramolecular rearrangement pathway) and by providing a suitable site for partial protonation (preventing overstabilization of the reaction intermediate which could lead to enzyme inactivation). The PLP hydroxyl group is also identified as an important participant in these reactions. Finally, the enzyme holds the various reaction components in place and is the source of acidic functional groups that can provide partial protonation.

Catalysis↗
PubMed2001

Interconversion of (S)-glutamate and (2S,3S)-3-methylaspartate: a distinctive B(12)-dependent carbon-skeleton rearrangement.

The interconversion of (S)-glutamate and (2S,3S)-3-methylaspartate catalyzed by B(12)-dependent glutamate mutase is discussed using results from high-level ab initio molecular orbital calculations. Evidence is presented regarding the possible role of coenzyme-B(12) in substrate activation and product formation via radical generation. Calculated electron paramagnetic resonance parameters support experimental evidence for the involvement of substrate-derived radicals and will hopefully aid the future detection of other important radical intermediates. The height of the rearrangement barrier for a fragmentation-recombination pathway, calculated with a model that includes neutral amino and carboxylic acid substituents in the migrating glycyl group, supports recent experimental evidence for the interconversion of (S)-glutamate and (2S,3S)-3-methylaspartate through such a pathway. Our calculations suggest that the enzyme may facilitate the rearrangement of (S)-glutamate through (partial) proton-transfer processes that control the protonation state of substituents in the migrating group.

Aspartic Acid↗
PubMed2001

Sodium 2-nitroethanaloximate forms a layered-framework structure.

In the title compound, [Na(C(2)H(3)N(2)O(3))], the Na(+) cation lies on a centre of inversion in space group P2(1)/m and all the atoms of the anion lie on a mirror plane. Na is octahedrally coordinated by four O and two N atoms from six different anions and each anion is coordinated to six different Na(+) cations, forming chains of confacial octahedra which link the anion layers. Within these layers, the individual anions are linked by both O-H...O and C-H...O hydrogen bonds.

Journal Article↗
PubMed2001

Medial geniculate, amygdalar and cingulate cortical training-induced neuronal activity during discriminative avoidance learning in rabbits with auditory cortical lesions.

This study addressed the neural mediation of discriminative avoidance learning, wherein rabbits step in a wheel apparatus in response to an acoustic conditional stimulus, the CS+, to avoid a foot shock, and they learn to ignore a different stimulus, the CS-, not followed by foot shock. Previously, muscimol-induced inactivation of the amygdala in the first session of training prevented learning during the inactivation and permanently blocked the development of discriminative training-induced neuronal activity (TIA) in the medial division of the medial geniculate nucleus (MGm). These results suggested that amygdalar neurons induce discriminative TIA in the MGm via basolateral (BL) amygdalar axonal projections to the auditory cortex. To test this hypothesis, the activity of neurons in the MGm was recorded during learning in rabbits with lesions of the auditory cortex. Recordings were also made in the lateral and BL amygdalar nuclei and in the cingulate cortex. In support of the hypothesis, discriminative learning in rabbits with lesions was impaired significantly during early training sessions 1-4; in these same sessions, discriminative TIA was abolished in the MGm, the BL nucleus, and the anterior cingulate cortex. The lesions also blocked posterior cingulate cortical discriminative TIA in training sessions 1-2 but spared TIA in sessions 3-7. Lateral amygdalar neurons showed gradual development of discrimination that was not significantly affected by the lesions. The results demonstrate a critical role of auditory cortex in early discriminative learning and in the production of early discriminative TIA in multiple areas.

Acoustic Stimulation↗
PubMed2001

Understanding the mechanism of B(12)-dependent diol dehydratase: a synergistic retro-push--pull proposal.

Ab initio molecular orbital theory is used to investigate the coenzyme B(12)-dependent reactions catalyzed by diol dehydratase. The key step in such reactions is believed to be a 1,2-hydroxyl migration, which occurs within free-radical intermediates. The barrier for this migration, if unassisted, is calculated to be too high to be consistent with the observed reaction rate. However, we find that "pushing" the migrating hydroxyl, through interaction with a suitable acid, is able to provide significant catalysis. This is denoted retro-push catalysis, the retro prefix signifying that the motion of the migrating group is in the direction opposite to the electron motion. Similarly, the "pulling" of the migrating group, through interaction of the spectator hydroxyl with an appropriate base, is found to substantially reduce the rearrangement barrier. Importantly, the combination of these two effects results in a barrier reduction that is notably greater than additive. This synergistic interplay of the push and the pull provides an attractive means of catalysis. Our proposed retro-push--pull mechanism leads to results that are consistent with isotope-labeling experiments, with experimental rate data, and with the crystal structure of the enzyme.

Catalysis↗
PubMed2001

Ester bond-containing tea polyphenols potently inhibit proteasome activity in vitro and in vivo.

It has been discovered that proteasome inhibitors are able to induce tumor growth arrest or cell death and that tea consumption is correlated with cancer prevention. Here, we show that ester bond-containing tea polyphenols, such as (-)-epigallocatechin-3-gallate (EGCG), potently and specifically inhibit the chymotrypsin-like activity of the proteasome in vitro (IC(50) = 86-194 nm) and in vivo (1-10 microm) at the concentrations found in the serum of green tea drinkers. Atomic orbital energy analyses and high performance liquid chromatography suggest that the carbon of the polyphenol ester bond is essential for targeting, thereby inhibiting the proteasome in cancer cells. This inhibition of the proteasome by EGCG in several tumor and transformed cell lines results in the accumulation of two natural proteasome substrates, p27(Kip1) and IkappaB-alpha, an inhibitor of transcription factor NF-kappaB, followed by growth arrest in the G(1) phase of the cell cycle. Furthermore, compared with their simian virus-transformed counterpart, the parental normal human fibroblasts were much more resistant to EGCG-induced p27(Kip1) protein accumulation and G(1) arrest. Our study suggests that the proteasome is a cancer-related molecular target of tea polyphenols and that inhibition of the proteasome activity by ester bond-containing polyphenols may contribute to the cancer-preventative effect of tea.

Calpain↗
PubMed2001

No unique role for nausea attributed to eating a food in the recalled acquisition of sensory aversion for that food.

Currently in the U.K., as in the U.S.A. 20 years ago, when students were asked to name a food that they did not like, more dislike for the food was attributed to nausea or vomiting after eating than to other gastrointestinal symptoms or to illness in other parts of the body. However, when members of the public first identified adverse symptoms and then attributed them to foods, and dislike for the food was first enquired about on a later occasion, there was no evidence for a unique role for a causal association with nausea in the human acquisition of food aversions. Furthermore, fear of the symptom was more prevalent than acquired sensory aversion when there was more precise recall of memories of the food being followed by nausea or vomiting and greater likelihood of there having been causal contingency rather than mere coincidence. Therefore, the more frequent invocation of nausea than of some other symptoms as the cause of a sensory aversion to a food may result from personal theory of the body, rather than from a veridically recalled occasion when nausea was contingent on eating the food---an event that must occur for aversion to arise from associative conditioning.

Adolescent↗
PubMed2001

Identification of lysine decarboxylase as a mammalian cell growth inhibitor in Eikenella corrodens: possible role in periodontal disease.

The pathogenesis of inflammatory periodontal disease was studied by examining the mechanism of HeLa and HL60 cell growth inhibition by cell-free saline-soluble extracts of Eikenella corrodens and bacterial plaque. Previous studies identified a protein (p80) as causing growth inhibition by E. corrodens extracts. After purification by two-dimensional SDS-PAGE, p80 was digested with protease lysC. Amino acid sequences were obtained and backtranslated for use as PCR primers. A 5840 nucleotide sequence containing a lysine decarboxylase gene was obtained from a Sau3 A1 genomic library of E. corrodens DNA. Lysine decarboxylase activity was present at physiologic pH in the E. corrodens extracts containing p80, and also in bacterial plaque. Both extracts caused growth inhibition by depleting lysine from cell culture media through conversion to cadaverine. Adding lysine, or immune goat IgG to a peptide derived from the active site sequence of E. corrodens lysine decarboxylase, retarded lysine depletion and growth inhibition. epsilon-Amino caproic acid specifically enhanced lysine decarboxylase activity at the low lysine concentration in HL60 cell culture media, and also increased the growth inhibition. Thus, lysine decarboxylases such as p80 inhibit growth by removing lysine from mammalian cell culture media. A new role for lysine decarboxylase activity in the microbial aetiology of periodontal disease is discussed.

Carboxy-Lyases↗
PubMed2001

Lesions in the central nucleus of the amygdala: discriminative avoidance learning, discriminative approach learning, and cingulothalamic training-induced neuronal activity.

The amygdala is critically involved in discriminative avoidance learning. Large lesions of the amygdala block discriminative avoidance learning and abolish cingulothalamic training-induced neuronal activity. These results indicated that amygdalar processing is critical for cingulothalamic plasticity. The larger lesions did not allow differentiation of the specific functioning of various amygdalar nuclei. Anatomical analysis showed that damage in the central (CE) nucleus of the amygdala was correlated with the severity of the behavioral deficit. The present study was carried out to determine whether smaller lesions, centered in the CE nucleus, would impair discriminative avoidance learning and block cingulothalamic plasticity. In addition, the possible role of the CE nucleus in appetitively motivated discriminative approach learning was examined for the first time. New Zealand White rabbits with CE nuclear lesions were first trained in the discriminative approach task. After attaining asymptotic performance, discriminative avoidance training sessions were alternated with continuing approach training sessions, one session each day. The rabbits with lesions were severely impaired in avoidance learning but showed no impairment of approach learning. Surprisingly, the attenuating effects of the lesions on cingulothalamic training-induced neuronal activity were more prevalent during approach learning than during avoidance learning. These results indicated that avoidance learning can be impaired by lesions centered in the CE nucleus that leave cingulothalamic plasticity largely intact and that the CE nucleus is involved in extra-cingulothalamic learning processes.

Amygdala↗
PubMed2001

Studies of the microvascular effects of adrenomedullin and related peptides.

Adrenomedullin (ADM) exerts potent vasoactive effects in the microvasculature. These activities have been most extensively studied in the cutaneous microcirculation. In this review we examine the knowledge gained to date of the ability of ADM to influence microvascular effects that include increased blood flow, microvascular permeability (leading to edema formation), neutrophil accumulation and cutaneous thermal hyperalgesia. ADM is structurally related to the vasodilator neuropeptide calcitonin gene-related peptide (CGRP). The peptides are considered to act via a family of receptor activity modifying proteins (RAMPs) that interact with a G-protein linked receptor, calcitonin receptor-like receptor (CRLR). A correlation of microvascular activity with effects mediated via CRLR and RAMP is discussed.

Adrenomedullin↗
PubMed2001

Adrenomedullin receptors: molecular identity and function.

Since its discovery in 1993 adrenomedullin (AM) has been the subject over 600 published articles. This multifunctional peptide has powerful vasodilator actions and recent evidence from AM gene-deleted mice suggest that AM plays an essential role in vascular development. However the lack of valid AM receptor clones and non-peptide receptor ligands has considerably slowed research progress on this important peptide. In this review we have focused on the proposition that the calcitonin receptor-like receptor (CRLR) is a receptor both for AM and the related vasoactive peptide calcitonin gene-related peptide (CGRP). The receptor activity modifying proteins (RAMPs) that are essential for defining CRLR pharmacology will also be discussed. We will describe how AM receptors have been reported to signal and be regulated and to consider whether further receptors for AM beyond CRLR/RAMP combinations might exist.

Adrenomedullin↗
PubMed2001

ACTIVE: a cognitive intervention trial to promote independence in older adults.

The Advanced Cognitive Training for Independent and Vital Elderly (ACTIVE) trial is a randomized, controlled, single-masked trial designed to determine whether cognitive training interventions (memory, reasoning, and speed of information processing), which have previously been found to be successful at improving mental abilities under laboratory or small-scale field conditions, can affect cognitively based measures of daily functioning. Enrollment began during 1998; 2-year follow-up will be completed by January 2002. Primary outcomes focus on measures of cognitively demanding everyday functioning, including financial management, food preparation, medication use, and driving. Secondary outcomes include health-related quality of life, mobility, and health-service utilization. Trial participants (n = 2832) are aged 65 and over, and at entry into the trial, did not have significant cognitive, physical, or functional decline. Because of its size and the carefully developed rigor, ACTIVE may serve as a guide for future behavioral medicine trials of this nature.

Activities of Daily Living↗
PubMed2001

Cloning and expression of a novel cysteine-rich secreted protein family member expressed in thyroid and pancreatic mesoderm within the chicken embryo.

We have isolated a new chicken gene that is a member of the cysteine-rich secreted protein family (CRISP). The CRISP family is composed of over 70 members that are found in many phyla of organisms, including: vertebrates, plants, fungi, yeast, and insects. Here we describe the cloning of a novel member of this family, SugarCrisp, and its expression pattern throughout chicken embryogenesis. We also describe its utility as a marker of thyroid and pancreatic mesoderm in the developing chicken embryo and its expression within the human and mouse in glandular tissue.

Amino Acid Sequence↗
PubMed2001

Denaturation and aggregation of myosin from two bovine muscle types.

The thermal behaviors of myosin from bovine vastus intermedius (VI, predominantly red muscle) and semimembranosus (SM, predominantly white muscle) at pH 6.05 (ultimate pH of VI muscle) and 5.50 (ultimate pH of SM muscle) were compared. Differential scanning microcalorimetry and turbidity measurements were used to monitor changes in myosin during heating from 25 to 80 degrees C at 1 degrees C/min. VI and SM myosin heavy chain isoforms were identified on gradient SDS-PAGE. Endotherms of VI myosin at pH 6.05 had three transition temperatures (T(m)) of 45, 53, and 57 degrees C, whereas at pH 5.50 two transitions were observed at 42 and 59 degrees C. SM myosin had two T(m) values of 46 and 58 degrees C at pH 6.05 and T(m) values of 43 and 62 degrees C at pH 5.5. SM myosin at its ultimate pH was less heat stable than VI myosin at its ultimate pH; however, when SM and VI myosin were compared at the same pH, VI myosin was less stable.

Animals↗
PubMed2001

Heat-induced gelation of chicken Pectoralis major myosin and beta-lactoglobulin.

The denaturation, aggregation, and rheological properties of chicken breast muscle myosin, beta-lactoglobulin (beta-LG), and mixed myosin/beta-LG solutions were studied in 0.6 M NaCl, 0.05 mM sodium phosphate buffer, pH 7.0, during heating. The endotherm of a mixture of myosin and beta-LG was identical to that expected if the endotherm of each protein was overlaid on the same axis. The maximum aggregation rate (AR(max)) increased, and the temperature at the AR(max) (T(max)) and initial aggregation temperature (T(o)) decreased as the concentration of both proteins was increased. The aggregation profile of <0.5% myosin was altered by the presence of 0.25% beta-LG. Addition of 0.5-3.0% beta-LG decreased storage moduli of 1% myosin between 55 and 75 degrees C, but increased storage moduli (G') when heated to 90 degrees C and after cooling. beta-LG had no effect on the gel point of > or =1.0% myosin, but enhanced gel strength when heated to 90 degrees C and after cooling. After cooling, the G' of 1% myosin/2%beta-LG gels was about 1.7 times greater than that of gels prepared from 2% myosin/1% beta-LG.

Animals↗
PubMed2001

Elisa to quantify hexanal-protein adducts in a meat model system.

Monoclonal antibodies (MAb) were produced to hexanal-bovine serum albumin conjugates. An indirect competitive ELISA was developed with a detection range of 1-50 ng of hexanal/mL. Hexanal conjugated to three different proteins was recognized, whereas free hexanal and the native proteins were not detected. The antibody cross-reacted with pentanal, heptanal, and 2-trans-hexenal conjugated to chicken serum albumin (CSA) with cross-reactivities of 37.9, 76.6, and 45.0%, respectively. There was no cross-reactivity with propanal, butanal, octanal, and nonanal conjugated to CSA. The hexanal content of a meat model system was determined using MAb and polyclonal antibody-based ELISAs and compared with analysis by a dynamic headspace gas chromatographic (HS-GC) method and a thiobarbituric acid reactive substances (TBARS) assay. Both ELISAs showed strong correlations with the HS-GC and TBARS methods. ELISAs may be a fast and simple alternative to GC for monitoring lipid oxidation in meat.

Aldehydes↗
PubMed2001

Arylamine N-acetyltransferase of Mycobacterium tuberculosis is a polymorphic enzyme and a site of isoniazid metabolism.

Arylamine N-acetyltransferases (NATs; E.C 2.3.1.5) N-acetylate arylhydralazine and arylamine substrates using acetyl coenzyme A. Human NAT2 acetylates and inactivates the antituberculosis drug, isoniazid (INH), and is polymorphic. We previously demonstrated that there is a homologue of human NAT2 in Mycobacterium tuberculosis, whose product N-acetylates INH in vitro. We now demonstrate that the nat gene is expressed in M. tuberculosis and M. bovis Bacille Calmette-Guerin (BCG), using reverse transcription-polymerase chain reaction and Western blotting. The NAT protein is active in M. bovis BCG in vivo, as detected by the presence of N-acetyl INH in M. bovis BCG lysates grown in INH. Sequence analysis of the M. tuberculosis nat coding region reveals a single nucleotide polymorphism in 18% of a random cohort of M. tuberculosis clinical isolates, conferring a G to R change. The recombinant mutant protein appears less stable than the wild type, and has an apparent affinity for INH of 10-fold less than the wild type. Modelling the change in M. tuberculosis NAT shows that the G to R change is close to the active site, and supports the experimental findings. Minimum inhibitory concentration data suggest that this polymorphism in nat is linked to low-level changes in the INH susceptibility of M. tuberculosis clinical isolates.

Acetylation↗