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Biomedical subjects

K E Smith

Publications and source records attributed to K E Smith.

At least 19 recordsLinked to original sources
PubMed1998

BstB7SI (R decreases CCGGY), a thermostable isoschizomer of Cfr10I, from a strain of Bacillus stearothermophilus isolated from oil-contaminated soil in Kuwait.

Isolates of thermophilic bacteria from desert soil in Kuwait, heavily contaminated with crude oil, have been screened for the presence of restriction endonuclease activity. One of the isolates (B7S), identified as Bacillus stearothermophilus, showed a high level of restriction endonuclease activity when a cell-free extract was incubated with lambda bacteriophage DNA at 65 degrees C. A type II restriction endonuclease (BstB7SI) has been partially purified from this isolate. BstB7SI recognises the six-base sequence RCCGGY (R = A or G; Y = T or C) and hydrolyses the phosphodiester bond in both strands of the DNA substrate between the first and second bases of the recognition sequence 5'-R decreases CCGGY-3'producing four-base 5' overhangs. BstB7SI is therefore an isoschizomer of the mesophilic prototype restriction endonuclease Cfr10I. BstB7SI has a pH optimum of 9.7, requires 10 mM MgCl2 and 75 mM NaCl for maximum activity, and retains full enzyme activity when incubated for 5 min at temperatures up to 70 degrees C.

Base Sequence↗
PubMed1998

Below-knee residual limb shape change measurement and visualization.

OBJECTIVE: The ability to measure and visualize shape change (deformation) of a residual limb within and between prostheses is an important step toward improved prosthetic fit assessment. The objective was to develop measurement and visualization methods for below-knee residual limb soft tissue shape change after donning and loading a prosthesis to detect small shape changes (30mm or less). DESIGN: Spiral X-ray computed tomography imaging was utilized to acquire 3D volumetric data of the below-knee residual limb and prosthesis in situ from poor- and a good-fitting prostheses without and with a load. A new sum projection depth-shaded cylindrical mapping technique to measure and visualize small changes in shape was developed. From the volumetric data, the relative displacement of small lead markers placed on the residual limb's skin surface were measured using multiplanar reconstruction images and cylindrical maps. Displacement measurements averaging 15mm or less were obtained. RESULTS: The precision and accuracy was 1mm and 2mm, respectively, when measuring the shape change or deformation of the skin surface from the sum projection cylindrical maps. The skin surface deformation was at least 7mm or greater when comparing marker locations between scans with the prostheses in situ. CONCLUSION: These new image-based measurement and visualization methods provide a feasible means for measuring and displaying lower extremity residual limb shape change within and between different prostheses with and without loading.

Adult↗
PubMed1998

The relation of change in maternal interactive styles to the developing social competence of full-term and preterm children.

This study evaluated the changing nature of mothers' interactive behaviors to understand alterations in children's social development across 6, 12, 24, and 40 months of age. Social skills were observed during daily activities and toy play in the home for medically high risk (HR; n = 73) and low risk (LR; n = 114) very low birthweight (VLBW) preterm and full-term (FT; n = 112) children. Variations in mothers' responses to children's changing capabilities predicted rates of change in children's social skills. For example, mothers who showed higher levels of maintaining measured across 6 to 40 months had children who displayed greater increases in initiating, but this was more apparent in daily activities than toy play and for the VLBW children compared to the FT children. Those VLBW children at the highest degrees of biological risk displayed faster gains in initiating than the other groups when their mothers provided even greater levels of support. Results demonstrate the importance of using methodologies that test more complex models of growth when evaluating parent-child relations.

Child Development↗
PubMed1997

Expression cloning of a rat hypothalamic galanin receptor coupled to phosphoinositide turnover.

The neuropeptide galanin is widely distributed throughout the central and peripheral nervous systems and participates in the regulation of processes such as nociception, cognition, feeding behavior, and insulin secretion. Multiple galanin receptors are predicted to underlie its physiological effects. We now report the isolation by expression cloning of a rat galanin receptor cDNA distinct from GALR1. The receptor, termed GALR2, was isolated from a rat hypothalamus cDNA library using a 125I-porcine galanin (125I-pGAL) binding assay. The GALR2 cDNA encoded a protein of 372 amino acids exhibiting 38% amino acid identity with rat GALR1. Binding of 125I-pGAL to transiently expressed GALR2 receptors was saturable (KD = 0.15 nM) and displaceable by galanin peptides and analogues in rank order: porcine galanin approximately M32 approximately M35 approximately M40 >/= galanin-(1-16) approximately M15 approximately [D-Trp2]galanin-(1-29) > C7 >> galanin-(3-29). This profile resembles that of the rat GALR1 receptor with the notable exception that [D-Trp2]galanin exhibited significant selectivity for GALR2 over GALR1. Activation of GALR2 receptors with porcine galanin and other galanin analogues increased inositol phospholipid turnover and intracellular calcium levels in stably transfected Chinese hamster ovary cells and generated calcium-activated chloride currents in Xenopus oocytes, suggesting that the rat GALR2 receptor is primarily coupled to the activation of phospholipase C.

Amino Acid Sequence↗
PubMed1997

The role of child-centered perspectives in a model of parenting.

The mediating role of mothers' child-centered perspectives was examined in a longitudinal study of 323 children. The conceptual model of parenting was tested to determine whether maternal perspectives mediated the relations between the parenting resources of social support, child-rearing history, and self-esteem and the child's developmental level with parenting behavior. This conceptual model was compared to alternative models using structural equation modeling. Results indicate that mothers' perspectives directly related to parenting behavior in two different contexts as well as mediated the relations between maternal resources and behavior. Maternal self-esteem also mediated the relation between social support and child-rearing history with child-centered perspectives. Results support the importance of examining child-centered perspectives as an influence on parental competence as well as the importance of examining how parenting resources interrelate with one another to impact parenting behavior.

Child Development↗
PubMed1997

Lower extremity residual limb slippage within the prosthesis.

OBJECTIVE: The ability to measure skin surface slippage relative to the internal wall of a lower extremity prosthesis under various loading conditions is important for evaluation of socket fit and function, and creation of finite element models. Skin surface three-dimensional (3D) shape measurements with tracking of fiducial displacements in situ under axial loading of the prosthesis have not previously been reported. Analysis of slippage within the prosthesis has been performed using a new experimental measurement method based on spiral x-ray computed tomography (SXCT) imaging. DESIGN: Small lead markers were placed on the residuum of an adult with a below-knee amputation, and SXCT scans were obtained with the prosthesis in situ under two static axial loading conditions (44.5N and 178N). The 3D scan data were used to assess slippage with three methods: gross displacement of the tibia and distal end of the residuum; relative displacement of markers; and distance measurements between markers. RESULTS: The markers affixed to the below-knee skin surface within the prosthesis were measured. The skin slipped from 2 to 6 mm relative to the internal prosthesis wall when an additional load of 133.5N was applied in the axial direction to the distal end of the prosthesis. The tibial remnant moved 10.0 mm distally relative to the prosthesis internal wall. CONCLUSION: This method provides a feasible means for measuring residuum skin slippage relative to the prosthesis and skin deformation relative to tibia within an in situ prosthesis under load.

Artificial Limbs↗
PubMed1997

Finite element modeling and experimental verification of lower extremity shape change under load.

Prediction and measurement of residuum shape change inside the prosthesis under various loading conditions is important for prosthesis design and evaluation. Residual limb surface measurements with the prosthesis in situ were used for construction of a finite element model (FEM). These surface measurements were obtained from volumetric computed tomography. A new experimental method for modeling the shape of the in situ lower residual limb was developed based on spiral X-ray computed tomography (SXCT) imaging. The p-version of the finite element method was used for estimating the material properties from known load and displacement data. A homogeneous, isotropic, linear constitutive model with accommodation of the constitutive soft and hard tissues of the residuum was evaluated with static axial loading applied to the in situ prosthesis and compared with experimental results obtained in a human volunteer. Two FEMs were created for similar coronal cross sections of the below knee residuum under two loading conditions. Agreement between observed (from SXCT) and predicted (from FEA) residual limb shape changes inside the prosthesis were maximized with a single modulus of elasticity for the residuum soft tissue of 0.06 MPa, consistent with previously published results. This methodology provides a framework to predict and objectively evaluate FEMs and determine residuum material properties by inverse methods.

Anthropometry↗
PubMed1997

Distribution of the neuropeptide Y Y2 receptor mRNA in rat central nervous system.

Our group has recently reported the expression cloning of the human neuropeptide Y Y2 receptor DNA and subsequently the cloning of the rat homologue. These studies have made it possible to localize the mRNA encoding this NPY receptor subtype in rat tissues. We have, thus, carried out in situ hybridization studies, using radiolabeled oligonucleotide probes to the rat Y2 receptor mRNA, to determine the distribution of Y2 mRNA in rat brain and limited peripheral ganglia. Probe specificity was confirmed by testing antisense and sense probes in transfected cells. In rat brain, hybridization signals obtained with the antisense probes were discrete and were restricted to neuronal profiles in specific subregions of the cortex, hippocampus, amygdala, thalamus, hypothalamus, mesencephalon and pons. Among the regions exhibiting the most intense labeling were the CA3 region of the hippocampus, the arcuate nucleus of the hypothalamus and layer 3 of the piriform cortex. Other regions containing labeled neurons included the medial amygdala, the centromedial thalamic nucleus, the dorsal raphe, the dorsal motor nucleus of the vagus and the trigeminal ganglion. The present results indicate that the mRNA encoding the Y2 receptor is discretely localized in the rat brain and that the distribution is generally consistent with previous radioligand-binding studies. This study should help clarify the relationship between the Y2 receptor distribution and functional studies of NPY receptor subtype classification and provides further evidence for the involvement of the Y2 receptor in multiple physiological processes.

Animals↗
PubMed1997

A structure-activity analysis of the cloned rat and human Y4 receptors for pancreatic polypeptide.

We cloned and expressed the rat Y4 receptor for pancreatic polypeptide (PP). Structure-activity profiles derived from 125I-PP binding assays and [cAMP] radioimmunoassays reveal a selective receptor interaction with rat PP vs. neuropeptide Y (NPY) or peptide YY (PYY). Rat and human Y4 receptor clones share 75% amino acid identity. Based on [cAMP] radioimmunoassay, the human Y4 receptor exhibits a less selective interaction with rat PP vs. NPY or PYY and a greater dependence on N-terminal PP residues, relative to rat Y4. Differences in sequence and structure-activity profiles suggest the rat be used with caution to model human Y4 receptor function.

Animals↗
PubMed1997

Predicting cognitive-language and social growth curves from early maternal behaviors in children at varying degrees of biological risk.

Growth modeling was used to examine the relation of early parenting behaviors (averaged across 6 and 12 months) with rates of change in children's cognitive-language and social response and initiating skills assessed at 6, 12, 24, and 40 months. Groups of full-term (n = 112) and very low birth weight children, divided into medically low (n = 114) and high risk (HR; n = 73), were included to evaluate whether children who vary in their rate of development are influenced in different ways by early parenting styles. Parenting behaviors that were sensitive to children's focus of interest and did not highly control or restrict their behaviors predicted greater increases and faster rates of cognitive-language and social development, with relations stronger for the HR versus the other two groups. These maternal behaviors may provide the support all infants need to establish an optimal early foundation for later development and the specific support HR children need to learn in spite of early attentional and organizational problems.

Adult↗
PubMed1997

The use of carrion as breeding sites by the blowfly Lucilia sericata and other Calliphoridae.

To examine the species composition of flies breeding in carrion in the field, the carcasses of mice and quail were exposed on sheep farms in the South West of England. Calliphora vicina was the dominant species of Diptera; 19,294 individuals emerged from 175 of the 241 infested carcasses recovered. Lucilia sericata emerged from only 39 of the infested carcasses, at a median of 10 adults per infested carcass. Other species of Lucilia present were L.ampullacea, L.caesar and L.illustris. The highest number of L.sericata emerged from carcasses placed in open pasture, the highest numbers of C.vicina emerged from carcasses in hedgerow, whereas the highest numbers of L.caesar, L.ampullacea and L.illustris emerged from carcasses in woodland. The duration of exposure of carcasses in the field was negatively related to the size of the adult L.sericata which emerged and, in woodland and hedgerow habitats, to the number of L.sericata which emerged. These data indicate that the larvae of L.sericata in carcasses experience significant levels of competition and that the intensity of this competition may be sufficient to reduce the numbers of L.sericata able to emerge successfully. The size distributions of female L.sericata which emerged from carcasses or which were caught as adults in the field showed only a small degree of overlap, suggesting that only a relatively small proportion of the wild L.sericata population emerge from carcass breeding sites. The results are discussed in relation to the development of new approaches to the control of blowfly strike of sheep.

Animals↗
PubMed1997

The potential for control of the blowfly Lucilia sericata using odour-baited targets.

A deterministic simulation model is used to explore the potential for control of the sheep blowfly, Lucilia sericata, using odour-baited targets. The simulations show that the ambient temperatures of 15 degrees C or 20 degrees C, 13% and 24% of the females present in a population would need to be killed per day, respectively, to prevent the population growing from its initial spring density during the summer blowfly season. In contrast, if both sexes could be sterilized, so that sterilized males were also able to disrupt the fertility of unsterilized females, only 6.5-13% of both sexes would need to be attracted and sterilized at 15 degrees C or 20 degrees C respectively. To examine whether these numbers of individuals could be attracted by targets baited with liver and sodium sulphide, mark-release-recapture studies in sheep pastures were used to quantify sticky target efficiencies. The percentage of the L.sericata population caught on any one day was shown to be positively related to average ambient temperature. At 15 degrees C each sticky target caught approximately 13% and at 20 degrees C, 41% of the females within a 20 m radius. The results indicate that, at ambient temperatures between 15 and 20 degrees C, if distributed at a density of approximately 5/ha, targets baited with liver and sodium sulphide could effectively suppress a population of L.sericata. Lower densities of targets would be required if they were able to sterilize both sexes rather than kill or if more attractive synthetic semiochemical baits could be developed.

Animals↗
PubMed1996

Decreased levels of CD44 protein and mRNA in prostate carcinoma. Correlation with tumor grade and ploidy.

BACKGROUND: CD44, a transmembrane protein, is associated with cell-cell and cell-matrix interaction and with tumor growth and metastasis. Expression of both standard form and variant isoforms of CD44 protein has been associated with aggressive behavior and metastasis in various tumors, but has not been characterized in prostate adenocarcinoma (PAC). METHODS: The expression of CD44 standard (CD44s) and splice variant v3, v4/5, v6, v7/8, and v10 proteins were studied in 109 PACs and correlated with tumor grade, DNA ploidy, and mRNA levels. Monoclonal antibodies against the various CD44 proteins were applied to microwave irradiated, formalin fixed, paraffin embedded sections. The DNA content of the tumors was evaluated by the Feulgen method with the CAS200 Image Analyzer. Total RNA exhibiting 18s and 28s bands was derived from two benign prostatic tissues and 5 PACs exhibiting decreased levels of CD44 protein by immunohistochemistry. The RNA was analyzed with reverse transcriptase polymerase chain reaction using CD44 specific primers. RESULTS: The basal cells of the benign prostatic acini revealed uniform membranous staining for CD44s, v3, and v6 in 95-97% of cases. Similar staining was observed for v4/5, v7/8, and v10 in 40%, 30%, and 2% of cases, respectively. Secretory epithelial cells of the benign prostatic acini showed predominant expression of CD44s (97% of cases). Staining for CD44 variant proteins (v3, v4/5, v6, v7/8, and v10) in this location ranged from 9-22% of cases. Approximately 70% of the PACs showed significant loss of CD44s expression, which correlated with high tumor grade (Gleason > or = 7) (P = 0.01) and aneuploid status (P = 0.002). In 93-98% of the PACS, there was a complete lack of membranous expression for all CD44 variant isoforms. The metastatic PACS did not show preferential expression of either the standard form or any variant isoform. The cDNA from the normal prostates yielded a prominent CD44 standard form polymerase chain reaction product at 482 base pair (bp) and variant isoforms at approximately 650 and 850 bp. No CD44 products could be amplified from the subset of five PAC cDNAs, even when present at four-fold excess. CONCLUSIONS: PACS exhibit down-regulation of CD44 protein expression, which correlates with high tumor grade and aneuploidy. v6 and v3 isoforms were preferentially expressed in the basal cells of benign prostatic acini. Based on a subset of cases, loss of CD44 protein expression is associated with decreased abundance of CD44 mRNA.

Adenocarcinoma↗
PubMed1996

A receptor subtype involved in neuropeptide-Y-induced food intake.

Neuropeptide Y (NPY) is a powerful stimulant of food intake and is proposed to activate a hypothalamic 'feeding' receptor distinct from previously cloned Y-type receptors. This receptor was first suggested to explain a feeding response to NPY and related peptides, including NPY2-36, that differed from their activities at the Y1 receptor. Here we report the expression cloning of a novel Y-type receptor from rat hypothalamus, which we name Y5. The complementary DNA encodes a 456-amino-acid protein with less than 35% overall identity to known Y-type receptors. The messenger RNA is found primarily in the central nervous system, including the paraventricular nucleus of the hypothalamus. The extent to which selected peptides can inhibit adenylate cyclase through the Y5 receptor and stimulate food intake in rats correspond well. Our data support the idea that the Y5 receptor is the postulated 'feeding' receptor, and may provide a new method for the study and treatment of obesity and eating disorders.

Amino Acid Sequence↗
PubMed1996

Distribution of a rat galanin receptor mRNA in rat brain.

In situ hybridization histochemistry has been employed to determine the distribution of the mRNA encoding a recently cloned rat galanin receptor (rGalR1). The galanin receptor mRNA has been found to be discretely localized in rat brain. The most intense hybridization signals were found over neurons in the nucleus of the lateral olfactory tract, in the ventral posterior hippocampus, and in the lateral external subdivision of the parabrachial nucleus. A number of other brain regions also contain significant hybridization signals, including the hypothalamus, brain stem and spinal cord. The localization of rGalR1 mRNA indicates that this receptor may play a role in the varied functions ascribed to GAL, among them feeding, cognition and modulation of sensory information.

Animals↗