Search PubMed⌕ Search

Biomedical subjects

P Harris

Publications and source records attributed to P Harris.

At least 37 records · Page 2Linked to original sources
PubMed2001

Crystallization and preliminary X-ray analysis of maltose O-acetyltransferase.

Maltose O-acetyltransferase (Mac) is a member of the hexapeptide-repeat family of enzymes, which contains proteins with left-handed parallel beta-helix architecture forming homotrimers. Diffraction data for four well diffracting crystal forms were collected. Crystal form I diffracted beyond 1.53 A resolution but was perfectly merohedrally twinned with an apparent space group P622. Crystal forms II and III (space groups R3 and C2, respectively) could be obtained under very similar conditions by adjusting the buffer pH differently. Crystal forms II and III had several monomers in the asymmetric unit and were difficult to derivatize. However, during soaking with trimethyl lead acetate, the form III crystals dissolved and crystals with a different habit and space group grew in their place (form IV). In three of the crystal forms, a ladder of peaks was visible in the native Patterson maps along the c axis. These peaks were interpreted as corresponding to the vectors between the beta-strands in the turns of the beta-helix. Crystal form IV is suitable for structure determination of Mac exploiting the anomalous scattering of lead.

Acetyltransferases↗
PubMed2001

Selenomethionine substitution of orotidine-5'-monophosphate decarboxylase causes a change in crystal contacts and space group.

Orotidine 5'-monophosphate decarboxylase (ODCase) catalyses the decarboxylation of orotidine 5'-monophosphate to uridine 5'-monophosphate, the last step in the de novo biosynthesis of uridine 5'-monophosphate. In order to determine the structure of ODCase from Escherichia coli by the multi-wavelength anomalous dispersion technique, both native and SeMet-substituted proteins have been produced and purified. During the production of SeMet ODCase, it was observed that SeMet was the only amino acid that it was necessary to add to the defined medium during expression. SeMet-substituted ODCase in complex with the inhibitor 1-(5'-phospho-beta-D-ribofuranosyl)barbituric acid crystallizes under similar conditions as the native enzyme. In contrast to the native enzyme, where the crystals belong to the orthorhombic space group P2(1)2(1)2(1), the SeMet-substituted enzyme crystallizes in the monoclinic space group P2(1), with a quadrupling of the volume of the asymmetric unit. Despite the drastic difference in symmetry, the overall crystal packing is effectively identical in the two crystal forms. The change in space group appears to originate in differences in the crystal contacts near the SeMet and Met residues. These differences can be rationalized in terms of SeMet's larger size and hydrophobicity.

Amino Acid Substitution↗
PubMed2001

Steady state visually evoked potential (SSVEP) topography in a graded working memory task.

The steady state visually evoked potential (SSVEP) elicited by a diffuse 13-Hz visual flicker was recorded from 64 scalp sites in 30 subjects performing a low and high demand version of an object working memory task. During the perceptual component of the task, the SSVEP amplitude was reduced at left and right parieto-occipital sites. During the hold or memory component of the task, the SSVEP amplitude exhibited a load-dependent increase at frontal and occipito-parietal sites, while the SSVEP latency exhibited a load-dependent reduction at central and left frontal sites. We suggest that SSVEP amplitude changes index cortical information processing modes in that perceptual processes are associated with an SSVEP amplitude reduction, while holding information in active short-term or working memory is associated with an SSVEP amplitude increase. We also discuss changes in SSVEP amplitude and latency in terms of changes in the behavior of cortico-cortico and thalamo-cortico loops that utilize cortical layer I. Such cortico-cortico and thalamo-cortical loops are also proposed to constitute a neurophysiological mechanism for holding information in working memory.

Adult↗
PubMed2001

Long-term risk stratification for survivors of acute coronary syndromes. Results from the Long-term Intervention with Pravastatin in Ischemic Disease (LIPID) Study. LIPID Study Investigators.

OBJECTIVES: We developed a prognostic strategy for quantifying the long-term risk of coronary heart disease (CHD) events in survivors of acute coronary syndromes (ACS). BACKGROUND: Strategies for quantifying long-term risk of CHD events have generally been confined to primary prevention settings. The Long-term Intervention with Pravastatin in Ischemic Disease (LIPID) study, which demonstrated that pravastatin reduces CHD events in ACS survivors with a broad range of cholesterol levels, enabled assessment of long-term prognosis in a secondary prevention setting. METHODS: Based on outcomes in 8,557 patients in the LIPID study, a multivariate risk factor model was developed for prediction of CHD death or nonfatal myocardial infarction. Prognostic indexes were developed based on the model, and low-, medium-, high- and very high-risk groups were defined by categorizing the prognostic indexes. RESULTS: In addition to pravastatin treatment, the independently significant risk factors included: total and high density lipoprotein cholesterol, age, gender, smoking status, qualifying ACS, prior coronary revascularization, diabetes mellitus, hypertension and prior stroke. Pravastatin reduced coronary event rates in each risk level, and the relative risk reduction did not vary significantly between risk levels. The predicted five-year coronary event rates ranged from 5% to 19% for those assigned pravastatin and from 6.4% to 23.6% for those assigned placebo. CONCLUSIONS: Long-term prognosis of ACS survivors varied substantially according to conventional risk factor profile. Pravastatin reduced coronary risk within all risk levels; however, absolute risk remained high in treated patients with unfavorable profiles. Our risk stratification strategy enables identification of ACS survivors who remain at very high risk despite statin therapy.

Aged↗
PubMed2001

Tuberin-dependent membrane localization of polycystin-1: a functional link between polycystic kidney disease and the TSC2 tumor suppressor gene.

The PKD1 gene accounts for 85% of autosomal dominant polycystic kidney disease (ADPKD), the most common human genetic disorder. Rats with a germline inactivation of one allele of the Tsc2 tumor suppressor gene developed early onset severe bilateral polycystic kidney disease, with similarities to the human contiguous gene syndrome caused by germline codeletion of PKD1 and TSC2 genes. Polycystic rat renal cells retained two normal Pkd1 alleles but were null for Tsc2 and exhibited loss of lateral membrane-localized polycystin-1. In tuberin-deficient cells, intracellular trafficking of polycystin-1 was disrupted, resulting in sequestration of polycystin-1 within the Golgi and reexpression of Tsc2 restored correct polycystin-1 membrane localization. These data identify tuberin as a determinant of polycystin-1 functional localization and, potentially, ADPKD severity.

Alleles↗
PubMed2001

Challenging behaviour in community services.

The implementation of community care in the UK has led to the requirement that services should be able to meet the needs of adults with intellectual disability (ID) and additional needs in terms of challenging behaviour. However, the extent to which people with challenging behaviour are present in the community and the extent to which community services can support them effectively still requires significant research. The present study examines the prevalence of challenging behaviour amongst adults with ID residing in three London boroughs and the issues which arise from service delivery to this client group. All service providers and general practitioners in the area were contacted and asked to identify any individuals with ID and challenging behaviour. All responses were screened, and then key staff were interviewed for information on a range of demographic factors and on the Checklist of Challenging Behaviour. The reliability of the instrument was also assessed. Four hundred and forty-eight individuals were identified from a total borough population of 670 000. There was consistency in the types of behaviour which were frequently identified across the three boroughs. There were significant levels of self-injury as well as a range of behaviours of the 'hard to engage' type. Most individuals had more then one challenging behaviour and some individuals with seriously aggressive behaviour used local community services. Twenty-five per cent of the sample lived at home with their families and 50% were in community residential services. The boroughs differed in their ability to manage those with challenging behaviour in that one borough had many more people placed out-of-borough. Significant numbers of individuals with challenging behaviour were living in the community. The range and number of behaviours suggest that staff need to be very skilled in supporting such individuals, and that effective planning and support are essential if people with challenging behaviour are to be maintained in community settings.

Activities of Daily Living↗
PubMed2001

Theory of mind development in deaf children: a nonverbal test of false-belief understanding.

Our aim in this study was to investigate whether previous findings pointing to a delay in deaf children's theory of mind development are replicated when linguistic demands placed on the deaf child are minimized in a nonverbal version of standard false-belief tasks. Twenty-four prelingually deaf, orally trained children born of hearing parents were tested with both a verbal and a nonverbal version of a false-belief task. Neither the younger (range: 4 years 7 months-6 years 5 months) nor the older (range: 6 years 9 months-11 years 11 months) children of the final sample of 21 children performed above chance in the verbal task. The nonverbal task significantly facilitated performance in children of all ages. Despite this facilitation, we observed a developmental delay: only the older group performed significantly above chance in the nonverbal false-belief task, even though the younger children were at the average age when hearing children normally pass standard false-belief tests. We discuss these findings in light of the hypothesis that language development and conversational competence are crucial to the acquisition of a theory of mind.

Journal Article↗
PubMed2001

Cardiac markers and point-of-care testing: a perfect fit.

Biochemical markers are of increasing importance in diagnostic strategies for ruling in and ruling out acute myocardial infarction (AMI), particularly when electrocardiographic (ECG) findings do not allow a diagnosis. Point-of-care testing (POCT) or "near-patient" testing allows for diagnostic assays to be performed at the site of patient care delivery. The biochemical markers that are commonly used by physicians to aid in the diagnosis of AMI are myoglobin, CK-MB, troponin I, and troponin T. Currently available POCT assays possess comparable diagnostic performance to laboratory-based cardiac marker assays. This provides an opportunity for POCT to evolve into the standard of care for evaluating the greater than 6 million American patients presenting with chest discomfort. Full acceptance of this relatively new technology will not be realized until users reach a comfort level where utilization of these devices is foolproof and they have faith in the results.

Biomarkers↗
PubMed2001

Proinflammatory cytokine expression in gastric tissue from children with Helicobacter pylori-associated gastritis.

BACKGROUND: Helicobacter pyloriinfection of the gastric mucosa in humans is usually acquired early in life. The chronic inflammation that ensues involves the increased production of inflammatory cytokines. Published data on production of these mediators by gastric mucosa of H. pylori-infected children are few. METHODS: Seventy-nine children, aged 5 to 18 years, referred for upper gastrointestinal endoscopy to four separate hospitals in Chile, were studied. The concentrations of interleukin (IL)-1beta, IL-6, IL-8, and tumor necrosis factor alpha were measured in homogenates of gastric mucosal biopsy specimens. Cytokine expression was confirmed by reverse transcription polymerase chain reaction. These data were correlated with the patients' clinical, histologic and sociodemographic status. RESULTS: Patient rate of colonization by H. pylori was inversely correlated with socioeconomic status (P < 0.005) and positively correlated with age (P < 0.0025). In gastric mucosa, concentrations of IL-1beta, IL-8, and tumor necrosis factor alpha were all significantly higher in H. pylori-positive patients than in H. pylori-negative patients and in patients who had histologic gastritis than in those with normal gastric mucosa. In patients with peptic ulcer disease, only IL-1beta and IL-8 concentrations were significantly elevated when compared with those of patients without ulcers. Interleukin-6 concentrations were comparable among the different groups analyzed. CONCLUSIONS: This study suggests that increased gastric mucosal production of the proinflammatory cytokines IL-1beta and IL-8 is probably involved in H. pylori-associated gastric damage in children and may be crucial in determining the different clinical outcomes.

Adolescent↗
PubMed2001

Arg389Gly beta 1-adrenoceptor polymorphism varies in frequency among different ethnic groups but does not alter response in vivo.

There are marked interethnic differences in beta 1-adrenoceptor-mediated responsiveness, with sensitivity decreased in African-Americans and increased in Chinese compared with Caucasians. Therefore, the frequency of a common naturally occurring polymorphism of the human beta 1-adrenoceptor gene (Arg389Gly), which has functional importance in vitro, was determined in 194 African-Americans, 316 Caucasian-Americans, 221 Hispanic-Americans and 142 Chinese. African-Americans were found to have a significantly lower frequency of the Arg389 allele than the other three ethnic groups (all P < 0.01). In the populations studied, the order of the distribution of the Arg389 allele was: Chinese (74%) > Caucasians (72%) > Hispanics (67%) > African-Americans (58%). To determine the functional significance of the Arg389Gly beta 1-adrenoceptor polymorphism, in-vivo heart rate responses to exercise were compared in healthy subjects homozygous for the Arg (n = 9) and Gly (n = 8) alleles. Heart rate response to exercise was not affected by genotype (P = 0.4). Although ethnic differences in the frequency of the beta 1-adrenoceptor Arg389Gly polymorphism exist, the polymorphism does not appear to have functional significance in healthy subjects and therefore may not contribute to ethnic differences in response to drugs acting through the beta 1-adrenoceptor.

Alleles↗
PubMed2001

Recovery from transport and acclimatisation of competition horses in a hot humid environment.

The aims of the present field-based study were to investigate changes in fit horses undergoing acclimatisation to a hot humid environment and to provide data on which to base recommendations for safe transport and acclimatisation. Six horses (age 7-12 years) were flown from Europe to Atlanta and underwent a 16 day period of acclimatisation. Exercise conditions during acclimatisation (wet bulb globe temperature index 27.6+/-0.0 [mean +/- s.e.]) were more thermally stressful compared with the European climate from which the horses had come (22.0+/-1.8, P<0.001). Following the flight, weight loss was 4.1+/-0.8% bodyweight and took around 7 days to recover. Water intake during the day was significantly increased (P<0.05) compared with night during acclimatisation. Daily mean exercise duration was 72+/-12 min and the majority of work was performed with a heart rate below 120 beats/min. Respiratory rate (fR) was increased (P<0.05) throughout acclimatisation compared with in Europe, but resting morning (AM) and evening (PM) rectal temperature (TREC), heart rate (fC) and plasma volume were unchanged. White blood cell (WBC) count was significantly increased at AM compared with in Europe on Days 4 and 10 of acclimatisation (P<0.01), but was not different by Day 16. In conclusion, horses exposed to hot humid environmental conditions without prior acclimatisation are able to accommodate these stresses and, with appropriate management, remain fit and clinically healthy, without significant risk of heat illness or heat-related disorders, provided they are allowed sufficient time to recover from transport, acclimatisation is undertaken gradually and they are monitored appropriately.

Acclimatization↗
PubMed2001

[Identification of mutation in the gene cystic fibrosis transmembrane regulator (CFTR) in Chilean patients with cystic fibrosis].

BACKGROUND: Cystic fibrosis (CF) is an autosomal recessive disease caused by mutations in the CFTR gene, that codes for a chloride channel located in the apical surface of epithelial cells. The main role of this protein is the regulation of chloride transport, and secondarily, of sodium and water to the extracellular space. More than 900 gene mutations have been described, and their relative frequency in different populations depends on their ethnic origin. AIM: To report the findings of Chilean patients with cystic fibrosis, in whom the presence of 20 common mutations was analyzed. PATIENTS AND METHODS: Fifty seven patients with established diagnosis or suspicion of CF were studied. The simultaneous identification of 20 mutations and the normal delta F508 allele was done using polymerase chain reactions with a commercial assay. RESULTS: Eight mutations were found. Fifty patients fulfilled diagnostic criteria proposed by the Consensus Panel of the CF Foundation and 66% of alleles were identified in this group. delta F508 mutation was found in 45%. We did not identify mutations in any of the remaining 7 patients. CONCLUSIONS: Our results suggest that the majority of undetected mutations are associated with atypical phenotypes or that some patients in this series could have other diseases. We recommend to include mutation analysis in the evaluation of Chilean patients with CF. It is useful to establish prognosis and genetic counselling.

Adolescent↗
PubMed2001

[Importance of the neuropsychological assessment of memory in the clinical practice].

Memory assessment is used to diagnose possible disturbances of this function and to relate the findings to their neurological implications and to the issues of clinical treatment and prognosis. Neuropsychological examination of memory function involves (a) the administration of objective procedures that are proven sensitive to the effects of brain injury, and specific to the changes due to functional impairment, and (b) the integration of statistical and observational findings that demonstrate a consistent pattern of results that are commonly seen with specific brain disorders. Analysis of the pattern of performance among the tests is key to a good understanding of the results; thus, the selection of tests used should sample a wide range of memory domains. This paper analyzes the most frequently measuring instruments in neuropsychological assessment of memory, as well as the main batteries and tests used to evaluate the existence or absence of mnesic deficits.

Humans↗
PubMed2000

Structural basis for the catalytic mechanism of a proficient enzyme: orotidine 5'-monophosphate decarboxylase.

Orotidine 5'-monophosphate decarboxylase (ODCase) catalyzes the decarboxylation of orotidine 5'-monophosphate, the last step in the de novo synthesis of uridine 5'-monophosphate. ODCase is a very proficient enzyme [Radzicka, A., and Wolfenden, R. (1995) Science 267, 90-93], enhancing the reaction rate by a factor of 10(17). This proficiency has been enigmatic, since it is achieved without metal ions or cofactors. Here we present a 2.5 A resolution structure of ODCase complexed with the inhibitor 1-(5'-phospho-beta-D-ribofuranosyl)barbituric acid. It shows a closely packed dimer composed of two alpha/beta-barrels with two shared active sites. The orientation of the orotate moiety of the substrate is unambiguously deduced from the structure, and previously proposed catalytic mechanisms involving protonation of O2 or O4 can be ruled out. The proximity of the OMP carboxylate group with Asp71 appears to be instrumental for the decarboxylation of OMP, either through charge repulsion or through the formation of a very short O.H.O hydrogen bond between the two carboxylate groups.

Amino Acid Sequence↗