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K Harris

Publications and source records attributed to K Harris.

At least 19 recordsLinked to original sources
PubMed2006

Are surgical stainless steel wires used for intracranial implantation of PrPsc a good model of iatrogenic transmission from contaminated surgical stainless steel instruments after cleaning?

Transmissible spongiform encephalopathies are a group of fatal, neurodegenerative diseases commonly known as prion diseases. Prion diseases can resist traditional inactivation strategies and may be iatrogenically transmitted by surgical instruments through the human population. These properties have led to the need for a suitable detection method of the prion infectious agent, and increased pressure regarding the development of anti-prion cleaning methodologies that would ensure the safety of surgical instruments. Although other techniques have been applied, the animal bioassay remains the 'gold standard' method for assessing infectivity. As the vast majority of surgical instruments are made of stainless steel, and in order to test this surface using the animal bioassay, the application of very thin surgical stainless steel wires has been widely adopted. These wires are easily inoculated and may be reimplanted into animals without the requirement for elution of the residual material. However, their comparability to the dimensions, shape and size of surgical instruments is questionable. This article shows how such contaminated wires (residual protein between 6.3 and 16.0 ng/mm(2)) can be cleaned more easily than flat metal surfaces (residual protein between 63.9 and 89.3 ng/mm(2)) under comparable conditions using recommended cleaning agents. These results indicate that the application of wires as a realistic means of assessing the removal or inactivation of the prion infectious agent from surgical instruments should be treated with caution.

Animals↗
PubMed2006

The sensitivity of approved Ninhydrin and Biuret tests in the assessment of protein contamination on surgical steel as an aid to prevent iatrogenic prion transmission.

Regulations recommend the routine application of biochemical tests, such as the Ninhydrin or Biuret tests, to confirm the efficacy of hospital sterile service department (SSD) washer-disinfector cycles in removing proteinaceous material, particularly with respect to prions. The effectiveness of these methods relies on both the effective sampling of the instruments and the sensitivity of the tests employed. Two commercially available contamination assessment tests were evaluated for their sensitivity to ME7 brain homogenate on surgical-grade stainless steel surfaces. Controls were visualized by the application of episcopic differential interference contrast/Epi-fluorecence microscopy (EDIC/EF) combined with the sensitive fluorescent reagent, SYPRO Ruby, which has been shown previously to rapidly visualize and assess low levels of contamination on medical devices. The Ninhydrin test displayed a minimum level of detection observed by 75% of volunteers (MLD(75)) of 9.25 microg [95% confidence interval (95% CI) 8.6-10.0 microg]. The Biuret test provided better sensitivity, with a MLD(75) of 6.7 microg (95% CI 5.4-8.2 microg). However, much lower concentrations of proteinaceous soiling (pg) were visualized using the EDIC/EF microscopy method. From these findings, it is clear that these approved colorimetric tests of cleaning are relatively insensitive. This investigation demonstrates how large amounts (up to 6.5 microg) of proteinaceous brain contamination could remain undetected and the instruments deemed clean using such methods. The application of more sensitive cleanliness evaluation methods should be applied to reduce the risk of iatrogenic transmission of prion disease in 'high-risk' instruments such as neurosurgical devices.

Biuret Reaction↗
PubMed2006

Validation of the use of organic acids and acidified sodium chlorite to reduce Escherichia coli O157 and Salmonella typhimurium in beef trim and ground beef in a simulated processing environment.

A study was conducted to determine if acidified sodium chlorite (1,200 ppm) and acetic and lactic acids (2 and 4%) were effective in reducing foodborne pathogens in beef trim prior to grinding in a simulated processing environment. The reduction of Salmonella Typhimurium and Escherichia coli O157:H7 at high (4.0 log CFU/g) and low (1.0 log CFU/g) inoculation doses was evaluated at various processing steps, including the following: (i) in trim just after treatment application, (ii) in ground beef just after grinding, (iii) in ground beef 24 h after refrigerated storage, (iv) in ground beef 5 days after refrigerated storage, and (v) in ground beef 30 days after frozen storage. All antimicrobial treatments reduced the pathogens on the trim inoculated with the lower inoculation dose to nondetectable numbers in the trim and in the ground beef. There were significant reductions of both pathogens in the trim and in the ground beef inoculated with the high inoculation doses. On the trim itself, E. coli O157:H7 and Salmonella Typhimurium were reduced by 1.5 to 2.0 log cycles, with no differences among all treatments. In the ground beef, the organic acids were more effective in reducing both pathogens than the acidified sodium chlorite immediately after grinding, but after 1 day of storage, there were no differences among treatments. Overall, in the ground beef, there was a 2.5-log reduction of E. coli O157:H7 and a 1.5-log reduction of Salmonella Typhimurium that was sustained over time in refrigerated and frozen storage. Very few sensory differences between the control samples and the treated samples were detected by a consumer panel. Thus, antimicrobial treatments did not cause serious adverse sensory changes. Use of these antimicrobial treatments can be a promising intervention available to ground beef processors who currently have few interventions in their process.

Acetic Acid↗
PubMed2005

Ultra-fast three dimensional imaging of hyperpolarized 13C in vivo.

OBJECTIVE: PASADENA, a chemical method of enhancing nuclear spin polarization has demonstrated 13C polarizations of order unity for the nascent products of molecular addition by parahydrogen. The extreme brevity of signal enhancement obtained by hyperpolarization requires improved 13C MR in vivo imaging techniques for their optimum utility. MATERIALS AND METHODS: 13C imaging sequences, including 13C 3D FIESTA, were compiled for a GE LX 1.5 T clinical MR scanner. Two water soluble 13C imaging agents were hyperpolarized utilizing parahydrogen and an automated polarizer. 13C polarization was quantified in flow phantoms and in rats with jugular vein catheters. RESULTS: Fast 3D FIESTA 13C MR imaging technique acquired sequential 3D images (3.66 s/acquisition) with superior SNR. Hyperpolarized 13C solutions and vascular phantoms achieved a maximum signal of 26,624+/-593. In vivo 13C MR images of the cardiopulmonary circulation showed maximum 13C signal of 2,402+/-158. 13C images acquired within 3.66 s showed signal enhancement over 10,000 compared to equilibrium polarization. CONCLUSION: 3D-FIESTA was effective for sub-second in vivo imaging of hyperpolarized 13C reagents produced in a custom-built parahydrogen polarizer. Application to 13C hyperpolarized by parahydrogen is demonstrated in vitro and in vivo.

Algorithms↗
PubMed2005

Plasma levobupivacaine concentrations following scalp block in patients undergoing awake craniotomy.

BACKGROUND: Levobupivacaine is an effective local anaesthetic agent for nerve blockade with less systemic toxicity than racemic bupivacaine. The safety and efficacy of levobupivacaine for scalp blockade during awake craniotomy have not been addressed previously. METHODS: Serial arterial plasma levobupivacaine concentrations following scalp blockade were measured to 2 h in 10 patients booked for awake craniotomy for epilepsy or tumour surgery. Bilateral scalp blockade providing surgical anaesthesia was achieved with a mean dose of 177 mg (2.5 mg kg(-1), range 1.6-3.2 mg kg(-1)) of levobupivacaine (0.5%, 5 mg ml(-1)) with epinephrine (5 microg ml(-1)) added immediately before the block insertion. RESULTS: The maximum measured plasma levobupivacaine concentration was 1.58 (0.44) microg ml(-1) [mean (SD)] with a mean time to peak plasma concentration of 12 (4) min. There were no episodes in any of the 10 patients of symptoms or signs suggestive of either CNS or CVS toxicity. CONCLUSIONS: This study demonstrated a relatively rapid rise of plasma levobupivacaine concentration without evidence of cardiovascular or central nervous system sequelae in a sample population of patients who may be particularly prone to perioperative seizures.

Adult↗
PubMed2005

Quantitative measures of cluster quality for use in extracellular recordings.

While the use of multi-channel electrodes (stereotrodes and tetrodes) has allowed for the simultaneous recording and identification of many neurons, quantitative measures of the quality of neurons in such recordings are lacking. In multi-channel recordings, each spike waveform is discriminated in a high-dimensional space, making traditional measures of unit quality inapplicable. We describe two measures of unit isolation quality, Lratio and Isolation Distance, and evaluate their performance using simulations and tetrode recordings. Both measures quantified how well separated the spikes of one cluster (putative neuron) were from other spikes recorded simultaneously on the same multi-channel electrode. In simulations and tetrode recordings, both Lratio and Isolation Distance discriminated well- and poorly-separated clusters. In data sets from the rodent hippocampus in which neurons were simultaneously recorded intracellularly and extracellularly, values of Isolation Distance and Lratio were related to the correct identification of spikes.

Animals↗
PubMed2002

Nociceptin/orphanin FQ modulation of ionic conductances in rat basal forebrain neurons.

Nociceptin/orphanin FQ (N/OFQ) is an endogenous opioid-like heptadecapeptide that plays an important role in a variety of physiological functions. N/OFQ and its receptor opioid receptor-like orphan receptor-1 are abundant in the diagonal band of Broca (DBB), a basal forebrain nucleus where the loss of cholinergic neurons is linked to memory and spatial learning deficits. In the whole animal, central injections of N/OFQ have been shown to disrupt spatial learning. In this study, we investigated the basis for these behavioral observations by examining the cellular effects of N/OFQ on chemically identified DBB neurons. Whole cell patch-clamp recordings were performed on enzymatically dissociated DBB neurons. Under voltage-clamp conditions, bath application of N/OFQ (10 pM-1 microM) resulted in a dose-dependent depression of whole cell currents. Single cell reverse transcription-polymerase chain reaction analysis identified cholinergic and fewer GABAergic cells to be N/OFQ-responsive. [Nphe(1)]nociceptin-(1-13)-NH(2) and CompB (J-113397) antagonized the N/OFQ response, but both compounds also displayed partial agonist activity. Using a combination of channel blockers we determined that the effects of N/OFQ were mediated via a suite of Ca(2+) (N- and L-type) and Ca(2+)-dependent K(+) (iberiotoxin-sensitive) conductances. In addition, biophysical analysis of voltage subtraction protocols revealed that N/OFQ reduces transient outward and the delayed rectifier K(+) currents. Because N-type and L-type Ca(2+) channels are important in the context of neurotransmitter release, our observations indicate that N/OFQ inhibition of Ca(2+)-dependent conductances in cholinergic neurons would be expected to result in depression of acetylcholine release, which may explain the behavioral actions of N/OFQ in the brain.

Acetylcholine↗
PubMed2001

Role of scaffolds in MAP kinase pathway specificity revealed by custom design of pathway-dedicated signaling proteins.

BACKGROUND: Signal transduction pathways with shared components must be insulated from each other to avoid the inappropriate activation of multiple pathways by a single stimulus. Scaffold proteins are thought to contribute to this specificity by binding select substrates. RESULTS: We have studied the ability of scaffold proteins to influence signaling by the yeast kinase Ste11, a MAPKKK molecule that participates in three distinct MAP kinase pathways: mating, filamentation, and HOG. We used protein fusions to force Ste11 to associate preferentially with a subset of its possible binding partners in vivo, including Ste5, Ste7, and Pbs2. Signaling became confined to a particular pathway when Ste11 was covalently attached to these scaffolds or substrates. This pathway bias was conferred upon both stimulus-activated and constitutively active forms of Ste11. We also used membrane-targeted derivatives of the mating pathway scaffold, Ste5, to show that stimulus-independent signaling initiated by this scaffold remained pathway specific. Finally, we demonstrate that loss of pathway insulation has a negative physiological consequence, as nonspecific activation of both the HOG and mating pathways interfered with proper execution of the mating pathway. CONCLUSIONS: The signaling properties of these kinase fusions support a model in which scaffold proteins dictate substrate choice and promote pathway specificity by presenting preferred substrates in high local concentration. Furthermore, insulation is inherent to scaffold-mediated signaling and does not require that signaling be initiated by pathway-specific stimuli or activator proteins. Our results give insight into the mechanisms and physiological importance of pathway insulation and provide a foundation for the design of customized signaling proteins.

MAP Kinase Kinase Kinases↗
PubMed2001

Initial sequencing and analysis of the human genome.

The human genome holds an extraordinary trove of information about human development, physiology, medicine and evolution. Here we report the results of an international collaboration to produce and make freely available a draft sequence of the human genome. We also present an initial analysis of the data, describing some of the insights that can be gleaned from the sequence.

Animals↗
PubMed2001

Contrast-enhanced coronary MR angiography: relationship between coronary artery delineation and blood T1.

Contrast-enhanced coronary angiography has become an important technique for magnetic resonance (MR) coronary artery imaging. However, the relationship between the quality of the coronary artery images and blood T1 has not yet been fully explored. In this paper, we assessed this relationship in an animal model by using a prototypical blood pool agent. With accumulated injections of this agent, the blood T1 would be maintained at different levels. The measured blood T1 values in vivo were 147 +/- 3, 82 +/- 6, 48 +/- 4, 40 +/- 3, and 30 +/- 8 msec (N = 7). Fixed and variable flip angle schemes were used in coronary artery imaging. The signal to noise ratios (SNR) of coronary arteries were measured and the image quality was assessed. It was found that blood T1 less than 80 msec might be desired. No statistically significant difference was observed between two flip angle schemes. There was better vessel definition using variable flip angle at blood T1 lower than 50 msec. Understanding this relationship may be beneficial to optimizing image protocol and/or design of blood pool contrast agents for contrast-enhanced coronary angiography.

Animals↗
PubMed2001

Three-dimensional MR pulmonary perfusion imaging and angiography with an injection of a new blood pool contrast agent B-22956/1.

Initial evaluation of a new blood pool agent, B-22956/1, for pulmonary imaging was performed in five domestic pigs with artificial embolism. Pre-embolism 3D pulmonary perfusion images were first acquired by injecting an extravascular agent, gadoteridol. The pulmonary arteries of the pigs were then occluded by the artificial emboli. Post-embolism perfusion scans were subsequently performed by injecting B-22956/1. Additional post-embolism high-spatial-resolution angiograms were also acquired. Parenchyma perfusion deficits were well depicted in the post-embolism perfusion maps. The post-embolism angiography clearly revealed the location and extent of the filling defects in the pulmonary vessels. Signal intensities of perfusion maps on the normal parenchyma were significantly improved (30%) by using B-22956/1, in comparison with perfusion images using gadoteridol (P < 0.01). Many pulmonary angiograms with approximately equal contrast could be obtained even at 22 minutes after the injection of B-22956/1. Our initial results indicate that blood pool agent B-22956/1 may provide opportunities for whole-lung-coverage perfusion mapping and additional high-resolution target angiograms after a single injection.

Animals↗
PubMed2001

Predictors of length of stay after coronary stenting.

BACKGROUND: Postprocedure length of stay (LOS) remains an important determinant of medical costs after coronary stenting. Variables that predict LOS in this setting have not been well characterized. METHODS: We evaluated 359 consecutive patients who underwent coronary stenting with antiplatelet therapy. Sequential multiple linear regression (MLR) models were constructed with use of 4 types of variables to predict log-transformed LOS: preprocedure, intraprocedure, and postprocedure factors and adverse outcomes. RESULTS: Preprocedure factors alone explained more than one third of the variability in postprocedure LOS (adjusted R(2) = 0.37). The addition of procedural variables added little to the model (adjusted R(2) = 0.39). Entering nonoutcome postprocedure variables significantly enhanced the predictive capacity of the model, explaining more than half the variability in postprocedure LOS (adjusted R(2) = 0.54). In the final model, addition of outcome variables increased its predictive capacity only slightly (adjusted R(2) = 0.61). In this model, significant preprocedure factors included: myocardial infarction (MI) within 24 hours, MI within 1 to 30 days, women with peripheral vascular disease, intravenous heparin, and chronic atrial fibrillation. High-risk intervention was the only significant intraprocedure variable. Significant postprocedure factors included periprocedure ischemia; cerebrovascular accident or transient ischemic attack; treatment with intravenous heparin or nitroglycerin or intra-aortic balloon pump; and need for blood transfusion. Significant adverse outcomes included contrast nephropathy, gastrointestinal bleeding, arrhythmia, vascular complication, and repeat angiography. CONCLUSION: This prediction model identifies a number of potentially reversible factors responsible for prolonging LOS and may enable the development of more accurate risk-adjusted methods with which to improve or compare care.

Coronary Disease↗
PubMed2001

The effects of an ACE inhibitor and a calcium antagonist on the progression of renal disease: the Nephros Study.

BACKGROUND: The renoprotective effect of ACE inhibition in chronic renal disease is well established but the studies on effects of calcium antagonists on progression of renal disease and on proteinuria have given varying results. METHODS: We conducted an open long-term randomized prospective multi-centre study comparing the combination of ramipril and felodipine ER (F) with either drug alone in non-diabetic renal disease. Included were patients with uncontrolled hypertension (diastolic blood pressure (DBP)) > or =95 mmHg on treatment with a diuretic and a beta-blocker. Fifty-one patients received the combination of R and F, 54 patients R, and 53 patients F. The treatment goal was a DBP <90 mmHg and a similar BP reduction in the three groups. Mean doses at the last visit were 5+5, 10 and 9 mg, respectively, after a mean treatment time of nearly 2 years. The progression of renal impairment was studied by serial measurements of serum creatinine, iohexol clearance, and albuminuria. RESULTS: The reduction in supine systolic (S) BP and DBP expressed as median values were -19.0/-14.5,-14.3/-15.0 and -13.5/-13.3 mmHg in the R+F, R, and F groups, respectively. There was no significant difference between the groups. When correction for the acute drug effect was performed the R+F group had a slower progression rate of the renal disease (loss of glomerular filtration rate (GFR) ml/min/year) compared with the F group (P<0.05) but not to the R group (P>0.20). There was a rise in albuminuria after 2 years in the F group (P<0.05), but no significant change was found in the other groups. CONCLUSIONS: In patients with non-diabetic renal disease the combination of an ACE inhibitor and a calcium antagonist in reduced doses used in addition to baseline therapy with beta-blockers and diuretics, tended to cause a better BP reduction as each drug per se. The R+F treatment also caused a slower progression of the renal disease compared with F alone. The combination treatment seems to afford better BP control and appears to be a favourable therapeutic option in patients with renal disease and hypertension.

Adult↗
PubMed2001

Inspired CO(2) and O(2) in sleeping infants rebreathing from bedding: relevance for sudden infant death syndrome.

Some infants sleep facedown for long periods with no ill effects, whereas others become hypoxemic. Rebreathing of expired air has been determined by CO(2) measurement; however, O(2) levels under such conditions have not been determined. To evaluate this and other factors influencing inspired gas concentrations, we studied 21 healthy infants during natural sleep while facedown on soft bedding. We measured gas exchange with the environment and bedding, ventilatory response to rebreathing, and concentrations of inspired CO(2) and O(2). Two important factors influencing inspired gas concentrations were 1) a variable seal between bedding and infants' faces and 2) gas gradients in the bedding beneath the infants, with O(2)-poor and CO(2)-rich air nearest to the face, fresher air distal to the face, and larger tidal volumes being associated with fresher inspired air. Minute ventilation increased significantly while rebreathing because of an increase in tidal volume, not frequency. The measured drop in inspired O(2) was significantly greater than the accompanying rise in inspired CO(2). This appears to be due to effects of the respiratory exchange ratio and differential tissue solubilities of CO(2) and O(2) during unsteady conditions.

Air↗
PubMed2001

Cellular mechanisms for amyloid beta-protein activation of rat cholinergic basal forebrain neurons.

The deposition of amyloid beta-protein (Abeta) in the brain and the loss of cholinergic neurons in the basal forebrain are two pathological hallmarks of Alzheimer's disease (AD). Although the mechanism of Abeta neurotoxicity is unknown, these cholinergic neurons display a selective vulnerability when exposed to this peptide. In this study, application of Abeta(25-35) or Abeta(1-40) to acutely dissociated rat neurons from the basal forebrain nucleus diagonal band of Broca (DBB), caused a decrease in whole cell voltage-activated currents in a majority of cells. This reduction in whole cell currents occurs through a modulation of a suite of potassium conductances including calcium-activated potassium (I(C)), the delayed rectifier (I(K)), and transient outward potassium (I(A)) conductances, but not calcium or sodium currents. Under current-clamp conditions, Abeta evoked an increase in excitability and a loss of accommodation in cholinergic DBB neurons. Using single-cell RT-PCR technique, we determined that Abeta actions were specific to cholinergic, but not GABAergic DBB neurons. Abeta effects on whole cell currents were occluded in the presence of membrane-permeable protein tyrosine kinase inhibitors, genistein and tyrphostin B-44. Our data indicate that the Abeta actions on specific potassium conductances are modulated through a protein tyrosine kinase pathway and that these effects are selective to cholinergic but not GABAergic cells. These observations provide a cellular basis for the selectivity of Abeta neurotoxicity toward cholinergic basal forebrain neurons that are at the epicenter of AD pathology.

Actins↗