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Biomedical subjects

P Nelson

Publications and source records attributed to P Nelson.

At least 55 records · Page 3Linked to original sources

Immobilized metal ion affinity chromatography on Co2+-carboxymethylaspartate-agarose Superflow, as demonstrated by one-step purification of lactate dehydrogenase from chicken breast muscle.

A rapid method for the purification of lactate dehydrogenase from whole chicken muscle extract in one chromatographic step is reported. The purification procedure can be accomplished in less than 1 h. A new type of immobilized metal ion affinity chromatography adsorbent is used that can be utilized at linear flow rates higher than 5 cm/min. The final preparation of the enzyme was with purity higher than 95% as ascertained by SDS-PAGE. Three immobilized metal ions (Ni2+, Zn2+ and Co2+) were compared for their binding properties towards the purified enzyme. The binding site of the enzyme for immobilized intermediate metal ions was determined after cleavage with CNBr and binding studies of the derivative peptides on immobilized Co2+. A peptide located on the N-terminus of the enzyme, implicated in the binding, has great potential as a purification tag in fusion proteins.

Amino Acid Sequence↗

A comparison of ocular blood flow in untreated primary open-angle glaucoma and ocular hypertension.

PURPOSE: To compare ocular blood flow in untreated primary open-angle glaucoma and ocular hypertension using scanning laser Doppler flowmetry and pulsatile ocular blood flow. METHOD: Fourteen ocular hypertensive subjects and 10 patients with primary open-angle glaucoma were matched for intraocular pressure, mean arterial blood pressure, and age. They had scanning laser Doppler flowmetry images taken centered on the optic disk. Pulsatile ocular blood flow readings were performed in sitting, standing, and supine positions. No subjects were receiving topical antiglaucoma treatment, systemic beta-blockers, calcium antagonists, or nitrates at the time of measurement. RESULTS: Laser Doppler flowmetry results showed a significant reduction in blood velocity, volume, and flow at the lamina cribrosa and the temporal neuroretinal rim in glaucoma compared to ocular hypertension (P < .05). No difference was found between the groups at the nasal neuroretinal rim or the nasal juxtapapillary retina. There was a significant increase in minimum velocity (P = .03) at the temporal juxtapapillary retina in glaucoma compared to ocular hypertension. The ocular pulse amplitude, pulse volume, and pulsatile ocular blood flow were significantly lower (P < .05) in the glaucoma group compared to ocular hypertension in sitting and standing positions. CONCLUSION: Having controlled for factors known to affect perfusion pressure, we found evidence of reduced ocular blood flow in primary open-angle glaucoma compared with ocular hypertension. Our findings indicate a reduction in choroidal and short posterior ciliary artery circulation in primary open-angle glaucoma. Whether these changes in blood flow are a cause or a consequence of glaucomatous optic atrophy is still unknown.

Aged↗

Dynamic excitations in membranes induced by optical tweezers.

We present the phenomenology of transformations in lipid bilayers that are excited by laser tweezers. A variety of dynamic instabilities and shape transformations are observed, including the pearling instability, expulsion of vesicles, and more exotic ones, such as the formation of passages. Our physical picture of the laser-membrane interaction is based on the generation of tension in the bilayer and loss of surface area. Although tension is the origin of the pearling instability, it does not suffice to explain expulsion of vesicles, where we observe opening of giant pores and creeping motion of bilayers. We present a quantitative theoretical framework to understand most of the observed phenomenology. The main hypothesis is that lipid is pulled into the optical trap by the familiar dielectric effect, is disrupted, and finally is repackaged into an optically unresolvable suspension of colloidal particles. This suspension, in turn, can produce osmotic pressure and depletion forces, driving the observed transformations.

Biophysical Phenomena↗

New measurements of DNA twist elasticity.

The symmetries of the DNA double helix require a new term in its linear response to stress: the coupling between twist and stretch. Recent experiments with torsionally constrained single molecules give the first direct measurement of this new material parameter. We extract its value from a recent experiment. Finally, we sketch the effect of constrained twist on entropic elasticity of DNA arising from the connection between Link, Twist, and Writhe.

DNA↗

Diminished linear growth during intermittent calcitriol therapy in children undergoing CCPD.

Daily calcitriol therapy has been reported to improve linear growth in children with renal bone disease, and 1,25-dihydroxyvitamin D is a key regultor of chondrocyte proliferation and differentiation. Whereas large intermittent doses of calcitriol can lower serum parathyroid hormone (PTH) levels and reverse the skeletal changes of secondary hyperparathyroidism, the impact of intermittent calcitriol therapy on linear growth in children is not known. Thus, we studied 16 pre-pubertal patients with bone biopsy-proven secondary hyperparathyroidism who completed a 12-month prospective clinical trial of intermittent calcitriol therapy. Biochemical results and growth data obtained during intermittent calcitriol therapy were compared to values determined during the preceding 12 months of daily calcitriol therapy in each study subject; changes in bone histology were assessed after one year of intermittent calcitriol therapy. Z-scores for height did not change during 12 months of daily calcitriol therapy. Although the skeletal lesions of secondary hyperparathyroidism improved in most patients, Z-scores for height decreased from -1.8 +/- 0.32 to -2.0 +/- 0.33, P < 0.01, during intermittent calcitriol therapy. The largest reductions were seen in patients who developed adynamic bone lesions after 12 months of treatment. Delta Z-scores for height correlated with serum PTH, r = 0.71, P < 0.01, and alkaline phosphatase levels, r = 0.67, P < 0.01, during intermittent calcitriol therapy but not during daily calcitriol therapy. The data suggest that high dose intermittent calcitriol therapy adversely affects linear growth, particularly in patients with the adynamic lesion. The higher doses of calcitriol or the intermittent schedule of calcitriol administration may directly inhibit chondrocyte activity within growth plate cartilage of children with end-stage renal disease.

Adolescent↗

Increases in salivary lysozyme and IgA concentrations and secretory rates independent of salivary flow rates following viewing of humorous videotape.

Several studies have shown increases in salivary, secretory IgA (sIgA) levels following exposure to humor and various relaxation strategies. However, the use of sIgA as a marker of immunocompetence has been questioned. In this study we investigated the potential of an alternative marker, salivary lysozyme (sLys) in addition to sIgA. sIgA and sLys concentrations and secretion rates were measured from saliva samples collected immediately prior to and alter viewing a humorous (situation comedy) and a control videotape (documentary). sLys concentrations and secretion rates were significantly increased after viewing of the humorous videotape (p = .0007 and p = .009, respectively) but not following viewing of the control video. Similarly, increases were obtained for sIgA (p = .01 and p = .05) following viewing of the humorous tape. sIgA concentration decreased following viewing of the control tape (p = .05). Salivary flow rates did not significantly change following the viewing of either the humorous or the control tapes. Changes following the humorous video were significantly different from changes following the control video for both sLys and sIgA levels (concentration, p = .007 and p = .005; secretory rate p = .02 and p = .03), but not for salivary flow rates. The results suggest an enhancement of the innate and specific mucosal immune parameters following exposure to humor, independent of salivary flow. Use of sLys may serve as a useful additional or alternative marker of the effects of humor on mucosal immunity.

Journal Article↗

Heart rate variability reflects severity of COPD in PiZ alpha1-antitrypsin deficiency.

BACKGROUND: Analysis of heart rate variability (HRV) is a powerful method of assessing severity of conditions affecting the autonomic nervous system. STUDY OBJECTIVE: To determine if HRV is decreased and if HRV reflects severity in COPD. DESIGN: Prospective determination of HRV from 24-h outpatient Holter recordings. PATIENTS: Eighteen individuals with PiZ alpha1-antitrypsin deficiency: 13 with COPD and 5 with normal FEV1. HRV was also determined in 18 matched normal control subjects. Approximately 3 years after the initial recording, all COPD subjects were contacted to determine current status. MEASUREMENTS: Indexes of heart rate (HR) and HRV were compared for groups of patients with and without COPD and their control subjects. RESULTS: Mean and minimum HRs were higher in COPD patients. Virtually all indexes of HRV were significantly decreased in COPD patients. No differences were found in HR or HRV between PiZ individuals with normal FEV1 and their age-and gender-matched control subjects. Patients who had a change in status (ie, death, lung transplant, listed for transplant) had significantly higher daytime HRs, lower values for HRV indexes reflecting mixed sympathetic and parasympathetic modulation of HR, and reduced daytime high-frequency spectral power, an index of cardiac vagal modulation. Significant correlations (r=0.48 to 0.88) were found between FEV1 and these and other indexes of HRV. Most other indexes of HRV also tended to be lower for the group whose status had changed. CONCLUSION: PiZ alpha1-antitrypsin deficiency COPD is associated with abnormal cardiac autonomic modulation. Indexes of HRV appear to reflect severity and may have prognostic value in COPD patients.

Adult↗

Lactation inhibits the potentiating effect of galanin upon the GnRH-induced LH release observed in diestrous-1 rat.

Recent demonstrations of no changes in hypothalamic gonadotropin releasing hormone (GnRH) gene expression and GnRH levels detected at the pituitary gland in diestrous and lactating rats, indicate that lactational hypogonadotropism in this species is not associated with inhibition of hypothalamic GnRH synthesis and secretion. Hypothalamic galanin potentiates GnRH effects on luteinizing hormone (LH) secretion in male and cycling rats. To explore the interaction between GnRH and galanin during lactation, we studied in vitro the effects of pulsatile stimulation with those peptides upon LH synthesis and secretion from rat pituitaries on diestrous 1 or day 10 of lactation. Hemipituitaries were separately incubated in 1 ml Dulbecco's Minimal Essential Medium supplemented with 1% penicillin-streptomycin and fetal calf serum, at 37 degrees C in 5% CO2-air. The hemipituitaries were stimulated during 12 h with hourly pulses, 6 min each, of (a) gonadotropin releasing hormone (GnRH 25 ng/pulse), (b) rat galanin (600 ng/pulse), (c) GnRH plus galanin, or (d) saline. Medium was collected before each pulse to determine LH by radioimmunoassay. After the 12 h pulsatile regime total RNA was extracted and both actin and beta-LH mRNA were determined by reverse transcriptase polymerase chain reaction. There was a significant stimulation of LH secretion by GnRH (ANOVA, p < 0.001) without significant differences between diestrous and lactation pituitaries. Galanin alone did not modify LH secretion but it potentiated the effect of GnRH upon pituitaries from diestrous (p = 0.036) but not lactating rats. Neither peptide alone or its combination modified pituitary beta-LH mRNA levels. Results show that galanin regulates differently the secretion and synthesis of LH at the pituitary level. The disappearance of galanin-induced potentiation of GnRH effects upon LH secretion during lactation might contribute to the hypogonadotropism of lactation in the rat.

Animals↗

Measuring the quality of chiropractic alternative care in a managed care environment.

Today's health care environment is more consumer driven with regard to quality issues, and consumers want open access to alternative care, which includes chiropractic care. Given the growing demand for quality, the majority of the top health care plans are requiring their providers to participate in a credentialing process and an on-site office review. These standards allow managed care organizations to make an informed, objective decision regarding the network selection process of chiropractic providers.

Chiropractic↗

Torsional directed walks, entropic elasticity, and DNA twist stiffness.

DNA and other biopolymers differ from classical polymers because of their torsional stiffness. This property changes the statistical character of their conformations under tension from a classical random walk to a problem we call the "torsional directed walk." Motivated by a recent experiment on single lambda-DNA molecules [Strick, T. R., Allemand, J.-F., Bensimon, D., Bensimon, A. & Croquette, V. (1996) Science 271, 1835-1837], we formulate the torsional directed walk problem and solve it analytically in the appropriate force regime. Our technique affords a direct physical determination of the microscopic twist stiffness C and twist-stretch coupling D relevant for DNA functionality. The theory quantitatively fits existing experimental data for relative extension as a function of overtwist over a wide range of applied force; fitting to the experimental data yields the numerical values C = 120 nm and D = 50 nm. Future experiments will refine these values. We also predict that the phenomenon of reduction of effective twist stiffness by bend fluctuations should be testable in future single-molecule experiments, and we give its analytic form.

Animals↗

A cationic lipid for rapid and efficient delivery of plasmid DNA into mammalian cells.

Cationic lipids are widely used for gene transfer into cultured eukaryotic cells. However, lipids with potent transfection activity are often associated with high levels of cytotoxicity, and also require serum-free conditions for optimal performance. These characteristics in many cases result in unsatisfactory transfection efficiency. In this report, we describe a new cationic amphiphile, N-t-butyl-N'-tetradecyl-3-tetradecylaminopropionamidine (Amidine). Amidine requires only 1-2 hour incubation intervals to produce maximal transfection efficiency, and can transfect cells in the presence of serum. Such characteristics significantly minimize cytotoxicity, and also provide time flexibility for researchers. We routinely obtain over 80% transfection efficiency as evidenced by use of an enhanced green fluorescence protein (EGFP) as the reporter. These studies demonstrate the utility of Amidine for rapid and efficient transfection of mammalian cells.

Animals↗

Dynamically stabilized pores in bilayer membranes.

Zhelev and Needham have recently created large, quasistable pores in artificial lipid bilayer vesicles. Initially created by electroporation, the pores remain open for up to several seconds before quickly snapping shut. This result is surprising, in light of the large line tension for holes in bilayer membranes and the rapid time scale for closure of large pores. We show how pores can be dynamically stabilized via a new feedback mechanism. We also explain quantitatively the observed sudden pore closure as a tangent bifurcation. Finally, we show how Zhelev and Needham's experiment can be used to measure accurately the pore line tension, an important material parameter. For their stearoyloleoylphosphatidylcholine/cholesterol mixture we obtain a line tension of 2.6 x 10(-6) dyn.

Biophysical Phenomena↗

Carbohydrate deficient glycoprotein syndrome type I: a cause of cerebellar vermis hypoplasia.

To report the first case of carbohydrate deficient glycoprotein syndrome Type I (CDG I) that has been identified in Australia and confirmed enzymatically to raise the awareness of paediatricians with regard to CDG I and its manifestations, implications and diagnostic investigations. Clinical and autopsy findings of an infant with CDG I are presented. The diagnosis of CDG I was suggested by the clinical findings and biochemical abnormalities and was confirmed by showing an abnormal transferrin isoform pattern. Subsequent studies showed a reduced level of phosphomannomutase in skin fibroblasts. Carbohydrate-deficient glycoprotein syndrome I is one of the many causes of cerebellar hypoplasia. It is an important disorder to identify because of the prognostic and genetic implications and may be underdiagnosed in Australia.

Cerebellum↗

An interdisciplinary health team training program for school staff in Minnesota.

This paper describes an interdisciplinary health team training program for school-based clinic staff in Minnesota. The project sought to improve team functioning, level of practice, and health care services at the school sites. Participants were interdisciplinary staff members from clinics in senior high, middle, and elementary schools. The program consisted of further development in team training knowledge and skills and educational sessions on issues identified by participants. Evaluations indicated participants reported greater knowledge and improved team functioning experiences from the team training. Gains also were shown in knowledge and skills in specific school topic areas such as violence, resiliency, working with resistant families, and self-care. The program could serve as a model for other interdisciplinary school health team training.

Curriculum↗

Regulation of RANTES and ICAM-1 expression in murine mesangial cells.

Chemokines and adhesion molecules play a pivotal role in leukocyte infiltration during tissue injury. RANTES (regulated upon activation, normal T cell expressed and secreted) is a monocyte chemoattractant that induces the expression of CD11/CD18 integrins on leukocytes for which intercellular adhesion molecule-1 (ICAM-1) is the ligand. Both RANTES and ICAM-1 can be expressed by mesangial cells (MC) in culture and in glomeruli during immune injury. In this study, the role of reactive oxygen species (ROS) in the activation of RANTES and ICAM-1 in murine MC was examined. Tumor necrosis factor alpha (TNF-alpha) and aggregated immunoglobulin (aggr. Ig) G, which enhance ROS formation in MC, increased mRNA transcripts of both RANTES and ICAM-1. Thiol-containing free-radical scavengers N-acetyl cysteine, dimethyl- and tetramethylthiourea, or pyrrolidinedithiocarbamate abrogated the increase in mRNA for RANTES and ICAM-1 in response to TNF-alpha or IgG. Hydroxy-methoxy acetophenone, an inhibitor of NADPH-dependent oxidase, also attenuated RANTES and ICAM-1 in response to TNF-alpha or IgG. ROS generated by addition of xanthine oxidase and hypoxanthine induced RANTES and ICAM-1 expression, whereas hydrogen peroxide caused no response. Because cAMP can interfere with gene activation in MC, the effects of 8-Br-cAMP, forskolin, and prostaglandin E2 on mRNA levels were examined for RANTES and ICAM-1. These agents attenuated the response to IgG aggregates and also to superoxide generation. Finally, the effect of glucocorticoids, which are frequently used in glomerular immune injury, was examined. Dexamethasone decreased mRNA for both RANTES and ICAM-1 after stimulation with aggr. IgG or TNF-alpha. Both forskolin and dexamethasone also reduced the amount of RANTES protein secreted by MC in response to aggr. IgG. Only dexamethasone decreased RANTES secretion in response to TNF-alpha stimulation. The inhibitory effects of cAMP and dexamethasone may explain the beneficial effects of cAMP mimetics, such as prostaglandin E2 and glucocorticoid administration on glomerular inflammatory processes.

Animals↗