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Heparin modulates the single channel kinetics of reconstituted AMPA receptors from rat brain.

Glutamate receptors specifically activated by alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) have been reported to interact with the highly sulfated glycosaminoglycan, heparin, and to subsequently express lower binding affinity for [3H]AMPA. The present study examined whether heparin also modifies the kinetic properties of single channel activity expressed by isolated AMPA receptors from rat forebrain. Upon application of 280 nM AMPA, the partially purified receptors reconstituted in lipid bilayers expressed bursting channel activity that was inhibited by dinitroquinoxaline-2-3,-dione (DNQX). Treating the receptors with heparin (10 microg/ml) produced no change in conductance but the mean burst length for 280 nM AMPA was nearly doubled. Heparin also prolonged the lifetime of open states of the individual ion channels 3-5-fold, perhaps by causing a decrease in the closing rate constant for channel gating. Heparin had no effect on the lifetime of the closed state or on the amplitude of currents. The single channel open time was voltage-dependent and an increase of applied voltage caused a decrease in the heparin effect on channel open times. While the lifetime of the open channel was increased 3-4 times by heparin at 20 mV, there was no significant change induced at 43 mV. The equivalent electric charge of the channel gate was increased by 40%. The heparin effects were specific as another polysaccharide, dextran, and a monomeric constituent of heparin, glucosamine 2,3-disulfate, failed to have any effect on the receptors. These findings suggest that heparin-containing extracellular matrix components can interact with AMPA receptors and influence their functional properties.

Animals↗

Sodium channels in primary sensory neurons: relationship to pain states.

Electrophysiological studies of dorsal root ganglion (DRG) neurons, and the results of PCR, Northern blot and in situ hybridization analyses have demonstrated the molecular diversity of Na+ channels that operate in sensory neurons. Several subtypes of alpha-subunit have been detected in DRG neurons and transcripts encoding all three beta-subunits are also present. Interestingly, one alpha subunit, Na(v)1.8, is selectively expressed in C-fibre and Adelta fibre associated sensory neurons that are predominantly involved in damage sensing. Another channel, Na(v).3, is selectively up regulated in a variety of models of neuropathic pain. In this review we focus on Na+ channels that are selectively expressed in DRG neurons as potential analgesic drug targets. In the absence of subtype specific inhibitors, the production of null mutant mice provides useful information on the specialized functions of particular Na+ channels. A refinement of this approach is to delete Na+ channel genes flanked by lox-P sites in the sensory ganglia of adult animals, using viruses to deliver the bacteriophage Cre recombinase enzyme.

Animals↗

Post-exercise potentiation of wall motion to identify myocardial viability.

Resting left ventricular wall motion abnormalities may represent myocardial scars or areas of viable muscle which are ischemic and may thus benefit from revascularization surgery. Improvement in wall motion in the 3-6 minute post-exercise recovery period as compared to the resting state was investigated as a possible criterion to predict myocardial viability in 168 patients with coronary artery disease. Of 125 patients with abnormal resting wall motion, 14 of 14 who improved regional function post-exercise also improved following coronary bypass surgery. However, wall motion improved postoperatively in an additional 55 patients who had not demonstrated post-exercise improvement. An increase in ejection fraction post-exercise occurred in patients with coronary disease as well as an additional group of 24 normal subjects. Thus in the post-exercise period, improvement in resting wall motion is a highly specific but insensitive predictor of myocardial viability.

Journal Article↗

Antidepressant-associated mania: a controlled comparison with spontaneous mania.

OBJECTIVE: Antidepressants have been associated with the induction of mania and rapid cycling. This study examined whether antidepressant-associated manic states differ in any way from spontaneous mania. METHOD: Forty-nine consecutive inpatients with antidepressant-associated manic states were compared with 49 matched inpatients with spontaneous mania in a blind, retrospective chart review. RESULTS: Across virtually every clinical measure examined, the patients with antidepressant-associated manic states experienced milder and more time-limited manic episodes than the patients with spontaneous mania. The patients with antidepressant-associated manic states were subject to frequent checking by nurses and hall restriction for a significantly shorter period of time than the patients with spontaneous mania. The patients with antidepressant-associated manic states also had significantly less severe levels of delusions, hallucinations, psychomotor agitation, and bizarre behavior, according to a standard rating instrument, than the patients with spontaneous mania. For further study the patients with antidepressant-associated mania were divided into subgroups taking four individual classes of antidepressant drugs: tricyclics (N = 19), fluoxetine (N = 13), monoamine oxidase inhibitors (MAOIs) (N = 8), and bupropion (N = 6); three patients taking combinations of drugs were not included in these analyses. The patients with MAOI- and bupropion-associated mania had a slightly lower overall rating of severity of psychopathology at admission than the subgroups with fluoxetine- and tricyclic-associated mania. CONCLUSIONS: Antidepressant-associated mania appears to be a milder and more time-limited syndrome than spontaneous mania and may represent a distinct clinical entity. MAOIs and bupropion may be associated with milder manic states than either tricyclic drugs or fluoxetine.

Adult↗

2H(e,e(')p)n reaction at high recoil momenta.

The 2H(e,e(')p)n cross section was measured in Hall A of the Thomas Jefferson National Accelerator Facility near the top of the quasielastic peak (x(Bj)=0.964) at a four-momentum transfer squared, Q(2)=0.665 (GeV/c) (2) (omega=0.368 GeV, W=2.057 GeV), and for recoil momenta up to 550 MeV/c. The measured cross section deviates by 1-2sigma from a state-of-the-art calculation at low recoil momenta. At high recoil momenta the cross section is well described by the same calculation; however, in this region, final-state interactions and interaction currents are predicted to be large, and alternative choices of nucleon-nucleon potential and nucleon current operator may result in significant spread in the calculations.

Journal Article↗

A test to measure lift capacity of physically impaired adults. Part 1--Development and reliability testing.

STUDY DESIGN: Two laboratory studies and one field study evaluated the safety and test-retest reliability of a new test of lift capacity. The first two studies were conducted in a carefully controlled laboratory setting. The first study investigated the safety and intra-rater reliability of the EPIC Lift Capacity test protocol with healthy adult subjects. The second study assessed the safety and inter-rater reliability of the test with disabled subjects. The third study was conducted in the field with 65 evaluators and investigated the safety and intra-rater reliability of the test with healthy adult subjects. OBJECTIVE: To assess the safety and reliability of a new test of lift capacity. SUMMARY OF BACKGROUND DATA: A new test of lift capacity has been developed. Test development occurred within the context of ergonomic standards and guide-lines of the major professional associations and public agencies that govern test development in the United States. METHODS: In study no. 1, 26 healthy subjects participated. In study no. 2, 14 disabled subjects participated. In study no. 3, 318 healthy subjects participated. After subjects underwent basic screening and warm-up, the EPIC Lift Capacity test was administered. One to 2 weeks later, the test was administered again. Correlations between the times of testing were calculated. RESULTS: No subjects were injured. Hamstring soreness the next day that resolved without complication was reported by some healthy subjects. None of the disabled subjects reported new symptoms. CONCLUSION: The safety and reliability of the EPIC Lift Capacity test was adequately demonstrated in a laboratory setting and across multiple field sites with evaluators who have varying types and degrees of professional preparation.

Adult↗

Serological evidence of HTLV-I and HTLV-II coinfections in HIV-1 positive patients in Belém, state of Pará, Brazil.

The occurrence of HTLV-I/II and HIV-1 coinfections have been shown to be frequent, probably in consequence of their similar modes of transmission. This paper presents the prevalence of coinfection of HTLV among HIV-1 infected and AIDS patients in Belém, State of Pará, Brazil. A group of 149 patients attending the AIDS Reference Unit of the State Department of Health was tested for the presence of antibodies to HTLV-I/II using an enzyme immunoassay and the positive reactions were confirmed with a Western blot that discriminates between HTLV-I and HTLV-II infections. Four patients (2.7%) were positive to HTLV-I, seven (4.7%) to HTLV-II and one (0.7%) showed an indeterminate pattern of reaction. The present results show for the first time in Belém not only the occurrence of HTLV-II/HIV-1 coinfections but also a higher prevalence of HTLV-II in relation to HTLV-I. Furthermore, it also enlarges the geographical limits of the endemic area for HTLV-II in the Amazon region of Brazil.

Adolescent↗

Sedation of critically ill patients during mechanical ventilation. A comparison of propofol and midazolam.

Propofol (P) and midazolam (M) are frequently given by continuous infusion for sedation in critically ill, mechanically ventilated patients. We compared these drugs with regard to: (1) time-to-awaken; (2) reproducibility of bedside assessments of level of sedation; (3) time-to-sedation; and (4) change in oxygen consumption (V O2) from awake to sedated state. Seventy-three patients were prospectively randomized to receive either P (n=37) or M (n=36). Wake-up times after stopping the drug were assessed by blinded and unblinded observers, by asking patients to perform simple tasks. Times to sedate were assessed by consensus agreement among nurses and investigators. Demographics and APACHE II scores were not different between P and M. The P group had a significantly narrower range of wake-up times with a higher likelihood of waking in less than 60 min. Blinded versus unblinded observations had excellent correlation. Average time to sedate and decrease in V O2 were not different. We conclude that in this patient population: (1) both P and M achieved optimal sedation in a large fraction of patients when administered by specified dosing protocols; (2) P had a faster, more reliable, wake-up time; (3) assessments of time-to-awaken were objective and reproducible; (4) time to sedation was not significantly different; (5) V O2 decreased similarly with both.

APACHE↗

Mls-1a-induced peripheral tolerance to host minor histocompatibility antigens in radiation bone marrow chimeras. Modification of T cell repertoire associated with active suppression and permanent presentation of host antigens.

Minor histocompatibility Ag (mHAg) can be responsible for the development of graft vs host reaction (GVHR) after bone marrow transplantation. In a mouse model, B10.D2 donor immunization against Mls-1a prevents lethal GVHR developed by CD4+ T cells against DBA mHAg in irradiated (DBA/2 x B10.D2)F1 hosts. Such F1 hosts become 100% chimeric and show long term survival (LS mice). The cellular mechanisms underlying the tolerance in LS mice was investigated. It was found that a state of tolerance can be induced in thymectomized F1 hosts. Although spleen cells from LS mice are able to initiate lethal GVHR in third-party H-2k-incompatible hosts, no GVHR is observed in secondary hosts incompatible for specific DBA/2 mHAg. Mixed lymphocyte experiments in vitro confirm that T cells from LS mice are unresponsive toward specific DBA/2 mHAg, although they are able to proliferate in response to H-2 or Mls-1a Ag. The responsiveness to Mls-1a correlates with the presence of V beta 6+ cells in LS mice, probably derived from mature T cells present in the donor inoculum. The tolerance in LS mice is not due to the lack of DBA/2 mHAg presentation; instead, permanent presentation of Ag (Ag I and Ag II) previously described as being responsible for lethal GVHR is consistently observed. A significant protection against GVHR is obtained by transferring normal B10.D2 cells together with spleen cells from LS mice, clearly indicating the contribution of active suppression in the state of tolerance; this is further confirmed by in vitro results obtained in limiting dilution assays. It is concluded that tolerance in chimeric LS mice 1) is due to a peripheral (thymus-independent) mechanism; 2) is specific for mHAg; 3) correlates with unresponsiveness of the repertoire to host mHAg, without alteration of the repertoire for H-2 and Mls-1a Ag; and 4) is associated with an active suppression and with a permanent presentation of at least two mHAg responsible for GVHR mortality.

Animals↗

Ten-year summary of human immunodeficiency virus serology in west Tennessee.

Since the isolation of human immunodeficiency virus (HIV), numerous laboratory tests have been developed to confirm the immunologic response to the virus. In 1991, the state of Tennessee mandated that clinical laboratories performing Western blot analyses for HIV report all new cases of confirmed HIV infection. This report is a summary of HIV confirmed cases evaluated by this facility from 1985 until July 1, 1995. During each year, the laboratory screens approximately 20,000 persons for HIV and all positive screens are confirmed by an FDA-approved Western blot. During the period described, the laboratory has confirmed HIV serologic positivity in 4,096 individuals, principally from Shelby County and surrounding Tennessee counties. Although these data are not complete, they clearly indicate that we have a continuing problem with this infectious disease. Based on the current rate of positive cases seen thus far in 1995, this facility may confirm nearly 900 new cases of HIV infection in this year alone.

Adolescent↗

Low-copy microsatellite markers for Pinus taeda L.

Eighteen low-copy and genomic microsatellite markers were tested for Mendelian inheritance and then assayed in 41 Pinus taeda L. samples drawn from five regions in the southern United States. The PCR products had multiple alleles, high levels of polymorphism, and little non-specific priming. Fifteen of the 18 markers were informative for a P. taeda three-generation RFLP (restriction fragment length polymorphism) pedigree, and a P. taeda population survey revealed three to 28 alleles per locus. The highest allele numbers and polymorphic information content (PIC) values were associated with complex repeat sequences and (or) with sequences consisting of the longer strings of perfect repeats. The abundance of low- to rare-frequency alleles also accounted for high PIC values in both types of markers. Low-copy microsatellites are useful for the large, complex pine genome, especially in the absence of entire gene sequences in public databases and with the low levels of polymorphism in markers developed from expressed sequence tags (ESTs).

Alleles↗

Altered glycosylation sites of the delta subunit of the acetylcholine receptor (AChR) reduce alpha delta association and receptor assembly.

We have used mutagenesis to investigate the potential N-glycosylation sites in the delta subunit of the mouse muscle acetylcholine receptor (AChR). Of the three sites, Asn76, Asn143, and Asn169, only the first two were glycosylated when the delta subunit was expressed in COS cells. Because the heterologously expressed delta subunit was similar in its properties to that expressed in C2 muscle cells, the sites of glycosylation are likely to be the same in both cases. In COS cells, mutations of the delta subunit that prevented glycosylation at either of the sites did not change its metabolic stability nor its steady-state level. These results are in contrast to those found previously for the alpha subunit, in which glycosylation at a single site metabolically stabilized the polypeptide (Blount, P., and Merlie, J. P. (1990) J. Cell Biol. 111, 2613-2622). Mutations of the delta subunit that prevented glycosylation, however, decreased its ability to form an alpha delta heterodimer when the alpha and delta subunit were expressed together. When all four subunits of the AChR (alpha, beta, delta, and epsilon) were coexpressed, mutation of the delta subunit to prevent glycosylation resulted in a reduced amount of fully assembled AChR and reduced surface AChR levels, consistent with the role of the heterodimer in the assembly reaction. These results suggest that glycosylation of the delta subunit at both Asn76 and Asn143 is needed for its efficient folding and/or its subsequent interaction with the alpha subunit.

Animals↗

The virtual terrorism response academy: training for high-risk, low-frequency threats.

The Virtual Terrorism Response Academy is a reusable virtual learning environment to prepare emergency responders to deal with high-risk, low-frequency events in general, terrorist attacks in particular. The principal learning strategy is a traditional one: apprenticeship. Trainees enter the Academy and travel through its halls, selecting different learning experiences under the guidance of instructors who are simultaneously master practitioners and master trainers. The mentors are real individuals who have been videotaped according to courseware designs; they are subsequently available at any time or location via broadband Internet or CD-ROM. The Academy features a Simulation Area where trainees are briefed on a given scenario, select appropriate resources (e.g., protective equipment and hazmat instruments), then enter a 3-dimensional space where they must deal with various situations. Simulations are done under the guidance of a master trainer who functions as a coach, asking questions, pointing out things, explaining his reasoning at various points in the simulation. This is followed by a debriefing and discussion of lessons that could be learned from the simulation and the trainee's decisions.

Computer Simulation↗

The role of 'shared representations' in social perception and empathy: an fMRI study.

Evidence suggests that we use the same mechanisms for both producing and perceiving actions. Such 'shared representations' may also underlie social perception and empathy. However, this idea raises some important and as yet unresolved questions: (i) how do we distinguish other-orientated empathic responses from a self-orientated reactions such as personal distress and (ii) what are the neural substrates underpinning these processes? We employed event-related functional magnetic resonance imaging (fMRI) to explore whether 'shared representations' were recruited to decode dynamic social stimuli in 12 healthy volunteers. We used an adapted version of the Profile of Non-Verbal Sensitivity (Rosenthal, H., Hall, J.A., DiMatteo, M.R., Rogers, P.L., Archer, D., (1979). Sensitivity to nonverbal communication: the PONS test. The Johns Hopkins University Press, Baltimore) which taps social perception using brief silent video clips. Participants chose one of two words that best described the state conveyed by the actor, or in the control condition using the same clips, the word describing which parts of the body were visible (non-social labelling). Off-line self-report measures of empathy and personal distress engendered by thoughts about others, were also given along with an experimentally-derived index of the degree of self-other overlap during social perception. Brain activation specific to the main experimental condition was found in the inferior frontal gyrus (BA44) and premotor areas (BA6) consistent with the use of 'shared representations'. Somatosensory areas such as the insula and supramarginal gyrus (BA40) were also activated suggesting that participants constructed a qualitative representation of the target state. Activity in the rostral anterior cingulate was associated with self-reports of personal distress and increased blood flow to the anterior cingulate (BA24) and inferior parietal cortex (BA40) was related to self-other overlap.

Adult↗

Minimum energy structure of hydridotris(pyrazolyl)borato iridium(V) tetrahydride is not a C(3upsilon) capped octahedron.

Recent synthesis and NMR spectroscopy of neutral Ir(V) complexes hydridotris(3,5-dimethylpyrazol-1-yl)borato tetrahydride (Tp*IrH(4)) and hydridotris(pyrazol-1-yl)borato tetrahydride (TpIrH(4)) have been interpreted as supporting face-capped octahedral structures (C(3upsilon)) with each of three Ir-H bonds trans to an Ir-N bond and the fourth hydride capping the IrH(3) face. Here, density functional geometry optimizations and coupled cluster calculations on hydridotris(pyrazol-1-yl)borato iridium tetrahydrogen find that a C(s) edge-bridged octahedral tetrahydride structure and a C(1) eta(2)-dihydrogen, dihydride structure are local minima and find that the C(3upsilon) structure is a local maximum (second-order saddle point). Several low energy transition states connecting the local minima have been located, and these minima can be used to simulate the experimental NMR spectra. A comparison of the experimental infrared spectrum of Tp*IrH(4) and the harmonic frequency calculations on the C(s), C(1), and C(3upsilon) structures also supports the assignment of the C(s)and C(1) structures as the observed ones.

Borates↗

Detection of HTLV-IIa blood donors in an urban area of the Amazon Region of Brazil (Belém, PA).

The human lymphotropic viruses type I (HTLV-I) and type II (HTLV-II) are members of a group of mammalian retroviruses with similar biological properties, and blood transfusion is an important route of transmission. HTLV-I is endemic in a number of different geographical areas and is associated with several clinical disorders. HTLV-II is endemic in several Indian groups of the Americas and intravenous drug abusers in North and South America, Europe and Southeast Asia. During the year of 1995, all blood donors tested positive to HTLV-I/II in the State Blood Bank (HEMOPA), were directed to a physician and to the Virus Laboratory at the Universidade Federal do Pará for counselling and laboratory diagnosis confirmation. Thirty-five sera were tested by an enzyme immune assay, and a Western blot that discriminates HTLV-I and HTLV-II infection. Two HTLV-II positive samples were submitted to PCR analysis of pX and env genomic region, and confirmed to be of subtype IIa. This is the first detection in Belém of the presence of HTLV-IIa infection among blood donors. This result emphasizes that HTLV-II is also present in urban areas of the Amazon region of Brazil and highlights the need to include screening tests that are capable to detect antibodies for both types of HTLV.

Adult↗

A computational analysis of FXa generation by TF:FVIIa on the surface of rat vascular smooth muscle cells.

A computational model was developed to investigate the contribution of classical mass transport and flow parameters to factor X (FX) activation by the tissue factor-factor VIIa complex (TF:VIIa) on one wall of a parallel-plate flow chamber. The computational results were compared to previously obtained experimental data for the generation of factor Xa (FXa) by TF:VIIa on the surface of cultured rat vascular smooth muscle cells. In this study, the complete steady-state convection-diffusion equation was solved using the commercial software package, FLUENT (Fluent Inc., Lebanon, New Hampshire). A user-defined subroutine interfaced with FLUENT implemented the surface reaction which was modeled using classical Michaelis-Menten reaction kinetics. The numerical solutions were obtained for 12 cases which used combinations of three wall shear rates and four reaction rates. The numerically obtained fluxes for a given reaction rate displayed a wall shear rate dependence which ranged from classical kinetic reaction control (no dependence) to pure diffusional control (maximum dependence). The experimental data, however, were not represented by numerical data generated using a single reaction rate. The three numerically obtained fluxes which corresponded most closely to the experimental fluxes were determined using three different Vmax values. This finding supports the hypothesis that there may be a direct effect of flow on the TF:VIIa complex or the cell membrane.

Algorithms↗

Outcomes evaluation in TBI Rehabilitation. Part I: overview and system principles.

Questions of how to measure outcomes are commonly heard in rehabilitation today. In this supplement we review the current state of the art of outcomes measurement in traumatic brain injury rehabilitation, discuss key issues, and identify steps that need to be taken. The underlying question is not simply that of outcomes measurement but how to infer effective and cost-effective treatment. The validity of such inferences rests on multiple features of the entire clinical information system. Requirements for valid, feasible, outcome-oriented clinical data systems are described. Effectiveness is inferred from variations in outcomes experienced by similar persons who received differing interventions. Outcome data systems need large sample sizes and technically adequate severity adjusters. A nationwide TBI data system is needed to identify reasonably homogeneous subgroups in different programs whose outcomes can be meaningfully compared. In alliance with payors and persons with disabilities, rehabilitation professionals can create outcome data systems that will provide the knowledge necessary to improve the effectiveness and cost-effectiveness of rehabilitation programs.

Activities of Daily Living↗