Influence of a light touch reference on cutaneous reflexes from the hand during standing.

Light touch of a stable reference reduces sway during standing. However, unexpected displacement of a light touch reference leads to short-latency reactions in ankle muscles consistent with a balance reaction, that are replaced by responses in arm muscles on subsequent trials. We anticipated that the excitability of sensorimotor pathways arising from finger cutaneous afferents would reflect these changes in behavior. We hypothesized that (1) interlimb cutaneous reflexes in muscles of the ipsilateral leg, derived from median nerve (MED) stimulation would be facilitated when touch was stable, but reduced when touch was unreliable, (2) intralimb MED reflexes in muscles of the homonymous arm would be facilitated when touch was unreliable and participants tracked the touch reference with arm movements, and (3) radial nerve (RAD) evoked reflexes would be unaffected, given that the RAD innervation territory is not involved in the light touch task. Cutaneous reflexes were evoked using a transcutaneous train of pulses (5 × 1.0 ms square-wave pulses; 300 Hz) and recorded using electromyography of muscles of the ipsilateral arm and leg. As hypothesized, interlimb MED reflexes recorded in soleus (SOL) were larger when touching the stable reference (mean ± SD % MVC; 4.78 ± 1.57) than when not touching a reference (1.00 ± 1.05) or when touching an unstable reference (1.07 ± 1.16). In addition, intralimb MED reflexes in anterior deltoid (AD) were larger when touching an unstable reference (4.50 ± 1.31), compared to touching a stable reference (1.34 ± 1.01) or not touching (1.50 ± 1.00). In contrast, interlimb RAD reflexes in SOL were larger when not touching (4.29 ± 4.34), compared with touching a stable (1.14 ± 1.84) or unstable reference (3.11 ± 4.15). These findings indicate that cutaneous reflexes from the hand are scaled with a rapid change in motor behavior when a touch reference becomes unstable, suggesting that spinal sensorimotor pathways are functionally reweighted based in part upon the reliability of tactile inputs.

Cortisol and stimulus-induced arousal level differentially impact memory for items and backgrounds.

Most studies examine the effects of stress on memory for visual information test memory for entire scenes. However, arousal levels may differentially influence memory for backgrounds as opposed to items. Participants encoded scenes that included a negative-high-arousal, negative-moderate-arousal, or neutral item on a neutral background. After a 30-minute (Experiment 1) or 48-hour delay (Experiment 2), participants underwent a stressor or control task while heart rate was recorded. A recognition memory task was then given with items and backgrounds presented separately. High-arousal images had a greater detriment in background memory than moderate-arousal images. Further, though there was evidence that change in cortisol level at retrieval was associated with impaired memory for items, it was not associated with detriments in background memory. Increased heart rate was associated with impaired memory for both items and backgrounds. This suggests that the level of sympathetic and cortisol reactivity differentially affects memory for items and backgrounds.

Advancing youth transportation safety in North Carolina through addressing alcohol-related motor vehicle collisions.

The aim of this work is to analyze trends in youth transportation fatalities and injuries in North Carolina (NC), assess the implementation of ignition interlock devices (IIDs) in the United States and abroad, discuss policy implications for IIDs, and highlight health equity considerations related to motor vehicle collisions (MVCs). MVCs cause the highest number of unintentional injury-related deaths for children and teenagers in NC, and policymakers should pay special attention to MVCs related to alcohol consumption. IIDs are effective in reducing collision rates and recidivism for driving under the influence of alcohol (DUI). Ignition interlock device requirements have been increasingly implemented globally over the past three decades. However, the adoption of stricter IID policies after first-time DUI offenses in NC and across the U.S. is a prudent public health measure to enhance transportation safety for both adults and children. Evidence-based interventions such as IIDs must also strive to address inequities in transportation safety, and the framing of proposed policies should reflect the tenets of cultural humility.

Evidence for multiple sources of diamond from primitive chondrites.

Fine-grained diamonds, the most abundant form of circumstellar dust isolated from primitive meteorites, have elemental and isotopic characteristics that are dependent on the host meteorite type. Carbon isotopic compositions vary from -32 to -38 per mil, and nitrogen associated with the diamond changes in overall abundance by over a factor of four from 0.2 to 0.9 weight percent, between ordinary and CM2-type chondrites. Although the ratio of carbon to nitrogen evolves in a distinctive way during combustion of diamond separates, metamorphic degassing of nitrogen is not the main cause of the differences in nitrogen content. The data suggest that intrinsic differences must have been inherited by the diamonds at the time of their formation and that the diamonds were distributed heterogeneously in the solar nebula during condensation. However, the hypothesis that a distinct nitrogen carrier remains hidden within the diamond cannot be ruled out.

A new type of meteoritic diamond in the enstatite chondrite abee.

Diamonds with delta(13)C values of -2 per mil and less than 50 parts per million (by mass) nitrogen have been isolated from the Abee enstatite chondrite by the same procedure used for concentrating Cdelta, the putative interstellar diamond found ubiquitously in primitive meteorites and characterized by delta(13)C values of -32 to -38 per mil, nitrogen concentrations of 2,000 to 12,500 parts per million, and delta(15)N values of -340 per mil. Because the Abee diamonds have typical solar system isotopic compositions for carbon, nitrogen, and xenon, they are presumably nebular in origin rather than presolar. Their discovery in an unshocked meteorite eliminates the possibility of origins normally invoked to account for diamonds in ureilites and iron meteorites and suggests a low-pressure synthesis. The diamond crystals are approximately 100 nanometers in size, are of an unusual lath shape, and represent approximately 100 parts per million of Abee by mass.

Terrestrial carbon and nitrogen isotopic ratios from cretaceous-tertiary boundary nanodiamonds.

One hypothesis for the origin of the nanometer-size diamonds found at the Cretaceous-Tertiary (K-T) boundary is that they are relict interstellar diamond grains carried by a postulated asteroid. The (13)C/(12)C and (15)N/(14)N ratios of the diamonds from two sites in North America, however, show that the diamonds are two component mixtures differing in carbon and nitrogen isotopic composition and nitrogen abundance. Samples from a site from Italy show no evidence for either diamond component. All the isotopic signatures obtained from the K-T boundary are material well distinguished from known meteoritic diamonds, particularly the fine-grain interstellar diamonds that are abundant in primitive chondrites. The K-T diamonds were most likely produced during the impact of the asteroid with Earth or in a plasma resulting from the associated fireball.

Paper spray mass spectrometry for the direct, semi-quantitative measurement of fentanyl and norfentanyl in complex matrices.

BACKGROUND: Fentanyl is a potent, synthetic opioid at the centre of an international health crisis that has seen thousands of fatal overdoses. Most analytical methods focus on the detection of fentanyl in blood and/or urine (i.e., post-drug use). Harm reduction (including pre-screening before drug use) may be an effective strategy against fentanyl overdoses. METHOD: Paper spray-mass spectrometry (PS-MS) is an inexpensive, direct sampling strategy where a small volume of sample (<10 µL) is spotted onto a piece of paper that is then wetted and connected to high voltage. Ions are emitted from the paper and enter a mass spectrometer for sensitive and selective semi-quantitation using labeled internal standards. RESULTS: We present the use of PS-MS for the direct measurement of fentanyl and norfentanyl using a custom PS interface, demonstrating that paper tip position and quality can significantly affect quantitative results. Furthermore, we observe comparable calibrations for fentanyl and norfentanyl (0.5 to 600 ng/mL) across a variety of complex matrices (methanol, diluted urine, analgesic slurry). Detection limits for fentanyl are as low as 0.049 ng/mL (0.4 pg total material) in methanol, and 0.66 ng/mL (5.3 pg total material) spiked in an analgesic slurry (illicit substance simulation). PS-MS was compared with liquid chromatography-MS for the analyses of real urine samples, with satisfactory results. CONCLUSION: PS-MS shows potential as a sensitive and selective direct measurement strategy for use in fentanyl harm reduction strategies, and may also be used for pre-screening in advance of or in combination with more conventional (i.e., chromatographic) analyses.

Microwaves effectively examine the extent and type of coking over acid zeolite catalysts.

Coking leads to the deactivation of solid acid catalyst. This phenomenon is a ubiquitous problem in the modern petrochemical and energy transformation industries. Here, we show a method based on microwave cavity perturbation analysis for an effective examination of both the amount and the chemical composition of cokes formed over acid zeolite catalysts. The employed microwave cavity can rapidly and non-intrusively measure the catalytically coked zeolites with sample full body penetration. The overall coke amount is reflected by the obtained dielectric loss (ε″) value, where different coke compositions lead to dramatically different absorption efficiencies (ε″/cokes' wt%). The deeper-dehydrogenated coke compounds (e.g., polyaromatics) lead to an apparently higher ε″/wt% value thus can be effectively separated from lightly coked compounds. The measurement is based on the nature of coke formation during catalytic reactions, from saturated status (e.g., aliphatic) to graphitized status (e.g., polyaromatics), with more delocalized electrons obtained for enhanced Maxwell-Wagner polarization.Catalyst deactivation by coke deposition is a major drawback in industrial processes. Here, the authors show a non-intrusive microwave cavity perturbation technique as a powerful tool to determine the nature and extent of coke accumulation in industrially-relevant zeolite catalysts.

Basal phosphorylation of mu opioid receptor is agonist modulated and Ca2+-dependent.

The mu opioid receptor was shown to be phosphorylated at a basal rate in the absence of agonist, measured in permeabilized HEK293 cells transfected with an epitope tagged mu receptor (EE-mu) [Arden, J., Segredo, V., Wang, Z., Lameh, J. and Sadee, W. (1995) J. Neurochem. 65, 1636-1645]. In the present study, basal phosphorylation was found to be Ca2+ dependent; however, several inhibitors of protein kinase C and Ca2+-calmodulin dependent kinases failed to affect basal mu receptor phosphorylation. Thus, the basal mu receptor phosphorylating activity differed from the main kinases involved in receptor regulation. The general kinase inhibitor H7 (100 microM) suppressed basal mu receptor phosphorylation. Pretreatment with the agonist morphine, followed by drug removal, resulted in a sustained increase of basal mu receptor phosphorylation. The gradual agonist dependent modulation of basal mu receptor phosphorylation suggests a novel regulatory mechanism which may play a role in narcotic tolerance and dependence.

Inverse PCR amplification of low-abundancy message of gamma delta T cell receptor genes.

The technique of inverse PCR permits the rapid amplification and identification of unknown DNA segments adjacent to well characterized core regions. In the field of immunology anchored PCR and inverse PCR are useful methods for examining junctional diversity and unknown variable gene segments of rearranged T cell receptor genes. We have applied an improved inverse PCR protocol to study the repertoire of gamma delta T cell receptor genes in the developing thymus of the mouse.

Influence of N and K fertilization and growth temperature on 13C/12C ratios of timothy (Phleum pratense L.).

Measurements have been reported of carbon isotope ratios of timothy grown at different temperatures and with varying nitrogen and potassium supplies. Both total plant tissue and extracted plant tissue have been analyzed. The 13C/12C ratios were found to vary both with temperature and with nutrient level; the highest values of δ13C were found under the most optimum growth conditions.

The origin of chondritic macromolecular organic matter: a carbon and nitrogen isotope study.

The N and C abundances and isotopic compositions of acid-insoluble carbonaceous material in thirteen primitive chondrites (five unequilibrated ordinary chondrites, three CM chondrites, three enstatite chondrites, a CI chondrite and a CR chondrite) have been measured by stepped combustion. While the range of C isotopic compositions observed is only delta 13C = 30%, the N isotopes range from delta 15N approximately -40 to 260%. After correction for metamorphism, presolar nanodiamonds appear to have made up a fairly constant 3-4 wt% of the insoluble C in all the chondrites studied. The apparently similar initial presolar nanodiamond to organic C ratios, and the correlations of elemental and isotopic compositions with metamorphic indicators in the ordinary and enstatite chondrites, suggest that the chondrites all accreted similar organic material. This original material probably most closely resembles that now found in Renazzo and Semarkona. These two meteorites have almost M-shaped N isotope release profiles that can be explained most simply by the super-position of two components, one with a composition between delta 15N = -20 and -40% and a narrow combustion interval, the other having a broader release profile and a composition of delta 15N approximately 260%. Although isotopically more subdued, the CI and the three CM chondrites all appear to show vestiges of this M-shaped profile. How and where the components in the acid-insoluble organics formed remains poorly constrained. The small variation in nanodiamond to organic C ratio between the chondrite groups limits the local synthesis of organic matter in the various chondrite formation regions to at most 30%. The most 15N-rich material probably formed in the interstellar medium, and the fraction of organic N in Renazzo in this material ranges from 40 to 70%. The isotopically light component may have formed in the solar system, but the limited range in nanodiamond to total organic C ratios in the chondrite groups is consistent with most of the organic material being, presolar.

Concurrence of multiple sclerosis and intracranial glioma. Report of a case and review of the literature.

We report a 39-year-old female patient known to have multiple sclerosis (MS), who later developed cerebral glioblastoma. The tumor was documented on the brain-magnetic resonance imaging (MRI) during the work-up for an apparent relapsing MS, and was subsequently confirmed pathologically by stereotactic biopsy and the postmortem brain examination. Our case, as well as others, re-emphasizes the need to evaluate the symptoms and brain MRI carefully, even in well-documented MS subjects. The concurrence of MS and intracranial glioma is uncommon. The possible relationship between the 2 diseases was discussed, and related literature reviewed.

Increased sensitivity to etomidate in the elderly: initial distribution versus altered brain response.

To determine the effect of aging on the pharmacokinetics and pharmacodynamics of etomidate, we administered etomidate (5 to 10 mg/min) by intravenous infusion to 21 healthy surgical patients, age 22 to 82 yr. Etomidate produced progressive slowing of the EEG to an easily recognized pattern (stage 3) that determined the dosage endpoint. Subsequent power-spectrum analysis of the EEG gave the median frequency. Median frequency values and simultaneous measurements of blood etomidate concentration were incorporated into a sigmoid Emax pharmacodynamic model that permitted an estimate of IC50, the blood etomidate concentration which produced a 50% reduction in the median frequency. The dose of etomidate required to reach the uniform EEG endpoint decreased significantly with increasing age (r2 = .68) as did the dose needed to produce maximal median frequency depression (r2 = .69). None of the parameters of the pharmacodynamic effect model, including IC50, correlated with age, suggesting that increased brain sensitivity in the elderly does not cause the age-related change in dose requirement. The initial distribution volume for etomidate decreased significantly with increasing age (r = .56), implying that a higher initial blood concentration in the elderly following any given dose of etomidate is part of the cause of the lower dose requirement in the elderly patient. A contracted initial distribution volume in the elderly may result from well described physiologic changes of age. Etomidate clearance also decreased with age. Age-dependent changes in etomidate pharmacokinetics rather than altered brain responsiveness may be the basis for the decreased etomidate dose requirement in the elderly.

Childhood asthma mortality: the Brooklyn experience and a brief review.

Pediatric asthma mortality is a perplexing and increasingly serious problem. Although many possible etiologic factors have been suggested, clear relationships are yet to be delineated. In addition to reviewing the literature, we studied the clinical and pathologic features of 14 local pediatric asthma deaths that occurred over a 7-year period. Thirteen of 14 children were African-American. Nine of 14 children (64.3) were older than 10 years of age, and 11 of 14 (78.6) were males. Based on a history of clinical features, 10 of the 14 children were characterized as severe asthmatics. Despite the fact that the majority of the children were regarded as severe asthmatics, only 1 of 14 had been evaluated with pulmonary function testing, and only 2 of 14 were receiving corticosteroids. Furthermore, only 2 of 14 were regarded as having good medical follow-up for their asthma. Ten of 14 children died suddenly secondary to asthma. One child was possibly abusing sympathomimetic inhalers, and none had evidence of toxic serum levels of theophylline. Six of 14 children (all adolescents) who died suddenly were negative on urine toxicology screening for cocaine, heroin, etc. Pathologic findings available for 10 children revealed mainly mucus plugging of the airways and collapse of various segments of the lungs, as well as pneumonia and pneumothorax in one child. In this group of children with sudden deaths (except for one child with pneumothorax), no other cause of death could be found.

Mutational analysis of third cytoplasmic loop domains in G-protein coupling of the HM1 muscarinic receptor.

We measured dose-response curves for carbachol stimulation of phosphatidyl inositol (PI) turnover with mutants of the Hm1 muscarinic cholinergic receptor having various deletions from amino acids 219 to 358 of the large third intracellular (i3) loop (208 to 366). These deletions had only small or no effects on the ability of Hm1 transfected into HEK 293 cells to stimulate PI turnover. This result indicates that only small regions of 9 to 11 amino acids adjacent to trans-membrane domains (TMDs) 5 and 6 can be directly involved in G protein coupling. Point mutations were constructed to test the role of charged amino acids in these junctions. A triple point mutation of Hm1 (E214 A/ E216K/ E221 K), which mimics the charge distribution in Hm2 (negatively coupled to cAMP) over the first 14 amino acids of i3, and a double point mutation in the N terminal junction, K359A/K361A, both failed to affect carbachol stimulated PI turnover. Therefore, charge distribution in the loop junctions appears to play a minor role in G protein coupling of Hm1 in HEK 293 cells.

Phosphorylation and agonist-specific intracellular trafficking of an epitope-tagged mu-opioid receptor expressed in HEK 293 cells.

We expressed the cloned mu-opioid receptor (muR) in high abundance (5.5 x 10(6) sites/cell) with an amino-terminal epitope tag (EYMPME) in human embryonic kidney 293 cells. The epitope-tagged receptor (EE-muR) was similar to the untagged mu R ligand binding and agonist-dependent inhibition of cyclic AMP accumulation. By confocal microscopy, the labeled receptor was shown to be largely confined to the plasma membrane. Pretreatment with morphine failed to affect the cellular distribution of the receptor as judged by immunofluorescence and tracer binding studies. In contrast, exposure to the mu-specific peptide agonist [D-Ala2, MePhe4, Gly-ol5]enkephalin (DAMGO) caused strong labeling of endocytic vesicles, indicating extensive agonist-induced cellular redistribution of EE-muR. Tracer binding studies suggested partial net internalization and a small degree of down-regulation caused by DAMGO. EE-muR-containing membranes were solubilized in detergent [3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate] and immunoprecipitated by an anti-epitope monoclonal antibody. Immunoblotting revealed a prominent band at approximately 70 kDa with weaker bands at approximately 65 kDa. EE-muR was labeled with [gamma-32P]ATP in permeabilized cells, immunoprecipitated, and analyzed by polyacrylamide gel electrophoresis autoradiography. A prominent band at 65-70 kDa indicated the presence of basal receptor phosphorylation occurring in the absence of agonist, which was enhanced approximately 1.8-fold with the addition of morphine. In conclusion, intracellular trafficking of the muR appears to depend on the agonist, with morphine and DAMGO having markedly different effects. Unlike other G protein-coupled receptors, basal phosphorylation is substantial, even in the absence of agonist.

Multifocal granular cell tumors of the gastrointestinal tract.

Granular cell tumors infrequently appear in the gastrointestinal tract. Lesions have been reported in all segments from the esophagus to the rectum, but no previous reports have identified simultaneous lesions in various segments. We describe a patient with granular cell tumors of the esophagus, stomach, appendix, and cecum. Our case emphasizes the need to evaluate the entire gastrointestinal tract when a single lesion is identified. The radiographic approach and pathological characteristics are discussed. Therapeutic alternatives are presented.