Peter B. Chase

Different mechanisms of increased proteolysis in atrophy induced by denervation or unweighting of rat soleus muscle

Mechanisms of accelerated proteolysis were compared in denervated and unweighted (by tail-cast suspension) soleus muscles. In vitro and in vivo proteolysis were more rapid and lysosomal latency was lower in denervated than in unweighted muscle. In vitro, lysosomotropic agents (eg, chloroquine, methylamine) did not lessen the increase in proteolysis caused by unweighting, but abolished the difference in proteolysis between denervated and unweighted muscle. Leucine methylester, an indicator of lysosome fragility, lowered latency more in denervated than in unweighted muscle. 3-Methyladenine, which inhibits phagosome formation, increased latency similarly in all muscles tested. Mersalyl, a thiol protease inhibitor, and 8-(diethylamino)octyl-3,4,5-trimethoxybenzoate hydrochloride (TMB-8), which antagonizes sarcoplasmic reticulum release of Ca2+, reduced accelerated proteolysis caused by unweighting without diminishing the faster proteolysis due to denervation. Calcium ionophore (A23187) increased proteolysis more so in unweighted than control muscles whether or not Ca2+ was present. Different mechanisms of accelerated proteolysis were studied further by treating muscles in vivo for 24 hours with chloroquine or mersalyl. Chloroquine diminished atrophy of the denervated but not the unweighted muscle, whereas mersalyl prevented atrophy of the unweighted but not of the denervated muscle, both by inhibiting in vivo proteolysis. These results suggest that (1) atrophy of denervated, but not of unweighted, soleus muscle involves increased lysosomal proteolysis, possibly caused by greater permeability of the lysosome, and (2) cytosolic proteolysis is important in unweighting atrophy, involving some role of Ca2(+)-dependent proteolysis and/or thiol proteases.

Effects of graded doses of epinephrine on both noninvasive and invasive measures of myocardial perfusion and blood flow during cardiopulmonary resuscitation

ObjectivesEpinephrine administered during cardiopulmonary resuscitation (CPR) is known to increase aortic diastolic and myocardial perfusion pressures, while enhancing myocardial blood flow. Optimal dosing of epinephrine during CPR is less certain. Interest in high-dose epinephrine use under such circumstances is increasing. The effect of different doses of epinephrine on simultaneously measured perfusion pressures, myocardial blood flow, cardiac output, and end-tidal CO2 (Pco2) (used as an indirect measure of cardiac output during CPR) is unknown. DesignProspective, sequential evaluation of no epinephrine, standard dose epinephrine, and high-dose epinephrine. SettingAn experimental resuscitation laboratory. SubjectsTwelve domestic swine. InterventionsMyocardial perfusion pressure, myocardial blood flow, cardiac output, and end-tidal Pco2 were studied after various doses of epinephrine were administered during prolonged CPR. After 3 mins of untreated ventricular fibrillation, each animal received 5 mins of CPR without epinephrine, 5 mins of CPR after standard dose epinephrine (0.02 mg/kg), and 5 mins of CPR after high-dose epinephrine (0.2 mg/kg). Cardiac output and regional myocardial blood flow values were measured with nonradioactive, colored microspheres. Measurements and Main ResultsMyocardial perfusion pressure (aortic diastolic minus right atrial diastolic) was significantly (p < .05) increased over baseline with high-dose epinephrine (35 ± 8 vs. 14 ± 4 mm Hg), but not with standard dose epinephrine (20 ± 5 vs. 14 ± 4 mm Hg). Epinephrines effect on myocardial blood flow was similar, increasing after the high dose (71 ± 21 vs. 20 ± 5 mL/min/100 g; p > .05), but not with the standard dose (23 ± 6 vs. 20 ± 5 mL/min/100 g). Cardiac output decreased significantly (p < .05) after high-dose epinephrine (7 ± 1 vs. 13 ± 1 mL/min/kg). Mean end-tidal Pco2 levels were lower after high-dose epinephrine (15 ± 2 vs. 20 ± 2 mm Hg; p < .05) but not after standard dose epinephrine (19 ± 2 vs. 20 ± 2 mm Hg). ConclusionsStandard dose epinephrine had minimal effect on myocardial perfusion pressure, myocardial blood flow, cardiac output, or end-tidal Pco2. High-dose epinephrine enhanced myocardial perfusion pressure and myocardial blood flow despite significantly decreasing cardiac output. (Crit Care Med 1993; 21:413–419)

Epidemiology of organophosphate pesticide poisoning in Taiwan.

Introduction. The nationwide epidemiology of organophosphate pesticide (OP) poisoning has never been reported in detail for Taiwan. Methods. This study retrospectively reviewed all human OP exposures reported to Taiwans Poison Control Centers (PCCs) from July 1985 through December 2006. Results. There were 4799 OP exposures. Most OP exposures were acute (98.37%) ingestions (74.50%) of a single OP (80.37%) to attempt suicide (64.72%) in adults (93.25%). Males were the most common gender (64.95%). Most patients (61.97%) received atropine and/or pralidoxime. The mortality rate for all 4799 OP exposures was 12.71%. Exposures to single OPs without co-intoxicants caused 524 deaths; of these, 63.36% were due to dimethyl OPs. Conclusion. Dimethyl OPs cause the majority of deaths in Taiwan.

Serum levels and urine detection of Centruroides sculpturatus venom in significantly envenomated patients.

Introduction. Envenomation by Centruroides sculpturatus can cause systemic signs and symptoms requiring treatment. The toxicokinetics of C. sculpturatus venom has not been described. Methods. Venom components were separated for cross-reactivity testing. Serum and urine collected from three patients envenomated by C. sculpturatus had venom levels determined by sandwich enzyme-linked immunosorbent assay (ELISA). Results. Western blot analysis indicated recognition of C. sculpturatus venom by Alacramyn®, an equine F(ab′)2 antivenom developed against Centruroides scorpion venoms, including C. sculpturatus. Serum venom levels in ng/mL with post-envenomation times in minutes (min) were as follows: 85-year-old woman = 8.2 (∼150), 2.8 (515), 1.6 (1,200); 14-month-old girl = 29.7 (∼50), 5.0 (729); 3-year-old girl = 11.1 (∼313), urine venom level of 9.0 (∼490). Conclusion. There is sufficient venom cross-antigenicity among different Centruroides species to allow this ELISA technique with Alacramyn® to determine serum and urine C. sculpturatus venom concentrations in envenomated patients.

Cardiovascular adjustments to rhythmic handgrip exercise: relationship to electromyographic activity and post-exercise hyperemia

SummaryThe purpose of this study was to examine the association among electromyographic (EMG) activity, recovery blood flow, and the magnitude of the autonomic adjustments to rhythmic exercise in humans. To accomplish this, 10 healthy subjects (aged 23–37 y) performed rhythmic handgrip exercise for 2 min at 5, 15, 25, 40, and 60% of maximal voluntary force. Heart rate and arterial blood pressure were measured at rest (control), during each level of exercise, and for 2 min following exercise (recovery). The rectified, filtered EMG activity of the exercising forearm was measured continuously during each level of exercise and was used as an index of the level of central command. Post-exercise hyperemia was calculated as the difference between the control and the average recovery (2 min) forearm blood flows (venous occlusion plethysmography) and was examined as a possible index of the stimulus for muscle chemoreflex activation. Heart rate, arterial pressure, forearm EMG activity, and post-exercise hyperemia all increased progressively with increasing exercise intensity. The magnitudes of the increases in heart rate and arterial pressure from control to exercise were directly related to both the level of EMG activity and the degree of post-exercise hyperemia across the five exercise intensities (Δheart rate vs EMG activity:r=0.99; Δarterial pressure vs EMG activity:r=0.99; Δheart rate vs hyperemia:r=0.99; and Δarterial pressure vs hyperemia:r=0.98; allp<0.01). Furthermore, the level of EMG activity was directly related (r=0.99) to the corresponding degree of hyperemia. These results are consistent with the hypothesis that central command is a primary mechanism by which tachycardia is mediated during submaximal, rhythmic exercise.

Evidence for platelet-activating factor receptor subtypes on human polymorphonuclear leukocyte membranes☆

Platelet-activating factor (PAF) is a potent phospholipid mediator that acts through specific cell surface receptors. The existence of PAF receptor subtypes has been suggested by functional and radioligand binding studies in a variety of cells and tissues. This report addresses this issue more directly and demonstrates differences between specific PAF receptors in human polymorphonuclear leukocytes (PMNs) and COS-7 cells transfected with the cloned human PAF receptor gene. The presence of more than one receptor in human PMNs is supported by three different studies. First, the Kd from the saturation isotherms for the binding of [3H]WEB 2086 on PMNs was 7-fold larger (Kd = 29.2 nM) than the kinetic Kd (4.2 nM). Second, the pseudo-Hill slope determined from the saturation experiments with PMNs was significantly lower than unity (0.69 +/- 0.05 SEM), and the saturation Kd values for transfected COS-7 (Kd = 9.6 nM) and PMN membranes were significantly different. These results contrasted with those for the transfected COS-7 cells, which showed a Kd from the saturation isotherms similar to that of the kinetic Kd (3.2 nM) and a pseudo-Hill slope that was not different from 1.0. Third, when the radiolabeled ligand [3H]WEB 2086 was increased in concentration from 10 to 50 nM in inhibition experiments with the human PMN membranes, the Ki increased, indicative of binding mainly to receptors with lower affinity. These results suggest that PAF receptor subtypes exist in human PMNs based on distinct radioligand binding characteristics from the human cloned PAF receptor.

Regional mapping of the human platelet-activating factor receptor gene (PTAFR) to 1p35→p34.3 by fluorescence in situ hybridization

The human platelet-activating factor cell-surface receptor (PTAFR) is a G protein-coupled receptor thought to contribute to many atopic and inflammatory diseases and, perhaps, to the growth of some neoplasms. Exploring the possibility that the PTAFR might be involved in the genetic predisposition to any disease requires knowledge of its chromosomal localization. In this paper we have used a 20-kb human genomic fragment containing the coding sequence of the cloned PTAFR to determine the regional chromosomal localization of the gene. Using fluorescence in situ hybridization, the localization of the human PTAFR gene was mapped to 1p35-->p34.3.

Differential physiological and behavioral cues observed in individuals smoking botanical marijuana versus synthetic cannabinoid drugs.

Abstract Context: Synthetic cannabinoid use has increased in many states, and medicinal and/or recreational marijuana use has been legalized in some states. These changes present challenges to law enforcement drug recognition experts (DREs) who determine whether drivers are impaired by synthetic cannabinoids or marijuana, as well as to clinical toxicologists who care for patients with complications from synthetic cannabinoids and marijuana. Our goal was to compare what effects synthetic cannabinoids and marijuana had on performance and behavior, including driving impairment, by reviewing records generated by law enforcement DREs who evaluated motorists arrested for impaired driving. Methods: Data were from a retrospective, convenience sample of de-identified arrest reports from impaired drivers suspected of using synthetic cannabinoids (n = 100) or marijuana (n = 33). Inclusion criteria were arrested drivers who admitted to using either synthetic cannabinoids or marijuana, or who possessed either synthetic cannabinoids or marijuana; who also had a DRE evaluation at the scene; and whose blood screens were negative for alcohol and other drugs. Exclusion criteria were impaired drivers arrested with other intoxicants found in their drug or alcohol blood screens. Blood samples were analyzed for 20 popular synthetic cannabinoids by using liquid chromatography-tandem mass spectrometry. Delta-9-tetrahydrocannabinol (THC) and THC-COOH were quantified by gas chromatography-mass spectrometry. Statistical significance was determined by using Fisher’s exact test or Student’s t-test, where appropriate, to compare the frequency of characteristics of those in the synthetic cannabinoid group versus those in the marijuana group. Results: 16 synthetic cannabinoid and 25 marijuana records met selection criteria; the drivers of these records were arrested for moving violations. Median age for the synthetic cannabinoid group (n = 16, 15 males) was 20 years (IQR 19–23 years). Median age for the marijuana group (n = 25, 21 males) was 20 years (IQR 19–24 years) (p = 0.46). In the synthetic cannabinoid group, 94% (15/16) admitted to using synthetic cannabinoids. In the marijuana group, 96% (24/25) admitted to using marijuana. Blood was available for testing in 96% (24/25) of the marijuana group; 21 of these 24 had quantitative levels of THC (mean + SD = 10.7 + 5 ng/mL) and THC-COOH (mean + SD = 57.8 + 3 ng/mL). Blood was available for testing in 63% (10/16) of the synthetic cannabinoid group, with 80% (8/10) of these positive for synthetic cannabinoids. Those in the synthetic cannabinoid group were more frequently confused (7/16 [44%] vs. 0/25 [0%], p ≤ 0.003) and disoriented (5/16 [31%] vs. 0/25 [0%], p ≤ 0.003), and more frequently had incoherent, slurred speech (10/16 [63%] vs. 3/25 [12%], p = 0.0014) and horizontal gaze nystagmus (8/16 [50%] vs. 3/25 [12%], p = 0.01) than those in the marijuana group. Conclusion: Drivers under the influence of synthetic cannabinoids were more frequently impaired with confusion, disorientation, and incoherent, slurred speech than drivers under the influence of marijuana in this population evaluated by DREs.

Response of Upper Limb Blood Flow to Handgrip Exercise After Blalock-Taussig Operation (for Tetralogy of Fallot) or Subclavian Flap Operation (for Aortic Isthmic Coarctation)

To evaluate the effects of long-term reductions in perfusion pressure on blood flow responses to increased functional demand, 5 patients (aged 12 to 26 years) without normal aortic to subclavian artery blood flow to 1 arm as a result of surgery to treat congenital heart disease were studied. Five age- and sex-matched healthy (control) subjects were also studied. In the patients, forearm blood flow was not different in the surgical and normal arms at rest (3.6 +/- 0.6 vs 4.0 +/- 0.7 ml/min/100 ml, respectively, mean +/- standard error, difference not significant) despite lower systolic blood pressure in the surgical arm (87 +/- 2 vs 115 +/- 2 mm Hg, p less than 0.05). The increases in heart rate, systolic blood pressure, forearm electromyographic activity (index of muscle fatigue) and postexercise forearm blood flow (index of muscle oxygen deficit) were not different in response to 2.5 minutes of submaximal rhythmic handgrip exercise (50% of maximal force) performed with the surgical versus the normal arms. Peak forearm blood flow elicited by combined ischemia and maximal isometric handgrip exercise was not significantly different in surgical and normal arms in the group as a whole (39 +/- 4 vs 43 +/- 3 ml/min/100 ml, difference not significant), although some bilateral deficit (20 to 38%) was observed in 2 patients. No bilateral differences were observed in the control subjects under any condition. The finding of normal physiologic adjustments to submaximal rhythmic handgrip exercise with the surgical arm suggests that oxygen delivery during exercise was adequate.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiology of the reported severity of cottonmouth (Agkistrodon piscivorus) snakebite.

Objective The goal of this study was to analyze trends in the annual rates of reported medical outcomes of cottonmouth (Agkistrodon piscivorus) snakebites in the United States, published in the annual reports of the American Association of Poison Control Centers in the course of 29 years. Methods This was a retrospective analysis of medical outcomes for cottonmouth snakebite victims who developed fatal, major, moderate, minor, or no effects. The annual rates for these medical outcomes were calculated by dividing the annual number of patients in each outcome category by the total annual number of people reported as being bitten by cottonmouths. Negative binomial regression was used to examine trends in annual rates. Results From 1985 through 2011, after controlling for the availability of CroFab, the annual incidence rate of cottonmouth snakebites causing no effect decreased significantly by 7.3%/year (incidence rate ratio [IRR] 0.927, 95% confidence interval [CI] 0.885–0.970), the incidence rate of minor outcomes did not change significantly (IRR 0.989, CI 0.974–1.006), the incidence rate of moderate outcomes increased significantly by 2.3%/year (IRR 1.023, CI 1.004–1.042), and the incidence rate of major outcomes did not change significantly (IRR 0.987, CI 0.935–1.041). One fatality was reported in 2011. Conclusions Annual rates of cottonmouth snakebites producing no effects decreased significantly, those producing minor outcomes did not change significantly, those producing moderate outcomes increased significantly, and those producing major outcomes did not change significantly, from 1985 through 2011.