A finite element model of electrode placement during stimulus evoked electromyographic monitoring of iliosacral screw insertion.

Pelvic ring fractures that occur as a result of substantial orthopedic trauma are frequently repaired using iliosacral screws to stabilize the fracture. Stimulus evoked electromyography, using pulsed current stimuli provided through the drill bit cathode, has been advocated to prevent nerve root injury during iliosacral screw insertion. Our objective was to examine the effects of anode location, drill bit position, and anatomical structure on the nerve monitoring technique. A three-dimensional finite element model was constructed from computed tomography data to evaluate the effectiveness of five anode locations at four stations of drill bit insertion. Results indicate that the anode location should be at the midline or on the side contralateral to drill bit insertion. Locating the anode at other positions, such that the nerve root is outside of the primary electromagnetic field, leads to an attenuated electromyographic response that will ultimately lead to the failure of the monitoring technique.

Optimization of multi-frequency techniques used for cell membrane capacitance estimation.

Measurements of cell membrane capacitance serve as an indicator of cell membrane surface area and thus have traditionally been used in stimulus-secretion coupling to monitor exocytosis and endocytosis of secretory vesicles. In order to accurately monitor secretion, high-resolution methods of tracking small (10/sup -15/ F) changes in baseline capacitance must be utilized. Most presently used techniques require assumptions that are not appropriate under all recording conditions or suffer from a low signal-to-noise ratio (SNR). This paper proposes to improve upon current estimation techniques by developing an optimized multi-frequency sinusoidal stimulus waveform for accurate capacitance estimations. The waveform is designed by jointly optimizing the phase and voltage of individual frequency components so as to minimize the theoretical bound on the variance of the capacitance estimate, while maintaining a preselected limit on the overall stimulus magnitude.

Perinatal herpes simplex infection presenting with pneumonia and pleural effusions.

An unusual case of perinatal herpes infection presenting with pneumonia and pleural effusions is described.

An optimized approach to membrane capacitance estimation using dual-frequency excitation.

We present an optimized solution to the problem of membrane impedance estimation when a patch-clamped cell is stimulated by a dual-frequency, sinusoidal excitation. The complete data set of raw whole-cell current samples is typically reduced, via digital lock-in detection, to measurements of the complex cell model admittance at the two stimulus frequencies. We describe a statistical model of both data sets and demonstrate that the admittance data adequately represent the essential features obtained from the raw data. The parameter estimates obtained by a nonlinear weighted least-squares solution (NWLS), which under normal recording conditions is equivalent to the maximum likelihood solution, essentially obtain the theoretical lower bound on variance established by the Cramér-Rao bound. Our software implementation of the NWLS solution produces estimates of the cell model parameters that are less noisy than other dual-frequency systems. Our system can be used 1) to measure slow changes in membrane capacitance-in the face of large, slow changes in membrane resistance, 2) to detect with confidence capacitance changes expected from the exocytosis of moderate-sized dense core granules, and 3) to reduce the cross-talk between transient changes in membrane conductance and membrane capacitance.

Single-cell measurements of quantal secretion induced by alpha-latrotoxin from rat adrenal chromaffin cells: dependence on extracellular Ca2+.

alpha-Latrotoxin (alpha-LT), from black widow spider venom, is a potent enhancer of the spontaneous quantal release of neurotransmitter from a variety of nerve terminals and clonal neurosecretory cells. Using electrochemical amperometry and estimation of membrane impedance by phase detection, we present evidence that alpha-LT induces exocytosis of catecholamines from rat adrenal chromaffin cells beginning as rapidly as 30 s after close application of the toxin. This release is largely dependent on adequate levels of extracellular Ca2+ ([Ca2+]o). Lowering [Ca2+]o from 2 mM to

Voltage-dependent Na+ and Ca2+ currents in human pancreatic islet beta-cells: evidence for roles in the generation of action potentials and insulin secretion.

We describe three voltage-dependent inward currents in human pancreatic beta-cells. First, a rapidly inactivating Na+ current, blocked by tetrodotoxin (TTX) is seen upon brief depolarization to or beyond -40 mV. Second, a transient, low-voltage-activated (LVA), amiloride-blockable Ca2+ current is seen upon depolarization to or beyond -55 mV; it inactivates within less than 1s of sustained depolarization to -40 mV. Third, a more sustained, high-voltage-activated (HVA) Ca2+ current, which shows variable sensitivity to dihydropyridines is seen upon depolarization to or beyond -40 mV, and thereafter slowly inactivates over a time course of many seconds. Our pharmacological evidence suggests that all three currents contribute to action potential initiation and upstroke when the background membrane potential (Vm) is equal or negative to -45 to -40 mV, a situation often induced by glucose concentrations (5-6 mM) in the range of those seen post-prandially. Consistent with this, TTX drastically reduces both transient and sustained insulin secretion in the presence of 5-6 mM glucose, but has little effect in 10 mM glucose, at which concentration cells rapidly depolarize to approximately -35 mV, a Vm sufficient to rapidly inactivate Na+ and LVA Ca2+ currents.

Coupling of exocytosis to depolarization in rat pancreatic islet beta-cells: effects of Ca2+, Sr2+ and Ba(2+)-containing extracellular solutions.

Using rat beta-cells we present evidence that Sr2+ and Ba2+, like Ca2+, support depolarization-induced increases in membrane capacitance which reflect insulin granule exocytosis. Even with identical total charge entry, Sr2+ and Ba2+ are 3-5 and 20-fold less effective than Ca2+ in supporting release. While exocytosis supported by Sr2+ is graded with cation entry and complete within 250ms of depolarization, exocytosis supported by Ba2+ begins abruptly after a threshold of charge entry and continues for many seconds. Ba(2+)-supported release continues in the presence of greatly enhanced cytosolic Ca2+ buffering, arguing against release of Ca2+ from stores as its principal action. These results suggest that Sr2+ and Ba2+ support exocytosis largely by binding to Ca(2+)-dependent release-activating sites, though with less affinity than Ca2+.

Pharmacology of volume regulation following hypotonicity-induced cell swelling in clonal N1E115 neuroblastoma cells.

When exposed to hypotonic solutions, clonal N1E115 neuroblastoma cells initially swell and later undergo a regulatory volume decrease (RVD). We studied the effects of a variety of transport inhibitors on the time course of cross-sectional area of N1E115 cells exposed to a solution of reduced osmolarity (pi = 186 mosm). Application to the bath of either: (i) blockers of net K efflux through K channels (e.g. isotonic KCl or 20 mM TEA); or (ii) blockers of net efflux through anion channels (e.g. isotonic methanesulfonate, 10 microM DIDS or 100 microM IAA-94) all prevent RVD. In contrast, ouabain (a Na+/K+ pump blocker), bumetanide (a Na+/K+/Cl- cotransporter blocker) and SITS (a HCO3-/Cl- exchange blocker) do not. These data support the involvement of these channels over pumps or exchangers in solute exit during RVD. Only variable block of RVD was achieved using blockers of stretch activated non-selective cation C+ (SA) channels (i.e., amiloride and gadolinium, Gd3+) or a membrane permeant Ca chelator (BAPTA-AM) suggesting that neither the opening of C+ (SA) channels nor a global rise in cytosolic Ca2+ is critical for triggering RVD.

Reliability of differential PCR for the detection of EGFR and MDM2 gene amplification in DNA extracted from FFPE glioma tissue.

A series of 43 human gliomas, consisting of 30 glioblastomas, 7 anaplastic astrocytomas, 3 low grade astrocytomas, 2 ependymomas, and 1 oligodendroglioma, was studied for amplification of the epidermal growth factor receptor (EGFR) and mouse double minute 2 (MDM2) genes. DNA extracted from formalin-fixed, paraffin-embedded tissue sections was analyzed by differential PCR and the results were compared with slot blot examination of DNA extracted from frozen tissue from the same neoplasms. Twelve glioblastomas (40%) showed amplification of the EGFR gene, and overexpression of EGFR was evident in each of these tumors as indicated by the immunoperoxidase technique. Two of the tumors with EGFR gene amplification also revealed amplification of the MDM2 gene, while one additional glioblastoma revealed MDM2 amplification only. A 100% concordance in the detection of amplification was observed between differential PCR and slot blot analysis; consequently, these results indicate that differential PCR using DNA extracted from archival tissue sections is a reliable method of demonstrating gene amplifications in glial tumors.

Low-grade astrocytomas arising from the pineal gland.

Two cases of low-grade astrocytoma arising from the pineal gland are described in this report. These rare lesions have only been reported on two previous occasions to arise distinctly from the pineal gland. Histologically, one tumor was consistent with a juvenile pilocytic astrocytoma, while the other was consistent with a fibrillary astrocytoma. The pathological features, clinical implications, and treatment of low-grade astrocytomas in the pineal region are discussed.

The microsurgical anatomy of the superior hypophyseal artery.

Interest in the anatomy of the proximal segment of the intracranial internal carotid artery has been kindled by the recognition that there are multiple potential sites of aneurysm formation in this region. These various aneurysm locations have characteristic hemodynamic and clinical features as well as surgical considerations. Recently recognized as a distinct clinical and anatomical entity are aneurysms that are hemodynamically related to the superior hypophyseal artery. Although aneurysms arising in proximity to the superior hypophyseal artery are not rare, the anatomy of this vessel arising from the medial or posteromedial aspect of the proximal internal carotid artery is poorly understood. We performed a cadaveric microsurgical anatomical study of 20 internal carotid arteries between the ophthalmic and posterior communicating arteries to develop a better understanding of the anatomical relationships of the superior hypophyseal artery. There were an average of 1.8 superior hypophyseal arteries arising from each carotid artery with an average diameter of 0.22 mm. The origin of the superior hypophyseal arteries was within 5 mm of the ophthalmic artery origin in 85% of the specimens. There were two distinct patterns of superior hypophyseal artery anatomy. In 42%, a large, dominant superior hypophyseal artery branched like a candelabra with smaller branches to the pituitary stalk, optic nerve, and chiasm. The average diameter of the larger branches was 0.3 mm. In the absence of a large dominant branch, two or three medial vessels were found. In one specimen, an incidental aneurysm was discovered at the origin of the superior hypophyseal artery on the medial aspect of the internal carotid artery at the origin of a large candelabra-like branch.(ABSTRACT TRUNCATED AT 250 WORDS)

Combined extracranial-intracranial bypass and intraoperative balloon occlusion for the treatment of intracavernous and proximal carotid artery aneurysms.

A protocol for the treatment of selected intracavernous and proximal internal carotid artery aneurysms is described. Intraoperative angiography is used together with intraoperative balloon occlusion of the internal carotid artery and electroencephalography to optimize the timing of an extracranial-intracranial bypass before occlusion of the carotid artery and to provide intraoperative documentation of graft patency. This protocol has been used successfully in seven patients with complex aneurysms that were unsuitable for other endovascular methods or a direct microsurgical approach. Six aneurysms were located in the cavernous sinus, and one was located on the supraclinoid portion of the internal carotid artery. There were no permanent complications; one patient had a brief episode of dysphasia, which resolved without sequelae.

cAMP-enhancing agents "permit" stimulus-secretion coupling in canine pancreatic islet beta-cells.

Isolated canine islets of Langerhans differ from isolated islets of other species (including rodents and man) in that elevated glucose concentrations are unable to stimulate insulin secretion. Here we demonstrate that addition to the perifusate of isobutylmethylxanthine (IBMX), forskolin or 8-CPT-cAMP, all of which enhance cytosolic cAMP, permits insulin secretion in response to glucose, leucine or tolbutamide. These cAMP enhancers increase secretogogue-induced electrical activity in beta-cells and restore depolarization-induced, Ca(2+)-dependent granule exocytosis measured as stepwise increases in membrane capacitance. We propose that the primary permissive action of cAMP is to tightly link Ca2+ entry to insulin granule release, while a secondary action is to tighten the link between glucose metabolism and cell depolarization.

The blood supply of the intracavernous cranial nerves: an anatomic study.

Cranial nerve deficits are the most common complications of cavernous sinus surgery. Often the deficit occurs despite anatomic preservation of the nerve, and ischemic injury is thought to be the cause. A better understanding of the blood supply of these nerves may help to prevent such complications. The authors performed a cadaveric microsurgical study of the intracavernous cranial nerves and their blood supply in 20 cavernous sinuses. The oculomotor nerve received branches from the inferolateral trunk or its equivalent in all specimens (100%). The proximal trochlear nerve received branches from the inferolateral trunk in 80% of the specimens and from the tentorial artery of the meningohypophyseal trunk in 20%. The distal half was supplied by the branches from the inferolateral trunk only. In the region of Dorello's canal, the proximal third of the abducens nerve received branches from the dorsal clival artery of the meningohypophyseal trunk. The middle and distal thirds received branches from the inferolateral trunk. The ophthalmic and proximal maxillary segments of the trigeminal nerve received branches from the inferolateral trunk. The distal maxillary segment was supplied by the artery of the foramen rotundum. In the majority of cases, the medial third of the Gasserian ganglion received branches from both the inferolateral trunk and the tentorial artery. The middle third of the ganglion received branches from either the inferolateral trunk or the middle meningeal artery. Our findings indicate the important role the intracavernous branches of the internal carotid artery play in the blood supply of the intracavernous cranial nerves, and stress the need to preserve these branches to prevent or minimize postoperative deficits.

Reliability of psychiatric and psychological diagnoses of mental retardation severity: judgments under naturally occurring conditions.

The reliability of diagnoses of mental retardation severity was examined through the comparison of psychiatric and psychological case reports found in client records. For a sample of 126 dually diagnosed clients, overall chance-corrected agreement was .47, ranging from .38 (for diagnoses of moderate mental retardation) to .55 (for diagnoses of borderline mental retardation). Results showed that the analysis of naturally occurring variation in diagnostic practices (including variability in professional judgment and assessment methods) may offer a more realistic appraisal of the reliability of diagnostic decisions in professional practice situations than do studies in which important sources of diagnostic error are controlled.

Electrophysiology of stimulus-secretion coupling in human beta-cells.

Herein, we review the applicability to human beta-cells of an electrophysiologically based hypothesis of the coupling of glucose metabolism to insulin secretion. According to this hypothesis, glucose metabolism leads to the generation of intracellular intermediates (including ATP), which leads to closure of ATP-sensitive K+ channels. Channel closure results in membrane depolarization, the onset of electrical activity, and voltage-dependent Ca2+ entry. The resultant rise in cytosolic Ca2+ leads to Ca(2+)-dependent exocytosis of insulin granules. We found that most of the published experimental evidence for human beta-cells supports this hypothesis. In addition, we present three other emerging lines of evidence in support of this hypothesis for human islet beta-cells: 1) the effects of pHi-altering maneuvers on insulin secretion and electrical activity; 2) preliminary identification of LVA and HVA single Ca2+ channel currents; and 3) validation of the feasibility of Cm measurements to track insulin granule exocytosis. On the basis of this last new line of evidence, we suggest that combinations of Cm measurements and electrical activity/membrane current measurements may help define the roles of diverse electrical activity patterns, displayed by human beta-cells, in stimulus-induced insulin secretion.

Stimulus-secretion coupling in beta-cells of transplantable human islets of Langerhans. Evidence for a critical role for Ca2+ entry.

With human islets isolated for transplantation, we examined the applicability to humans of a metabolic fuel hypothesis of glucose transduction and a Ca2+ hypothesis of depolarization-secretion coupling, both previously proposed for rodent islet beta-cells. We report that several features of human beta-cell physiology are well accounted for by these hypotheses. With whole-islet perifusion, we demonstrated that insulin secretion induced by glucose, tolbutamide, or elevated K+ is dependent on extracellular Ca2+. Insulin release induced by these secretagogues is enhanced by the dihydropyridine Ca2+ channel agonist BAYk8644 and depressed by the dihydropyridine Ca(2+)-channel antagonist nifedipine. All of the aforementioned secretagogues provoke increases in cytosolic free Ca2+, which are dependent on extracellular Ca2+ and are altered by the dihydropyridine drugs. Individual beta-cells in the islet display diminished resting membrane conductance, graded depolarization, and complex electrical patterns, including bursts of action potentials in response to stimulatory concentrations of glucose or tolbutamide. Individual islet beta-cells display voltage-dependent Ca2+ currents that are activated at membrane potentials traversed during the excursion of the action potential. In most cells, the Ca2+ currents are enhanced by BAYk8644 and depressed by nifedipine at concentrations that have parallel effects on secretagogue-induced increases in cytosolic Ca2+ and insulin secretion. These survey studies should provide the basis for more detailed investigations of the relationship of voltage-dependent ionic currents to electrical activity patterns and of electrical activity patterns to granule exocytosis in single human beta-cells.

Effects of metabolic inhibition by sodium azide on stimulus-secretion coupling in B cells of human islets of Langerhans.

Sodium azide (NaN3), a reversible inhibitor of mitochondrial respiration, blocks glucose-induced electrical activity and insulin secretion in human pancreatic islet B cells. Here we show that brief (10-15 min) application followed by removal of 3 mM NaN3 results in transient overshoot of electrical activity and insulin secretion even at substimulatory levels of glucose (3-5 mM). In addition, application of NaN3, even at very low [Ca2+]o, reversibly increases cytosolic Ca2+ to levels usually associated with substantial insulin release. These results suggest that (i) metabolic inhibition may reset B cell stimulus-secretion coupling and (ii) a rise in free cytosolic Ca2+, by itself, is not sufficient to trigger insulin secretion.

A comparison of performance measures of an added-purpose task versus a single-purpose task for upper extremities.

Occupational therapists often employ multidimensional tasks. These tasks, referred to as added-purpose tasks, provide a person with necessary exercise, yet direct the focus of attention toward another objective or outcome. The assumption is that the additional purpose will result in improved task performance, provided it is sufficiently distracting or meaningful to the person. The present study examined the effects of an added-purpose task compared with a single-purpose task on performance, as measured by the number of repetitions, task duration, and exercise heart rate. Thirty subjects performed either the added-purpose task or the single-purpose task three times during a 2-week period. Each session was terminated when the subject reported that he or she was exercising at a "very hard" rate on a measure of perceived exertion. A multiple analysis of variance for repeated measures indicated no significant difference between the performance of the subjects in the added-purpose versus the single-purpose task group on any of the dependent measures. Solicitation of patients' assessment of the value and meaningfulness of the rehabilitative task has practical importance.

Effects of cocaine, exercise, and resting conditions on plasma corticosterone and catecholamine concentrations in the rat.

Cocaine and exercise are both known as stressors, but little is known about the combined effects of these two treatments. In this study, rats under the influence of cocaine (12.5 mg/kg, intraperitoneally [IP]) or saline were exposed to a variety of resting conditions, as well as exercise (running, 26 m/min, 10% grade, for 30 minutes), to evaluate the amount of stress imposed by these conditions as determined by the changes in the plasma concentrations of corticosterone (C) and catecholamines (norepinephrine [NE], epinephrine [E], dopamine [DA]). After injection of saline, resting near the operating treadmill for 30 minutes caused the concentration of C to increase from 0.07 +/- 0.03 to 0.30 +/- 0.05 microgram/mL (P less than .05), compared to the increase to only 0.15 +/- 0.04 micrograms/mL after resting in a cage. This increase due to proximity to the treadmill subsided after 50 minutes. After cocaine, the 30-minute resting values were 0.70 +/- 0.15 (treadmill) and 0.55 +/- 0.13 (cage) (P less than .05), and did not subside after 50 minutes. Cocaine also increased levels of E, NE, and DA above those in saline under all rest conditions. With exercise, the value for C in saline increased to 0.61 +/- 0.18, but, in cocaine, the value went to 0.93 +/- 0.05 (P less than .05). The concentrations of E (946 +/- 74 v 603 +/- 101 pg/mL, cocaine v saline) and NE (1,027 +/- 102 v 440 +/- 153, cocaine v saline) during exercise also were exaggerated by cocaine treatment (P less than .05).(ABSTRACT TRUNCATED AT 250 WORDS)