Prevention of murine experimental allergic encephalomyelitis: cooperative effects of cyclosporine and 1 α, 25-(OH)2D3

The hormone 1 α,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) has immune modulatory activities in vitro and in vivo, and can prevent or delay the onset of experimental or spontaneous autoimmune diseases. At therapeutical doses, however, hypercalcemic side effects are found. The present experiments examined the effects of combined treatment with subtherapeutic doses of cyclosporine A (CsA) and 1,25(OH)₂D₃ on the evolution of experimental autoimmune encephalomyelitis (EAE) in SJL mice. 1,25(OH)₂D₃ at 5 μg/kg body weight (given by i.p. injection every 2 days) prevented the appearance of paralysis in 70% of the treated mice. The treatment with 1,25(OH)₂D₃ at 2 μg/kg/2 days alone had less substantial protective effects (22% disease-free animals versus 5% in the control group). However, when this subtherapeutic dose was associated to treatment with a daily dose of CsA (2 or 5 mg/kg/day), which by itself was subtherapeutic (24 and 50% disease-free animals, respectively), the association of both drugs led to near-total protection (86% disease-free animals when combined with the highest dose of CsA). When an alternate day administration schedule (CsA at 10 mg/kg and 1,25(OH)₂D₃ at 2 μg/kg, each given on alternate days from day −3 to +19 after disease induction) was used, all treated mice were completely protected clinically and histologically. The two drugs also showed additive effects on serum osteocalcin and urinary calcium and desoxypyridinoline excretion, but not on serum calcium concentration. Our experiments demonstrate that 1,25(OH)₂D₃ might be a potential dose-reducing agent for CsA in immunosuppressive therapy.     

Formation of glutathione conjugates during oxidation of eugenol by microsomal fractions of rat liver and lung

Rat hepatic and pulmonary microsomes catalyzed the formation of at least three distinct glutathione conjugates with eugenol (4-allyl-2-methoxyphenol). These three conjugates were identical with the products obtained from the chemical reaction of synthetic eugenol quinone methide and glutathione. The microsomal reaction was dependent on NADPH and oxygen and was inhibited by cytochrome P450 inhibitors such as metyrapone, 2-diethylaminoethyl-2,2'-diphenylvalerate (SKF 525-A), alpha-naphthoflavone and piperonyl butoxide. The enzyme responsible for eugenol oxidation was inducible with 3-methylcholanthrene but not phenobarbital pretreatment. The rate of formation of conjugates was not affected by the presence of glutathione-depleted cytosol which contained active glutathione transferase, even at low glutathione concentrations, suggesting that conjugation occurs nonenzymatically with an electrophilic metabolite of eugenol. Covalent binding to microsomal protein was observed using [3H]eugenol. Cumene hydroperoxide catalyzed the formation of these same glutathione conjugates via the formation of a quinone methide-like intermediate which was detected by spectroscopic means. Our results suggest that eugenol is oxidized by cytochrome P450 to a reactive quinone methide intermediate which can then covalently modify protein or conjugate with glutathione.     

Prevalence of anabolic steroid use by male and female adolescents

This study was performed to determine the prevalence of anabolic steroid use among male and female high school students. A total of 1,010 questionnaires were distributed in five high schools in a relatively affluent school district (Group I, 510 surveys with a 92.4% response rate) and five high schools in a relatively less affluent school district (Group II, 500 surveys with an 86.0% response rate). Three percent of all high school students reported using anabolic steroids: 5.0% of males and 1.4% of females. Among Group I students, 5.9% of all athletes reported using anabolic steroids, whereas only 1.5% of all Group II athletes admitted using them. A 10.2% usage of anabolic steroids among Group I male athletes was observed, but only 2.8% of Group II male athletes reported their use. The median age for anabolic steroid consumption was 17 yr. The primary source of anabolic steroids was the black market (85.2%), while physicians directly supplied anabolic steroids to only 7.4% of the adolescents. It appears that within the high school adolescent population, Group I male athletes are the primary consumers of these drugs, with a reported 10.2% prevalence.     

Electrophysiologic evaluation of the facial nerve in Bell's palsy. A review

Facial nerve paralysis is the most common mononeuropathy and idiopathic facial paralysis (Bell's palsy) the most common seventh nerve disease electromyographers may be asked to evaluate. The electrophysiologic method of choice to assess the facial nerve is side-to-side evoked amplitude comparison with the affected side expressed as a percentage of the nonaffected side. This examination should be performed on days 3, 5, 7, 9, 11 and 13 after onset of paralysis. If the percentage of surviving axons falls below 10% within the first 14 days, an incomplete recovery is suggested. Electromyography may assist in prognosticating a functional return, determining neural conduction across the site of injury and following reinervation in the recovery period. The persistence or early return of an absent R1 component of the blink reflex may qualitatively suggest a satisfactory functional outcome in facial paralysis. Supramaximally exciting the facial nerve at the stylomastoid foramen and comparing the clinical response on the affected and nonaffected side, maximum stimulation test, can also predict eventual seventh nerve return. Observing a minimal twitch, utilizing the nerve excitability test or measuring the facial nerve latency have yielded poor correlations with functional return and are of limited usefulness in the prognostication of acute facial palsies. Trigeminal somatosensory evoked potentials can be employed to evaluate the status of the trigeminal nerve as approximately 50% of patients with Bell's palsy also have lesions involving the fifth nerve. Side-to-side amplitude comparison and electromyography are the two most valuable electrophysiologic methods of assessing facial nerve functioning.     

Axilla to elbow radial nerve conduction

Numerous techniques that evaluate radial nerve conduction from the axilla or supraclavicular fossa to the elbow have been reported. A shortcoming of most protocols is determining the precise radial nerve length as it proceeds along the spiral groove. The present study dissected out and measured directly eight cadaver radial nerves from the axilla to the elbow. These values were compared with a new surface tape measurement technique from axilla to elbow across the bicep muscle, obstetrical calipers over this region, and a surface determination approximating the course of the radial nerve posteriorly in the spiral groove. The anterior surface tape-measuring procedure compared most favorably with the actual anatomic length. Nerve conduction velocities were then calculated in 20 volunteers using all 3 techniques and compared with the median nerve in the arm. The anterior and posterior tape measurements yielded a conduction velocity of 72.5 +/- 4.7 and 86.6 +/- 7.0 m/s, respectively, whereas the caliper resulted in 65.7 +/- 3.9 m/s. We conclude that proximal radial nerve length assessment employing an anterior tape measurement from axilla to elbow across the bicep musculature is precise and compares favorably with the actual anatomic length of the radial nerve.     

Comparative study of the continuous wavelet transform, derivative and partial least squares methods applied to the overlapping spectra for the simultaneous quantitative resolution of ascorbic acid and acetylsalicylic acid in effervescent tablets

The simultaneous spectrophotometric determination of ascorbic acid (AA) and acetylsalicylic acid (ASA) in effervescent tablets in the presence of the overlapping spectra was accomplished by the continuous wavelet transform (CWT), derivative spectrophotometry (DS) and partial least squares (PLS) approaches without using any chemical pre-treatment. CWT and DS calibration equations for AA and ASA were obtained by measuring the CWT and DS amplitudes corresponding to zero-crossing points of spectra obtained by plotting continuous wavelet coefficients and first-derivative absorbance values versus the wavelengths, respectively. The PLS calibration was constructed by using the concentration set and its full absorbance data consisting of 850 points from 220 to 305 nm in the range of 210-310 nm. These three methods were tested by analyzing the synthetic mixtures of the above drugs and they were applied to the real samples containing two commercial pharmaceutical preparations of subjected drugs. A comparative study was carried out by using the experimental results obtained from three analytical methodologies and precise and accurate results were obtained.     

Propagated insertional activity: a model of positive sharp wave generation

In this study we utilized a dual monopolar needle recording technique to assess propagated electromyographic insertional activity from the same single muscle fiber in order to characterize different categories of insertional activity. A total of six combinations of insertional activity were identified. Only two fundamental types of single muscle-fiber insertional discharge configurations were generated: biphasic initially-negative and monophasic positive. The propagated waveforms corresponding to these two insertional discharges were primarily triphasic initially-positive and, only rarely, monophasic positive. The monophasic positive insertional activity generated at the inserting electrode site is postulated to arise from a depolarization zone adjacent to a needle-induced peri-electrode membrane crush. The monophasic positive discharge was utilized as a model for positive sharp wave generation. It is postulated that the majority of positive sharp waves are initiated at the inserting electrode adjacent to a needle-induced zone of muscle membrane crush in contrast to the previous supposition that positive sharp waves are blocked fibrillation potentials.     

Resistance of Akt kinases to dephosphorylation through ATP-dependent conformational plasticity

Phosphorylation of a threonine residue (T308 in Akt1) in the activation loop of Akt kinases is a prerequisite for deregulated Akt activity frequently observed in neoplasia. Akt phosphorylation in vivo is balanced by the opposite activities of kinases and phosphatases. Here we describe that targeting Akt kinase to the cell membrane markedly reduced sensitivity of phosphorylated Akt to dephosphorylation by protein phosphatase 2A. This effect was amplified by occupancy of the ATP binding pocket by either ATP or ATP-competitive inhibitors. Mutational analysis revealed that R273 in Akt1 and the corresponding R274 in Akt2 are essential for shielding T308 in the activation loop against dephosphorylation. Thus, occupancy of the nucleotide binding pocket of Akt kinases enables intramolecular interactions that restrict phosphatase access and sustain Akt phosphorylation. This mechanism provides an explanation for the "paradoxical" Akt hyperphosphorylation induced by ATP-competitive inhibitor, A-443654. The lack of phosphatase resistance further contributes insight into the mechanism by which the human Akt2 R274H missense mutation may cause autosomal-dominant diabetes mellitus.