Does methocarbamol affect fatigue markers in the low-back electromyogram?

Many low-back patients undergo electromyography (EMG)-based evaluations of muscle performance but present to the clinic after being prescribed muscle relaxants. The question that needed to be addressed was, do centrally acting muscle relaxants (methocarbamol; Robaxin®) affect the EMG spectral indices of muscle fatigue that are often used to assess muscle performance. Participants performed an isometric spine extension protocol involving a 30 s fatigue exertion trial, then 1 min rest, and finally a 10 s long repeat exertion trial, at a 60% maximum voluntary contraction (MVC) level of exertion. Seven men were tested on two separate days (approximately 3–7 days apart), one day while medicated (six doses) with Robaxin and on another while not medicated. Specifically, the following parameters were studied in the bilateral multifidus (L5), lower erector spinae (L3) and upper erector spinae (T9): the slope of median power frequencies (MPFs) over the duration of the trial and the initial y-intercept of the MPF. The results generally suggest that methocarbamol (Robaxin) does not have any significant affect on the EMG median power frequency of the extensors during a fatiguing contraction followed by a repeat exertion, at least in normal people (one exception was observed—one side of multifidus at L5). However, given that this appears to be the first study of its kind, and that a relatively small number of subjects were used in this study, further investigation is needed to make a definitive conclusion about the effects of this drug on the several features of the electromyogram, over a broad spectrum of the clinical population performing a wider variety of tasks.     

Effect of multiple-load training on the force-velocity relationship

The effect of training with a combination of different loads (multiple-load training) on the force-velocity and force-power relationships was examined with training programs that included maximal isometric contraction (Fmax) and concentric contraction of the elbow flexor muscles. Twenty-one male college students were placed into 3 equal training groups (G(30 + 60), G(30 + 100), and G(30 + 60 + 100)) and performed multiple-load training 3 days per week for 8 weeks. The training load was a set fraction of the maximal isometric strength (% Fmax). The G(30 + 60) group performed 6 repetitions of elbow flexion at 30 and 60% Fmax. The G(30 + 100) group performed 6 repetitions at 30% Fmax and six 5-second Fmax loads. The G(30 + 60 + 100) group performed 4 repetitions at 30 and 60% Fmax and four 5-second Fmax loads. After training, Fmax and maximal velocity significantly increased (p < 0.05) in all 3 training groups. The increases in maximal power were significantly (p < 0.05) different between the G(30 + 60 + 100) group (52.9%) and the G(30 + 100) group (24.2%). These results suggest that multiple-load training programs with 4-6 repetitions are effective for improving muscle power and velocity of the elbow flexors.     

不同频率的振动训练对大鼠早期膝骨关节炎关节软骨的修复作用及其JNK/NF-κB、SOX9机制*

目的: 探讨不同频率的振动训练对大鼠早期膝骨关节炎关节软骨的修复作用及其JNK/NF-κB、SOX9的机制。方法: 成年雄性SD大鼠48只,随机分为6组(n=8):模型对照组(MC组),高频振动1组(GP₁组,频率60 Hz),高频振动2组(GP₂组,频率40 Hz)、中频振动组(ZP组,频率20 Hz)和低频振动组(DP组,频率10 Hz),正常对照组(NC组)。除正常组外,各组大鼠经第1、4、7日双后腿膝关节腔注射2%木瓜蛋白酶溶液和L-半胱氨酸混合液6周后建立早期膝骨关节炎模型后,对振动组大鼠双膝进行4周,每天 40 min,振幅2~5 mm,每周振动5 d的训练。4周后,检测双膝关节股骨外侧髁关节软骨HE染色、番红O染色和Mankin评分形态学观察,股骨内侧髁关节软骨RT-qPCR检测JNK、NF-κB p65、SOX9 mRNA,Western blot 检测JNK、NF-κB p65、SOX9蛋白表达。结果: 与NC比较,其余各组Mankin评分均显著增高(P＜0.01),与MC相比,各振动组Mankin评分均显著降低(P＜0.05),其JNK、NF-κB p65mRNA和蛋白质表达显著降低(P＜0.01),SOX9mRNA和蛋白质表达显著升高(P＜0.01);与高频组相比,低频组的Mankin评分,JNK、NF-κB p65mRNA和蛋白质表达均显著降低(P＜0.05,P＜0.01),而SOX9mRNA和蛋白质表达则显著升高(P＜0.05,P＜0.01)。结论: 不同频率的振动训练对早期膝骨关节炎关节软骨可表现出不同程度的修复效应,低频振动训练软骨修复优于高频振动。可能通过下调关节软骨JNK/NF-κB表达,提高SOX9活性来调控胶原合成。     

基底颜色对两种沙蜥体色变异的影响

生存在不同基底颜色环境下的爬行动物种群通常表现出丰富的体色地理变异, 其体色变化的潜在机制具有多样性。变色沙蜥(Phrynocephalus versicolor)和草原沙蜥(P. frontalis)具有较近的遗传关系, 曾被认为与荒漠沙蜥(P. przewalskii)组成同一系统发育种组。本文应用光纤光谱仪(AvaSpec-2048), 通过记录沙蜥背部体表12个部位的皮肤光反射率, 定量比较在黑化环境下的深色变色沙蜥与非黑化环境下的浅色草原沙蜥自然体色变异, 研究其种群体色变异是否具有时间可逆性, 并探讨基底颜色对沙蜥体色的影响机制。研究结果表明, 黑化生境下的变色沙蜥体色显著深于非黑化枯黄色生境下的草原沙蜥。此外, 对黑化与非黑化样本开展的生境互换移植围栏实验, 即把枯黄色生境中非黑化的草原沙蜥移植于黑色的基底环境中饲养, 把黑化生境中黑化的变色沙蜥移植于枯黄色生境中饲养。结果表明, 饲养1周后黑化群体背部6个检测部位的光反射率显著变大, 其他部位均无显著变化; 而非黑化群体只有左后肢和背部右上方2个部位的皮肤光反射率发生显著变化, 其他部位反射率无显著变化。结果表明, 变色沙蜥体色变异能力比草原沙蜥强, 体色表型可能已经在两个近缘沙蜥物种中稳定遗传, 基底生境颜色的短期变化在统计学上能引起肉眼难以识别的轻微的体色变异, 个体发育相关的一些遗传因素可能对体色变异起控制 作用。     

Reliability of burst superimposed technique to assess central activation failure during fatiguing contraction.

Recording a superimposed electrically-induced contraction at the limit of endurance during voluntary contraction is used as an indicator of failure of muscle activation by the central nervous system and discards the existence of peripheral muscle fatigue. We questioned on the reliability of this method by using other means to explore peripheral muscle failure. Fifteen normal subjects sustained handgrip at 60% of maximal voluntary contraction (MVC) until exhaustion. During sustained contraction, the power spectrum analysis of the flexor digitorum surface electromyogram allowed us to calculate the leftward shift of median frequency (MF). A superimposed 60 Hz 3 s pulse train (burst superimposition) was delivered to the muscle when force levelled off close to the preset value. Immediately after the fatigue trial had ended, the subject was asked to perform a 5 s 60% MVC and we measured the peak contractile response to a 60 Hz 3 s burst stimulation. Recordings of the compound evoked muscle action potential (M-wave) allowed us to explore an impairment of neuromuscular propagation. A superimposed contraction was measured in 7 subjects in their two forearms, whereas it was absent in the 8 others. Despite these discrepancies, all subjects were able to reproduce a 3 s 60% MVC immediately after the fatigue trial ended and there was no post-fatigue decrease of contraction elicited by the 60 Hz 3 s burst stimulation, as well as no M-wave decrease in amplitude and conduction time. Thus, there was no indication of peripheral muscle fatigue. MF decrease was present in all individuals throughout the fatiguing contraction and it was not correlated with the magnitude of superimposed force. These observations indicate that an absence of superimposed electrically-induced muscle contraction does not allow us to conclude the existence of a sole peripheral muscle fatigue in these circumstances.     

Loss of caveolin-1 expression is associated with disruption of muscarinic cholinergic activities in the urinary bladder

Caveolin-1 (Cav1), a structural protein of caveolae, plays cell- and context-dependent roles in signal transduction pathway regulation. We have generated a knockout mouse homozygous for a null mutation of the Cav1 gene. Cav1 knockout mice exhibited impaired urinary bladder contractions in vivo during cystometry. Contractions of male bladder strips were evoked with electric and pharmacologic stimulation (5–40 Hz, 1–10 μM carbachol, 10 mM ,β-methylene ATP, 100 mM KCl). Acetylcholine (ACh) and norepinephrine (NE) release from bladder strips were measured with a radiochemical method by incubating the strips with ¹⁴C-choline and ³H-NE prior to electric stimulation, whereas ATP release was measured using the luciferin-luciferase assay with a luminometer. A 60–75% decline in contractility was observed when Cav1 knockout muscle strips were stimulated with electric current or carbachol, compared to wildtype muscle strips. No difference in contractility was noted when contractions were evoked either by the purinergic agonist ,β-methylene ATP, or by extracellular potassium. To investigate the relative contribution of non-cholinergic activity to bladder contractility, the amplitude of the electric stimulation-evoked contractions was compared in the presence of the muscarinic antagonist atropine (1 μM). While the non-muscarinic (purinergic) response was unaltered, muscarinic cholinergic response was principally disrupted in Cav1 knockout mice. The loss of Cav1 gene expression was also associated with a 70% reduction in ACh release. NE and ATP release was not altered. It is concluded that the loss of caveolin-1 is associated with disruption of M₃ muscarinic cholinergic activity in the bladder. Both pre-junctional (acetylcholine neurotransmitter release from neuromuscular junctions) and post-junctional (M₃ receptor-mediated signal transduction in bladder smooth muscles) mechanisms are disrupted, resulting in impaired bladder contraction.     

The effect of two piglet teeth resection procedures on the welfare of sows in farrowing crates. Part 2

Recent EU legislation discourages the practice of resecting piglets’ needle teeth. However, the effect of leaving piglets’ teeth intact on the welfare of sows in farrowing crates is poorly understood. Therefore, the aim of the study was to compare the effects of clipping and grinding piglets’ needle teeth, compared to leaving them intact, on the welfare of sows in farrowing crates.

Six days pre-partum 60 multiparous sows were assigned to one of three treatments. Litters had their teeth clipped (C), ground (G) or left intact (I) at birth. Sows’ teats were inspected for lesions pre-partum (day −3) and on days 1, 4, 11, 18 and 27 post-partum. Instantaneous scan samples (5 min intervals) of sow behaviour were carried out during three 2 h periods on days 1, 4, 8, 14, 21 and 26. On days 1, 4 and 11 all piglets were removed from the crate for 60 min. On re-introduction of the piglets, sow maternal behaviour was recorded continuously for 20 min.

The number of sows with teat lesions tended to differ between treatments on days 11 (P = 0.06) and 18 (P = 0.10). There was an interactive effect between treatment and day on sow dog-sitting behaviour throughout lactation (P < 0.001) and a tendency for an interactive effect on posture-changing behaviour (P = 0.08). On day 21, I sows were dog-sitting in more observations than C sows (P < 0.05) and on day 26 in more observations than C and G sows (P < 0.001). There was an interaction between treatment and day in the latency of sows to suckle their piglets following 60 min separation (P < 0.05). On day 4, I sows had a shorter latency to suckle than C and G sows (P < 0.05). There was an effect of treatment on the number of sows that terminated bouts of post-suckling udder massage (P < 0.05). On day 4, more I than C sows terminated post-suckling udder massage (P = 0.01). Finally, there was an effect of treatment on the time spent lying in the ventral posture in the observations of maternal behaviour (P < 0.05). C sows spent less time lying in the ventral posture than G and I sows (P < 0.05).

There were indications that leaving the teeth intact and to a lesser extent grinding caused injury and disturbance to sows. In farrowing crates, leaving piglets’ teeth intact cannot be recommended.     

Reliability of EMG spectral parameters in repeated measurements of back muscle fatigue.

The change in median frequency of the power spectrum of the electromyographic (EMG) signal may be used as a measure of muscle fatigue. The reliability of the median frequency parameters was investigated for EMG-recording sites at L1 and L5 right and left on the erector spinae. The reliability of subjective fatigue ratings of the back muscles (Borg CR-10 scale) and of maximal trunk extension torque (MVC) was also investigated. Eleven subjects with healthy backs performed a 45-s isometric trunk extension at 80% of MVC twice a day, on three different days. Two-factor analysis of variance was made to obtain the different variances from which the standard error of measurement (SEM) and the intra class correlation coefficient (ICC) were calculated. The SEM within-day was somewhat lower than that between-days. Both were about the same at all four electrode sites. The 95% confidence interval for the studied variables was for the initial median frequency +/- 10 Hz, for the slope +/- 0.4-0.5%/s, for the MVC +/- 36 Nm and for the Borg ratings +/- 1.6. We conclude that, with the presently used method, changes or differences within these limits should be regarded as normal variability. The slope may be of limited value because of its large variability. Whether the low intraclass correlation coefficient for the EMG parameters in the presently studied test group implies a low potential in discriminating subjects with back pain can not be decisively concluded.     

High-precision satellite positioning system as a new tool to study the biomechanics of human locomotion

New Global Positioning System (GPS) receivers allow now to measure a location on earth at high frequency (5 Hz) with a centimetric precision using phase differential positioning method. We studied whether such technique was accurate enough to retrieve basic parameters of human locomotion. Eight subjects walked on an athletics track at four different imposed step frequencies (70–130 steps/min) plus a run at free pace. Differential carrier phase localization between a fixed base station and the mobile antenna mounted on the walking person was calculated. In parallel, a triaxial accelerometer, attached to the low back, recorded body accelerations. The different parameters were averaged for 150 consecutive steps of each run for each subject (total of 6000 steps analyzed). We observed a perfect correlation between average step duration measured by accelerometer and by GPS (r=0.9998, N=40). Two important parameters for the calculation of the external work of walking were also analyzed, namely the vertical lift of the trunk and the velocity variation per step. For an average walking speed of 4.0 km/h, average vertical lift and velocity variation were, respectively, 4.8 cm and 0.60 km/h. The average intra-individual step-to-step variability at a constant speed, which includes GPS errors and the biological gait style variation, were found to be 24.5% (coefficient of variation) for vertical lift and 44.5% for velocity variation. It is concluded that GPS technique can provide useful biomechanical parameters for the analysis of an unlimited number of strides in an unconstrained free-living environment.     

Comparison of surface electromyography and myotonometric measurements during voluntary isometric contractions

Objectives: Muscle stiffness increases during muscle contraction. The purpose of this study was to determine the strength of the correlation between myotonometric measurements of muscle stiffness and surface electromyography (sEMG) measurements during various levels of voluntary isometric contractions of the biceps brachii muscle. Subjects: Eight subjects (four female; four male), with mean age of 30.6±8.23 years, volunteered to participate in this study. Methods: Myotonometer and sEMG measurements were taken simultaneously from the right biceps brachii muscle. Data were obtained: (1) at rest, (2) while the subject held a 15 lb (6.8 kg) weight isometrically and, (3) during a maximal voluntary isometric contraction. Myotonometer force–displacement curves (amount of tissue displacement to a given unit of force applied perpendicular to the muscle) were compared with sEMG measurements using Pearson’s product–moment correlation coefficients. Results: Myotonometer and sEMG measurement correlations ranged from −0.70 to −0.90. The strongest correlations to sEMG were from Myotonometer force measurements between 1.00 and 2.00 kg. Conclusions: Myotonometer and sEMG measurements were highly correlated. Tissue stiffness, as measured by the Myotonometer, appears capable of assessing changes in muscle activation levels.     

Kinetic disposition of an aqueous formulation of alphacypermethrin applied to the dorsal mid-line of sheep with long wool and its effect on lice

A group of 5 adult Merino sheep with fleeces about 70 mm long (7-months growth of wool) was treated with a topical formulation of the synthetic pyrethroid insecticide, alphacypermethrin, applied to the dorsal mid-line. Insecticide concentrations at the tip, middle and base of wool staples collected from meridians along the back, upper and lower flanks were measured at intervals from 1 to 98 days after treatment. Some movement of the alphacypermethrin from the back to the lower body occurred within 24h after treatment, but despite careful application of the insecticide there was wide variation in the concentration between and within meridians. The majority of the alphacypermethrin remained close to the dorsal mid-line and near the tip of the staple. There were significant differences in the concentration between the tip, middle and base segments of the staples in the back and lower flank meridians (P < 0.05). Despite exposure of the sheep to normal weathering, there was no significant difference in the concentration of alphacypermethrin between samples collected at day 1 or day 98 after treatment (P > 0.05). Numbers of pyrethroid-susceptible lice surviving exposure in vitro for 20 h differed significantly between samples collected at different times after treatment (P < 0.05). The numbers of lice surviving in samples collected within 28 days after treatment tended to be lower than in those collected from 28 to 98 days but, in some samples, regardless of time after treatment, lice survived for 20 h in wool taken from parts of the fleece that contained high concentrations of alphacypermethrin.     

Mechano-electric interactions in heterogeneous myocardium: development of fundamental experimental and theoretical models

The heart is structurally and functionally a highly non-homogenous organ, yet its main function as a pump can only be achieved by the co-ordinated contraction of millions of ventricular cells. This apparent contradiction gives rise to the hypothesis that ‘well-organised’ inhomogeneity may be a pre-requisite for normal cardiac function. Here, we present a set of novel experimental and theoretical tools for the study of this concept. Heterogeneity, in its most condensed form, can be simulated using two individually controlled, mechanically interacting elements (duplex). We have developed and characterised three different types of duplexes: (i) biological duplex, consisting of two individually perfused biological samples (like thin papillary muscles or a trabeculae), (ii) virtual duplex, made-up of two interacting mathematical models of cardiac muscle, and (iii) hybrid duplex, containing a biological sample that interacts in real-time with a virtual muscle. In all three duplex types, in-series or in-parallel mechanical interaction of elements can be studied during externally isotonic, externally isometric, and auxotonic modes of contraction and relaxation. Duplex models, therefore, mimic (patho-)physiological mechano-electric interactions in heterogeneous myocardium at the multicellular level, and in an environment that allows one to control mechanical, electrical and pharmacological parameters. Results obtained using the duplex method show that: (i) contractile elements in heterogeneous myocardium are not ‘independent’ generators of tension/shortening, as their ino- and lusitropic characteristics change dynamically during mechanical interaction—potentially matching microscopic contractility to macroscopic demand, (ii) mechanical heterogeneity contributes differently to action potential duration (APD) changes, depending on whether mechanical coupling of elements is in-parallel or in-series, which may play a role in mechanical tuning of distant tissue regions, (iii) electro-mechanical activity of mechanically interacting contractile elements is affected by their activation sequence, which may optimise myocardial performance by smoothing intrinsic differences in APD. In conclusion, we present a novel set of tools for the experimental and theoretical investigation of cardiac mechano-electric interactions in healthy and/or diseased heterogeneous myocardium, which allows for the testing of previously inaccessible concepts.     

Isolation and characterization of an estrogen binding protein which may integrate the plethora of estrogenic actions in non-reproductive organs

A putative estrogen receptor (pER) from mouse liver has been characterized. The heterodimer protein (81–84 kDa) consists of two covalently bound subunits (61–67 and 17–27 kDa) with following characteristics: sedimentation constant — 4.9 S; IP — 4.8; dissociation constant (K_d) for estradiol-17β binding — 0.7 nmol; binding sites — 0.746 pmol/mg protein; relative binding affinity — estradiol-17β — 100, estrone — 80 and estriol — 30; specificity — does not bind, other natural steroids, synthetic estrogens, antiestrogens and bioflavonoids. Importantly, immunosuppressants, neuroleptic and carcinogens influence ³H-estradiol-17β binding to pER. Interestingly, pER is a serine phosphatase and this may have relevancy to estrogen action in Alzheimer's disease. The polyclonal anti-pER antibody does not react with estrogen receptors (ER). ER antibody does not react with pER. Remarkably, anti-pER antibody reacts with calcineurin, a brain phosphatase and anti-calcineurin antibody reacts with pER. Immunohistochemical analyses showed that pER is undetectable in reproductive organs (except ovary). It is localized on the plasma or the nuclear membranes in some, in cytoplasm and/or nucleus in other cells of non-reproductive organs (skeletal, neural, vascular, hair and retina), and in tumors (mammary, endometrial and prostate cancers, and prostatic hyperplasia). The information presented justifies the proposition that pER may mediate the estrogenic actions in non-reproductive organs.     

Nontransferrin-bound Iron in Serum of Patients Receiving Bone Marrow Transplants

Nontransferrin-bound iron (NTBI) and other parameters of iron status were measured in 40 patients undergoing bone marrow transplantation (BMT) prior to conditioning therapy (between day −10 and −7), at the time of BMT (day 0), and 2 weeks later (day +14). Serum iron and transferrin saturation values were normal before conditioning therapy. At day 0 serum iron values were high and median transferrin saturation was 98% (changes in the values of both serum iron and transferrin saturation, p < .0001). Transferrin saturation values were still elevated 2 weeks posttransplant (day +14 vs. baseline values, p = .0001). Starting at low NTBI levels pretransplant (median 0.4 , range 0–4.2 , controls: ≤ 0.4 ), all patients revealed high levels on day 0 (median 4.0 , range 1.9–6.9 , p < .0001) and 2 weeks posttransplant (median 2.7 , range 0–6.2 , p < .0001). These observations indicate that the plasma iron pool in patients undergoing BMT increases to a level at which the normal ability to sequestrate iron becomes exhausted and considerable amounts of NTBI appear in serum. This “free” form of iron can mediate the production of reactive oxygen species and may cause organ toxicity in the early posttransplantation period. © 1997 Elsevier Science Inc.     

The Calcitonin Gene-Related Peptide Activates Both cAMP and NO Pathways to Induce Relaxation of Circular Smooth Muscle Cells of Guinea-Pig Ileum

Rekik, M., M. Delvaux, J. Frexinos, and L. Bueno. Calcitonin gene-related peptide activates both cAMP and NO pathways to induce relaxation of circular smooth muscle cells of guinea-pig ileum. Peptides 18(10) 1517–1522, 1997.—The direct effects and the intracellular pathways of rCGRP were investigated on smooth muscle cells (SMC) isolated by enzymatic digestion from the circular and longitudinal layers of guinea-pig ileum. In circular SMC, rCGRP inhibited CCK8-induced contraction in a concentration-dependent manner (Cmax = 100 μM and EC₅₀ = 0.7 ± 0.4 nM). Preincubation of SMC with 1 μM Rp-cAMPs, a cAMP antagonist, abolished the relaxing effect of rCGRP; moreover, preincubation of SMC with 100 μM L-NAME, an inhibitor of NOS, inhibited the relaxing effect of rCGRP. hCGRP(8-37), a selective antagonist of rCGRP receptors, inhibited the rCGRP-induced relaxation in a concentration dependent manner whereas the vasoactive intestinal polypeptide (VIP) antagonist had no significant effect. In longitudinal SMC, rCGRP-induced relaxation was abolished by Rp-cAMPs, whereas L-NAME had no effect. In conclusion, rCGRP triggers different intracellular pathways to induce relaxation of circular or longitudinal intestinal SMC; cAMP is involved in cells from both layers while nitric oxide (NO) is involved only in relaxation of circular SMC.     

Serum malondialdehyde: possible use for the clinical management of chronic hepatitis C patients

Serum lipid peroxidation products are increased in inflammatory liver disease and, as we previously reported, also in chronic hepatitis C. We have performed a specific assay of malondialdehyde, the reported most abundant product of lipid peroxidation, in serum of twenty four chronic hepatitis C patients, before, during, and after interferon treatment. Liver biopsies were performed in each patient before and after interferon treatment. The results show higher serum malondialdehyde values in chronic hepatitis C patients than healthy subjects (n = 68) before interferon treatment (p < .001). Mean value of serum malondialdehyde levels after interferon treatment was significantly lower than before it (p < .002). Associating the histopathological findings in each of the 48 biopsies performed, with serum malondialdehyde and alanine aminotransferase activity levels, of the sample obtained the same day of biopsy, a much better correspondence with the histopathological severity was observed for malondialdehyde concentration than for alanine aminotransferase activity. These levels decreased significantly after interferon treatment. However, when the patients were grouped in responding (group I; n = 9) and non-responding (group II; n = 15) to interferon treatment, according to the histopathological findings before and after interferon, the values of group I before interferon treatment were significantly higher than group II (p < .03). Thus, a potential predictive value could be ascribed to the serum malondialdehyde levels before interferon treatment in these patients. We propose the utility of the specific assay of malondialdehyde for the clinical management of chronic hepatitis C patients.     

High-performance liquid chromatographic determination of oxodipine enantiomers, a new 1,4-dihydropyridine, applied to stereoselectivity studies in man and dog

A specific and reproducible HPLC method using a Chiral-AGP column and UV detection was developed for the evaluation of the pharmacokinetic profile of oxodipine enantiomers in dog and man. Each enantiomer was determined in plasma in the concentration range 1–400 ng/ml using the internal standard calibration method with linear regression analysis. After extraction of oxodipine and the internal standard at alkaline pH with diethyl ether—n-hexane (50:50, v/v), this method permitted the determination of each enantiomer at levels down to 10 ng/ml in dog plasma and 25 ng/ml in human plasma with sufficient accuracy (relative error <11%, n = 6) and precision (coefficient of variation <16%, n = 6). The extracted plasma volume was 500 μl and after evaporation of the organic phase, the dry residue was dissolved in 100 μl of water—2-propanol; an aliquot of 80 μl was injected into the HPLC system.     

Comparative analysis of structure, expression and PSD95-binding capacity of Lrfn, a novel family of neuronal transmembrane proteins

Leucine-rich repeat and fibronectin III domain-containing (Lrfn) has five members in mouse and human (Lrfn1, Lrfn2, Lrfn3, Lrfn4, Lrfn5), and homologues in other vertebrates. Lrfn proteins share leucine-rich repeat (LRR)–immunoglobulin-like (Ig)–fibronectin type III (Fn)–transmembrane domain structure, which is also found in LRR–Ig–Fn superfamily proteins. Mouse Lrfn genes were expressed at adult stage predominantly in the brain. In the course of development, expression of Lrfn1, Lrfn3, and Lrfn4 started from immature neural cells, whereas that of Lrfn2 and Lrfn5 was limited to mature ones. Lrfn1–5 commonly encode glycoproteins spanning the plasma membrane, with their N-terminus located on the extracellular side. C-termini of Lrfn1, Lrfn2 and Lrfn4 were bound by PDZ domains of postsynaptic protein PSD95, re-distributing PSD95 to cell periphery where the Lrfn proteins were detected. These results suggest that Lrfn proteins are neuronal components with a role in the developing or mature vertebrate nervous system.