Evidence that the Na+-K+ leak/pump ratio contributes to the difference in endurance between fast- and slow-twitch muscles.

Science.gov (United States)

Clausen, T; Overgaard, K; Nielsen, O B

2004-02-01

Muscles containing predominantly fast-twitch (type II) fibres [ext. dig. longus (EDL)] show considerably lower contractile endurance than muscles containing mainly slow-twitch (type I) fibres (soleus). To assess whether differences in Na+-K+ fluxes and excitability might contribute to this phenomenon, we compared excitation-induced Na+-K+ leaks, Na+ channels, Na+-K+ pump capacity, force and compound action potentials (M-waves) in rat EDL and soleus muscles. Isolated muscles were mounted for isometric contractions in Krebs-Ringer bicarbonate buffer and exposed to direct or indirect continuous or intermittent electrical stimulation. The time-course of force decline and concomitant changes in Na+-K+ exchange and M-waves were recorded. During continuous stimulation at 60-120 Hz, EDL showed around fivefold faster rate of force decline than soleus. This was associated with a faster loss of excitability as estimated from the area and amplitude of the M-waves. The net uptake of Na+ and the release of K+ per action potential were respectively 6.5- and 6.6-fold larger in EDL than in soleus, which may in part be due to the larger content of Na+ channels in EDL. During intermittent stimulation with 1 s 60 Hz pulse trains, EDL showed eightfold faster rate of force decline than soleus. The considerably lower contractile endurance of fast-twitch compared with slow-twitch muscles reflects differences in the rate of excitation-induced loss of excitability. This is attributed to the much larger excitation-induced Na+ influx and K+ efflux, leading to a faster rise in [K+]o in fast-twitch muscles. This may only be partly compensated by the concomitant activation of the Na+-K+ pumps, in particular in fibres showing large passive Na+-K+ leaks or reduced content of Na+-K+ pumps. Thus, endurance depends on the leak/pump ratio for Na+ and K+.

Toxicity of the sulfhydryl-containing radioprotector dithiothreitol

International Nuclear Information System (INIS)

Held, K.D.; Biaglow, J.E.

1987-01-01

The toxicity of the sulfhydryl-containing radioprotector dithiothreitol (DTT) has been studied in Chinese hamster V79 cells growing in monolayer. Under the conditions used here DTT causes a biphasic toxic response in which low concentrations of the drug (0.5 to 1.0 mM) are more toxic than are lower (0.2 mM) or higher (10 mM) concentrations. This response is similar to that seen by others with other sulfhydryl compounds. This DTT-induced toxicity is prevented by catalase, glutathione, and lowered temperatures. The toxicity is enhanced by some metal chelators (EDTA) but prevented by others (desferal). Metals (copper and iron) can either enhance or decrease the toxicity depending on their concentration and whether the exposure is in medium or in buffered salt solution. The results suggest a complex chain of chemical reactions and interactions with a role of H/sub 2/O/sub 2/ and perhaps . OH in this DTT toxicity. This is discussed

Ischemia Increases the Twitch Latent Period in the Soleus and Extensor Carpi Radialis Longus Muscles from Adult Rats.

Science.gov (United States)

Morales, Camilo; Fierro, Leonardo

2017-10-01

Complete ischemia and reperfusion effects on twitch force (∫(F·t)), twitch latent period (TLP), maximal rate of rise of twitch tension (δF/δt) max , and twitch maximum relaxation rate (TMRR) were assessed. We divided 36 adult rats into four groups; two control groups (n = 9), a group undergoing 1 hour of ischemia followed by 1 hour of reperfusion (n = 9), and one group exposed to 2 hours of ischemia followed by 1 hour of reperfusion (n = 9). We have induced twitch contractions every 10 minutes in the soleus and the extensor carpi radialis longus (ECRL). Twitch contractions were recorded and then analyzed for ∫(F·t), TLP, (δF/δt) max , and TMRR. During 1 hour and 40 minutes of ischemia, TLP increased to 179 ± 24% (p values.

Effects of superoxide dismutase, dithiothreitol and formate ion on the inactivation of papain by hydroxyl and superoxide radicals in aerated solutions

International Nuclear Information System (INIS)

Lin, W.S.; Armstrong, D.A.

1978-01-01

Losses in enzyme activity and sulphydryl content have been studied in aerated papain solutions containing formate, superoxide dismutase and dithiothreitol. Both formate and dithiothreitol converted .OH to .0 2 -, whereas superoxide dismutase completely suppressed the inactivation by .0 2 -. Using results from all systems, the fraction of .0 2 - reactions with papain that caused inactivation of the enzyme was 0.33+-0.07. The results also showed that the fraction of .OH reactions, which cause inactivation of papain, is significantly higher in aerated than in oxygen-free solutions. (author)

Characterization of a half-molecular fragment obtained by reduction of human α2-macroglobulin with dithiothreitol

DEFF Research Database (Denmark)

Sjöberg, B.; Pap, S.; Kjems, Jørgen

1985-01-01

A half-molecular fragment of α2-macroglobulin has been prepared by reducing and alkylating the inter-subunit disulfide bonds in the tetrameric α2-macroglobulin molecule with 1 mM dithiothreitol (40 min) and 3 mM iodoacetamide (40 min). Further purification was made by gel chromatography...

Effect of tibial bone resection on the development of fast- and slow-twitch skeletal muscles in foetal sheep.

Science.gov (United States)

West, J M; Williams, N A; Luff, A R; Walker, D W

2000-04-01

To determine if longitudinal bone growth affects the differentiation of fast- and slow-twitch muscles, the tibial bone was sectioned at 90 days gestation in foetal sheep so that the lower leg was permanently without structural support. At 140 days (term is approximately 147 days) the contractile properties of whole muscles, activation profiles of single fibres and ultrastructure of fast- and slow-twitch muscles from the hindlimbs were studied. The contractile and activation profiles of the slow-twitch soleus muscles were significantly affected by tibial bone resection (TIBX). The soleus muscles from the TIBX hindlimbs showed: (1) a decrease in the time to peak of the twitch responses from 106.2 +/- 10.7 ms (control, n = 4) to 65.1 +/- 2.48 ms (TIBX, n = 5); (2) fatigue profiles more characteristic of those observed in the fast-twitch muscles: and (3) Ca2+ - and Sr2+ -activation profiles of skinned fibres similar to those from intact hindlimbs at earlier stages of gestation. In the FDL, TIBX did not significantly change whole muscle twitch contraction time, the fatigue profile or the Ca2+ - and Sr2+ -activation profiles of skinned fibres. Electron microscopy showed an increased deposition of glycogen in both soleus and FDL muscles. This study shows that the development of the slow-twitch phenotype is impeded in the absence of the physical support normally provided by the tibial bone. We suggest that longitudinal stretch is an important factor in allowing full expression of the slow-twitch phenotype.

The role of Six1 in muscle progenitor cells and the establishment of fast-twitch muscle fibres

OpenAIRE

Nord, Hanna

2014-01-01

Myogenesis is the process of skeletal muscle tissue formation where committed muscle progenitor cells differentiate into skeletal muscle fibres. Depending on the instructive cues the muscle progenitor cells receive they will differentiate into specific fibre types with different properties. The skeletal muscle fibres can be broadly classified as fast-twitch fibres or slow-twitch fibres, based on their contractile speed. However, subgroups of fast- and slow-twitch fibres with different metabol...

Time-resolved X-ray diffraction studies of frog skeletal muscle isometrically twitched by two successive stimuli using synchrotron radiation

International Nuclear Information System (INIS)

Tanaka, Hidehiro; Kobayashi, Takakazu; Wakabayashi, Katsuzo

1986-01-01

In order to clarify the delay between muscular structural changes and mechanical responses, the intensity changes of the equatorial and myosin layer-line reflections were studied by a time-resolved X-ray diffraction technique using synchrotron radiation. The muscle was stimulated at 12-13 0 C by two successive stimuli at an interval during which the second twitch started while tension was still being exerted by the muscle. At the first twitch, the intensity changes of the 1,0 and 1,1 equatorial reflections reached 65 and 200% of the resting values, and further changes to 55 and 220% were seen at the second twitch, respectively. Although the second twitch decreased not only the time to peak tension but also that to the maximum intensity changes of the equatorial reflections, the delay between the intensity changes and the development of tension at the first twitch were still observed at the second twitch. On the other hand, the intensities of the 42.9 nm off-meridional and the 21.5 nm meridional myosin reflections decreased at the first twitch to the levels found when a muscle was isometrically tetanized, and no further decrease in their intensities was observed at the second twitch. These results indicate that a certain period of time is necessary for myosin heads to contr0116e to tension development after their arrival in the vicinity of the thin filaments during contraction. (Auth.)

Identification of an operon, Pil-Chp, that controls twitching motility and virulence in Xylella fastidiosa.

Science.gov (United States)

Cursino, Luciana; Galvani, Cheryl D; Athinuwat, Dusit; Zaini, Paulo A; Li, Yaxin; De La Fuente, Leonardo; Hoch, Harvey C; Burr, Thomas J; Mowery, Patricia

2011-10-01

Xylella fastidiosa is an important phytopathogenic bacterium that causes many serious plant diseases, including Pierce's disease of grapevines. Disease manifestation by X. fastidiosa is associated with the expression of several factors, including the type IV pili that are required for twitching motility. We provide evidence that an operon, named Pil-Chp, with genes homologous to those found in chemotaxis systems, regulates twitching motility. Transposon insertion into the pilL gene of the operon resulted in loss of twitching motility (pilL is homologous to cheA genes encoding kinases). The X. fastidiosa mutant maintained the type IV pili, indicating that the disrupted pilL or downstream operon genes are involved in pili function, and not biogenesis. The mutated X. fastidiosa produced less biofilm than wild-type cells, indicating that the operon contributes to biofilm formation. Finally, in planta the mutant produced delayed and less severe disease, indicating that the Pil-Chp operon contributes to the virulence of X. fastidiosa, presumably through its role in twitching motility.

Matching of sarcoplasmic reticulum and contractile properties in rat fast- and slow-twitch muscle fibres.

Science.gov (United States)

Trinh, Huong H; Lamb, Graham D

2006-07-01

1. The twitch characteristics (fast-twitch or slow-twitch) of skeletal muscle fibres are determined not only by the contractile apparatus properties of the fibre, but also by the time-course of Ca2+ release and re-uptake by the sarcoplasmic reticulum (SR). The present study examined, in individual fibres from non-transforming muscle of the rat, whether particular SR properties are matched to the contractile apparatus properties of the fibre, in particular in the case of fibres with fast-twitch contractile apparatus located in a slow-twitch muscle, namely the soleus. 2. Force was recorded in single, mechanically skinned fibres from extensor digitorum longus (EDL), gastrocnemius, peroneus longus and soleus muscles. Using repeated cycles in which the SR was emptied of all releasable Ca2+ and then reloaded, it was possible to determine the relative amount of Ca2+ present in the SR endogenously, the maximum SR capacity and the rate of Ca2+ loading. The sensitivity of the contractile apparatus to Ca2+ and Sr2+ was used to classify the fibres as fast-twitch (FT), slow-twitch (ST) or mixed (fibres examined) and thereby identify the likely troponin C and myosin heavy chain types present. 3. There was no significant difference in SR properties between the groups of FT fibres obtained from the four different muscles, including soleus. Despite some overlap in the SR properties of individual fibres between the FT and ST groups, the properties of the FT fibres in all four muscles studied were significantly different from those of the ST and mixed fibres. 4. In general, in FT fibres the SR had a larger capacity and the endogenous Ca2+ content was a relatively lower percentage of maximum compared with ST fibres. Importantly, in terms of their SR properties, FT fibres from soleus muscle more closely resembled FT fibres from other muscles than they did ST fibres from soleus muscle.

X-ray and fast neutron-induced mutations in Arabidopsis thaliana, and the effect of dithiothreitol upon the mutant spectrum

International Nuclear Information System (INIS)

Dellaert, L.M.W.

1980-01-01

The author discusses the genetic effects of X-ray and fast neutron seed-irradiation of Arabidopsis thaliana (L.) Heynh., and the influence of a pre-irradiation treatment with the radio-protector dithiothreitol (DTT). (Auth.)

L-carnitine pretreatment protects slow-twitch skeletal muscles in a rat model of ischemia-reperfusion injury.

Science.gov (United States)

Demirel, Mert; Kaya, Burak; Cerkez, Cem; Ertunc, Mert; Sara, Yildirim

2013-10-01

Ischemia-reperfusion (I/R) injury negatively affects the outcome of surgical interventions for amputated or severely traumatized extremities. This study aimed to evaluate the protective role of l-carnitine on the contractile properties of fast-twitch (extensor digitorum longus [EDL]) and slow-twitch (soleus [SOL]) skeletal muscles following I/R-induced injury in a rat model. Rats were divided into 4 groups (1) saline pretreatment, (2) l-carnitine pretreatment, (3) saline pretreatment and I/R, and (4) l-carnitine pretreatment and I/R. Twitch and tetanic contractions in the EDL and SOL muscles in each group were recorded. Additionally, a fatigue protocol was performed in these muscles. Twitch and tetanic contraction amplitudes were lower in the EDL and SOL muscles in which I/R was induced (P contraction amplitude in the SOL muscles following I/R (P muscles. l-Carnitine pretreatment did not alter the fatigue response in any of the muscles.

Twitch interpolation technique in testing of maximal muscle strength

DEFF Research Database (Denmark)

Bülow, P M; Nørregaard, J; Danneskiold-Samsøe, B

1993-01-01

The aim was to study the methodological aspects of the muscle twitch interpolation technique in estimating the maximal force of contraction in the quadriceps muscle utilizing commercial muscle testing equipment. Six healthy subjects participated in seven sets of experiments testing the effects...

Developmental changes in the activation properties and ultrastructure of fast- and slow-twitch muscles from fetal sheep.

Science.gov (United States)

West, J M; Barclay, C J; Luff, A R; Walker, D W

1999-04-01

At early stages of muscle development, skeletal muscles contract and relax slowly, regardless of whether they are destined to become fast- or slow-twitch. In this study, we have characterised the activation profiles of developing fast- and slow-twitch muscles from a precocial species, the sheep, to determine if the activation profiles of the muscles are characteristically slow when both the fast- and slow-twitch muscles have slow isometric contraction profiles. Single skinned muscle fibres from the fast-twitch flexor digitorum longus (FDL) and slow-twitch soleus muscles from fetal (gestational ages 70, 90, 120 and 140 days; term 147 days) and neonatal (8 weeks old) sheep were used to determine the isometric force-pCa (pCa = -log10[Ca2+]) and force-pSr relations during development. Fast-twitch mammalian muscles generally have a greatly different sensitivity to Ca2+ and Sr2+ whereas slow-twitch muscles have a similar sensitivity to these divalent cations. At all ages studied, the force-pCa and force-pSr relations of the FDL muscle were widely separated. The mean separation of the mid-point of the curves (pCa50-pSr50) was approximately 1.1. This is typical of adult fast-twitch muscle. The force-pCa and force-pSr curves for soleus muscle were also widely separated at 70 and 90 days gestation (pCa50-pSr50 approximately 0.75); between 90 days and 140 days this separation decreased significantly to approximately 0.2. This leads to a paradoxical situation whereby at early stages of muscle development the fast muscles have contraction dynamics of slow muscles but the slow muscles have activation profiles more characteristic of fast muscles. The time course for development of the FDL and soleus is different, based on sarcomere structure with the soleus muscle developing clearly defined sarcomere structure earlier in gestation than the FDL. At 70 days gestation the FDL muscle had no clearly defined sarcomeres. Force (N cm-2) increased almost linearly between 70 and 140 days

Prdm1a and miR-499 act sequentially to restrict Sox6 activity to the fast-twitch muscle lineage in the zebrafish embryo.

Science.gov (United States)

Wang, XinGang; Ono, Yosuke; Tan, Swee Chuan; Chai, Ruth JinFen; Parkin, Caroline; Ingham, Philip W

2011-10-01

Sox6 has been proposed to play a conserved role in vertebrate skeletal muscle fibre type specification. In zebrafish, sox6 transcription is repressed in slow-twitch progenitors by the Prdm1a transcription factor. Here we identify sox6 cis-regulatory sequences that drive fast-twitch-specific expression in a Prdm1a-dependent manner. We show that sox6 transcription subsequently becomes derepressed in slow-twitch fibres, whereas Sox6 protein remains restricted to fast-twitch fibres. We find that translational repression of sox6 is mediated by miR-499, the slow-twitch-specific expression of which is in turn controlled by Prdm1a, forming a regulatory loop that initiates and maintains the slow-twitch muscle lineage.

Cyclosporin A preferentially attenuates skeletal slow-twitch muscle regeneration

Directory of Open Access Journals (Sweden)

Miyabara E.H.

2005-01-01

Full Text Available Calcineurin, a Ca2+/calmodulin-dependent phosphatase, is associated with muscle regeneration via NFATc1/GATA2-dependent pathways. However, it is not clear whether calcineurin preferentially affects the regeneration of slow- or fast-twitch muscles. We investigated the effect of a calcineurin inhibitor, cyclosporin A (CsA, on the morphology and fiber diameter of regenerating slow- and fast-twitch muscles. Adult Wistar rats (259.5 ± 9 g maintained under standard conditions were treated with CsA (20 mg/kg body weight, ip for 5 days, submitted to cryolesion of soleus and tibialis anterior (TA muscles on the 6th day, and then treated with CsA for an additional 21 days. The muscles were removed, weighed, frozen, and stored in liquid nitrogen. Cryolesion did not alter the body weight gain of the animals after 21 days of regeneration (P = 0.001 and CsA significantly reduced the body weight gain (15.5%; P = 0.01 during the same period. All treated TA and soleus muscles showed decreased weights (17 and 29%, respectively, P < 0.05. CsA treatment decreased the cross-sectional area of both soleus and TA muscles of cryoinjured animals (TA: 2108 ± 930 vs 792 ± 640 µm²; soleus: 2209 ± 322 vs 764 ± 439 m²; P < 0.001. Histological sections of both muscles stained with Toluidine blue revealed similar regenerative responses after cryolesion. In addition, CsA was able to minimize these responses, i.e., centralized nuclei and split fibers, more efficiently so in TA muscle. These results indicate that calcineurin preferentially plays a role in regeneration of slow-twitch muscle.

Twitching motility and biofilm formation are associated with tonB1 in Xylella fastidiosa

OpenAIRE

Cursino, Luciana; Li, Yaxin; Zaini, Paulo A.; De La Fuente, Leonardo; Hoch, Harvey C.; Burr, Thomas J.

2017-01-01

A mutation in the Xylella fastidiosa tonB1 gene resulted in loss of twitching motility and in significantly less biofilm formation as compared with a wild type. The altered motility and biofilm phenotypes were restored by complementation with a functional copy of the gene. The mutation affected virulence as measured by Pierce's disease symptoms on grapevines. The role of TonB1 in twitching and biofilm formation appears to be independent of the characteristic iron-uptake function of this prote...

TWITCH PARAMETERS IN TRANSVERSAL AND LONGITUDINAL BICEPS BRACHII RESPONSE

Directory of Open Access Journals (Sweden)

Boštjan Šimunič

2010-01-01

Full Text Available Assessment of the contractile properties of skeletal muscles is continuing to be an important issue and a difficult task methodologically. Longitudinal direction of skeletal muscle contraction blurs intrinsic muscle belly contractile properties with many factors. This study evaluates and explains contractile properties such as: delay time (Td, contraction time (Tc, half relaxation time (Tr and maximal amplitude (Dm extracted from twitch transversal response and compare them with torque response. In fifteen healthy males (age 23.7 ± 3.4 years isometric twitch transversal and torque responses were simultaneously recorded during graded electrically elicited contractions in the biceps brachii muscle. The amplitude of electrical stimulation was increased in 5 mA steps from a threshold up to a maximal response. The muscles’ belly transversal response was measured by a high precision mechanical displacement sensor while elbow joint torque was calculated from force readings. Results indicate a parabolic relation between the transversal displacement and the torque Dm. A significantly shorter Tc was found in transversal response without being correlated to torque Tc (r = -0.12; > 0.05. A significant correlation was found between torque Tc and the time occurrence of the second peak in the transversal response (r = 0.83; < 0.001. Electrical stimulation amplitude dependant variation of the Tc was notably different in transversal than in torque response. Td was similar at submaximal and maximal responses but larger in transversal at just above threshold contractions. Tr has a similar linear trend in both responses, however, the magnitude and the slope are much larger in the transversal response. We could conclude that different mechanisms affect longitudinal and transversal twitch skeletal muscle deformations. Contractile properties extracted from the transversal response enable alternative insights into skeletal muscle contraction mechanics.

Effect of ADP on slow-twitch muscle fibres of the rat: implications for muscle fatigue.

Science.gov (United States)

Macdonald, W A; Stephenson, D G

2006-05-15

Slow-twitch mechanically skinned fibres from rat soleus muscle were bathed in solutions mimicking the myoplasmic environment but containing different [ADP] (0.1 microm to 1.0 mm). The effect of ADP on sarcoplasmic reticulum (SR) Ca2+-content was determined from the magnitude of caffeine-induced force responses, while temporal changes in SR Ca2+-content allowed determination of the effective rates of the SR Ca2+-pump and of the SR Ca2+-leak. The SR Ca2+-pump rate, estimated at pCa (-log10[Ca2+]) 7.8, was reduced by 20% as the [ADP] was increased from 0.1 to 40 microm, with no further alteration when the [ADP] was increased to 1.0 mm. The SR Ca2+-leak rate constant was not altered by increasing [ADP] from 0.1 to 40 microm, but was increased by 26% when the [ADP] was elevated to 1.0 mm. This ADP-induced SR Ca2+-leak was insensitive to ruthenium red but was abolished by 2,5-di(tert-butyl)-1,4-hydroquinone (TBQ), indicating that the leak pathway is via the SR Ca2+-pump and not the SR Ca2+-release channel. The decrease in SR Ca2+-pump rate and SR Ca2+-leak rate when [ADP] was increased led to a 40% decrease in SR Ca2+-loading capacity. Elevation of [ADP] had only minor direct effects on the contractile apparatus of slow-twitch fibres. These results suggest that ADP has only limited depressing effects on the contractility of slow-twitch muscle fibres. This is in contrast to the marked effects of ADP on force responses in fast-twitch muscle fibres and may contribute to the fatigue-resistant nature of slow-twitch muscle fibres.

The protective effect of hypoxia and dithiothreitol on X-ray-induced genetic damage in Arabidopsis

International Nuclear Information System (INIS)

Sree Ramulu, K.; Veen, J.H. van der

1987-01-01

A study was made on the protective effect of hypoxia and dithiothreitol (DTT) on X-ray-induced ovule sterility and embryonic lethality in Arabidopsis. Both hypoxia and DTT gave a pronounced and additive reduction of radiation-induced genetic damage. The reduction was significantly higher for ovule sterility than for embryonic lethals. It is suggested that non-fertilized ovules contain a higher ratio of strand breaks/other damage than embryonic lethals do, for hypoxia and DTT are known specifically to give a reduction of strand breaks. (Auth.)

Twitching motility and biofilm formation are associated with tonB1 in Xylella fastidiosa.

Science.gov (United States)

Cursino, Luciana; Li, Yaxin; Zaini, Paulo A; De La Fuente, Leonardo; Hoch, Harvey C; Burr, Thomas J

2009-10-01

A mutation in the Xylella fastidiosa tonB1 gene resulted in loss of twitching motility and in significantly less biofilm formation as compared with a wild type. The altered motility and biofilm phenotypes were restored by complementation with a functional copy of the gene. The mutation affected virulence as measured by Pierce's disease symptoms on grapevines. The role of TonB1 in twitching and biofilm formation appears to be independent of the characteristic iron-uptake function of this protein. This is the first report demonstrating a functional role for a tonB homolog in X. fastidiosa.

Differences between glycogen biogenesis in fast- and slow-twitch rabbit muscle

DEFF Research Database (Denmark)

Cussó, R; Lerner, L R; Cadefau, J

2003-01-01

Skeletal muscle glycogen is an essential energy substrate for muscular activity. The biochemical properties of the enzymes involved in de novo synthesis of glycogen were analysed in two types of rabbit skeletal muscle fiber (fast- and slow-twitch). Glycogen concentration was higher in fast...

Can fast-twitch muscle fibres be selectively recruited during lengthening contractions? Review and applications to sport movements.

Science.gov (United States)

Chalmers, Gordon R

2008-01-01

Literature examining the recruitment order of motor units during lengthening (eccentric) contractions was reviewed to determine if fast-twitch motor units can be active while lower threshold slow-twitch motor units are not active. Studies utilizing surface electromyogram (EMG) amplitude, single motor unit activity, spike amplitude-frequency analyses, EMG power spectrum, mechanomyographic, and phosphocreatine-to-creatine ratio (PCr/Cr) techniques were reviewed. Only single motor unit and PCr/Cr data were found to be suitable to address the goals of this review. Nine of ten single motor unit studies, examining joint movement velocities up to 225 degrees/s and forces up to 53% of a maximum voluntary contraction, found that the size principle of motor unit recruitment applied during lengthening contractions. Deviation from the size principle was demonstrated by one study examining movements within a small range of low velocities and modest forces, although other studies examining similar low forces and lengthening velocities reported size-ordered recruitment. The PCr/Cr data demonstrated the activation of all fibre types in lengthening maximal contractions. Most evidence indicates that for lengthening contractions of a wide range of efforts and speeds, fast-twitch muscle fibres cannot be selectively recruited without activity of the slow-twitch fibres of the same muscle.

Properties of slow- and fast-twitch muscle fibres in a mouse model of amyotrophic lateral sclerosis.

Science.gov (United States)

Atkin, Julie D; Scott, Rachel L; West, Jan M; Lopes, Elizabeth; Quah, Alvin K J; Cheema, Surindar S

2005-05-01

This investigation was undertaken to determine if there are altered histological, pathological and contractile properties in presymptomatic or endstage diseased muscle fibres from representative slow-twitch and fast-twitch muscles of SOD1 G93A mice in comparison to wildtype mice. In presymptomatic SOD1 G93A mice, there was no detectable peripheral dysfunction, providing evidence that muscle pathology is secondary to motor neuronal dysfunction. At disease endstage however, single muscle fibre contractile analysis demonstrated that fast-twitch muscle fibres and neuromuscular junctions are preferentially affected by amyotrophic lateral sclerosis-induced denervation, being unable to produce the same levels of force when activated by calcium as muscle fibres from their age-matched controls. The levels of transgenic SOD1 expression, aggregation state and activity were also examined in these muscles but there no was no preference for muscle fibre type. Hence, there is no simple correlation between SOD1 protein expression/activity, and muscle fibre type vulnerability in SOD1 G93A mice.

On dithiothreitol (DTT as a measure of oxidative potential for ambient particles: evidence for the importance of soluble ewline transition metals

Directory of Open Access Journals (Sweden)

J. G. Charrier

2012-10-01

Full Text Available The rate of consumption of dithiothreitol (DTT is increasingly used to measure the oxidative potential of particulate matter (PM, which has been linked to the adverse health effects of PM. While several quinones are known to be very reactive in the DTT assay, it is unclear what other chemical species might contribute to the loss of DTT in PM extracts. To address this question, we quantify the rate of DTT loss from individual redox-active species that are common in ambient particulate matter. While most past research has indicated that the DTT assay is not sensitive to metals, our results show that seven out of the ten transition metals tested do oxidize DTT, as do three out of the five quinones tested. While metals are less efficient at oxidizing DTT compared to the most reactive quinones, concentrations of soluble transition metals in fine particulate matter are generally much higher than those of quinones. The net result is that metals appear to dominate the DTT response for typical ambient PM_2.5 samples. Based on particulate concentrations of quinones and soluble metals from the literature, and our measured DTT responses for these species, we estimate that for typical PM_2.5 samples approximately 80% of DTT loss is from transition metals (especially copper and manganese, while quinones account for approximately 20%. We find a similar result for DTT loss measured in a small set of PM_2.5 samples from the San Joaquin Valley of California. Because of the important contribution from metals, we also tested how the DTT assay is affected by EDTA, a chelator that is sometimes used in the assay. EDTA significantly suppresses the response from both metals and quinones; we therefore recommend that EDTA should not be included in the DTT assay.

The relationship between passive stiffness and evoked twitch properties: the influence of muscle CSA normalization

International Nuclear Information System (INIS)

Ryan, E D; Thompson, B J; Sobolewski, E J; Herda, T J; Costa, P B; Walter, A A; Cramer, J T

2011-01-01

Passive stiffness measurements are often used as a clinical tool to examine a muscle's passive lengthening characteristics. The purpose of this study was to examine the relationship between passive stiffness and evoked twitch properties prior to and following normalization of passive stiffness to muscle cross-sectional area (CSA). Ten healthy volunteers (mean ± SD age = 23 ± 3 year) performed passive range of motion, evoked twitch, and muscle CSA assessments of the plantar flexor muscles. Passive stiffness was determined from the slope of the final 5° of the angle–torque curve. Peak twitch torque (PTT) and rate of torque development (RTD) were determined via transcutaneous electrical stimulation, and muscle CSA was assessed using a peripheral quantitative computed tomography scanner. Pearson product moment correlation coefficients (r) were used to assess the relationships between passive stiffness and PTT and RTD and normalized passive stiffness (passive stiffness . muscle CSA −1 ) and PTT and RTD. Significant positive relationships were observed between passive stiffness and PTT (P = 0.003, r = 0.828) and RTD (P = 0.003, r = 0.825). There were no significant relationships between normalized passive stiffness and PTT (P = 0.290, r = 0.372) or RTD (P = 0.353, r = 0.329) demonstrating that stiffness did not account for a significant portion of the variance in twitch properties. Passive stiffness was largely influenced by the amount of muscle tissue in this study. Future studies that examine muscle stiffness and its relationship with performance measures, among different populations, and following various interventions may consider normalizing stiffness measurements to muscle CSA

Calcium-Enhanced Twitching Motility in Xylella fastidiosa Is Linked to a Single PilY1 Homolog.

Science.gov (United States)

Cruz, Luisa F; Parker, Jennifer K; Cobine, Paul A; De La Fuente, Leonardo

2014-12-01

The plant-pathogenic bacterium Xylella fastidiosa is restricted to the xylem vessel environment, where mineral nutrients are transported through the plant host; therefore, changes in the concentrations of these elements likely impact the growth and virulence of this bacterium. Twitching motility, dependent on type IV pili (TFP), is required for movement against the transpiration stream that results in basipetal colonization. We previously demonstrated that calcium (Ca) increases the motility of X. fastidiosa, although the mechanism was unknown. PilY1 is a TFP structural protein recently shown to bind Ca and to regulate twitching and adhesion in bacterial pathogens of humans. Sequence analysis identified three pilY1 homologs in X. fastidiosa (PD0023, PD0502, and PD1611), one of which (PD1611) contains a Ca-binding motif. Separate deletions of PD0023 and PD1611 resulted in mutants that still showed twitching motility and were not impaired in attachment or biofilm formation. However, the response of increased twitching at higher Ca concentrations was lost in the pilY1-1611 mutant. Ca does not modulate the expression of any of the X. fastidiosa PilY1 homologs, although it increases the expression of the retraction ATPase pilT during active movement. The evidence presented here suggests functional differences between the PilY1 homologs, which may provide X. fastidiosa with an adaptive advantage in environments with high Ca concentrations, such as xylem sap. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Time-resolved x-ray diffraction from frog skeletal muscle during an isotonic twitch under a small load

International Nuclear Information System (INIS)

Sugi, Haruo; Amemiya, Yoshiyuki; Hashizume, Hiroo.

1978-01-01

A time-resolved x-ray diffraction technique was used to study the time course of change in the intensity ratio Isub(1,0)/Isub(1,1) during isotonic twitch (initial sarcomere, 2.4 μm) under a small load and to determine the kinetic properties of the crossbridges responsible for muscle contraction. Isotonic twitches in four other preparations with an initial sarcomere of 2.4 μm and in two with an initial sarcomere of 2.3 μm and 2.2 μm, respectively, were examined. In each case, the intensity ratio started to decrease at stimulation, reached a minimum value of 0.8 - 1.0 within the first 20 - 30% of the shortening phase, and maintained this value until the beginning of the relaxation phase. Gradual recovery of the intensity ratio to the resting value was seen during the relaxation phase. During the recovery phase, the intensity ratio appeared to exhibit oscillatory changes. Though the extent of shortening was reduced by about 30% at the end of each experiment, the duration of the shortening phase remained almost unchanged in all the preparations examined. The time course of change in the intensity ratio was also examined during an isometric twitch in four preparations (sarcomere, 2.4 μm) with the tibial end connected to a strain gauge. The extent of internal shortening of muscle fibres against the tendons and the recording system during an isometric twitch or a tetanus at low temperatures was estimated. The intensity ratio decreased to a minimum value of 0.5 - 0.6 during the rising phase of isometric tension and started to return to the resting value after the beginning of relaxation. In both isotonic and isometric twitches, a decrease in the intensity ratio resulted from both a decrease in the 1,0 intensity and an increase in the 1,1 intensity. (J.P.N.)

Train-of-four recovery precedes twitch recovery during reversal with sugammadex in pediatric patients: A retrospective analysis.

Science.gov (United States)

Vieira Carlos, Ricardo; Luis Abramides Torres, Marcelo; de Boer, Hans Donald

2018-04-01

After reversal of a rocuronium-induced neuromuscular blockade with sugammadex, the recovery of train-of-four ratio to 0.9 is faster than recovery of first twitch of the train-of-four to 90% in adults. These findings after reversal of neuromuscular blockade with sugammadex have not yet been investigated in pediatric patients. The aim of this retrospective analysis was to investigate the relationship of the recovery of first twitch of the train-of-four height and train-of-four ratio after reversal of rocuronium-induced neuromuscular blockade with sugammadex in pediatric patients. Patients ASA I-III, aged 2-11 years, and who underwent abdominal and/or perineal surgery were included in the analysis. After extracting the necessary data from the hospital database, the patients were divided into 2 groups based on the dose of sugammadex received: group A: 2 mg.kg -1 for reversal of moderate neuromuscular blockade and group B: 4 mg.kg -1 for reversal of deep neuromuscular blockade. The relationship of the recovery of first twitch of the train-of-four height and train-of-four ratio in these 2 groups were analyzed. Data from 43 pediatric patients aged 2-11 years could be analyzed. The first twitch of the train-of-four height at the recovery of train-of-four ratio to 0.9 in group B was statistically significantly lower compared with group A. This height 3 and 5 minutes after the train-of-four ratio reached 0.9 showed no statistically significant differences between groups. The results were in line with the results found in adults and showed that the train-of-four ratio recovered to 0.9 was faster than first twitch of the train-of-four height recovered to the same level. © 2018 John Wiley & Sons Ltd.

American football and other sports injuries may cause migraine/persistent pain decades later and can be treated successfully with electrical twitch-obtaining intramuscular stimulation (ETOIMS).

Science.gov (United States)

Chu, J; McNally, S; Bruyninckx, F; Neuhauser, D

2017-04-01

Autonomous twitch elicitation at myofascial trigger points from spondylotic radiculopathies-induced denervation supersensitivity can provide favourable pain relief using electrical twitch-obtaining intramuscular stimulation (ETOIMS). To provide objective evidence that ETOIMS is safe and efficacious in migraine and persistent pain management due to decades-old injuries to head and spine from paediatric American football. An 83-year-old mildly hypertensive patient with 25-year history of refractory migraine and persistent pain self-selected to regularly receive fee-for-service ETOIMS 2/week over 20 months. He had 180 sessions of ETOIMS. Pain levels, blood pressure (BP) and heart rate/pulse were recorded before and immediately after each treatment alongside highest level of clinically elicitable twitch forces/session, session duration and intervals between treatments. Twitch force grades recorded were from 1 to 5, grade 5 twitch force being strongest. Initially, there was hypersensitivity to electrical stimulation with low stimulus parameters (500 µs pulse-width, 30 mA stimulus intensity, frequency 1.3 Hz). This resolved with gradual stimulus increments as tolerated during successive treatments. By treatment 27, autonomous twitches were noted. Spearman's correlation coefficients showed that pain levels are negatively related to twitch force, number of treatments, treatment session duration and directly related to BP and heart rate/pulse. Treatment numbers and session durations directly influence twitch force. At end of study, headaches and quality of life improved, hypertension resolved and antihypertensive medication had been discontinued. Using statistical process control methodology in an individual patient, we showed long-term safety and effectiveness of ETOIMS in simultaneous diagnosis, treatment, prognosis and prevention of migraine and persistent pain in real time obviating necessity for randomised controlled studies.

Inositol 1,4,5-trisphosphate-sensitive Ca2+ release in rat fast- and slow-twitch skinned muscle fibres.

Science.gov (United States)

Talon, S; Huchet-Cadiou, C; Léoty, C

1999-11-01

Inositol 1,4,5-trisphosphate (InsP3), an intracellular messenger, induces Ca2+ release in various types of cells, particularly smooth muscle cells. Its role in skeletal muscle, however, is controversial. The present study shows that the application of InsP3 to rat slow- and fast-twitch saponin-skinned fibres induced contractile responses that were not related to an effect of InsP3 on the properties of the contractile proteins. The amplitude of the contractures was dependent upon the Ca(2+)-loading period, and was larger in slow- than in fast-twitch muscle. In both types of skeletal muscle, these responses, unlike caffeine contractures, were not inhibited by ryanodine (100 microM), but were abolished by heparin (20 micrograms.ml-1). In soleus muscle, the concentration of heparin required to inhibit the response by 50% (IC50) was 5.7 micrograms.ml-1, a similar value to that obtained previously in smooth muscle. Furthermore, the results show that in slow-twitch muscle, the InsP3 contractures have a "bell-shaped" dependency on the intracellular Ca2+ concentration. These results show that InsP3 receptors should be present in skeletal muscle. Thus, it is possible that InsP3 participates in the regulation of sarcoplasmic reticulum Ca2+ release in skeletal muscle, particularly in slow-twitch fibres.

Observation of the molecular organization of calcium release sites in fast- and slow-twitch skeletal muscle with nanoscale imaging.

Science.gov (United States)

Jayasinghe, Isuru D; Munro, Michelle; Baddeley, David; Launikonis, Bradley S; Soeller, Christian

2014-10-06

Localization microscopy is a fairly recently introduced super-resolution fluorescence imaging modality capable of achieving nanometre-scale resolution. We have applied the dSTORM variation of this method to image intracellular molecular assemblies in skeletal muscle fibres which are large cells that critically rely on nanoscale signalling domains, the triads. Immunofluorescence staining in fixed adult rat skeletal muscle sections revealed clear differences between fast- and slow-twitch fibres in the molecular organization of ryanodine receptors (RyRs; the primary calcium release channels) within triads. With the improved resolution offered by dSTORM, abutting arrays of RyRs in transverse view of fast fibres were observed in contrast to the fragmented distribution on slow-twitch muscle that were approximately 1.8 times shorter and consisted of approximately 1.6 times fewer receptors. To the best of our knowledge, for the first time, we have quantified the nanometre-scale spatial association between triadic proteins using multi-colour super-resolution, an analysis difficult to conduct with electron microscopy. Our findings confirm that junctophilin-1 (JPH1), which tethers the sarcoplasmic reticulum ((SR) intracellular calcium store) to the tubular (t-) system at triads, was present throughout the RyR array, whereas JPH2 was contained within much smaller nanodomains. Similar imaging of the primary SR calcium buffer, calsequestrin (CSQ), detected less overlap of the triad with CSQ in slow-twitch muscle supporting greater spatial heterogeneity in the luminal Ca2+ buffering when compared with fast twitch muscle. Taken together, these nanoscale differences can explain the fundamentally different physiologies of fast- and slow-twitch muscle. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Blood flow response to electrically induced twitch and tetanic lower-limb muscle contractions.

NARCIS (Netherlands)

Janssen, T.W.; Hopman, M.T.E.

2003-01-01

OBJECTIVES: To compare the effect of electric stimulation (ES)-induced twitch with tetanic leg muscle contractions on blood flow responses and to assess blood flow responses in the contralateral inactive leg. DESIGN: Intervention with within-subject comparisons. SETTING: University research

EXERCISE, MANUAL THERAPY AND POSTURAL RE-EDUCATION FOR UNCONTROLLED EAR TWITCHING AND RELATED IMPAIRMENTS AFTER WHIPLASH INJURY: A CASE REPORT.

Science.gov (United States)

Flanders, Kelsey; Feldner, Heather

2017-10-01

Whiplash Associated Disorders and the interventions used to remediate them are well documented in physical therapy literature. However, specific interventions for spasms of the neck musculature that also involve constant ear twitching have yet to be addressed. The purpose of this case report is twofold. First, to describe comprehensive physical therapy management and outcomes for a subject with uncontrolled ear twitching and related musculoskeletal impairments, and second, to discuss the physical therapist's approach to evidence-based care when faced with a paucity of literature addressing physical therapy interventions for subjects with uncontrolled ear twitching. The subject was a 14-year-old female who sustained a right anterolateral whiplash injury when struck in the head by a volleyball seven months prior to physical therapy. Beginning five months after that injury, she experienced uncontrolled and constant superior/inferior movement of her right ear (hereafter described in this report as a twitch) in addition to facial and cervical pain from her initial injury. She was unable to participate in high school athletics due to her pain. A multimodal treatment approach including exercise, manual therapy, and postural reeducation was utilized during the subject's episode of care. After eight treatment sessions, the subjects's cervical range of motion and upper extremity strength improved. The reported frequency of ear twitching decreased, as did reports of neck and shoulder pain. In addition, her Neck Disability Index improved from a score of 22, indicating moderate disability, to 9, indicating mild disability and she was able to return to sport activity. With limited research to direct intervention, clinical reasoning was utilized to formulate an effective therapeutic intervention. A combination of manual therapy, exercise, and postural reeducation intervention was effective for this subject and could assist in guiding interventions for similarly unique clinical

Coexistence of potentiation and fatigue in skeletal muscle

Directory of Open Access Journals (Sweden)

D.E. Rassier

2000-05-01

Full Text Available Twitch potentiation and fatigue in skeletal muscle are two conditions in which force production is affected by the stimulation history. Twitch potentiation is the increase in the twitch active force observed after a tetanic contraction or during and following low-frequency stimulation. There is evidence that the mechanism responsible for potentiation is phosphorylation of the regulatory light chains of myosin, a Ca2+-dependent process. Fatigue is the force decrease observed after a period of repeated muscle stimulation. Fatigue has also been associated with a Ca2+-related mechanism: decreased peak Ca2+ concentration in the myoplasm is observed during fatigue. This decrease is probably due to an inhibition of Ca2+ release from the sarcoplasmic reticulum. Although potentiation and fatigue have opposing effects on force production in skeletal muscle, these two presumed mechanisms can coexist. When peak myoplasmic Ca2+ concentration is depressed, but myosin light chains are relatively phosphorylated, the force response can be attenuated, not different, or enhanced, relative to previous values. In circumstances where there is interaction between potentiation and fatigue, care must be taken in interpreting the contractile responses.

Reduced phrenic motoneuron recruitment during sustained inspiratory threshold loading compared to single-breath loading: a twitch interpolation study

Directory of Open Access Journals (Sweden)

Mathieu Raux

2016-11-01

Full Text Available In humans, inspiratory constraints engage cortical networks involving the supplementary motor area. Functional magnetic resonance imaging (fMRI shows that the spread and intensity of the corresponding respiratory-related cortical activation dramatically decrease when a discrete load becomes sustained. This has been interpreted as reflecting motor cortical reorganisation and automatisation, but could proceed from sensory and/or affective habituation. To corroborate the existence of motor reorganisation between single-breath and sustained inspiratory loading (namely changes in motor neurones recruitment, we conducted a diaphragm twitch interpolation study based on the hypothesis that motor reorganisation should result in changes in the twitch interpolation slope. Fourteen healthy subjects (age: 21 – 40 years were studied. Bilateral phrenic stimulation was delivered at rest, upon prepared and targeted voluntary inspiratory efforts (vol, upon unprepared inspiratory efforts against a single-breath inspiratory threshold load (single-breath, and upon sustained inspiratory efforts against the same type of load (continuous. The slope of the relationship between diaphragm twitch transdiaphragmatic pressure and the underlying transdiaphragmatic pressure was –1.1 ± 0.2 during vol, –1.5 ± 0.7 during single-breath, and -0.6 ± 0.4 during continuous (all slopes expressed in percent of baseline.percent of baseline-1 all comparisons significant at the 5% level. The contribution of the diaphragm to inspiration, as assessed by the gastric pressure to transdiaphragmatic pressure ratio, was 31 ± 17 % during vol, 22 ± 16 % during single-breath (p=0.13, and 19 ± 9 % during continuous (p = 0.0015 vs. vol. This study shows that the relationship between the amplitude of the transdiaphragmatic pressure produced by a diaphragm twitch and its counterpart produced by the underlying diaphragm contraction is not unequivocal. If twitch interpolation is interpreted as

Fast-twitch glycolytic skeletal muscle is predisposed to age-induced impairments in mitochondrial function

DEFF Research Database (Denmark)

Jacobs, Robert A; Díaz, Víctor; Soldini, Lavinia

2013-01-01

The etiology of mammalian senescence is suggested to involve the progressive impairment of mitochondrial function; however, direct observations of age-induced alterations in actual respiratory chain function are lacking. Accordingly, we assessed mitochondrial function via high-resolution respirom......The etiology of mammalian senescence is suggested to involve the progressive impairment of mitochondrial function; however, direct observations of age-induced alterations in actual respiratory chain function are lacking. Accordingly, we assessed mitochondrial function via high......-resolution respirometry and mitochondrial protein expression in soleus, quadricep, and lateral gastrocnemius skeletal muscles, which represent type 1 slow-twitch oxidative muscle (soleus) and type 2 fast-twitch glycolytic muscle (quadricep and gastrocnemius), respectively, in young (10-12 weeks) and mature (74-76 weeks......) mice. Electron transport through mitochondrial complexes I and III increases with age in quadricep and gastrocnemius, which is not observed in soleus. Mitochondrial coupling efficiency during respiration through complex I also deteriorates with age in gastrocnemius and shows a tendency (p = .085...

Juxtaposition of the changes in intracellular calcium and force during staircase potentiation at 30 and 37°C

Science.gov (United States)

Vandenboom, Rene

2014-01-01

Ca2+ entry during the action potential stimulates muscle contraction. During repetitive low frequency stimulation, skeletal muscle undergoes staircase potentiation (SP), a progressive increase in the peak twitch force induced by each successive stimulus. Multiple mechanisms, including myosin regulatory light chain phosphorylation, likely contribute to SP, a temperature-dependent process. Here, we used the Ca2+-sensitive fluorescence indicators acetoxymethyl (AM)-furaptra and AM-fura-2 to examine the intracellular Ca2+ transient (ICT) and the baseline Ca2+ level at the onset of each ICT during SP at 30 and 37°C in mouse lumbrical muscle. The stimulation protocol, 8 Hz for 8 s, resulted in a 27 ± 3% increase in twitch force at 37°C and a 7 ± 2% decrease in twitch force at 30°C (P < 0.05). Regardless of temperature, the peak rate of force production (+df/dt) was higher in all twitches relative to the first twitch (P < 0.05). Consistent with the differential effects of stimulation on twitch force at the two temperatures, raw ICT amplitude decreased during repetitive stimulation at 30°C (P < 0.05) but not at 37°C. Cytosolic Ca2+ accumulated during SP such that baseline Ca2+ at the onset of ICTs occurring late in the train was higher (P < 0.05) than that of those occurring early in the train. ICT duration increased progressively at both temperatures. This effect was not entirely proportional to the changes in twitch duration, as twitch duration characteristically decreased before increasing late in the protocol. This is the first study identifying a changing ICT as an important, and temperature-sensitive, modulator of muscle force during repetitive stimulation. Moreover, we extend previous observations by demonstrating that contraction-induced increases in baseline Ca2+ coincide with greater +df/dt but not necessarily with higher twitch force. PMID:25422504

Neuromuscular blockade of slow twitch muscle fibres elevates muscle oxygen uptake and energy turnover during submaximal exercise in humans

DEFF Research Database (Denmark)

Krustrup, Peter; Secher, Niels; Relu, Mihai U.

2008-01-01

We tested the hypothesis that a greater activation of fast-twitch (FT) fibres during dynamic exercise leads to a higher muscle oxygen uptake (VO2 ) and energy turnover as well as a slower muscle on-kinetics. Subjects performed one-legged knee-extensor exercise for 10 min at an intensity of 30 W...... without (CON) and with (CUR) arterial injections of the non-depolarizing neuromuscular blocking agent cisatracurium. In CUR, creatine phosphate (CP) was unaltered in slow twitch (ST) fibres and decreased (P fibres, whereas in CON, CP decreased (P ... at a contraction frequency of 1 Hz, and that the muscle VO2 kinetics is slowed by FT fibre activation....

A Reduction in Selenoprotein S Amplifies the Inflammatory Profile of Fast-Twitch Skeletal Muscle in the mdx Dystrophic Mouse.

Science.gov (United States)

Wright, Craig Robert; Allsopp, Giselle Larissa; Addinsall, Alex Bernard; McRae, Natasha Lee; Andrikopoulos, Sofianos; Stupka, Nicole

2017-01-01

Excessive inflammation is a hallmark of muscle myopathies, including Duchenne muscular dystrophy (DMD). There is interest in characterising novel genes that regulate inflammation due to their potential to modify disease progression. Gene polymorphisms in Selenoprotein S ( Seps1 ) are associated with elevated proinflammatory cytokines, and in vitro SEPS1 is protective against inflammatory stress. Given that SEPS1 is highly expressed in skeletal muscle, we investigated whether the genetic reduction of Seps1 exacerbated inflammation in the mdx mouse. F1 male mdx mice with a heterozygous Seps1 deletion ( mdx : Seps1 -/+ ) were generated. The mdx:Seps1 -/+ mice had a 50% reduction in SEPS1 protein expression in hindlimb muscles. In the extensor digitorum longus (EDL) muscles, mRNA expression of monocyte chemoattractant protein 1 ( Mcp-1 ) ( P = 0.034), macrophage marker F4/80 ( P = 0.030), and transforming growth factor-β1 ( Tgf-β1 ) ( P = 0.056) were increased in mdx:Seps1 -/+ mice. This was associated with a reduction in muscle fibre size; however, ex vivo EDL muscle strength and endurance were unaltered. In dystrophic slow twitch soleus muscles, SEPS1 reduction had no effect on the inflammatory profile nor function. In conclusion, the genetic reduction of Seps1 appears to specifically exacerbate the inflammatory profile of fast-twitch muscle fibres, which are typically more vulnerable to degeneration in dystrophy.

Determination of protein by resonance light scattering technique using dithiothreitol-sodium dodecylbenzene sulphonate as probe

Science.gov (United States)

Wu, Lihang; Mu, Dan; Gao, Dejiang; Deng, Xinyu; Tian, Yuan; Zhang, Hanqi; Yu, Aimin

2009-02-01

The resonance light scattering (RLS) spectra of bovine serum albumin (BSA)-dithiothreitol (DTT)-sodium dodecylbenzene sulphonate (SDBS) and its analytical application were investigated. The RLS intensity of this system can be effectively enhanced in the presence of BSA. Based on the enhanced RLS intensity, a simple assay for BSA was developed. The experimental results indicate that the enhanced RLS intensity is proportional to the concentration of BSA in the range from 1.0 × 10 -8 to 7.5 × 10 -7 mol L -1 with the determination limit of 5.0 × 10 -9 mol L -1. The effects of pH, concentration of SDBS and DTT on the RLS enhancement were discussed. Most metal ions have little interference on the determination of BSA. Some synthetic and real samples were analyzed, and the results obtained were in good agreement with those obtained by Bradford method.

Autonomic components of Complex Regional Pain Syndrome (CRPS) are favourably affected by Electrical Twitch-Obtaining Intramuscular Stimulation (ETOIMS): effects on blood pressure and heart rate.

Science.gov (United States)

Chu, Jennifer; Bruyninckx, Frans; Neuhauser, Duncan V

2017-07-01

Favourable pain relief results on evoking autonomous twitches at myofascial trigger points with Electrical Twitch Obtaining Intramuscular Stimulation (ETOIMS). To document autonomic nervous system (ANS) dysfunction in Complex Regional Pain Syndrome (CRPS) from blood pressure (BP) and pulse/heart rate changes with ETOIMS. A patient with persistent pain regularly received serial ETOIMS sessions of 60, 90, 120 or ≥150 min over 24 months. Outcome measures include BP: systolic, diastolic, pulse pressure and pulse/heart rate, pre-session/immediate-post-session summed differences (SDPPP index), and pain reduction. His results were compared with that of two other patients and one normal control. Each individual represented the following maximal elicitable twitch forces (TWF) graded 1-5: maximum TWF2: control subject; maximum TWF3: CRPS patient with suspected ANS dysfunction; and maximum TWF4 and TWF5: two patients with respective slow-fatigue and fast-fatigue twitches who during ETOIMS had autonomous twitching at local and remote myotomes simultaneously from denervation supersensitivity. ETOIMS results between TWFs were compared using one-way analysis of variance test. The patients showed immediate significant pain reduction, BP and pulse/heart rate changes/reduction(s) except for diastolic BP in the TWF5 patient. TWF2 control subject had diastolic BP reduction with ETOIMS but not with rest. Linear regression showed TWF grade to be the most significant variable in pain reduction, more so than the number of treatments, session duration and treatment interval. TWF grade was the most important variable in significantly reducing outcome measures, especially pulse/heart rate. Unlike others, the TWF3 patient had distinctive reductions in SDPPP index. Measuring BP and pulse/heart rate is clinically practical for alerting ANS dysfunction maintained CRPS. SDPPP index (≥26) and pulse/heart rate (≥8) reductions with almost every ETOIMS treatment, plus inability to evoke

Incubating Isolated Mouse EDL Muscles with Creatine Improves Force Production and Twitch Kinetics in Fatigue Due to Reduction in Ionic Strength

Science.gov (United States)

Head, Stewart I.; Greenaway, Bronwen; Chan, Stephen

2011-01-01

Background Creatine supplementation can improve performance during high intensity exercise in humans and improve muscle strength in certain myopathies. In this present study, we investigated the direct effects of acute creatine incubation on isolated mouse fast-twitch EDL muscles, and examined how these effects change with fatigue. Methods and Results The extensor digitorum longus muscle from mice aged 12–14 weeks was isolated and stimulated with field electrodes to measure force characteristics in 3 different states: (i) before fatigue; (ii) immediately after a fatigue protocol; and (iii) after recovery. These served as the control measurements for the muscle. The muscle was then incubated in a creatine solution and washed. The measurement of force characteristics in the 3 different states was then repeated. In un-fatigued muscle, creatine incubation increased the maximal tetanic force. In fatigued muscle, creatine treatment increased the force produced at all frequencies of stimulation. Incubation also increased the rate of twitch relaxation and twitch contraction in fatigued muscle. During repetitive fatiguing stimulation, creatine-treated muscles took 55.1±9.5% longer than control muscles to lose half of their original force. Measurement of weight changes showed that creatine incubation increased EDL muscle mass by 7%. Conclusion Acute creatine application improves force production in isolated fast-twitch EDL muscle, and these improvements are particularly apparent when the muscle is fatigued. One likely mechanism for this improvement is an increase in Ca2+ sensitivity of contractile proteins as a result of ionic strength decreases following creatine incubation. PMID:21850234

Incubating isolated mouse EDL muscles with creatine improves force production and twitch kinetics in fatigue due to reduction in ionic strength.

Directory of Open Access Journals (Sweden)

Stewart I Head

Full Text Available BACKGROUND: Creatine supplementation can improve performance during high intensity exercise in humans and improve muscle strength in certain myopathies. In this present study, we investigated the direct effects of acute creatine incubation on isolated mouse fast-twitch EDL muscles, and examined how these effects change with fatigue. METHODS AND RESULTS: The extensor digitorum longus muscle from mice aged 12-14 weeks was isolated and stimulated with field electrodes to measure force characteristics in 3 different states: (i before fatigue; (ii immediately after a fatigue protocol; and (iii after recovery. These served as the control measurements for the muscle. The muscle was then incubated in a creatine solution and washed. The measurement of force characteristics in the 3 different states was then repeated. In un-fatigued muscle, creatine incubation increased the maximal tetanic force. In fatigued muscle, creatine treatment increased the force produced at all frequencies of stimulation. Incubation also increased the rate of twitch relaxation and twitch contraction in fatigued muscle. During repetitive fatiguing stimulation, creatine-treated muscles took 55.1±9.5% longer than control muscles to lose half of their original force. Measurement of weight changes showed that creatine incubation increased EDL muscle mass by 7%. CONCLUSION: Acute creatine application improves force production in isolated fast-twitch EDL muscle, and these improvements are particularly apparent when the muscle is fatigued. One likely mechanism for this improvement is an increase in Ca(2+ sensitivity of contractile proteins as a result of ionic strength decreases following creatine incubation.

The Major Outer Membrane Protein MopB Is Required for Twitching Movement and Affects Biofilm Formation and Virulence in Two Xylella fastidiosa strains.

Science.gov (United States)

Chen, Hongyu; Kandel, Prem P; Cruz, Luisa F; Cobine, Paul A; De La Fuente, Leonardo

2017-11-01

MopB is a major outer membrane protein (OMP) in Xylella fastidiosa, a bacterial plant pathogen that causes losses on many economically important crops. Based on in silico analysis, the uncharacterized MopB protein of X. fastidiosa contains a β-barrel structure with an OmpA-like domain and a predicted calcium-binding motif. Here, MopB function was studied by mutational analysis taking advantage of the natural competence of X. fastidiosa. Mutants of mopB were constructed in two different X. fastidiosa strains, the type strain Temecula and the more virulent WM1-1. Deletion of the mopB gene impaired cell-to-cell aggregation, surface attachment, and biofilm formation in both strains. Interestingly, mopB deletion completely abolished twitching motility. Electron microscopy of the bacterial cell surface revealed that mopB deletion eliminated type IV and type I pili formation, potentially caused by destabilization of the outer membrane. Both mopB mutants showed reduced virulence using tobacco (Nicotiana tabacum) as a host under greenhouse conditions. These results suggest that MopB has pleiotropic functions in biofilm formation and twitching motility and is important for virulence of X. fastidiosa.

Altered fast- and slow-twitch muscle fibre characteristics in female mice with a (S248F) knock-in mutation of the brain neuronal nicotinic acetylcholine receptor.

Science.gov (United States)

Cannata, David J; Finkelstein, David I; Gantois, Ilse; Teper, Yaroslav; Drago, John; West, Jan M

2009-01-01

We generated a mouse line with a missense mutation (S248F) in the gene (CHRNA4) encoding the alpha4 subunit of neuronal nicotinic acetylcholine receptor (nAChR). Mutant mice demonstrate brief nicotine induced dystonia that resembles the clinical events seen in patients with the same mutation. Drug-induced dystonia is more pronounced in female mice, thus our aim was to determine if the S248F mutation changed the properties of fast- and slow-twitch muscle fibres from female mutant mice. Reverse transcriptase-PCR confirmed CHRNA4 gene expression in the brain but not skeletal muscles in normal and mutant mice. Ca(2+) and Sr(2+) force activation curves were obtained using skinned muscle fibres prepared from slow-twitch (soleus) and fast-twitch (EDL) muscles. Two significant results were found: (1) the (pCa(50) - pSr(50)) value from EDL fibres was smaller in mutant mice than in wild type (1.01 vs. 1.30), (2) the percentage force produced at pSr 5.5 was larger in mutants than in wild type (5.76 vs. 0.24%). Both results indicate a shift to slow-twitch characteristics in the mutant. This conclusion is supported by the identification of the myosin heavy chain (MHC) isoforms. Mutant EDL fibres expressed MHC I (usually only found in slow-twitch fibres) as well as MHC IIa. Despite the lack of spontaneous dystonic events, our findings suggest that mutant mice may be having subclinical events or the mutation results in a chronic alteration to muscle neural input.

A Reduction in Selenoprotein S Amplifies the Inflammatory Profile of Fast-Twitch Skeletal Muscle in the mdx Dystrophic Mouse

Directory of Open Access Journals (Sweden)

Craig Robert Wright

2017-01-01

Full Text Available Excessive inflammation is a hallmark of muscle myopathies, including Duchenne muscular dystrophy (DMD. There is interest in characterising novel genes that regulate inflammation due to their potential to modify disease progression. Gene polymorphisms in Selenoprotein S (Seps1 are associated with elevated proinflammatory cytokines, and in vitro SEPS1 is protective against inflammatory stress. Given that SEPS1 is highly expressed in skeletal muscle, we investigated whether the genetic reduction of Seps1 exacerbated inflammation in the mdx mouse. F1 male mdx mice with a heterozygous Seps1 deletion (mdx:Seps1−/+ were generated. The mdx:Seps1−/+ mice had a 50% reduction in SEPS1 protein expression in hindlimb muscles. In the extensor digitorum longus (EDL muscles, mRNA expression of monocyte chemoattractant protein 1 (Mcp-1 (P=0.034, macrophage marker F4/80 (P=0.030, and transforming growth factor-β1 (Tgf-β1 (P=0.056 were increased in mdx:Seps1−/+ mice. This was associated with a reduction in muscle fibre size; however, ex vivo EDL muscle strength and endurance were unaltered. In dystrophic slow twitch soleus muscles, SEPS1 reduction had no effect on the inflammatory profile nor function. In conclusion, the genetic reduction of Seps1 appears to specifically exacerbate the inflammatory profile of fast-twitch muscle fibres, which are typically more vulnerable to degeneration in dystrophy.

"Positive people always win" : en studie av hur kvinnor som livestreamar på Twitch.tv upplever interaktionen med sina tittare i kanalchatten

OpenAIRE

Thoresen, Josef; Elfwendahl, Sofia

2017-01-01

I denna studie har vi undersökt kvinnor som livestreamar sitt datorspelande på hemsidan Twitch.tv och deras erfarenheter gällande chattkommentarer på deras livesändningar. Vi har främst fokuserat på negativa chattkommentarer och hur dessa påverkar kvinnorna, då tidigare forskning redan har visat på att kvinnor utsätts för fler trakasserier än män, både generellt på internet och på Twitch.tv (Nakandala, Ciampaglia, Su & Ahn. 2016). Vidare ville vi också se hur kvinnorna hanterar dessa nega...

Interaction of Mechanical Load with Growth Hormone (GH) and Insulin-Like Growth Factor I (IGF-I) on Slow-Twitch Skeletal Muscle and Bone

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Linderman, Jon K.; Gosselink, Kristin L.; Wang, Tommy J.; Mukku, Venkat R.; Grindeland, Richard E.

1994-01-01

Exogenous humoral growth factors, combined with increased mechanical loading, reportedly induce hypertrophy of fast-, but not slow-twitch skeletal muscles, and have little effect in attenuating atrophy of slow-twitch muscle associated with exposure to microgravity in animals with intact neuroendocrine systems. These observations suggest that anabolic adjuvants and muscle tension do not interact to stimulate growth or maintenance of slow-twitch skeletal muscle. The purpose of the present study was to determine whether a chronic increase in mechanical loading (synergistic ablation) or hindlimb unweighting (hindlimb suspension) interact with exogenous GH and IGF-I (Genentech, So San Francisco, CA) in the slow-twitch soleus muscles of female rats (approx. 250 g). Bilateral ablation of the plantaris and gastrocnemius muscles induced 38% and 40% increases in the absolute (mg/pair) and relative (mg/100 g body weight) weights of the soleus, respectively (p less than or = 0.05), in ambulatory rats. GH and IGF-I interacted with chronic loading to increase absolute soleus mass an additional 20% (p less than or = 0.05), and mixed and myofibrillar protein contents an additional 12% and 7%, respectively (NS). In contrast, hindlimb suspension (HLS) resulted in 20% and 18% decreases in the absolute and relative weights of the soleus, respectively (p less than or = 0.05); GH and IGF-I did not spare loss of soleus mass or protein content in HLS rats. HLS decreased tibial plate thickness approx. 11% (p less than or = 0.05), but not weights of the tibia or femus. GH and IGF-I increased tibial plate thickness approx. 30% (p less than or = 0.05), in ambulatory and HLS rats, and increased femur and tibial weights 12% (p less than or = 0.05) and 8% (NS), respectively, in ambulatory rats, but had no effect in HLS rats. Results of the present investigation suggest that GH and IGF-I can stimulate hypertrophy of slow-twitch skeletal muscle when chronically overloaded, but can also stimulate

In situ hybridisation of a large repertoire of muscle-specific transcripts in fish larvae: the new superficial slow-twitch fibres exhibit characteristics of fast-twitch differentiation.

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Chauvigné, F; Ralliere, C; Cauty, C; Rescan, P Y

2006-01-01

Much of the present information on muscle differentiation in fish concerns the early embryonic stages. To learn more about the maturation and the diversification of the fish myotomal fibres in later stages of ontogeny, we investigated, by means of in situ hybridisation, the developmental expression of a large repertoire of muscle-specific genes in trout larvae from hatching to yolk resorption. At hatching, transcripts for fast and slow muscle protein isoforms, namely myosins, tropomyosins, troponins and myosin binding protein C were present in the deep fast and the superficial slow areas of the myotome, respectively. During myotome expansion that follows hatching, the expression of fast isoforms became progressively confined to the borders of the fast muscle mass, whereas, in contrast, slow muscle isoform transcripts were uniformly expressed in all the slow fibres. Transcripts for several enzymes involved in oxidative metabolism such as citrate synthase, cytochrome oxidase component IV and succinate dehydrogenase, were present throughout the whole myotome of hatching embryos but in later stages became concentrated in slow fibre as well as in lateral fast fibres. Surprisingly, the slow fibres that are added externally to the single superficial layer of the embryonic (original) slow muscle fibres expressed not only slow twitch muscle isoforms but also, transiently, a subset of fast twitch muscle isoforms including MyLC1, MyLC3, MyHC and myosin binding protein C. Taken together these observations show that the growth of the myotome of the fish larvae is associated with complex patterns of muscular gene expression and demonstrate the unexpected presence of fast muscle isoform-expressing fibres in the most superficial part of the slow muscle.

YouTube Live and Twitch: A Tour of User-Generated Live Streaming Systems

OpenAIRE

Pires , Karine; SIMON , Gwendal

2015-01-01

International audience; User-Generated live video streaming systems are services that allow anybody to broadcast a video stream over the Internet. These Over-The-Top services have recently gained popularity, in particular with e-sport, and can now be seen as competitors of the traditional cable TV. In this paper, we present a dataset for further works on these systems. This dataset contains data on the two main user-generated live streaming systems: Twitch and the live service of YouTube. We ...

A quantitative description of tubular system Ca2+ handling in fast‐ and slow‐twitch muscle fibres

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Cully, Tanya R.; Edwards, Joshua N.; Murphy, Robyn M.

2016-01-01

Key points Current methods do not allow a quantitative description of Ca2+ movements across the tubular (t‐) system membrane without isolating the membranes from their native skeletal muscle fibre.Here we present a fluorescence‐based method that allows determination of the t‐system [Ca2+] transients and derivation of t‐system Ca2+ fluxes in mechanically skinned skeletal muscle fibres. Differences in t‐system Ca2+‐handling properties between fast‐ and slow‐twitch fibres from rat muscle are resolved for the first time using this new technique.The method can be used to study Ca2+ handling of the t‐system and allows direct comparisons of t‐system Ca2+ transients and Ca2+ fluxes between groups of fibres and fibres from different strains of animals. Abstract The tubular (t‐) system of skeletal muscle is an internalization of the plasma membrane that maintains a large Ca2+ gradient and exchanges Ca2+ between the extracellular and intracellular environments. Little is known of the Ca2+‐handling properties of the t‐system as the small Ca2+ fluxes conducted are difficult to resolve with conventional methods. To advance knowledge in this area we calibrated t‐system‐trapped rhod‐5N inside skinned fibres from rat and [Ca2+]t‐sys, allowing confocal measurements of Ca2+‐dependent changes in rhod‐5N fluorescence during rapid changes in the intracellular ionic environment to be converted to [Ca2+] transients in the t‐system ([Ca2+]t‐sys (t)). Furthermore, t‐system Ca2+‐buffering power was determined so that t‐system Ca2+ fluxes could be derived from [Ca2+]t‐sys (t). With this new approach, we show that rapid depletion of sarcoplasmic reticulum (SR) Ca2+ induced a robust store‐operated Ca2+ entry (SOCE) in fast‐ and slow‐twitch fibres, reducing [Ca2+]t‐sys to fibre types. Abruptly introducing internal solutions with 1 mm Mg2+ and [Ca2+]cyto (28 nm–1.3 μm) to Ca2+‐depleted fibres generated t‐system Ca2+ uptake rates

A quantitative description of tubular system Ca(2+) handling in fast- and slow-twitch muscle fibres.

Science.gov (United States)

Cully, Tanya R; Edwards, Joshua N; Murphy, Robyn M; Launikonis, Bradley S

2016-06-01

Current methods do not allow a quantitative description of Ca(2+) movements across the tubular (t-) system membrane without isolating the membranes from their native skeletal muscle fibre. Here we present a fluorescence-based method that allows determination of the t-system [Ca(2+) ] transients and derivation of t-system Ca(2+) fluxes in mechanically skinned skeletal muscle fibres. Differences in t-system Ca(2+) -handling properties between fast- and slow-twitch fibres from rat muscle are resolved for the first time using this new technique. The method can be used to study Ca(2+) handling of the t-system and allows direct comparisons of t-system Ca(2+) transients and Ca(2+) fluxes between groups of fibres and fibres from different strains of animals. The tubular (t-) system of skeletal muscle is an internalization of the plasma membrane that maintains a large Ca(2+) gradient and exchanges Ca(2+) between the extracellular and intracellular environments. Little is known of the Ca(2+) -handling properties of the t-system as the small Ca(2+) fluxes conducted are difficult to resolve with conventional methods. To advance knowledge in this area we calibrated t-system-trapped rhod-5N inside skinned fibres from rat and [Ca(2+) ]t-sys , allowing confocal measurements of Ca(2+) -dependent changes in rhod-5N fluorescence during rapid changes in the intracellular ionic environment to be converted to [Ca(2+) ] transients in the t-system ([Ca(2+) ]t-sys (t)). Furthermore, t-system Ca(2+) -buffering power was determined so that t-system Ca(2+) fluxes could be derived from [Ca(2+) ]t-sys (t). With this new approach, we show that rapid depletion of sarcoplasmic reticulum (SR) Ca(2+) induced a robust store-operated Ca(2+) entry (SOCE) in fast- and slow-twitch fibres, reducing [Ca(2+) ]t-sys to fibre types. Abruptly introducing internal solutions with 1 mm Mg(2+) and [Ca(2+) ]cyto (28 nm-1.3 μm) to Ca(2+) -depleted fibres generated t-system Ca(2+) uptake rates dependent on [Ca(2

Expression of multiple slow myosin heavy chain genes reveals a diversity of zebrafish slow twitch muscle fibres with differing requirements for Hedgehog and Prdm1 activity.

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Elworthy, Stone; Hargrave, Murray; Knight, Robert; Mebus, Katharina; Ingham, Philip W

2008-06-01

The zebrafish embryo develops a series of anatomically distinct slow twitch muscle fibres that characteristically express genes encoding lineage-specific isoforms of sarcomeric proteins such as MyHC and troponin. We show here that different subsets of these slow fibres express distinct members of a tandem array of slow MyHC genes. The first slow twitch muscle fibres to differentiate, which are specified by the activity of the transcription factor Prdm1 (also called Ubo or Blimp1) in response to Hedgehog (Hh) signalling, express the smyhc1 gene. Subsequently, secondary slow twitch fibres differentiate in most cases independently of Hh activity. We find that although some of these later-forming fibres also express smyhc1, others express smyhc2 or smyhc3. We show that the smyhc1-positive fibres express the ubo (prdm1) gene and adopt fast twitch fibre characteristics in the absence of Prdm1 activity, whereas those that do not express smyhc1 can differentiate independently of Prdm1 function. Conversely, some smyhc2-expressing fibres, although independent of Prdm1 function, require Hh activity to form. The adult trunk slow fibres express smyhc2 and smyhc3, but lack smyhc1 expression. The different slow fibres in the craniofacial muscles variously express smyhc1, smyhc2 and smyhc3, and all differentiate independently of Prdm1.

Myosin phosphorylation improves contractile economy of mouse fast skeletal muscle during staircase potentiation.

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Bunda, Jordan; Gittings, William; Vandenboom, Rene

2018-01-30

Phosphorylation of the myosin regulatory light chain (RLC) by skeletal myosin light chain kinase (skMLCK) potentiates rodent fast twitch muscle but is an ATP-requiring process. Our objective was to investigate the effect of skMLCK-catalyzed RLC phosphorylation on the energetic cost of contraction and the contractile economy (ratio of mechanical output to metabolic input) of mouse fast twitch muscle in vitro (25°C). To this end, extensor digitorum longus (EDL) muscles from wild-type (WT) and from skMLCK-devoid (skMLCK -/- ) mice were subjected to repetitive low-frequency stimulation (10 Hz for 15 s) to produce staircase potentiation of isometric twitch force, after which muscles were quick frozen for determination of high-energy phosphate consumption (HEPC). During stimulation, WT muscles displayed significant potentiation of isometric twitch force while skMLCK -/- muscles did not (i.e. 23% versus 5% change, respectively). Consistent with this, RLC phosphorylation was increased ∼3.5-fold from the unstimulated control value in WT but not in skMLCK -/- muscles. Despite these differences, the HEPC of WT muscles was not greater than that of skMLCK -/- muscles. As a result of the increased contractile output relative to HEPC, the calculated contractile economy of WT muscles was greater than that of skMLCK -/- muscles. Thus, our results suggest that skMLCK-catalyzed phosphorylation of the myosin RLC increases the contractile economy of WT mouse EDL muscle compared with skMLCK -/- muscles without RLC phosphorylation. © 2018. Published by The Company of Biologists Ltd.

Dithiothreitol activity by particulate oxidizers of SOA produced from photooxidation of hydrocarbons under varied NOx levels

Science.gov (United States)

Jiang, Huanhuan; Jang, Myoseon; Yu, Zechen

2017-08-01

When hydrocarbons (HCs) are atmospherically oxidized, they form particulate oxidizers, including quinones, organic hydroperoxides, and peroxyacyl nitrates (PANs). These particulate oxidizers can modify cellular materials (e.g., proteins and enzymes) and adversely modulate cell functions. In this study, the contribution of particulate oxidizers in secondary organic aerosols (SOAs) to the oxidative potential was investigated. SOAs were generated from the photooxidation of toluene, 1,3,5-trimethylbenzene, isoprene, and α-pinene under varied NOx levels. Oxidative potential was determined from the typical mass-normalized consumption rate (reaction time t = 30 min) of dithiothreitol (DTTt), a surrogate for biological reducing agents. Under high-NOx conditions, the DTTt of toluene SOA was 2-5 times higher than that of the other types of SOA. Isoprene DTTt significantly decreased with increasing NOx (up to 69 % reduction by changing the HC / NOx ratio from 30 to 5). The DTTt of 1,3,5-trimethylbenzene and α-pinene SOA was insensitive to NOx under the experimental conditions of this study. The significance of quinones to the oxidative potential of SOA was tested through the enhancement of DTT consumption in the presence of 2,4-dimethylimidazole, a co-catalyst for the redox cycling of quinones; however, no significant effect of 2,4-dimethylimidazole on modulation of DTT consumption was observed for all SOA, suggesting that a negligible amount of quinones was present in the SOA of this study. For toluene and isoprene, mass-normalized DTT consumption (DTTm) was determined over an extended period of reaction time (t = 2 h) to quantify their maximum capacity to consume DTT. The total quantities of PANs and organic hydroperoxides in toluene SOA and isoprene SOA were also measured using the Griess assay and the 4-nitrophenylboronic acid assay, respectively. Under the NOx conditions (HC / NOx ratio: 5-36 ppbC ppb-1) applied in this study, the amount of organic hydroperoxides was

Oxidative potential of ambient water-soluble PM2.5 in the southeastern United States: contrasts in sources and health associations between ascorbic acid (AA and dithiothreitol (DTT assays

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T. Fang

2016-03-01

Full Text Available The ability of certain components of particulate matter to induce oxidative stress through the generation of reactive oxygen species (ROS in vivo may be one mechanism accounting for observed linkages between ambient aerosols and adverse health outcomes. A variety of assays have been used to measure this so-called aerosol oxidative potential. We developed a semi-automated system to quantify oxidative potential of filter aqueous extracts utilizing the dithiothreitol (DTT assay and report here the development of a similar semi-automated system for the ascorbic acid (AA assay. Approximately 500 PM2.5 filter samples collected in contrasting locations in the southeastern US were analyzed for a host of aerosol species, along with AA and DTT activities. We present a detailed contrast in findings from these two assays. Water-soluble AA activity was higher in summer and fall than in winter, with highest levels near heavily trafficked highways, whereas DTT activity was higher in winter compared to summer and fall and more spatially homogeneous. AA activity was nearly exclusively correlated with water-soluble Cu (r  =  0.70–0.94 at most sites, whereas DTT activity was correlated with organic and metal species. Source apportionment models, positive matrix factorization (PMF and a chemical mass balance method with ensemble-averaged source impact profiles (CMB-E, suggest a strong contribution from traffic emissions and secondary processes (e.g., organic aerosol oxidation or metals mobilization by secondary acids to both AA and DTT activities in urban Atlanta. In contrast, biomass burning was a large source for DTT activity, but insignificant for AA. AA activity was not correlated with PM2.5 mass, while DTT activity co-varied strongly with mass (r  =  0.49–0.86 across sites and seasons. Various linear models were developed to estimate AA and DTT activities for the central Atlanta Jefferson Street site, based on the CMB-E sources. The

Effects of resistance training on fast- and slow-twitch muscles in rats

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M Umnova

2010-09-01

Full Text Available The purpose of this study was to investigate the effect of resistance training (RT on muscle strength, the dependence of that on the fast-twitch (FT and slow-twitch (ST fibers hypertrophy, nuclear domain size, synthesis and degradation rate of contractile proteins and on the expression of myosin isoforms’. 16 weeks old Wistar rats were trained on a vertical treadmill for six days a week during six weeks. The power of exercise increased 4.9% per session. In RT group the mass of studied muscles increased about 10%, hindlimb grip strength increased from 5.20±0.27 N/100g bw to the 6.05±0.29 N/100g bw (p<0.05. Cross-sectional area and number of myonuclei of FT and ST fibers in plantaris (Pla and soleus (Sol muscles increased, myonuclear domain size did not change significantly. RT increased the MyHC IId isoforms relative content and decreased that of IIb and IIa isoforms in Pla muscle, in Sol muscle increased only IIa isoform. In Pla muscle the relative content of myosin light chain (MyLC 1slow and 2slow isoforms decreased and that of MyLC 2fast isoforms increased during RT. MyLC 3 and MyLC 2 ratio did not change significantly in Pla but increased in Sol muscle by 14.3±3.4�0(p<0.01. The rat RT programme caused hypertrophy of FT and ST muscle fibers, increase of myonuclear number via fusion of satellite cells with damaged fibers or formation of new muscle fibers as a result of myoblast fusion and myotubes formation, maintaining myonuclear domain size.

An Active Learning Mammalian Skeletal Muscle Lab Demonstrating Contractile and Kinetic Properties of Fast- and Slow-Twitch Muscle

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Head, S. I.; Arber, M. B.

2013-01-01

The fact that humans possess fast and slow-twitch muscle in the ratio of approximately 50% has profound implications for designing exercise training strategies for power and endurance activities. With the growth of exercise and sport science courses, we have seen the need to develop an undergraduate student laboratory that demonstrates the basic…

Transcriptional co-activator PGC-1 alpha drives the formation of slow-twitch muscle fibres.

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Lin, Jiandie; Wu, Hai; Tarr, Paul T; Zhang, Chen-Yu; Wu, Zhidan; Boss, Olivier; Michael, Laura F; Puigserver, Pere; Isotani, Eiji; Olson, Eric N; Lowell, Bradford B; Bassel-Duby, Rhonda; Spiegelman, Bruce M

2002-08-15

The biochemical basis for the regulation of fibre-type determination in skeletal muscle is not well understood. In addition to the expression of particular myofibrillar proteins, type I (slow-twitch) fibres are much higher in mitochondrial content and are more dependent on oxidative metabolism than type II (fast-twitch) fibres. We have previously identified a transcriptional co-activator, peroxisome-proliferator-activated receptor-gamma co-activator-1 (PGC-1 alpha), which is expressed in several tissues including brown fat and skeletal muscle, and that activates mitochondrial biogenesis and oxidative metabolism. We show here that PGC-1 alpha is expressed preferentially in muscle enriched in type I fibres. When PGC-1 alpha is expressed at physiological levels in transgenic mice driven by a muscle creatine kinase (MCK) promoter, a fibre type conversion is observed: muscles normally rich in type II fibres are redder and activate genes of mitochondrial oxidative metabolism. Notably, putative type II muscles from PGC-1 alpha transgenic mice also express proteins characteristic of type I fibres, such as troponin I (slow) and myoglobin, and show a much greater resistance to electrically stimulated fatigue. Using fibre-type-specific promoters, we show in cultured muscle cells that PGC-1 alpha activates transcription in cooperation with Mef2 proteins and serves as a target for calcineurin signalling, which has been implicated in slow fibre gene expression. These data indicate that PGC-1 alpha is a principal factor regulating muscle fibre type determination.

Postactivation Potentiation of the Plantar Flexors Does Not Directly Translate to Jump Performance in Female Elite Young Soccer Players.

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Prieske, Olaf; Maffiuletti, Nicola A; Granacher, Urs

2018-01-01

High-intensity muscle actions have the potential to temporarily improve muscle contractile properties (i.e., postactivation potentiation, PAP) thereby inducing acute performance enhancements. There is evidence that balance training can improve performance during strength exercises. Taking these findings together, the purpose of this study was to examine the acute effects of a combined balance and strength (B+S) exercise vs. a strength only (S) exercise on twitch contractile properties, maximum voluntary strength, and jump performance in young athletes. Female elite young soccer players ( N = 12) aged 14-15 years conducted three experimental conditions in randomized order: S included 3 sets of 8-10 dynamic leg extensions at 80% of the 1-repetition maximum, B+S consisted of 3 sets of 40 s double-leg stances on a balance board prior to leg extensions (same as S), and a resting control period. Before and 7 min after exercise, participants were tested for their electrically-evoked isometric twitches (i.e., twitch peak torque, twitch rate of torque development) and maximal voluntary contraction (MVC) torque of the plantar flexor muscles. Additionally, countermovement (CMJ) and drop jump (DJ) performances (i.e., CMJ/DJ height, DJ ground contact time) were assessed. Significant effects of condition on twitch contractile properties ( p jump performance outputs ( p jump performance. It is concluded that PAP effects in the plantar flexors may not directly translate to improved jump performance in female elite young soccer players. Therefore, the observed gains in jump performance following B+S are most likely related to neuromuscular changes (e.g., intramuscular coordination) rather than improved contractile properties.

Type IV Pili are required for virulence, twitching motility, and biofilm formation of acidovorax avenae subsp. Citrulli.

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Bahar, Ofir; Goffer, Tal; Burdman, Saul

2009-08-01

Acidovorax avenae subsp. citrulli is the causal agent of bacterial fruit blotch (BFB), a threatening disease of watermelon, melon, and other cucurbits. Despite the economic importance of BFB, relatively little is known about basic aspects of the pathogen's biology and the molecular basis of its interaction with host plants. To identify A. avenae subsp. citrulli genes associated with pathogenicity, we generated a transposon (Tn5) mutant library on the background of strain M6, a group I strain of A. avenae subsp. citrulli, and screened it for reduced virulence by seed-transmission assays with melon. Here, we report the identification of a Tn5 mutant with reduced virulence that is impaired in pilM, which encodes a protein involved in assembly of type IV pili (TFP). Further characterization of this mutant revealed that A. avenae subsp. citrulli requires TFP for twitching motility and wild-type levels of biofilm formation. Significant reductions in virulence and biofilm formation as well as abolishment of twitching were also observed in insertional mutants affected in other TFP genes. We also provide the first evidence that group I strains of A. avenae subsp. citrulli can colonize and move through host xylem vessels.

Postactivation Potentiation of the Plantar Flexors Does Not Directly Translate to Jump Performance in Female Elite Young Soccer Players

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Olaf Prieske

2018-03-01

Full Text Available High-intensity muscle actions have the potential to temporarily improve muscle contractile properties (i.e., postactivation potentiation, PAP thereby inducing acute performance enhancements. There is evidence that balance training can improve performance during strength exercises. Taking these findings together, the purpose of this study was to examine the acute effects of a combined balance and strength (B+S exercise vs. a strength only (S exercise on twitch contractile properties, maximum voluntary strength, and jump performance in young athletes. Female elite young soccer players (N = 12 aged 14–15 years conducted three experimental conditions in randomized order: S included 3 sets of 8–10 dynamic leg extensions at 80% of the 1-repetition maximum, B+S consisted of 3 sets of 40 s double-leg stances on a balance board prior to leg extensions (same as S, and a resting control period. Before and 7 min after exercise, participants were tested for their electrically-evoked isometric twitches (i.e., twitch peak torque, twitch rate of torque development and maximal voluntary contraction (MVC torque of the plantar flexor muscles. Additionally, countermovement (CMJ and drop jump (DJ performances (i.e., CMJ/DJ height, DJ ground contact time were assessed. Significant effects of condition on twitch contractile properties (p < 0.05, d = 1.1 and jump performance outputs (p < 0.05, 1.1 ≤ d ≤ 1.2 were found. Post-hoc tests revealed that S compared to control produced larger PAP for twitch peak torques by trend (p = 0.07, d = 1.8, 33 vs. 21% and significantly larger PAP for twitch rate of torque development (p < 0.05, d = 2.4, 55 vs. 43%. Following B+S compared to control, significant improvements in CMJ height (p < 0.01, d = 1.9, 3% and DJ contact time were found (p < 0.01, d = 2.0, 10%. This study revealed protocol-specific acute performance improvements. While S resulted in significant increases in twitch contractile properties, B+S produced

Dithiothreitol activity by particulate oxidizers of SOA produced from photooxidation of hydrocarbons under varied NOx levels

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H. Jiang

2017-08-01

Full Text Available When hydrocarbons (HCs are atmospherically oxidized, they form particulate oxidizers, including quinones, organic hydroperoxides, and peroxyacyl nitrates (PANs. These particulate oxidizers can modify cellular materials (e.g., proteins and enzymes and adversely modulate cell functions. In this study, the contribution of particulate oxidizers in secondary organic aerosols (SOAs to the oxidative potential was investigated. SOAs were generated from the photooxidation of toluene, 1,3,5-trimethylbenzene, isoprene, and α-pinene under varied NOx levels. Oxidative potential was determined from the typical mass-normalized consumption rate (reaction time t =  30 min of dithiothreitol (DTTt, a surrogate for biological reducing agents. Under high-NOx conditions, the DTTt of toluene SOA was 2–5 times higher than that of the other types of SOA. Isoprene DTTt significantly decreased with increasing NOx (up to 69 % reduction by changing the HC ∕ NOx ratio from 30 to 5. The DTTt of 1,3,5-trimethylbenzene and α-pinene SOA was insensitive to NOx under the experimental conditions of this study. The significance of quinones to the oxidative potential of SOA was tested through the enhancement of DTT consumption in the presence of 2,4-dimethylimidazole, a co-catalyst for the redox cycling of quinones; however, no significant effect of 2,4-dimethylimidazole on modulation of DTT consumption was observed for all SOA, suggesting that a negligible amount of quinones was present in the SOA of this study. For toluene and isoprene, mass-normalized DTT consumption (DTTm was determined over an extended period of reaction time (t =  2 h to quantify their maximum capacity to consume DTT. The total quantities of PANs and organic hydroperoxides in toluene SOA and isoprene SOA were also measured using the Griess assay and the 4-nitrophenylboronic acid assay, respectively. Under the NOx conditions (HC ∕ NOx ratio: 5–36 ppbC ppb−1 applied in

Accumulation of ceramide in slow-twitch muscle contributes to the development of insulin resistance in the obese JCR:LA-cp rat.

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Fillmore, Natasha; Keung, Wendy; Kelly, Sandra E; Proctor, Spencer D; Lopaschuk, Gary D; Ussher, John R

2015-06-01

What is the central question of this study? The aim was to determine whether the accumulation of ceramide contributes to skeletal muscle insulin resistance in the JCR obese rat. What is the main finding and its importance? Our main new finding is that ceramides accumulate only in slow-twitch skeletal muscle in the JCR obese rat and that reducing ceramide content in this muscle type by inhibition of serine palmitoyl transferase-1 halts the progression of insulin resistance in this rat model predisposed to early development of type 2 diabetes. Our findings highlight the importance of assessing insulin signalling/sensitivity and lipid intermediate accumulation in different muscle fibre types. It has been postulated that insulin resistance results from the accumulation of cytosolic lipid metabolites (i.e. diacylglycerol/ceramide) that impede insulin signalling and impair glucose homeostasis. De novo ceramide synthesis is catalysed by serine palmitoyl transferase-1. Our aim was to determine whether de novo ceramide synthesis plays a role during development of insulin resistance in the JCR:LA-cp obese rat. Ten-week-old JCR:LA-cp obese rats were supplemented with either vehicle or the serine palmitoyl transferase-1 inhibitor l-cycloserine (360 mg l(-1) ) in their drinking water for a 2 week period, and glycaemia was assessed by meal tolerance testing. Treatment of JCR:LA-cp obese rats with l-cycloserine improved their plasma glucose and insulin levels during a meal tolerance test. Examination of muscle lipid metabolites and protein phosphorylation patterns revealed differential signatures in slow-twitch (soleus) versus fast-twitch muscle (gastrocnemius), in that ceramide levels were increased in soleus but not gastrocnemius muscles of JCR:LA-cp obese rats. Likewise, improved glycaemia in l-cycloserine-treated JCR:LA-cp obese rats was associated with enhanced Akt and pyruvate dehydrogenase signalling in soleus but not gastrocnemius muscles, probably as a result of l

Validation of jump squats as a practical measure of post-activation potentiation.

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Nibali, Maria L; Chapman, Dale W; Robergs, Robert A; Drinkwater, Eric J

2013-03-01

To determine if post-activation potentiation (PAP) can augment sports performance, it is pertinent that researchers be confident that any enhancement in performance is attributable to the PAP phenomenon. However, obtaining mechanistic measures of PAP in the daily training environment of highly trained athletes is impractical. We sought to validate jump squats as a practical measure with ecological validity to sports performance against a mechanistic measure of PAP. We assessed the evoked muscle twitch properties of the knee extensors and jump squat kinetics of 8 physically trained males in response to a 5-repetition-maximum back squat conditioning stimulus (CS). Evoked muscle twitch, followed by 3 jump squats, was assessed before and at 4, 8, and 12 min post CS. Time intervals were assessed on separate occasions using a Latin square design. Linear regression was used to determine the relationship between post-pre changes in kinetic variables and muscle twitch peak force (Ft) and twitch rate of force development (RFDt). Large correlations were observed for both concentric relative and absolute mean power and Ft (r = 0.50 ± 0.30) and RFDt (r = 0.56 ± 0.27 and r = 0.58 ± 0.26). Concentric rate of force development (RFD) showed moderate correlations with Ft (r = 0.45 ± 0.33) and RFDt (r = 0.49 ± 0.32). Small-to-moderate correlations were observed for a number of kinetic variables (r = -0.42-0.43 ± 0.32-0.38). Jump squat concentric mean power and RFD are valid ecological measures of muscle potentiation, capable of detecting changes in athletic performance in response to the PAP phenomenon.

Paradoxical Acceleration of Dithiothreitol-Induced Aggregation of Insulin in the Presence of a Chaperone

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Boris Kurganov

2010-11-01

Full Text Available The kinetics of dithiothreitol (DTT-induced aggregation of human recombinant insulin and the effect of α-crystallin, a representative of the family of small heat shock proteins, on the aggregation process have been studied using dynamic light scattering technique. Analysis of the distribution of the particles by size measured in the course of aggregation showed that the initial stage of the aggregation process was the stage of formation of the start aggregates with a hydrodynamic radius (Rh of about 90 nm. When studying the effect of α-crystallin on the rate of DTT-induced aggregation of insulin, it was demonstrated that low concentrations of α-crystallin dramatically accelerated the aggregation process, whereas high concentrations of α-crystallin suppressed insulin aggregation. In the present study, at the molar stoichiometric ratio (insulin:α-crystallin less than 1:0.5, a pronounced accelerating effect of α-crystallin was observed; whereas a ratio exceeding the value of 1:0.6 caused suppression of insulin aggregation. The mechanisms underlying the dual effect of α-crystallin have been proposed. It is assumed that heterogeneous nucleation occurring on the surface of the α-crystallin particle plays the key role in the paradoxical acceleration of insulin aggregation by α-crystallin that may provide an alternative biologically significant pathway of the aggregation process.

Neuromuscular blockade of slow twitch muscle fibres elevates muscle oxygen uptake and energy turnover during submaximal exercise in humans.

Science.gov (United States)

Krustrup, Peter; Secher, Niels H; Relu, Mihai U; Hellsten, Ylva; Söderlund, Karin; Bangsbo, Jens

2008-12-15

We tested the hypothesis that a greater activation of fast-twitch (FT) fibres during dynamic exercise leads to a higher muscle oxygen uptake (VO2 ) and energy turnover as well as a slower muscle on-kinetics. Subjects performed one-legged knee-extensor exercise for 10 min at an intensity of 30 W without (CON) and with (CUR) arterial injections of the non-depolarizing neuromuscular blocking agent cisatracurium. In CUR, creatine phosphate (CP) was unaltered in slow twitch (ST) fibres and decreased (P fibres, whereas in CON, CP decreased (P fibres, respectively. From 127 s of exercise, muscle VO2 was higher (P muscle VO2 response was slower (P muscle homogenate CP was lowered (P muscle lactate production was similar in CUR and CON (37.8 +/- 4.1 versus 35.2 +/- 6.2 mmol). Estimated total muscle ATP turnover was 19% higher (P fibres are less efficient than ST fibres in vivo at a contraction frequency of 1 Hz, and that the muscle VO2 kinetics is slowed by FT fibre activation.

Changing oxidoreduction potential to improve water-soluble yellow pigment production with Monascus ruber CGMCC 10910.

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Huang, Tao; Tan, Hailing; Lu, Fangju; Chen, Gong; Wu, Zhenqiang

2017-11-21

Monascus pigments are widely used in the food and pharmaceutical industries due to their safety to human health. Our previous study found that glucose concentration induced extracellular oxidoreduction potential (ORP) changes could influence extracellular water-soluble yellow pigment production by Monascus ruber CGMCC 10910 in submerged fermentation. In this study, H 2 O 2 and dithiothreitol (DTT) were used to change the oxidoreduction potential for investigating the effects of oxidative or reductive substances on Monascus yellow pigment production by Monascus ruber CGMCC 10910. The extracellular ORP could be controlled by H 2 O 2 and DTT. Both cell growth and extracellular water-soluble yellow pigment production were enhanced under H 2 O 2 -induced oxidative (HIO) conditions and were inhibited under dithiothreitol-induced reductive conditions. By optimizing the amount of H 2 O 2 added and the timing of the addition, the yield of extracellular water-soluble yellow pigments significantly increased and reached a maximum of 209 AU, when 10 mM H 2 O 2 was added on the 3rd day of fermentation with M. ruber CGMCC 10910. Under HIO conditions, the ratio of NADH/NAD+ was much lower than that in the control group, and the expression levels of relative pigment biosynthesis genes were up-regulated; moreover, the activity of glucose-6-phosphate dehydrogenase (G6PDH) was increased while 6-phosphofructokinase (PFK) activity was inhibited. Oxidative conditions induced by H 2 O 2 increased water-soluble yellow pigment accumulation via up-regulation of the expression levels of relative genes and by increasing the precursors of pigment biosynthesis through redirection of metabolic flux. In contrast, reductive conditions induced by dithiothreitol inhibited yellow pigment accumulation. This experiment provides a potential strategy for improving the production of Monascus yellow pigments.

Differentiation of the intracellular structure of slow- versus fast-twitch muscle fibers through evaluation of the dielectric properties of tissue

Science.gov (United States)

Sanchez, B.; Li, J.; Bragos, R.; Rutkove, S. B.

2014-05-01

Slow-twitch (type 1) skeletal muscle fibers have markedly greater mitochondrial content than fast-twitch (type 2) fibers. Accordingly, we sought to determine whether the dielectric properties of these two fiber types differed, consistent with their distinct intracellular morphologies. The longitudinal and transverse dielectric spectrum of the ex vivo rat soleus (a predominantly type 1 muscle) and the superficial layers of rat gastrocnemius (predominantly type 2) (n = 15) were measured in the 1 kHz-10 MHz frequency range and modeled to a resistivity Cole-Cole function. Major differences were especially apparent in the dielectric spectrum in the 1 to 10 MHz range. Specifically, the gastrocnemius demonstrated a well-defined, higher center frequency than the soleus muscle, whereas the soleus muscle showed a greater difference in the modeled zero and infinite resistivities than the gastrocnemius. These findings are consistent with the fact that soleus tissue has larger and more numerous mitochondria than gastrocnemius. Evaluation of tissue at high frequency could provide a novel approach for assessing intracellular structure in health and disease.

Differentiation of the intracellular structure of slow- versus fast-twitch muscle fibers through evaluation of the dielectric properties of tissue

International Nuclear Information System (INIS)

Sanchez, B; Li, J; Rutkove, S B; Bragos, R

2014-01-01

Slow-twitch (type 1) skeletal muscle fibers have markedly greater mitochondrial content than fast-twitch (type 2) fibers. Accordingly, we sought to determine whether the dielectric properties of these two fiber types differed, consistent with their distinct intracellular morphologies. The longitudinal and transverse dielectric spectrum of the ex vivo rat soleus (a predominantly type 1 muscle) and the superficial layers of rat gastrocnemius (predominantly type 2) (n = 15) were measured in the 1 kHz–10 MHz frequency range and modeled to a resistivity Cole–Cole function. Major differences were especially apparent in the dielectric spectrum in the 1 to 10 MHz range. Specifically, the gastrocnemius demonstrated a well-defined, higher center frequency than the soleus muscle, whereas the soleus muscle showed a greater difference in the modeled zero and infinite resistivities than the gastrocnemius. These findings are consistent with the fact that soleus tissue has larger and more numerous mitochondria than gastrocnemius. Evaluation of tissue at high frequency could provide a novel approach for assessing intracellular structure in health and disease. (paper)

The effect of dithiothreitol on radiation-induced genetic damage in Arabidopsis thaliana (L) Heynh

International Nuclear Information System (INIS)

Dellaert, L.M.W.

1980-01-01

A study was made on the effect of dithiothreitol (DTT; present during irradiation) on M 1 ovule sterility, M 2 embryonic lethals, M 2 chlorophyll mutants and M 2 viable mutants induced with fast neutrons or X-rays in Arabidopsis thaliana. DTT provides considerable protection against both fast-neutron and X-ray induced genetic damage. However, a higher protection was observed against M 1 ovule sterility, than against embryonic lethals, chlorophylls and viable mutants. This implies a significant DTT-induced spectral shift (0.01 < p < 0.05), i.e. a shift in the relative frequencies of the different genetic parameters. This spectral shift is explained on the basis of a specific DTT protection against radiation-induced strand breaks, and by differences in the ratio strand breaks/base damage for the genetic parameters concerned, i.e. a higher ratio for ovule sterility than for the other parameters. The induction of the genetic damage by ionizing radiation, either with or without DTT, is described by a mathematical model, which includes both strand breaks and base damage. The model shows that the resolving power of a test for a 'mutation'spectral shift depends on the relative values of the strandbreak reduction factor of -SH compounds and on the ratio strand breaks/base damage of the genetic parameters. For each genetic parameter the DTT damage reduction factor (DRF) is calculated per irradiation dose, and in addition the average (over-all doses) ratio strand breaks/base damage. (orig.)

The effects of tetracaine on charge movement in fast twitch rat skeletal muscle fibres.

Science.gov (United States)

Hollingworth, S; Marshall, M W; Robson, E

1990-02-01

1. The effects of tetracaine, a local anaesthetic that inhibits muscle contraction, on membrane potential and intramembrane charge movements were investigated in fast twitch rat muscle fibres (extensor digitorum longus). 2. The resting membrane potentials of surface fibres from muscles bathed in isotonic Ringer solution containing 2 mM-tetracaine were well maintained, but higher concentrations of tetracaine caused a time-dependent fall of potential. Muscle fibres bathed in hypertonic solutions containing 2 mM-tetracaine were rapidly depolarized. In both isotonic and hypertonic solutions, the depolarizing effect of tetracaine could not be reversed. 3. Charge movement measurements were made using the middle-of-the-fibre voltage clamp technique. The voltage dependence of charge movements measured in cold isotonic solutions was well fitted by a Boltzmann distribution (Q(V) = Qmax/(1 + exp(-(V-V)/k] where Qmax = 37.3 +/- 2.8 nC muF-1, V = -17.9 +/- 1.2 mV and k = 12.6 +/- 0.8 mV (n = 6, 2 degrees C; means +/- S.E. of means). Similar values were obtained when 2 mM-tetracaine was added to the isotonic bathing fluid (Qmax = 40.6 +/- 2.3 nC microF-1, V = -14.1 +/- 1.3 mV, k = 15.3 +/- 0.8 mV; n = 8, 2 degrees C). 4. Charge movements measured around mechanical threshold in muscle fibres bathed in hypertonic solutions were reduced when 2 mM-tetracaine was added to the bathing fluid. The tetracaine-sensitive component of charge was well fitted with an unconstrained Boltzmann distribution which gave: Qmax = 7.5 nC microF-1, V = -46.5 mV, k = 5.5 mV. The e-fold rise of the foot of the curve was 9.3 mV.

Observation of the molecular organization of calcium release sites in fast- and slow-twitch skeletal muscle with nanoscale imaging

OpenAIRE

Jayasinghe, Isuru D.; Munro, Michelle; Baddeley, David; Launikonis, Bradley S.; Soeller, Christian

2014-01-01

Localization microscopy is a fairly recently introduced super-resolution fluorescence imaging modality capable of achieving nanometre-scale resolution. We have applied the dSTORM variation of this method to image intracellular molecular assemblies in skeletal muscle fibres which are large cells that critically rely on nanoscale signalling domains, the triads. Immunofluorescence staining in fixed adult rat skeletal muscle sections revealed clear differences between fast- and slow-twitch fibres...

Cycle Training Increased GLUT4 and Activation of mTOR in Fast Twitch Muscle Fibers

Science.gov (United States)

Stuart, Charles A.; Howell, Mary E.A.; Baker, Jonathan D.; Dykes, Rhesa J.; Duffourc, Michelle M.; Ramsey, Michael W.; Stone, Michael H.

2009-01-01

Purpose To determine if cycle training of sedentary subjects would increase the expression of the principle muscle glucose transporters, six volunteers completed six weeks of progressively increasing intensity stationary cycle cycling. Methods In vastus lateralis muscle biopsies, changes in expression of GLUT1, GLUT4, GLUT5, and GLUT12 were compared using quantitative immunoblots with specific protein standards. Regulatory pathway components were evaluated by immunoblots of muscle homogenates and immunohistochemistry of microscopic sections. Results GLUT1 was unchanged, GLUT4 increased 66%, GLUT12 increased 104%, and GLUT5 decreased 72%. A mitochondrial marker (cytochrome c) and regulators of mitochondrial biogenesis (PGC-1α and phospho-AMPK) were unchanged, but the muscle hypertrophy pathway component, phospho-mTOR increased 83% after the exercise program. In baseline biopsies, GLUT4 by immunohistochemical techniques was 37% greater in Type I (slow twitch, red) muscle fibers, but the exercise training increased GLUT4 expression in Type II (fast twitch, white) fibers by 50%, achieving parity with the Type I fibers. Baseline phospho-mTOR expression was 50% higher in Type II fibers and increased more in Type II fibers (62%) with training, but also increased in Type I fibers (34%). Conclusion Progressive intensity stationary cycle training of previously sedentary subjects increased muscle insulin-responsive glucose transporters (GLUT4 and GLUT12) and decreased the fructose transporter (GLUT5). The increase in GLUT4 occurred primarily in Type II muscle fibers and this coincided with activation of the mTOR muscle hypertrophy pathway. There was little impact on Type I fiber GLUT4 expression and no evidence of change in mitochondrial biogenesis. PMID:20010125

Quantifying Ca2+ release and inactivation of Ca2+ release in fast- and slow-twitch muscles.

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Barclay, C J

2012-12-01

The aims of this study were to quantify the Ca(2+) release underlying twitch contractions of mammalian fast- and slow-twitch muscle and to comprehensively describe the transient inactivation of Ca(2+) release following a stimulus. Experiments were performed using bundles of fibres from mouse extensor digitorum longus (EDL) and soleus muscles. Ca(2+) release was quantified from the amount of ATP used to remove Ca(2+) from the myoplasm following stimulation. ATP turnover by crossbridges was blocked pharmacologically (N-benzyl-p-toluenesulphonamide for EDL, blebbistatin for soleus) and muscle heat production was used as an index of Ca(2+) pump ATP turnover. At 20°C, Ca(2+) release in response to a single stimulus was 34 and 84 μmol (kg muscle)(-1) for soleus and EDL, respectively, and increased with temperature (30°C: soleus, 61 μmol kg(-1); EDL, 168 μmol kg(-1)). Delivery of another stimulus within 100 ms of the first produced a smaller Ca(2+) release. The maximum magnitude of the decrease in Ca(2+) release was greater in EDL than soleus. Ca(2+) release recovered with an exponential time course which was faster in EDL (mean time constant at 20°C, 32.1 ms) than soleus (65.6 ms) and faster at 30°C than at 20°C. The amounts of Ca(2+) released and crossbridge cycles performed are consistent with a scheme in which Ca(2+) binding to troponin-C allowed an average of ∼1.7 crossbridge cycles in the two muscles.

Slow-tonic muscle fibers and their potential innervation in the turtle, Pseudemys (Trachemys) scripta elegans.

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Callister, Robert J; Pierce, Patricia A; McDonagh, Jennifer C; Stuart, Douglas G

2005-04-01

fiber type, with the distribution of slow-tonic fibers paralleling that of the SO fibers. In the five test muscles, fiber cross-sectional area was usually ranked Fg > FOG > SO, and slow-twitch always > slow-tonic. In terms of weighted cross-sectional area, which provides a coarse-grain measure of each fiber type's potential contribution to whole muscle force, all five muscles exhibited a higher Fg and lower SO contribution to cross-sectional area than suggested by their corresponding fiber-type prevalence. This was also the case for the slow-twitch vs. slow-tonic fibers. We conclude that slow-tonic fibers are widespread in turtle muscle. The weighted cross-sectional area evidence suggested, however, that their contribution to force generation is minor except in highly oxidative muscles, with a special functional role, like TeC4. There is discussion of: 1) the relationship between the present results and previous work on homologous neck and hindlimb muscles in other nonmammalian species, and 2) the potential motoneuronal innervation of slow-tonic fibers in turtle hindlimb muscles. Copyright 2005 Wiley-Liss, Inc.

Swimming, swarming, twitching, and chemotactic responses of Cupriavidus metallidurans CH34 and Pseudomonas putida mt2 in the presence of cadmium.

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Shamim, Saba; Rehman, Abdul; Qazi, Mahmood Hussain

2014-04-01

To use of microorganisms for bioremediation purposes, the study of their motility behavior toward metals is essential. In the present study, Cupriavidus metallidurans CH34 and Pseudomonas putida mt2 were used as cadmium (Cd)-resistant and -sensitive bacteria, respectively, to evaluate the effects of Cd on their motility behaviors. Potassium morpholinopropane sulfonate (MOPS) buffer was used to observe the motility behavior of both isolates. Movement of mt2 was less in MOPS buffer compared with CH34, likely reflecting the mono-flagellated nature of mt2 and the peritrichous nature of CH34. The swimming, swarming, twitching, and chemotaxis behaviors of mt2 were greater in the presence of glucose than that of Cd. mt2 exhibited negative motility behaviors when exposed to Cd, but the opposite effect was seen in CH34. Cd was found to be a chemorepellent for mt2 but a chemoattractant for CH34, suggesting that CH34 is a potential candidate for metal (Cd) bioremediation.

AMPK alpha1 activation is required for stimulation of glucose uptake by twitch contraction, but not by H2O2, in mouse skeletal muscle

DEFF Research Database (Denmark)

Jensen, Thomas Elbenhardt; Schjerling, Peter; Viollet, Benoit

2008-01-01

into muscle by certain stimuli. In contrast, no clear function has yet been determined for alpha(1) AMPK in skeletal muscle, possibly due to alpha-AMPK isoform signaling redundancy. By applying low-intensity twitch-contraction and H(2)O(2) stimulation to activate alpha(1) AMPK, but not alpha(2) AMPK......, in wildtype and alpha-AMPK transgenic mouse muscles, this study aimed to define conditions where alpha(1) AMPK is required to increase muscle glucose uptake. METHODOLOGY/PRINCIPAL FINDINGS: Following stimulation with H(2)O(2) (3 mM, 20 min) or twitch-contraction (0.1 ms pulse, 2 Hz, 2 min), signaling and 2......-deoxyglucose uptake were measured in incubated soleus muscles from wildtype and muscle-specific kinase-dead AMPK (KD), alpha(1) AMPK knockout or alpha(2) AMPK knockout mice. H(2)O(2) increased the activity of both alpha(1) and alpha(2) AMPK in addition to Akt phosphorylation, and H(2)O(2)-stimulated glucose...

A low redox potential affects monoclonal antibody assembly and glycosylation in cell culture.

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Dionne, Benjamin; Mishra, Neha; Butler, Michael

2017-03-20

Glycosylation and intracellular assembly of monoclonal antibodies (MAbs) is important for glycan profile consistency. To better understand how these factors may be influenced by a lower redox potential, an IgG1-producing NS0 cell line was grown in the presence of varying concentrations of dithiothreitol (DTT). Cultures were monitored for growth and culture redox potential (CRP) with glycan heterogeneity determined using a HILIC-HPLC method. Macroheterogeneity was unchanged in all conditions whereas the Galactosylation Index (GI) decreased by as much as 50% in cultures with lower CRP or higher dithiothreitol levels. This shift in GI is reflected in more agalactosylated and asialylated species being produced. The MAb assembly pathway was determined using radioactive isotope 35 S incorporated into nascent IgG1 molecules. The assembly pathway for this IgG1 was shown to progress via HC→HC 2 →HC 2 LC→HC 2 LC 2 in all conditions tested and autoradiographs highlighted that the ratio of heavy chain dimer to heavy chain monomer increased over time with increasing DTT concentrations. This increase and correspondingly lower GI values may be due to disruption of the disulfide bonds at higher levels of assembly. A change in the assembly pathway may alter the final IgG glycan pattern and lead to control mechanisms that influence glycan profiles of MAbs. Copyright © 2017. Published by Elsevier B.V.

Improved recovery of post-thaw motility and vitality of human spermatozoa cryopreserved in the presence of dithiothreitol.

Science.gov (United States)

Rao, B; David, G

1984-10-01

Semen was collected in the laboratory from nine healthy donors. The concentrations and the percentages of live and motile spermatozoa in all semen samples were within the normal range. Each sample was diluted with citrate-egg yolk-glycerol medium with and without 5 mM dithiothreitol (DTT). Samples were frozen in liquid nitrogen vapor (-70 degrees C) for 7 min and subsequently stored in liquid nitrogen. The effect of DTT in cryopreservation of sperm was determined by comparing percentage of motile and live spermatozoa between controls and DTT-treated post-thaw samples. Percentage of motile spermatozoa was determined by two techniques, laser Doppler velocimetry (LDV) and light microscopy. The percentage of live spermatozoa was measured by microscopic evaluation after staining with eosin-nigrosin. It was shown that the addition of DTT to the freezing medium significantly improved the recovery of motile and live spermatozoa in the post-thaw samples. The mean motility recovery, as measured by LDV, was 44.9% in the controls as compared to 73.9% in the DTT-treated samples. Similarly the mean recovery of live spermatozoa in the controls and DTT-treated samples was 66.5 and 86.6%, respectively. Based on these results, a new hypothesis implicating lipid peroxidation in cryoinjury is proposed. It is also suggested that the use of DTT in the freezing medium may offer an advantage over the commonly used techniques of human sperm cryopreservation.

Caffeine and length dependence of staircase potentiation in skeletal muscle.

Science.gov (United States)

Rassier, D E; Tubman, L A; MacIntosh, B R

1998-01-01

Skeletal muscle sensitivity to Ca2+ is greater at long lengths, and this results in an optimal length for twitch contractions that is longer than optimal length for tetanic contractions. Caffeine abolishes this length dependence of Ca2+ sensitivity. Muscle length (ML) also affects the degree of staircase potentiation. Since staircase potentiation is apparently caused by an increased Ca2+ sensitivity of the myofilaments, we tested the hypothesis that caffeine depresses the length dependence of staircase potentiation. In situ isometric twitch contractions of rat gastrocnemius muscle before and after 10 s of 10-Hz stimulation were analyzed at seven different lengths to evaluate the length dependence of staircase potentiation. In the absence of caffeine, length dependence of Ca2+ sensitivity was observed, and the degree of potentiation after 10-Hz stimulation showed a linear decrease with increased length (DT = 1.47 - 0.05 ML, r2 = 0.95, where DT is developed tension). Length dependence of Ca2+ sensitivity was decreased by caffeine when caffeine was administered in amounts estimated to result in 0.5 and 0.75 mM concentrations. Furthermore, the negative slope of the relationship between staircase potentiation and muscle length was diminished at the lower caffeine dose, and the slope was not different from zero after the higher dose (DT = 1.53 - 0.009 ML, r2 = 0.43). Our study shows that length dependence of Ca2+ sensitivity in intact skeletal muscle is diminished by caffeine. Caffeine also suppressed the length dependence of staircase potentiation, suggesting that the mechanism of this length dependence may be closely related to the mechanism for length dependence of Ca2+ sensitivity.

THE EFFECT OF STIMULUS ANTICIPATION ON THE INTERPOLATED TWITCH TECHNIQUE

Directory of Open Access Journals (Sweden)

Duane C. Button

2008-12-01

Full Text Available The objective of this study was to investigate the effect of expected and unexpected interpolated stimuli (IT during a maximum voluntary contraction on quadriceps force output and activation. Two groups of male subjects who were either inexperienced (MI: no prior experience with IT tests or experienced (ME: previously experienced 10 or more series of IT tests received an expected or unexpected IT while performing quadriceps isometric maximal voluntary contractions (MVCs. Measurements included MVC force, quadriceps and hamstrings electromyographic (EMG activity, and quadriceps inactivation as measured by the interpolated twitch technique (ITT. When performing MVCs with the expectation of an IT, the knowledge or lack of knowledge of an impending IT occurring during a contraction did not result in significant overall differences in force, ITT inactivation, quadriceps or hamstrings EMG activity. However, the expectation of an IT significantly (p < 0.0001 reduced MVC force (9.5% and quadriceps EMG activity (14.9% when compared to performing MVCs with prior knowledge that stimulation would not occur. While ME exhibited non-significant decreases when expecting an IT during a MVC, MI force and EMG activity significantly decreased 12.4% and 20.9% respectively. Overall, ME had significantly (p < 0.0001 higher force (14.5% and less ITT inactivation (10.4% than MI. The expectation of the noxious stimuli may account for the significant decrements in force and activation during the ITT

The age related slow and fast contributions to the overall changes in tibialis anterior contractile features disclosed by maximal single twitch scan.

Science.gov (United States)

Orizio, Claudio; Cogliati, Marta; Bissolotti, Luciano; Diemont, Bertrand; Gobbo, Massimiliano; Celichowski, Jan

2016-01-01

This work aimed to verify if maximal electrically evoked single twitch (STmax) scan discloses the relative functional weight of fast and slow small bundles of fibres (SBF) in determining the contractile features of tibialis anterior (TA) with ageing. SBFs were recruited by TA main motor point stimulation through 60 increasing levels of stimulation (LS): 20 stimuli at 2Hz for each LS. The lowest and highest LS provided the least ST and STmax, respectively. The scanned STmax was decomposed into individual SBF STs. They were identified when twitches from adjacent LS were significantly different and then subtracted from each other. Nine young (Y) and eleven old (O) subjects were investigated. Contraction time (CT) and STarea/STpeak (A/PT) were calculated per each SBF ST. 143 and 155 SBF STs were obtained in Y and O, respectively. Y: CT and A/PT range: 45-105ms and 67-183mNs/mN, respectively. Literature data set TA fast fibres at 34% so, from the arrays of CT and A/PT, 65ms and 100mNs/mN were identified as the upper limit for SBF fast ST classification. O: no SBF ST could be classified as fast. STmax scan reveals age-related changes in the relative contribution of fast and slow SBFs to the overall muscle mechanics. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Juxtaposition of the changes in intracellular calcium and force during staircase potentiation at 30 and 37?C

OpenAIRE

Smith, Ian C.; Vandenboom, Rene; Tupling, A. Russell

2014-01-01

Ca2+ entry during the action potential stimulates muscle contraction. During repetitive low frequency stimulation, skeletal muscle undergoes staircase potentiation (SP), a progressive increase in the peak twitch force induced by each successive stimulus. Multiple mechanisms, including myosin regulatory light chain phosphorylation, likely contribute to SP, a temperature-dependent process. Here, we used the Ca2+-sensitive fluorescence indicators acetoxymethyl (AM)-furaptra and AM-fura-2 to exam...

Dysregulated corticostriatal activity in open-field behavior and the head-twitch response induced by the hallucinogen 2,5-dimethoxy-4-iodoamphetamine.

Science.gov (United States)

Rangel-Barajas, Claudia; Estrada-Sánchez, Ana María; Barton, Scott J; Luedtke, Robert R; Rebec, George V

2017-02-01

The substituted amphetamine, 2,5-dimethoxy-4-iodoamphetamine (DOI), is a hallucinogen that has been used to model a variety of psychiatric conditions. Here, we studied the effect of DOI on neural activity recorded simultaneously in the primary motor cortex (M1) and dorsal striatum of freely behaving FvB/N mice. DOI significantly decreased the firing rate of individually isolated neurons in M1 and dorsal striatum relative to pre-drug baseline. It also induced a bursting pattern of activity by increasing both the number of spikes within a burst and burst duration. In addition, DOI increased coincident firing between simultaneously recorded neuron pairs within the striatum and between M1 and dorsal striatum. Local field potential (LFP) activity also increased in coherence between M1 and dorsal striatum after DOI in the low frequency gamma band (30-50 Hz), while corticostriatal coherence in delta, theta, alpha, and beta activity decreased. We also assessed corticostriatal LFP activity in relation to the DOI-induced head-twitch response (HTR), a readily identifiable behavior used to assess potential treatments for the conditions it models. The HTR was associated with increased delta and decreased theta power in both M1 and dorsal striatum. Together, our results suggest that DOI dysregulates corticostriatal communication and that the HTR is associated with this dysregulation. Copyright © 2016 Elsevier Ltd. All rights reserved.

An in vivo model for studying the dynamics of intracellular free calcium changes in slow- and fast-twitch muscle fibres.

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Bátkai, S; Rácz, I B; Ivanics, T; Tóth, A; Hamar, J; Slaaf, D W; Reneman, R S; Ligeti, L

1999-10-01

The understanding of the regulation of the free cytosolic [Ca2+] ([Ca2+]i) in skeletal muscle is hampered by the lack of techniques for quantifying free [Ca2+]i in muscle fibres in situ. We describe a model for studying the dynamics of free [Ca2+]i in the fast-twitch extensor digitorum longus (EDL) and the slow-twitch soleus (SOL) muscles of the rat in vivo using caffeine superfusion to induce changes in free [Ca2+]i. We assumed that differences in sensitivity between the two muscle types for this substance reflect differences in intracellular Ca2+ handling in the fibres of which these muscles consist. The Indo-1 ratiometric method, using intravital microscopy with incident light, was adapted to measure free [Ca2+]i in vivo. Fluorescence images were collected by means of a digital camera. Caffeine superfusion at 37 degrees C for 2 min, at concentrations of 1, 2, 5, 10 or 20 mmol/l, induced a concentration-dependent increase in free [Ca2+]i and revealed differences in caffeine sensitivity between the muscle types, with the SOL being more sensitive. In a separate set of experiments the contracture threshold, as assessed by topical application of caffeine, was determined in both muscle types. EDL had a higher threshold for developing contracture than SOL. These finding are in agreement with previous in vitro studies. We may conclude that the dynamics of free [Ca2+]i can be assessed reliably in intact mammalian muscle in vivo.

Potentiation by substance P of contractions of the isolated vas deferens of the mouse elicited by electric field stimulation and by drugs

International Nuclear Information System (INIS)

Blackwell, M.; James, T.A.; Starr, M.S.

1978-01-01

Isolated vasa deferentia from the mouse were opened longitudinally and suspended in Krebs solution at 37 0 C in an organ bath. Contractions of the muscle were elicited by electric field stimulation, noradrenaline (10 -6 M) and acetylcholine (10 -6 M). Continued transmural stimulation evoked a biphasic response comprising a rapid twitch followed by about 10 s later by a smaller, sustained rise in muscle tone. The amplitudes of nerve-mediated and drug-induced responses were considerably potentiated by substances P (SP) in the dose range 10 -12 to 10 -7 M. Higher concentrations of SP were directly spasmogenic. The sensitizing property of SP was dose-dependent and was usually well maintained, but always disappeared quickly on washing the preparation. In some experiments SP facilitated the twitch, but not the subsequent phase of the electrically-induced contraction or the response to externally applied noradrenaline. Phentolamine (10 -6 M) failed to block this effect of SP, but itself potentiated the nerve-mediated twitch, and completely abolished the sustained secondary contraction. Desmethylimipramine (10 -6 M) enhanced the delayed contraction but not the immediate contraction. The uptake of tritiated noradrenaline (3 x 10 -7 M) by vasa was inhibited by desmethylimipramine (10 -6 M) and increased by nialamide (3 x 10 -5 M), but was not modified by SP (10 -6 M). Nerve-mediated release of accumulated radioactivity was accelerated by phentolamine, but not by SP or desmethylimipramine. These findings suggest that SP sensitizes the muscle cells to depolarizing stimuli but that it has no facilitatory effect on sympathetic neural elements. (author)

Use of standard laboratory methods to obviate routine dithiothreitol treatment of blood samples with daratumumab interference.

Science.gov (United States)

Lintel, Nicholas J; Brown, Debra K; Schafer, Diane T; Tsimba-Chitsva, Farai M; Koepsell, Scott A; Shunkwiler, Sara M

2017-01-01

Daratumumab is an antibody currently used in the treatment of patients with refractory multiple myeloma. Blood samples from patients being treated with daratumumab may show panreactivity during pre-transfusion testing. To facilitate the provision of blood components for such patients, it is recommended that a baseline phenotype or genotype be established prior to starting treatment with daratumumab. If patient red blood cells (RBCs) require phenotyping after the start of daratumumab treatment, dithiothreitol (DTT) treatment of the patient's RBCs should be performed. The medical charts of four patients treated with daratumumab were reviewed. The individual number of doses ranged from 1 to 14; patient age ranged from 55 to 78 years; two men and two women were included in the review. Type and screen data were obtained from samples collected over 33 encounters with a range of 1 to 13 encounters per patient. All samples were tested initially by automated solid-phase testing. Any reactivity with solid phase led to tube testing with either low-ionic-strength saline, polyethylene glycol, or both. If incubation failed to eliminate the reactivity, the sample was sent to a reference laboratory for DTT treatment and phenotyping. Of the 33 samples tested, 23 (69.7%) samples had reactivity in solid-phase testing. In 8 of the 10 samples that did not react in solid-phase, testing was conducted more than four half-lives after the last dose of daratumumab. Of the 23 that had reactivity in solid-phase, 16 (69.6%) samples demonstrated loss of reactivity using common laboratory methods. For the seven patients whose sample reactivity was not initially eliminated, six were provided with phenotypically matched blood based on prior molecular testing. Only one sample was sent out for DTT treatment. These results suggest that daratumumab interference with pre-transfusion testing can be addressed using common laboratory methods. This finding could save time and money for laboratories that do

Oxidative shift in tissue redox potential increases beat-to-beat variability of action potential duration.

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Kistamás, Kornél; Hegyi, Bence; Váczi, Krisztina; Horváth, Balázs; Bányász, Tamás; Magyar, János; Szentandrássy, Norbert; Nánási, Péter P

2015-07-01

Profound changes in tissue redox potential occur in the heart under conditions of oxidative stress frequently associated with cardiac arrhythmias. Since beat-to-beat variability (short term variability, SV) of action potential duration (APD) is a good indicator of arrhythmia incidence, the aim of this work was to study the influence of redox changes on SV in isolated canine ventricular cardiomyocytes using a conventional microelectrode technique. The redox potential was shifted toward a reduced state using a reductive cocktail (containing dithiothreitol, glutathione, and ascorbic acid) while oxidative changes were initiated by superfusion with H2O2. Redox effects were evaluated as changes in "relative SV" determined by comparing SV changes with the concomitant APD changes. Exposure of myocytes to the reductive cocktail decreased SV significantly without any detectable effect on APD. Application of H2O2 increased both SV and APD, but the enhancement of SV was the greater, so relative SV increased. Longer exposure to H2O2 resulted in the development of early afterdepolarizations accompanied by tremendously increased SV. Pretreatment with the reductive cocktail prevented both elevation in relative SV and the development of afterdepolarizations. The results suggest that the increased beat-to-beat variability during an oxidative stress contributes to the generation of cardiac arrhythmias.

Sarcoplasmic reticulum function in slow- and fast-twitch skeletal muscles from mdx mice.

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Divet, Alexandra; Huchet-Cadiou, Corinne

2002-08-01

The aim of the present study was to establish whether alterations in sarcoplasmic reticulum function are involved in the abnormal Ca(2+) homeostasis of skeletal muscle in mice with muscular dystrophy ( mdx). The properties of the sarcoplasmic reticulum and contractile proteins of fast- and slow-twitch muscles were therefore investigated in chemically skinned fibres isolated from the extensor digitorum longus (EDL) and soleus muscles of normal (C57BL/10) and mdx mice at 4 and 11 weeks of development. Sarcoplasmic reticulum Ca(2+) uptake, estimated by the Ca(2+) release following exposure to caffeine, was significantly slower in mdx mice, while the maximal Ca(2+) quantity did not differ in either type of skeletal muscle at either stage of development. In 4-week-old mice spontaneous sarcoplasmic reticulum Ca(2+) leakage was observed in EDL and soleus fibres and this was more pronounced in mdx mice. In addition, the maximal Ca(2+)-activated tension was smaller in mdx than in normal fibres, while the Ca(2+) sensitivity of the contractile apparatus was not significantly different. These results indicate that mdx hindlimb muscles are affected differently by the disease process and suggest that a reduced ability of the Ca(2+)-ATPase to load Ca(2+) and a leaky sarcoplasmic reticulum membrane may be involved in the altered intracellular Ca(2+) homeostasis.

Influence of fast and slow alkali myosin light chain isoforms on the kinetics of stretch-induced force transients of fast-twitch type IIA fibres of rat.

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Andruchov, Oleg; Galler, Stefan

2008-03-01

This study contributes to understand the physiological role of slow myosin light chain isoforms in fast-twitch type IIA fibres of skeletal muscle. These isoforms are often attached to the myosin necks of rat type IIA fibres, whereby the slow alkali myosin light chain isoform MLC1s is much more frequent and abundant than the slow regulatory myosin light chain isoform MLC2s. In the present study, single-skinned rat type IIA fibres were maximally Ca(2+) activated and subjected to stepwise stretches for causing a perturbation of myosin head pulling cycles. From the time course of the resulting force transients, myosin head kinetics was deduced. Fibres containing MLC1s exhibited slower kinetics independently of the presence or absence of MLC2s. At the maximal MLC1s concentration of about 75%, the slowing was about 40%. The slowing effect of MLC1s is possibly due to differences in the myosin heavy chain binding sites of the fast and slow alkali MLC isoforms, which changes the rigidity of the myosin neck. Compared with the impact of myosin heavy chain isoforms in various fast-twitch fibre types, the influence of MLC1s on myosin head kinetics of type IIA fibres is much smaller. In conclusion, the physiological role of fast and slow MLC isoforms in type IIA fibres is a fine-tuning of the myosin head kinetics.

High efficiency electrotransformation of Lactococcus lactis spp. lactis cells pretreated with lithium acetate and dithiothreitol

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Filioussis George

2007-03-01

Full Text Available Abstract Background A goal for the food industry has always been to improve strains of Lactococcus lactis and stabilize beneficial traits. Genetic engineering is used extensively for manipulating this lactic acid bacterium, while electropolation is the most widely used technique for introducing foreign DNA into cells. The efficiency of electrotransformation depends on the level of electropermealization and pretreatment with chemicals which alter cell wall permeability, resulting in improved transformation efficiencies is rather common practice in bacteria as in yeasts and fungi. In the present study, treatment with lithium acetate (LiAc and dithiothreitol (DTT in various combinations was applied to L. lactis spp. lactis cells of the early-log phase prior to electroporation with plasmid pTRKH3 (a 7.8 kb shuttle vector, suitable for cloning into L. lactis. Two strains of L. lactis spp. lactis were used, L. lactis spp. lactis LM0230 and ATCC 11454. To the best of our knowledge these agents have never been used before with L. lactis or other bacteria. Results Electrotransformation efficiencies of up to 105 transformants per μg DNA have been reported in the literature for L. lactis spp.lactis LM0230. We report here that treatment with LiAc and DDT before electroporation increased transformation efficiency to 225 ± 52.5 × 107 transformants per μg DNA, while with untreated cells or treated with LiAc alone transformation efficiency approximated 1.2 ± 0.5 × 105 transformants per μg DNA. Results of the same trend were obtained with L. lactis ATCC 11454, although transformation efficiency of this strain was significantly lower. No difference was found in the survival rate of pretreated cells after electroporation. Transformation efficiency was found to vary directly with cell density and that of 1010 cells/ml resulted in the highest efficiencies. Following electrotransformation of pretreated cells with LiAc and DDT, pTRKH3 stability was examined

Twitching motility of bacteria with type-IV pili: Fractal walks, first passage time, and their consequences on microcolonies

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Bisht, Konark; Klumpp, Stefan; Banerjee, Varsha; Marathe, Rahul

2017-11-01

A human pathogen, Neisseria gonorrhoeae (NG), moves on surfaces by attaching and retracting polymeric structures called Type IV pili. The tug-of-war between the pili results in a two-dimensional stochastic motion called twitching motility. In this paper, with the help of real-time NG trajectories, we develop coarse-grained models for their description. The fractal properties of these trajectories are determined and their influence on first passage time and formation of bacterial microcolonies is studied. Our main observations are as follows: (i) NG performs a fast ballistic walk on small time scales and a slow diffusive walk over long time scales with a long crossover region; (ii) there exists a characteristic persistent length lp*, which yields the fastest growth of bacterial aggregates or biofilms. Our simulations reveal that lp*˜L0.6 , where L ×L is the surface on which the bacteria move; (iii) the morphologies have distinct fractal characteristics as a consequence of the ballistic and diffusive motion of the constituting bacteria.

Calcium Increases Xylella fastidiosa Surface Attachment, Biofilm Formation, and Twitching Motility

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Cruz, Luisa F.; Cobine, Paul A.

2012-01-01

Xylella fastidiosa is a plant-pathogenic bacterium that forms biofilms inside xylem vessels, a process thought to be influenced by the chemical composition of xylem sap. In this work, the effect of calcium on the production of X. fastidiosa biofilm and movement was analyzed under in vitro conditions. After a dose-response study with 96-well plates using eight metals, the strongest increase of biofilm formation was observed when medium was supplemented with at least 1.0 mM CaCl2. The removal of Ca by extracellular (EGTA, 1.5 mM) and intracellular [1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA/AM), 75 μM] chelators reduced biofilm formation without compromising planktonic growth. The concentration of Ca influenced the force of adhesion to the substrate, biofilm thickness, cell-to-cell aggregation, and twitching motility, as shown by assays with microfluidic chambers and other assays. The effect of Ca on attachment was lost when cells were treated with tetracycline, suggesting that Ca has a metabolic or regulatory role in cell adhesion. A double mutant (fimA pilO) lacking type I and type IV pili did not improve biofilm formation or attachment when Ca was added to the medium, while single mutants of type I (fimA) or type IV (pilB) pili formed more biofilm under conditions of higher Ca concentrations. The concentration of Ca in the medium did not significantly influence the levels of exopolysaccharide produced. Our findings indicate that the role of Ca in biofilm formation may be related to the initial surface and cell-to-cell attachment and colonization stages of biofilm establishment, which rely on critical functions by fimbrial structures. PMID:22194297

Differential expression of FGF receptors and of myogenic regulatory factors in primary cultures of satellite cells originating from fast (EDL) and slow (Soleus) twitch rat muscles.

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Martelly, I; Soulet, L; Bonnavaud, S; Cebrian, J; Gautron, J; Barritault, D

2000-11-01

In the rat, the fast and slow twitch muscles respectively Extensor digitorum longus (EDL) and Soleus present differential characteristics during regeneration. This suggests that their satellite cells responsible for muscle growth and repair represent distinct cellular populations. We have previously shown that satellite cells dissociated from Soleus and grown in vitro proliferate more readily than those isolated from EDL muscle. Fibroblast growth factors (FGFs) are known as regulators of myoblast proliferation and several studies have revealed a relationship between the response of myoblasts to FGF and the expression of myogenic regulatory factors (MRF) of the MyoD family by myoblasts. Therefore, we presently examined the possibility that the satellite cells isolated from EDL and Soleus muscles differ in the expression of FGF receptors (FGF-R) and of MRF expression. FGF-R1 and -R4 were strongly expressed in proliferating cultures whereas FGF-R2 and R3 were not detected in these cultures. In differentiating cultures, only -R1 was present in EDL satellite cells while FGF-R4 was also still expressed in Soleus cells. Interestingly, the unconventional receptor for FGF called cystein rich FGF receptor (CFR), of yet unknown function, was mainly detected in EDL satellite cell cultures. Soleus and EDL satellite cell cultures also differed in the expression MRFs. These results are consistent with the notion that satellite cells from fast and slow twitch muscles belong to different types of myogenic cells and suggest that satellite cells might play distinct roles in the formation and diversification of fast and slow fibres.

The influence of D2O, perchlorate, and variation in temperature on the potential-dependent contractile function of frog skeletal muscle

International Nuclear Information System (INIS)

Foulks, J.G.; Morishita, L.

1985-01-01

D 2 O and perchlorate manifest opposing effects on the contractile function of skeletal muscle (amplitude of twitches and maximum K contractures, potential dependence of contraction and inactivation), and when combined the influence of one may effectively antagonize that of the other. The ratio of perchlorate concentrations required to produce effects of equal intensity, (e.g., twitch enhancement and restoration of maximum K contractures in media lacking divalent cations or containing a depressant concentration of a cationic amphipath) in H 2 O and D 2 O solutions was generally rather constant. These findings are compatible with the view that both agents can influence contractile function by virtue of their effects on solvent structure. In the absence of divalent cations, the effects of reduced temperature resemble those of D 2 O whereas the effects of increased temperature resemble those of the chaotropic anion. However, in other media, variation in temperature was found to result in additional nonsolvent effects so that low temperature could oppose rather than enhance the effects of D 2 O. These observations are discussed in terms of a model which postulates a role for solvent influences on the kinetics of two separate potential-dependent conformational transitions of membrane proteins which mediate the activation and inactivation of contraction in skeletal muscle

Partial fast-to-slow conversion of regenerating rat fast-twitch muscle by chronic low-frequency stimulation.

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Pette, Dirk; Sketelj, Janez; Skorjanc, Dejan; Leisner, Elmi; Traub, Irmtrud; Bajrović, Fajko

2002-01-01

Chronic low-frequency stimulation (CLFS) of rat fast-twitch muscles induces sequential transitions in myosin heavy chain (MHC) expression from MHCIIb --> MHCIId/x --> MHCIIa. However, the 'final' step of the fast-to-slow transition, i.e., the upregulation of MHCI, has been observed only after extremely long stimulation periods. Assuming that fibre degeneration/regeneration might be involved in the upregulation of slow myosin, we investigated the effects of CLFS on extensor digitorum longus (EDL) muscles regenerating after bupivacaine-induced fibre necrosis. Normal, non-regenerating muscles responded to both 30- and 60-day CLFS with fast MHC isoform transitions (MHCIIb --> MHCIId --> MHCIIa) and only slight increases in MHCI. CLFS of regenerating EDL muscles caused similar transitions among the fast isoforms but, in addition, caused significant increases in MHCI (to approximately 30% relative concentration). Stimulation periods of 30 and 60 days induced similar changes in the regenerating bupivacaine-treated muscles, indicating that the upregulation of slow myosin was restricted to regenerating fibres, but only during an early stage of regeneration. These results suggest that satellite cells and/or regenerating fast rat muscle fibres are capable of switching directly to a slow program under the influence of CLFS and, therefore, appear to be more malleable than adult fibres.

Temperature-dependent changes in the viscoelasticity of intact resting mammalian (rat) fast- and slow-twitch muscle fibres.

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Mutungi, G; Ranatunga, K W

1998-04-01

1. The tension and sarcomere length responses induced by ramp stretches (at amplitudes of 1-3 % fibre length (Lo) and speeds of 0.01-12 Lo s-1) were examined at different temperatures (range, 10-35 degrees C) in resting intact muscle fibre bundles isolated from the soleus (a slow-twitch muscle) and extensor digitorum longus (a fast-twitch muscle) of the rat. Some observations are also presented on the effects of chemical skinning on passive viscoelasticity at 10 degrees C. 2. As previously reported, the tension response to a ramp stretch, in different preparations and under various conditions, could be resolved into a viscous (P1), a viscoelastic (P2) and an elastic (P3) component and showed characteristic differences between slow and fast muscle fibres. 3. Chemical skinning of the muscle fibres led to a decrease in the amplitude of all three tension components. However, the fast-slow fibre differences remained after skinning. For example, the viscosity coefficient derived from P1 tension data decreased from 0.84 +/- 0.06 before skinning to 0.44 +/- 0.06 kN s m-2 after skinning in fast fibres; the corresponding values in slow fibres were 2.1 +/- 0.08 and 0.87 +/- 0.09 kN s m-2, respectively. 4. Increasing the experimental temperature from 10 to 35 degrees C led to a decrease in all the tension components in both fast and slow muscle fibre bundles. The decrease of P1 (viscous) tension was such that the viscosity coefficient calculated using P1 data was reduced from 0.84 +/- 0.1 to 0.43 +/- 0.05 kN s m-2 in fast fibres and from 2.0 +/- 0.1 to 1.0 +/- 0.1 kN s m-2 in slow fibres (Q10 of approximately 1.3 in both). 5. In both fast and slow muscle fibre preparations, the plateau tension of the viscoelastic component (P2) decreased by 60-80 % as the temperature was increased from 10 to 35 degrees C giving P2 tension a Q10 of approximately 1.4 in slow fibres and approximately 1.7 in the fast fibres. Additionally, the relaxation time of the viscoelasticity decreased from

Denervation in murine fast-twitch muscle: short-term physiological changes and temporal expression profiling.

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Raffaello, Anna; Laveder, Paolo; Romualdi, Chiara; Bean, Camilla; Toniolo, Luana; Germinario, Elena; Megighian, Aram; Danieli-Betto, Daniela; Reggiani, Carlo; Lanfranchi, Gerolamo

2006-03-13

Denervation deeply affects muscle structure and function, the alterations being different in slow and fast muscles. Because the effects of denervation on fast muscles are still controversial, and high-throughput studies on gene expression in denervated muscles are lacking, we studied gene expression during atrophy progression following denervation in mouse tibialis anterior (TA). The sciatic nerve was cut close to trochanter in adult CD1 mice. One, three, seven, and fourteen days after denervation, animals were killed and TA muscles were dissected out and utilized for physiological experiments and gene expression studies. Target cDNAs from TA muscles were hybridized on a dedicated cDNA microarray of muscle genes. Seventy-one genes were found differentially expressed. Microarray results were validated, and the expression of relevant genes not probed on our array was monitored by real-time quantitative PCR (RQ-PCR). Nuclear- and mitochondrial-encoded genes implicated in energy metabolism were consistently downregulated. Among genes implicated in muscle contraction (myofibrillar and sarcoplasmic reticulum), genes typical of fast fibers were downregulated, whereas those typical of slow fibers were upregulated. Electrophoresis and Western blot showed less pronounced changes in myofibrillar protein expression, partially confirming changes in gene expression. Isometric tension of skinned fibers was little affected by denervation, whereas calcium sensitivity decreased. Functional studies in mouse extensor digitorum longus muscle showed prolongation in twitch time parameters and shift to the left in force-frequency curves after denervation. We conclude that, if studied at the mRNA level, fast muscles appear not less responsive than slow muscles to the interruption of neural stimulation.

Unchanged content of oxidative enzymes in fast-twitch muscle fibers and V˙O2 kinetics after intensified training in trained cyclists

DEFF Research Database (Denmark)

Christensen, Peter Møller; Gunnarsson, Thomas Gunnar Petursson; Thomassen, Martin

2015-01-01

perturbation during INT. Pulmonary V˙O2 kinetics was determined in eight trained male cyclists (V˙O2-max: 59 ± 4 (means ± SD) mL min(-1) kg(-1)) during MOD (205 ± 12 W ~65% V˙O2-max) and INT (286 ± 17 W ~85% V˙O2-max) exercise before and after a 7-week HIT period (30-sec sprints and 4-min intervals) with a 50...... DW(-1) min(-1)) of CS (56 ± 8 post-HIT vs. 59 ± 10 pre-HIT), HAD (27 ± 6 vs. 29 ± 3) and PFK (340 ± 69 vs. 318 ± 105) and the capillary to fiber ratio (2.30 ± 0.16 vs. 2.38 ± 0.20) was unaltered following HIT. V˙O2 kinetics was unchanged with HIT and the speed of the primary response did not differ...... of oxidative enzymes in fast-twitch fibers, and did not change V˙O2 kinetics....

Dithiothreitol activation of the insulin receptor/kinase does not involve subunit dissociation of the native α2β2 insulin receptor subunit complex

International Nuclear Information System (INIS)

Sweet, L.J.; Wilden, P.A.; Pessin, J.E.

1986-01-01

The subunit composition of the dithiothreitol- (DTT) activated insulin receptor/kinase was examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and gel filtration chromatography under denaturing or nondenaturing conditions. Pretreatment of 32 P-labeled insulin receptors with 50 mM DTT followed by gel filtration chromatography in 0.1% SDS demonstrated the dissociation of the α 2 β 2 insulin receptor complex (M/sub r/ 400,000) into the monomeric 95,000 β subunit. In contrast, pretreatment of the insulin receptors with 1-50 mM DTT followed by gel filtration chromatography in 0.1% Triton X-100 resulted in no apparent alteration in mobility compared to the untreated insulin receptors. Resolution of this complex by nonreducing SDS-polyacrylamide gel electrophoresis and autoradiography demonstrated the existence of the α 2 β 2 heterotetrameric complex with essentially no αβ heterodimeric or free monomeric β subunit species present. This suggests that the insulin receptor can reoxidize into the M/sub r/ 400,000 complex after the removal of DTT by gel filtration chromatography. To prevent reoxidation, the insulin receptors were pretreated with 50 mM DTT. Under the conditions the insulin receptors migrated as the M/sub r/ 400,000 α 2 β 2 complex. These results demonstrate that treatment of the insulin receptors with high concentrations of DTT, followed by removal of DTT by gel filtration, results in reoxidation of the reduced α 2 β 2 insulin receptor complex. Further, these results document that although the DTT stimulation of the insulin receptor/kinase does involve reduction of the insulin receptor subunits, it does not result in dissociation of the native α 2 β 2 insulin receptor subunit complex

Percent voluntary inactivation and peak force predictions with the interpolated twitch technique in individuals with high ability of voluntary activation

International Nuclear Information System (INIS)

Herda, Trent J; Walter, Ashley A; Hoge, Katherine M; Stout, Jeffrey R; Costa, Pablo B; Ryan, Eric D; Cramer, Joel T

2011-01-01

The purpose of this study was to examine the sensitivity and peak force prediction capability of the interpolated twitch technique (ITT) performed during submaximal and maximal voluntary contractions (MVCs) in subjects with the ability to maximally activate their plantar flexors. Twelve subjects performed two MVCs and nine submaximal contractions with the ITT method to calculate percent voluntary inactivation (%VI). Additionally, two MVCs were performed without the ITT. Polynomial models (linear, quadratic and cubic) were applied to the 10–90% VI and 40–90% VI versus force relationships to predict force. Peak force from the ITT MVC was 6.7% less than peak force from the MVC without the ITT. Fifty-eight percent of the 10–90% VI versus force relationships were best fit with nonlinear models; however, all 40–90% VI versus force relationships were best fit with linear models. Regardless of the polynomial model or the contraction intensities used to predict force, all models underestimated the actual force from 22% to 28%. There was low sensitivity of the ITT method at high contraction intensities and the predicted force from polynomial models significantly underestimated the actual force. Caution is warranted when interpreting the % VI at high contraction intensities and predicted peak force from submaximal contractions

Changes in contractile properties and action potentials of motor units in the rat medial gastrocnemius muscle during maturation.

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Dobrzynska, Z; Celichowski, J

2016-02-01

The early phase of development of muscles stops following the disappearance of embryonic and neonatal myosin and the elimination of polyneuronal innervation of muscle fibres with the formation of motor units (MUs), but later the muscle mass still considerably increases. It is unknown whether the three types are visible among newly formed MUs soon after the early postnatal period and whether their proportion is similar to that in adult muscle. Moreover, the processes responsible for MU-force regulation by changes in motoneuronal firing rate as well as properties of motor unit action potentials (MUAPs) during maturation are unknown. Three groups of Wistar rats were investigated - 1 month old, 2 months old and the adult, 9 months old. The basic contractile properties and action potentials of MUs in the medial gastrocnemius (MG) muscle were analysed. The three types of MUs were distinguishable in all age groups, but higher proportion of slow MUs was noticed in young rats (29%, 18% and 11% in 1, 2 and 9 months rats, respectively). The fatigue index for fast fatigable MUs in 1 month old rats was about 2 times higher than in 9 months old rats. The twitch time parameters of fast MUs were shortened during the maturation; for these units, the force-frequency curves in young rats were shifted towards lower frequencies, which suggested that fast motoneurons of young animals generate lower firing rates. Higher twitch-to-tetanus ratios noted for the three MU types in young rats suggested the smaller role of rate coding in force regulation processes, and the higher role of MU recruitment in young rats. No significant differences in MUAP parameters between two groups of young and adult animals were observed. Concluding, the maturation process evokes deeper changes in fast MUs than in slow ones.

Myosin Binding Protein-C Slow Phosphorylation is Altered in Duchenne Dystrophy and Arthrogryposis Myopathy in Fast-Twitch Skeletal Muscles.

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Ackermann, Maegen A; Ward, Christopher W; Gurnett, Christina; Kontrogianni-Konstantopoulos, Aikaterini

2015-08-19

Myosin Binding Protein-C slow (sMyBP-C), encoded by MYBPC1, comprises a family of regulatory proteins of skeletal muscles that are phosphorylated by PKA and PKC. MYBPC1 missense mutations are linked to the development of Distal Arthrogryposis-1 (DA-1). Although structure-function details for this myopathy are evolving, function is undoubtedly driven by sequence variations and post-translational modifications in sMyBP-C. Herein, we examined the phosphorylation profile of sMyBP-C in mouse and human fast-twitch skeletal muscles. We used Flexor Digitorum Brevis (FDB) isolated from young (~2-months old) and old (~14-months old) wild type and mdx mice, and human Abductor Hallucis (AH) and gastrocnemious muscles carrying the DA-1 mutations. Our results indicate both constitutive and differential phosphorylation of sMyBP-C in aged and diseased muscles. We report a 7-35% reduction in the phosphorylation levels of select sites in old wild type and young or old mdx FDB mouse muscles, compared to young wild type tissue. Similarly, we observe a 30-70% decrease in the phosphorylation levels of all PKA and PKC phospho-sites in the DA-1 AH, but not gastrocnemius, muscle. Overall, our studies show that the phosphorylation pattern of sMyBP-C is differentially regulated in response to age and disease, suggesting that phosphorylation plays important roles in these processes.

Maximal Voluntary Activation of the Elbow Flexors Is under Predicted by Transcranial Magnetic Stimulation Compared to Motor Point Stimulation Prior to and Following Muscle Fatigue

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Edward W. J. Cadigan

2017-09-01

Full Text Available Transcranial magnetic (TMS and motor point stimulation have been used to determine voluntary activation (VA. However, very few studies have directly compared the two stimulation techniques for assessing VA of the elbow flexors. The purpose of this study was to compare TMS and motor point stimulation for assessing VA in non-fatigued and fatigued elbow flexors. Participants performed a fatigue protocol that included twelve, 15 s isometric elbow flexor contractions. Participants completed a set of isometric elbow flexion contractions at 100, 75, 50, and 25% of maximum voluntary contraction (MVC prior to and following fatigue contractions 3, 6, 9, and 12 and 5 and 10 min post-fatigue. Force and EMG of the bicep and triceps brachii were measured for each contraction. Force responses to TMS and motor point stimulation and EMG responses to TMS (motor evoked potentials, MEPs and Erb's point stimulation (maximal M-waves, Mmax were also recorded. VA was estimated using the equation: VA% = (1−SITforce/PTforce × 100. The resting twitch was measured directly for motor point stimulation and estimated for both motor point stimulation and TMS by extrapolation of the linear regression between the superimposed twitch force and voluntary force. MVC force, potentiated twitch force and VA significantly (p < 0.05 decreased throughout the elbow flexor fatigue protocol and partially recovered 10 min post fatigue. VA was significantly (p < 0.05 underestimated when using TMS compared to motor point stimulation in non-fatigued and fatigued elbow flexors. Motor point stimulation compared to TMS superimposed twitch forces were significantly (p < 0.05 higher at 50% MVC but similar at 75 and 100% MVC. The linear relationship between TMS superimposed twitch force and voluntary force significantly (p < 0.05 decreased with fatigue. There was no change in triceps/biceps electromyography, biceps/triceps MEP amplitudes, or bicep MEP amplitudes throughout the fatigue protocol at

Attenuated fatigue in slow twitch skeletal muscle during isotonic exercise in rats with chronic heart failure.

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Morten Munkvik

Full Text Available During isometric contractions, slow twitch soleus muscles (SOL from rats with chronic heart failure (chf are more fatigable than those of sham animals. However, a muscle normally shortens during activity and fatigue development is highly task dependent. Therefore, we examined the development of skeletal muscle fatigue during shortening (isotonic contractions in chf and sham-operated rats. Six weeks following coronary artery ligation, infarcted animals were classified as failing (chf if left ventricle end diastolic pressure was >15 mmHg. During isoflurane anaesthesia, SOL with intact blood supply was stimulated (1s on 1s off at 30 Hz for 15 min and allowed to shorten isotonically against a constant afterload. Muscle temperature was maintained at 37°C. In resting muscle, maximum isometric force (F(max and the concentrations of ATP and CrP were not different in the two groups. During stimulation, F(max and the concentrations declined in parallel sham and chf. Fatigue, which was evident as reduced shortening during stimulation, was also not different in the two groups. The isometric force decline was fitted to a bi-exponential decay equation. Both time constants increased transiently and returned to initial values after approximately 200 s of the fatigue protocol. This resulted in a transient rise in baseline tension between stimulations, although this effect which was less prominent in chf than sham. Myosin light chain 2s phosphorylation declined in both groups after 100 s of isotonic contractions, and remained at this level throughout 15 min of stimulation. In spite of higher energy demand during isotonic than isometric contractions, both shortening capacity and rate of isometric force decline were as well or better preserved in fatigued SOL from chf rats than in sham. This observation is in striking contrast to previous reports which have employed isometric contractions to induce fatigue.

U-14C-lactate-to-glycogen conversion and glycogen resynthesis rates in Type I and Type II human vastus lateralis muscle determined from biopsy samples following supramaximal and submaximal exhaustive one-leg cycling: an in vitro versus in vivo comparison

International Nuclear Information System (INIS)

Thompson, J.L.

1987-01-01

To determine the in vitro lactate-to-glycogen conversion potential of human muscle, samples were incubated in U- 14 C-lactate. Because evidence existed suggesting that lactate-to-glycogen conversion occurred at a faster rate in Type II muscle in vivo glycogen resynthesis was calculated by the difference in muscle glycogen concentrations over the initial half-hour recovery period in the FT (Type II, fast-twitch) and ST (Type I, slow-twitch) muscle fiber pools from two of the original eight subjects

Connexin 39.9 Protein Is Necessary for Coordinated Activation of Slow-twitch Muscle and Normal Behavior in Zebrafish*

Science.gov (United States)

Hirata, Hiromi; Wen, Hua; Kawakami, Yu; Naganawa, Yuriko; Ogino, Kazutoyo; Yamada, Kenta; Saint-Amant, Louis; Low, Sean E.; Cui, Wilson W.; Zhou, Weibin; Sprague, Shawn M.; Asakawa, Kazuhide; Muto, Akira; Kawakami, Koichi; Kuwada, John Y.

2012-01-01

In many tissues and organs, connexin proteins assemble between neighboring cells to form gap junctions. These gap junctions facilitate direct intercellular communication between adjoining cells, allowing for the transmission of both chemical and electrical signals. In rodents, gap junctions are found in differentiating myoblasts and are important for myogenesis. Although gap junctions were once believed to be absent from differentiated skeletal muscle in mammals, recent studies in teleosts revealed that differentiated muscle does express connexins and is electrically coupled, at least at the larval stage. These findings raised questions regarding the functional significance of gap junctions in differentiated muscle. Our analysis of gap junctions in muscle began with the isolation of a zebrafish motor mutant that displayed weak coiling at day 1 of development, a behavior known to be driven by slow-twitch muscle (slow muscle). We identified a missense mutation in the gene encoding Connexin 39.9. In situ hybridization found connexin 39.9 to be expressed by slow muscle. Paired muscle recordings uncovered that wild-type slow muscles are electrically coupled, whereas mutant slow muscles are not. The further examination of cellular activity revealed aberrant, arrhythmic touch-evoked Ca2+ transients in mutant slow muscle and a reduction in the number of muscle fibers contracting in response to touch in mutants. These results indicate that Connexin 39.9 facilitates the spreading of neuronal inputs, which is irregular during motor development, beyond the muscle cells and that gap junctions play an essential role in the efficient recruitment of slow muscle fibers. PMID:22075003

Length dependence of staircase potentiation: interactions with caffeine and dantrolene sodium.

Science.gov (United States)

Rassier, D E; MacIntosh, B R

2000-04-01

In skeletal muscle, there is a length dependence of staircase potentiation for which the mechanism is unclear. In this study we tested the hypothesis that abolition of this length dependence by caffeine is effected by a mechanism independent of enhanced Ca2+ release. To test this hypothesis we have used caffeine, which abolishes length dependence of potentiation, and dantrolene sodium, which inhibits Ca2+ release. In situ isometric twitch contractions of rat gastrocnemius muscle before and after 20 s of repetitive stimulation at 5 Hz were analyzed at optimal length (Lo), Lo - 10%, and Lo + 10%. Potentiation was observed to be length dependent, with an increase in developed tension (DT) of 78 +/- 12, 51 +/- 5, and 34 +/- 9% (mean +/- SEM), at Lo - 10%, Lo, and Lo + 10%, respectively. Caffeine diminished the length dependence of activation and suppressed the length dependence of staircase potentiation, giving increases in DT of 65+/-13, 53 +/- 11, and 45 +/- 12% for Lo - 10%, Lo, and Lo + 10%, respectively. Dantrolene administered after caffeine did not reverse this effect. Dantrolene alone depressed the potentiation response, but did not affect the length dependence of staircase potentiation, with increases in DT of 58 +/- 17, 26 +/- 8, and 18 +/- 7%, respectively. This study confirms that there is a length dependence of staircase potentiation in mammalian skeletal muscle which is suppressed by caffeine. Since dantrolene did not alter this suppression of the length dependence of potentiation by caffeine, it is apparently not directly modulated by Ca2+ availability in the myoplasm.

Effect of endoplasmic reticulum stress on the response of HeLa cells to carbon ion radiation

International Nuclear Information System (INIS)

Xia Jiefang; Wang Zhuanzi; Wei Wei; Dang Bingrong; Li Wenjian

2015-01-01

To investigate the effect of endoplasmic reticulum stress on HeLa cells to "1"2C"6"+ ion irradiation, HeLa cells were pretreated with 2.5 mmol/L dithiothreitol and irradiated by "1"2C"6"+ ions with different doses. The results showed that, compared with IR alone, dithiothreitol combined with carbon ion irradiation caused HeLa cell survival decreased, and the apoptosis increased. Moreover, dithiothreitol and carbon ion radiation combination treatment led to a significant increase of G_2/M phase, and autophagy was activated obviously in combination treatment group. The results imply that continuous endoplasmic reticulum stress can change the response of HeLa cells to "1"2C"6"+ irradiation, and dithiothreitol may affect HeLa cells through the autophagy cell death pathway. (authors)

β-Adrenergic modulation of skeletal muscle contraction: key role of excitation-contraction coupling.

Science.gov (United States)

Cairns, Simeon P; Borrani, Fabio

2015-11-01

Our aim is to describe the acute effects of catecholamines/β-adrenergic agonists on contraction of non-fatigued skeletal muscle in animals and humans, and explain the mechanisms involved. Adrenaline/β-agonists (0.1-30 μm) generally augment peak force across animal species (positive inotropic effect) and abbreviate relaxation of slow-twitch muscles (positive lusitropic effect). A peak force reduction also occurs in slow-twitch muscles in some conditions. β2 -Adrenoceptor stimulation activates distinct cyclic AMP-dependent protein kinases to phosphorylate multiple target proteins. β-Agonists modulate sarcolemmal processes (increased resting membrane potential and action potential amplitude) via enhanced Na(+) -K(+) pump and Na(+) -K(+) -2Cl(-) cotransporter function, but this does not increase force. Myofibrillar Ca(2+) sensitivity and maximum Ca(2+) -activated force are unchanged. All force potentiation involves amplified myoplasmic Ca(2+) transients consequent to increased Ca(2+) release from sarcoplasmic reticulum (SR). This unequivocally requires phosphorylation of SR Ca(2+) release channels/ryanodine receptors (RyR1) which sensitize the Ca(2+) -induced Ca(2+) release mechanism. Enhanced trans-sarcolemmal Ca(2+) influx through phosphorylated voltage-activated Ca(2+) channels contributes to force potentiation in diaphragm and amphibian muscle, but not mammalian limb muscle. Phosphorylation of phospholamban increases SR Ca(2+) pump activity in slow-twitch fibres but does not augment force; this process accelerates relaxation and may depress force. Greater Ca(2+) loading of SR may assist force potentiation in fast-twitch muscle. Some human studies show no significant force potentiation which appears to be related to the β-agonist concentration used. Indeed high-dose β-agonists (∼0.1 μm) enhance SR Ca(2+) -release rates, maximum voluntary contraction strength and peak Wingate power in trained humans. The combined findings can explain how adrenaline

Synthesis, Characterization and DNA Cleavage of Copper(II ...

African Journals Online (AJOL)

Purpose: To study deoxyribonucleic acid (DNA) shearing capability of copper(II) complex of dithiothreitol (DTT) and to fevaluate its potential application in cancer therapy. Methods: A parrot green complex was synthesized by grinding copper acetate monohydrate and DTT in 1:2 molar ratio in a mortar until no fumes of acetic ...

Influence of lecithin liposomes on chlorophyllase-catalyzed chlorophyll hydrolysis. Comparison of intramembraneous and solubilized Phaeodactylum chlorophyllase

NARCIS (Netherlands)

Terpstra, Willemke

1980-01-01

1. 1. Chlorophyllase-catalyzed chlorophyll hydrolysis is greatly enhanced by the addition of divalent cations (Mg2+) combined with a reducing agent (dithiothreitol, ascorbate). A similar effect is obtained by the addition of lecithin. In the presence of lecithin, dithiothreitol has only slight or no

The Action of Botulinum Toxin at the Neuromuscular Junction

Science.gov (United States)

1980-12-22

fast - twitch " (gastrocnemius) and " slow - twitch " (soleus) muscles ... muscle fibers -"_re not significantly affected by the toxin. It is interesting to note that, although fast - twitch and slow - twitch mucles were...Duchen LW: An electron microscopic study of the changes induced by borulinum o::in in the motor end-plates of slow and fast skeletal muscle fibres of

Oxidative potential of secondary organic aerosols produced from photooxidation of different hydrocarbons using outdoor chamber under ambient sunlight

Science.gov (United States)

Jiang, Huanhuan; Jang, Myoseon; Sabo-Attwood, Tara; Robinson, Sarah E.

2016-04-01

The oxidative potential of various secondary organic aerosols (SOA) was measured using dithiothreitol (DTT) assay to understand how organic aerosols react with cellular materials. SOA was produced via the photooxidation of four different hydrocarbons (toluene, 1,3,5-trimethylbenzene, isoprene and α-pinene) in the presence of NOx using a large outdoor photochemical smog chamber. The DTT consumption rate was normalized by the aerosol mass, which is expressed as DTTmass. Toluene SOA and isoprene SOA yielded higher DTTmass than 1,3,5-trimethylbenzene SOA or α-pinene SOA. In order to discover the correlation between the molecular structure and oxidative potential, the DTT responses of selected model compounds were also measured. Among them, conjugated aldehydes, quinones, and H2O2 showed considerable DTT response. To investigate the correlation between DTT response and cell responses in vitro, the expression of biological markers, i.e. IL-6, IL-8, and HMOX-1 were studied using small airway epithelial cells. Higher cellular expression of IL-8 was observed with toluene SOA exposure compared to 1,3,5-trimethylbenzene SOA exposure, which aligned with the results from DTT assay. Our study also suggests that within the urban atmosphere, the contribution of toluene SOA and isoprene SOA to the oxidative potential of ambient SOA will be more significant than that of α-pinene SOA.

INTERACTION OF VERAPAMIL AND LITHIUM AT THE NEUROMUSCULAR JUNCTION ON RAT ISOLATED MUSCLE-HEMIDIAPHRAGM

Directory of Open Access Journals (Sweden)

H. R. Sadeghipour

1998-08-01

Full Text Available It has been reported that cither lithium or verapamil can potentiate the neuromuscular blocking activity of certain neuromuscular blockers. In the present investigation, possible interaction of verapamil with lithium has been described. The dose ■ response effects of verapamil and lithium on diaphragmatic contractility were assessed in vitro. Mechanical responses of the muscle to indirect (nerve and direct (muscle electrical stimulation were recorded. Verapamil depressed rat diaphragm twitch tensions induced by nerve stimulation in a dose - dependent manner with the 50 percent depression of the original twitch tensions (ICSQ by 5.6 xlO^mmol/l."nThe IC50 of verapamil for direct stimulation of the muscle was LI x W'5 mmol II. Partial replacement of sodium chloride by lithium chloride (0.5, 1.5 and 5 mmol /1 in the medium did not change the depressant effect of verapamil on muscle twitches induced by direct (muscle or indirect (nerve electrical stimulation.

Models of disuse - A comparison of hindlimb suspension and immobilization

Science.gov (United States)

Fitts, R. H.; Metzger, J. M.; Riley, D. A.; Unsworth, B. R.

1986-01-01

The effects of 1 and 2 weeks of hindlimb suspension (HS) on the contractile properties of fast- and slow-twitch skeletal muscles of male Sprague Dawley rats are studied and compared with hindlimb immobilization (HI) data. The optimal length and contractile properties of the slow-twitch soleus, fast-twitch extensor digitorum longus, and the vastus lateralis are measured. It is observed that HS and HI affect slow-twitch muscles; isometric twitch duration in the slow-twitch soleus is decreased. Soleus muscle mass and peak tetanic tension declines with disuse. A major difference in the influence of HS and HI on the maximal speed of soleus muscle shortening, V(max) is detected; HS produced a twofold increase in V(max) compared to control data and HI had no significant effect on V(max). The relation between V(max) and myosin concentration is analyzed. The data reveal that HS modifies slow-twitch muscle yielding hybrid fibers with elevated shortening velocities and this change may be dependent on the elimination of load-bearing contractions.

Limitations in Using Chemical Oxidative Potential to Understand Oxidative Stress from Particulate Matter

Science.gov (United States)

Chan, A. W. H.; Wang, S.; Wang, X.; Kohl, L.; Chow, C. W.

2017-12-01

Particulate matter (PM) in the atmosphere is known to cause adverse cardiorespiratory health effects. It has been suggested that the ability of PM to generate oxidative stress leads to a proinflammatory response. In this work, we study the biological relevance of using a chemical oxidative potential (OP) assay to evaluate proinflammatory response in airway epithelial cells. Here we study the OPs of laboratory secondary organic aerosol (SOA) and metal mixtures, ambient PM from India, ash from the 2016 Alberta wildfires, and diesel exhaust particles. We use SOA derived from naphthalene and from monoterpenes as model systems for SOA. We measure OP using the dithiothreitol (DTT) assay, and cytosolic reactive oxygen species (ROS) production in BEAS-2B cell culture was measured using CellROX assay. We found that both SOA and copper show high OPs individually, but the OP of the combined SOA/copper mixture, which is more atmospherically relevant, was lower than either of the individual OPs. The reduced activity is attributed to chelation between metals and organic compounds using proton nuclear magnetic resonance. There is reasonable association between DTT activity and cellular ROS production within each particle type, but weak association across different particle types, suggesting that particle composition plays an important role in distinguishing between antioxidant consumption and ROS production. Our results highlight that while oxidative potential is a useful metric of PM's ability to generate oxidative stress, the chemical composition and cellular environment should be considered in understanding health impacts of PM.

Polymorphism of myofibrillar proteins of rabbit skeletal-muscle fibres. An electrophoretic study of single fibres.

OpenAIRE

Salviati, G; Betto, R; Danieli Betto, D

1982-01-01

Rabbit predominantly fast-twitch-fibre and predominantly slow-twitch-fibre skeletal muscles of the hind limbs, the psoas, the diaphragm and the masseter muscles were fibre-typed by one-dimensional polyacrylamide-gel electrophoresis of the myofibrillar proteins of chemically skinned single fibres. Investigation of the distribution of fast-twitch-fibre and slow-twitch-fibre isoforms of myosin light chains and the type of myosin heavy chains, based on peptide 'maps' published in Cleveland. Fisch...

Changes of contractile responses due to simulated weightlessness in rat soleus muscle

Science.gov (United States)

Elkhammari, A.; Noireaud, J.; Léoty, C.

1994-08-01

Some contractile and electrophysiological properties of muscle fibers isolated from the slow-twitch soleus (SOL) and fast-twitch extensor digitorum longus (EDL) muscles of rats were compared with those measured in SOL muscles from suspended rats. In suspendede SOL (21 days of tail-suspension) membrane potential (Em), intracellular sodium activity (aiNa) and the slope of the relationship between Em and log [K]o were typical of fast-twitch muscles. The relation between the maximal amplitude of K-contractures vs Em was steeper for control SOL than for EDL and suspended SOL muscles. After suspension, in SOL muscles the contractile threshold and the inactivation curves for K-contractures were shifted to more positive Em. Repriming of K-contractures was unaffected by suspencion. The exposure of isolated fibers to perchlorate (ClO4-)-containing (6-40 mM) solutions resulted ina similar concentration-dependent shift to more negative Em of activation curves for EDL and suspended SOL muscles. On exposure to a Na-free TEA solution, SOL from control and suspended rats, in contrast to EDL muscles, generated slow contractile responses. Suspended SOL showed a reduced sensitivity to the contracture-producing effect of caffeine compared to control muscles. These results suggested that the modification observed due to suspension could be encounted by changes in the characteristics of muscle fibers from slow to fast-twitch type.

Charge Movement in a Fast Twitch Skeletal Muscle from Rat

OpenAIRE

Simon, B. J.; Beam, K. G.

1983-01-01

Voltage-dependent charge movement in the rat omohyoid muscle was investigated using the three microelectrode voltage clamp technique. The charge that moved during a depolarization from the holding potential (-90 mV) to the test potential, V, increased with increasing V, saturating around 0 mV. The charge vs. voltage relationship was well fitted by Q = Qmax/{1 + exp[-(V - V)/k]}, with Qmax = 28.5 nC/μF, V = -34.2 mV, and k = 8.7 mV. Repolarization of the fiber from the test potential back to t...

An in vitro model to test relative antioxidant potential: Ultraviolet-induced lipid peroxidation in liposomes

International Nuclear Information System (INIS)

Pelle, E.; Maes, D.; Padulo, G.A.; Kim, E.K.; Smith, W.P.

1990-01-01

Since antioxidants have been shown to play a major role in preventing some of the effects of aging and photoaging in skin, it is important to study this phenomenon in a controlled manner. This was accomplished by developing a simple and reliable in vitro technique to assay antioxidant efficacy. Inhibition of peroxidation by antioxidants was used as a measure of relative antioxidant potential. Liposomes, high in polyunsaturated fatty acids (PUFA), were dispersed in buffer and irradiated with ultraviolet (UV) light. Irradiated liposomes exhibited a significantly higher amount of hydroperoxides than liposomes containing antioxidants in a dose- and concentration-dependent manner. Lipid peroxidation was determined spectrophotometrically by an increase in thiobarbituric acid reacting substances. To further substantiate the production of lipid peroxides, gas chromatography was used to measure a decrease in PUFA substrate. In order of decreasing antioxidant effectiveness, the following results were found among lipophilic antioxidants: BHA greater than catechin greater than BHT greater than alpha-tocopherol greater than chlorogenic acid. Among hydrophilic antioxidants, ascorbic acid and dithiothreitol were effective while glutathione was ineffective. In addition, ascorbic acid was observed to act synergistically with alpha-tocopherol, which is in agreement with other published reports on the interaction of these two antioxidants. Although peroxyl radical scavengers seem to be at a selective advantage in this liposomal/UV system, these results demonstrate the validity of this technique as an assay for measuring an antioxidant's potential to inhibit UV-induced peroxidation

Immunohistochemical evidence for expression of fast-twitch type sarco(endo)plasmic reticulum Ca2+ ATPase (SERCA1) in German shepherd dogs with dilated cardiomyopathy myocardium.

Science.gov (United States)

Summerfield, Nuala; Peters, Mary E; Hercock, Carol A; Mobasheri, Ali; Young, Iain S

2010-04-01

Dilated cardiomyopathy (DCM) is one of the most common acquired canine heart diseases. It is particularly common in large and giant breed dogs. Although a great deal is known about the clinical progression and manifestations of the disease, the underlying cellular and molecular mechanisms remain poorly understood. One widely held belief is that calcium-handling abnormalities are critically involved in the disease process. This study investigates the changes in expression of the sarco(endo)plasmic reticulum calcium ATPase (SERCA) isoforms in DCM myocardium from German shepherd dogs. Affected tissue samples were obtained from German shepherd dogs with DCM, euthanized for intractable congestive heart failure while normal myocardial tissue samples were obtained from German shepherd dogs, euthanized for non-cardiovascular reasons. Tissue microarrays containing normal and DCM myocardium samples were prepared, immunostained with SERCA1 and SERCA2 antibodies and analyzed. We were able to demonstrate, for the first time, that while there is little change in the expression of the cardiac isoform (SERCA2), there is clear expression of the fast-twitch skeletal muscle isoform SERCA1 in the myocardium of dogs diagnosed with DCM. We propose that SERCA1 expression is evidence of a natural adaptive response to the impaired Ca2+ handling thought to occur in German shepherd dogs with DCM and heart failure. Copyright 2010 Elsevier B.V. All rights reserved.

The role of Sox6 in zebrafish muscle fiber type specification.

Science.gov (United States)

Jackson, Harriet E; Ono, Yosuke; Wang, Xingang; Elworthy, Stone; Cunliffe, Vincent T; Ingham, Philip W

2015-01-01

The transcription factor Sox6 has been implicated in regulating muscle fiber type-specific gene expression in mammals. In zebrafish, loss of function of the transcription factor Prdm1a results in a slow to fast-twitch fiber type transformation presaged by ectopic expression of sox6 in slow-twitch progenitors. Morpholino-mediated Sox6 knockdown can suppress this transformation but causes ectopic expression of only one of three slow-twitch specific genes assayed. Here, we use gain and loss of function analysis to analyse further the role of Sox6 in zebrafish muscle fiber type specification. The GAL4 binary misexpression system was used to express Sox6 ectopically in zebrafish embryos. Cis-regulatory elements were characterized using transgenic fish. Zinc finger nuclease mediated targeted mutagenesis was used to analyse the effects of loss of Sox6 function in embryonic, larval and adult zebrafish. Zebrafish transgenic for the GCaMP3 Calcium reporter were used to assay Ca2+ transients in wild-type and mutant muscle fibres. Ectopic Sox6 expression is sufficient to downregulate slow-twitch specific gene expression in zebrafish embryos. Cis-regulatory elements upstream of the slow myosin heavy chain 1 (smyhc1) and slow troponin c (tnnc1b) genes contain putative Sox6 binding sites required for repression of the former but not the latter. Embryos homozygous for sox6 null alleles expressed tnnc1b throughout the fast-twitch muscle whereas other slow-specific muscle genes, including smyhc1, were expressed ectopically in only a subset of fast-twitch fibers. Ca2+ transients in sox6 mutant fast-twitch fibers were intermediate in their speed and amplitude between those of wild-type slow- and fast-twitch fibers. sox6 homozygotes survived to adulthood and exhibited continued misexpression of tnnc1b as well as smaller slow-twitch fibers. They also exhibited a striking curvature of the spine. The Sox6 transcription factor is a key regulator of fast-twitch muscle fiber differentiation

Length dependence of force generation exhibit similarities between rat cardiac myocytes and skeletal muscle fibres.

Science.gov (United States)

Hanft, Laurin M; McDonald, Kerry S

2010-08-01

According to the Frank-Starling relationship, increased ventricular volume increases cardiac output, which helps match cardiac output to peripheral circulatory demand. The cellular basis for this relationship is in large part the myofilament length-tension relationship. Length-tension relationships in maximally calcium activated preparations are relatively shallow and similar between cardiac myocytes and skeletal muscle fibres. During twitch activations length-tension relationships become steeper in both cardiac and skeletal muscle; however, it remains unclear whether length dependence of tension differs between striated muscle cell types during submaximal activations. The purpose of this study was to compare sarcomere length-tension relationships and the sarcomere length dependence of force development between rat skinned left ventricular cardiac myocytes and fast-twitch and slow-twitch skeletal muscle fibres. Muscle cell preparations were calcium activated to yield 50% maximal force, after which isometric force and rate constants (k(tr)) of force development were measured over a range of sarcomere lengths. Myofilament length-tension relationships were considerably steeper in fast-twitch fibres compared to slow-twitch fibres. Interestingly, cardiac myocyte preparations exhibited two populations of length-tension relationships, one steeper than fast-twitch fibres and the other similar to slow-twitch fibres. Moreover, myocytes with shallow length-tension relationships were converted to steeper length-tension relationships by protein kinase A (PKA)-induced myofilament phosphorylation. Sarcomere length-k(tr) relationships were distinct between all three cell types and exhibited patterns markedly different from Ca(2+) activation-dependent k(tr) relationships. Overall, these findings indicate cardiac myocytes exhibit varied length-tension relationships and sarcomere length appears a dominant modulator of force development rates. Importantly, cardiac myocyte length

De novo synthesis of purine nucleotides in different fiber types of rat skeletal muscle

International Nuclear Information System (INIS)

Tullson, P.C.; John-Alder, H.; Hood, D.A.; Terjung, R.L.

1986-01-01

The contribution of de novo purine nucleotide synthesis to nucleotide metabolism in skeletal muscles is not known. The authors have determined rates of de novo synthesis in soleus (slow-twitch red), red gastrocnemius (fast-twitch red), and white gastrocnemius (fast-twitch white) using the perfused rat hindquarter. 14 C glycine incorporation into ATP was linear after 1 and 2 hours of perfusion with 0.2 mM added glycine. The intracellular (I) and extracellular (E) specific activity of 14 C glycine was determined by HPLC of phenylisothiocyanate derivatives of neutralized PCA extracts. The rates of de novo synthesis when expressed relative to muscle ATP content show slow and fast-twitch red muscles to be similar and about twice as great as fast-twitch white muscles. This could represent a greater turnover of the adenine nucleotide pool in more oxidative red muscle types

The GLUT4 density in slow fibres is not increased in athletes. How does training increase the GLUT4 pool originating from slow fibres?

DEFF Research Database (Denmark)

Gaster, M; Franch, J; Beck-Nielsen, H

2001-01-01

% of the fraction in the control group. Thus, GLUT4 originating from slow-twitch fibres was increased by 30% (Pincreases slow-twitch fibre GLUT4 expression by means of an elevated slow-twitch fibre mass in human skeletal muscle.......The influence of training on GLUT4 expression in slow- and fast-twitch skeletal muscle fibres was studied in male endurance-trained athletes and control subjects. The trained state was ensured by elevated maximal oxygen uptake (29%), as well as citrate synthase (60%) and 3-hydroxy......-acyl-CoA dehydrogenase (38%) activities in muscle biopsy samples of the vastus lateralis. GLUT4 densities in slow- and fast-twitch fibres were measured by the use of a newly developed, sensitive method combining immunohistochemistry with morphometry, and no effect of training was found. GLUT4 density was higher in slow...

Tetanic Ca2+ transient differences between slow- and fast-twitch mouse skeletal muscle fibres: a comprehensive experimental approach.

Science.gov (United States)

Calderón, Juan C; Bolaños, Pura; Caputo, Carlo

2014-12-01

One hundred and eighty six enzymatically dissociated murine muscle fibres were loaded with Mag-Fluo-4 AM, and adhered to laminin, to evaluate the effect of modulating cytosolic Ca(2+) buffers and sarcoendoplasmic reticulum Ca(2+) ATPase (SERCA), mitochondria, and Na(+)/Ca(2+) exchanger (NCX) on the differential tetanic Ca(2+) transient kinetics found in different fibre types. Tetanic Ca(2+) transients were classified as morphology type I (MT-I) or type II (MT-II) according to their shape. The first peak of the MT-I (n = 25) and MT-II (n = 23) tetanic Ca(2+) transients had an amplitude (∆F/F) of 0.41 ± 0.03 and 0.83 ± 0.05 and a rise time (ms) of 1.35 and 0.98, respectively. MT-I signals had a time constant of decay (τ1, ms) of 75.9 ± 4.2 while MT-II transients showed a double exponential decay with time constants of decay (τ1 and τ2, ms) of 18.3 ± 1.4 and 742.2 ± 130.3. Sarcoendoplasmic reticulum Ca(2+) ATPase inhibition demonstrated that the decay phase of the tetanic transients mostly rely on SERCA function. Adding Ca(2+) chelators in the AM form to MT-I fibres changed the morphology of the initial five peaks to a MT-II one, modifying the decay phase of the signal in a dose-dependent manner. Mitochondria and NCX function have a minor role in explaining differences in tetanic Ca(2+) transients among fibre types but still help in removing Ca(2+) from the cytosol in both MT-I and MT-II fibres. Cytoplasmic Ca(2+) buffering capacity and SERCA function explain most of the different kinetics found in tetanic Ca(2+) transients from different fibre types, but mitochondria and NCX have a measurable role in shaping tetanic Ca(2+) responses in both slow and fast-twitch muscle fibre types. We provided experimental evidence on the mechanisms that help understand the kinetics of tetanic Ca(2+) transients themselves and explain kinetic differences found among fibre types.

A comparative study of charge movement in rat and frog skeletal muscle fibres.

Science.gov (United States)

Hollingworth, S; Marshall, M W

1981-12-01

1. The middle of the fibre voltage--clamp technique (Adrian & Marshall, 1977), modified where necessary for electrically short muscle fibres, has been used to measure non-linear charge movements in mammalian fast twitch (rat extensor digitorum longus), mammalian slow twitch (rat soleus) and frog (sartorius) muscles. 2. The maximum amount of charge moved in mammalian fast twitch muscle at 2 degrees C in hypertonic solution, was 3--5 times greater than in slow twitch muscle. The voltage distribution of fast twitch charge was 10--15 mV more positive when compared to slow twitch. 3. In both mammalian muscle types hypertonic Ringer solution negatively shifted the voltage distribution of charge some 6 mV. The steepness of charge moved around mechanical threshold was unaffected by hypertonicity. 4. The amount of charge in frog sartorius fibres at 2 degrees C in hypertonic solution was about half of that in rat fast twitch muscle; the voltage distribution of the frog charge was similar to rat soleus muscle. 5. Warming between 2 and 15 degrees C had no effect on either the amount of steady-state distribution of charge in mammalian or frog muscles. 6. At 2 degrees C, the kinetics of charge movement in fast and slow twitch mammalian muscles were similar and 2--3 times faster than frog muscle at the same temperature. In fast and slow mammalian fibres at 2 degrees C similar times were taken to shift the same fractions of the total amount of charge. The Q10 of charge movement kinetics was between 1.2 and 2.0 in the three muscles studied.

Inotropic responses of the frog ventricle to adenosine triphosphate and related changes in endogenous cyclic nucleotides.

Science.gov (United States)

Flitney, F W; Singh, J

1980-07-01

1. A study has been made of a well documented but poorly understood response of the isolated frog ventricle to treatment with exogenous adenosine 5' triphosphate (ATP). Measurements of membrane potential, isometric twitch tension and levels of endogenous 3',5'-cyclic nucleotides have been made at various times during the ATP-induced response. 2. ATP elicits a characteristic triphasic response, which comprises an initial, abrupt increase in contractility, rising to a maximum within a few beats (first phase); followed by a period when the twitch amplitude falls, sometimes to below the control level (second phase); and superceded by a more slowly developing and longer-lasting increase in contractile force (third phase). The response is unaffected by atropine, propranolol or phentolamine. However, the prostaglandin synthetase inhibitor indomethacin depresses the first phase and entirely suppresses the third phase. 3. The inotropic effects of ATP are accompanied by changes in the shape of the action potential. These effects are dose-related. The duration of the action potential (D-30mV) and its positive overshoot (O) are increased during all phases of the response, for [ATP]o's up to 10(-5) M. However, at higher [ATP]o's, D-30mV and O ar both reduced during the second phase (but not the first or third phase), when isometric twitch tension is also depressed. The relationship between action potential duration and twitch tension (P) for different [ATP]o's is linear for all three phases of the response, but the slopes of the curves (delta P/delta D) are markedly different, indicating that the sensitivity of the contractile system to membrane depolarization is not constant, but varies continuously throughout the response. 4. ATP has a potent stimulatory effect on the metabolism of endogenous 3',5'-cyclic nucleotides. The time courses of the changes in adenosine 3','5-cyclic monophosphate (3',5'-cyclic AMP) and guanosine 3',5'-cyclic monophosphate (3',5'-cyclic GMP) are

Measuring Dynamic Kidney Function in an Undergraduate Physiology Laboratory

Science.gov (United States)

Medler, Scott; Harrington, Frederick

2013-01-01

Most undergraduate physiology laboratories are very limited in how they treat renal physiology. It is common to find teaching laboratories equipped with the capability for high-resolution digital recordings of physiological functions (muscle twitches, ECG, action potentials, respiratory responses, etc.), but most urinary laboratories still rely on…

Tributyltin interacts with mitochondria and induces cytochrome c release.

Science.gov (United States)

Nishikimi, A; Kira, Y; Kasahara, E; Sato, E F; Kanno, T; Utsumi, K; Inoue, M

2001-01-01

Although triorganotins are potent inducers of apoptosis in various cell types, the critical targets of these compounds and the mechanisms by which they lead to cell death remain to be elucidated. There are two major pathways by which apoptotic cell death occurs: one is triggered by a cytokine mediator and the other is by a mitochondrion-dependent mechanism. To elucidate the mechanism of triorganotin-induced apoptosis, we studied the effect of tributyltin on mitochondrial function. We found that moderately low doses of tributyltin decrease mitochondrial membrane potential and induce cytochrome c release by a mechanism inhibited by cyclosporine A and bongkrekic acid. Tributyltin-induced cytochrome c release is also prevented by dithiols such as dithiothreitol and 2,3-dimercaptopropanol but not by monothiols such as GSH, N-acetyl-L-cysteine, L-cysteine and 2-mercaptoethanol. Further studies with phenylarsine oxide agarose revealed that tributyltin interacts with the adenine nucleotide translocator, a functional constituent of the mitochondrial permeability transition pore, which is selectively inhibited by dithiothreitol. These results suggest that, at low doses, tributyltin interacts selectively with critical thiol residues in the adenine nucleotide translocator and opens the permeability transition pore, thereby decreasing membrane potential and releasing cytochrome c from mitochondria, a series of events consistent with established mechanistic models of apoptosis. PMID:11368793

Role of calpain in eccentric contraction-induced proteolysis of Ca2+-regulatory proteins and force depression in rat fast-twitch skeletal muscle.

Science.gov (United States)

Kanzaki, Keita; Watanabe, Daiki; Kuratani, Mai; Yamada, Takashi; Matsunaga, Satoshi; Wada, Masanobu

2017-02-01

The aim of this study was to examine the in vivo effects of eccentric contraction (ECC) on calpain-dependent proteolysis of Ca 2+ -regulatory proteins and force production in fast-twitch skeletal muscles. Rat extensor digitorum longus muscles were exposed to 200 repeated ECC in situ and excised immediately [recovery 0 (REC0)] or 3 days [recovery 3 (REC3)] after cessation of ECC. Calpain inhibitor (CI)-treated rats were intraperitoneally injected with MDL-28170 before ECC and during REC3. Tetanic force was markedly reduced at REC0 and remained reduced at REC3. CI treatment ameliorated the ECC-induced force decline but only at REC3. No evidence was found for proteolysis of dihydropyridine receptor (DHPR), junctophilin (JP)1, JP2, ryanodine receptor (RyR), sarcoplasmic reticulum Ca 2+ -ATPase (SERCA)1a, or junctional face protein-45 at REC0. At REC3, ECC resulted in decreases in DHPR, JP1, JP2, RyR, and SERCA1a. CI treatment prevented the decreases in DHPR, JP1, and JP2, whereas it had little effect on RyR and SERCA1a. These findings suggest that DHPR, JP1, and JP2, but not RyR and SERCA1a, undergo calpain-dependent proteolysis in in vivo muscles subjected to ECC and that impaired function of DHPR and/or JP might cause prolonged force deficits with ECC. NEW & NOTEWORTHY Calpain-dependent proteolysis is one of the contributing factors to muscle damage that occurs with eccentric contraction (ECC). It is unclear, however, whether calpains account for proteolysis of Ca 2+ -regulatory proteins in in vivo muscles subjected to ECC. Here, we provide evidence that dihydropyridine receptor and junctophilin, but not ryanodine receptor and sarcoplasmic reticulum Ca 2+ -ATPase, undergo calpain-dependent proteolysis. Copyright © 2017 the American Physiological Society.

Impediment in upper airway stabilizing forces assessed by phrenic nerve stimulation in sleep apnea patients

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Vérin E

2005-09-01

Full Text Available Abstract Background The forces developed during inspiration play a key role in determining upper airway stability and the occurrence of nocturnal breathing disorders. Phrenic nerve stimulation applied during wakefulness is a unique tool to assess Upper airway dynamic properties and to measure the overall mechanical effects of the inspiratory process on UA stability. Objectives To compare the flow/pressure responses to inspiratory and expiratory twitches between sleep apnea subjects and normal subjects. Methods Inspiratory and expiratory twitches using magnetic nerve stimulation completed in eleven untreated sleep apnea subjects and ten normal subjects. Results In both groups, higher flow and pressure were reached during inspiratory twitches. The two groups showed no differences in expiratory twitch parameters. During inspiration, the pressure at which flow-limitation occurred was more negative in normals than in apneic subjects, but not reaching significance (p = 0.07. The relationship between pharyngeal pressure and flow adequately fitted with a polynomial regression model providing a measurement of upper airway critical pressure during twitch. This pressure significantly decreased in normals from expiratory to inspiratory twitches (-11.1 ± 1.6 and -15.7 ± 1.0 cm H2O respectively, 95% CI 1.6–7.6, p Conclusion Inspiratory-related upper airway dilating forces are impeded in sleep apnea patients.

Women at Altitude: Voluntary Muscle Exercise Performance with and Without a-Adrenergic Receptor Blockage

Science.gov (United States)

1999-02-01

proportion of active muscle volume occupied by slow - twitch fibers (a consequence of women having a smaller, fast - twitch fiber cross-sectional area (11,27...oxidative metabolism and in the ratio of slow -to- fast twitch fiber area must be considered with caution, however, since the proportion of slow fatiguing...ventilatory acclimatization to 4300m. Respir.Physiol. 70: 195-204,1987. 27. Nygaard, E. Skeletal muscle fibre characteristics in young women. Acta

Effects of trace elements and mono- and dithiols on mitochondrial monoamine oxidase of rats

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Revis, N.; Horton, C.

1978-01-01

The effects of several trace elements on mitochondrial monoamine oxidase (MAO) were studied. Elements were studied at a concentration of 1 mM; only mercury, cadmium, and copper were significantly effective in reducing the activity of this enzyme. Of several thiols tested, only dithiothreitol could reverse the inhibition of MAO by these elements. Evidence is also presented in this report to show that cysteine, homocysteine, and reduced glutathione inhibit this MAO, whereas dithiothreitol or dithioerythritol evoke stimulatory responses.

Recruitment order of quadriceps motor units: femoral nerve vs. direct quadriceps stimulation.

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Rodriguez-Falces, Javier; Place, Nicolas

2013-12-01

To investigate potential differences in the recruitment order of motor units (MUs) in the quadriceps femoris when electrical stimulation is applied over the quadriceps belly versus the femoral nerve. M-waves and mechanical twitches were evoked using femoral nerve stimulation and direct quadriceps stimulation of gradually increasing intensity from 20 young, healthy subjects. Recruitment order was investigated by analysing the time-to-peak twitch and the time interval from the stimulus artefact to the M-wave positive peak (M-wave latency) for the vastus medialis (VM) and vastus lateralis (VL) muscles. During femoral nerve stimulation, time-to-peak twitch and M-wave latency decreased consistently (P  0.05). For the VM muscle, M-wave latency decreased with increasing stimulation level for both femoral nerve and direct quadriceps stimulation, whereas, for the VL muscle, the variation of M-wave latency with stimulus intensity was different for the two stimulation geometries (P recruitment order during direct quadriceps stimulation was more complex, depending ultimately on the architecture of the peripheral nerve and its terminal branches below the stimulating electrodes for each muscle. For the VM, MUs were orderly recruited for both stimulation geometries, whereas, for the VL muscle, MUs were orderly recruited for femoral nerve stimulation, but followed no particular order for direct quadriceps stimulation.

Myosin content of individual human muscle fibers isolated by laser capture microdissection.

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Stuart, Charles A; Stone, William L; Howell, Mary E A; Brannon, Marianne F; Hall, H Kenton; Gibson, Andrew L; Stone, Michael H

2016-03-01

Muscle fiber composition correlates with insulin resistance, and exercise training can increase slow-twitch (type I) fibers and, thereby, mitigate diabetes risk. Human skeletal muscle is made up of three distinct fiber types, but muscle contains many more isoforms of myosin heavy and light chains, which are coded by 15 and 11 different genes, respectively. Laser capture microdissection techniques allow assessment of mRNA and protein content in individual fibers. We found that specific human fiber types contain different mixtures of myosin heavy and light chains. Fast-twitch (type IIx) fibers consistently contained myosin heavy chains 1, 2, and 4 and myosin light chain 1. Type I fibers always contained myosin heavy chains 6 and 7 (MYH6 and MYH7) and myosin light chain 3 (MYL3), whereas MYH6, MYH7, and MYL3 were nearly absent from type IIx fibers. In contrast to cardiomyocytes, where MYH6 (also known as α-myosin heavy chain) is seen solely in fast-twitch cells, only slow-twitch fibers of skeletal muscle contained MYH6. Classical fast myosin heavy chains (MHC1, MHC2, and MHC4) were present in variable proportions in all fiber types, but significant MYH6 and MYH7 expression indicated slow-twitch phenotype, and the absence of these two isoforms determined a fast-twitch phenotype. The mixed myosin heavy and light chain content of type IIa fibers was consistent with its role as a transition between fast and slow phenotypes. These new observations suggest that the presence or absence of MYH6 and MYH7 proteins dictates the slow- or fast-twitch phenotype in skeletal muscle. Copyright © 2016 the American Physiological Society.

Dihydrotestosterone treatment rescues the decline in protein synthesis as a result of sarcopenia in isolated mouse skeletal muscle fibres.

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Wendowski, Oskar; Redshaw, Zoe; Mutungi, Gabriel

2017-02-01

Sarcopenia, the progressive decline in skeletal muscle mass and function with age, is a debilitating condition. It leads to inactivity, falls, and loss of independence. Despite this, its cause(s) and the underlying mechanism(s) are still poorly understood. In this study, small skeletal muscle fibre bundles isolated from the extensor digitorum longus (a fast-twitch muscle) and the soleus (a slow-twitch muscle) of adult mice of different ages (range 100-900 days old) were used to investigate the effects of ageing and dihydrotestosterone (DHT) treatment on protein synthesis as well as the expression and function of two amino acid transporters; the sodium-coupled neutral amino acid transporter (SNAT) 2, and the sodium-independent L-type amino-acid transporter (LAT) 2. At all ages investigated, protein synthesis was always higher in the slow-twitch than in the fast-twitch muscle fibres and decreased with age in both fibre types. However, the decline was greater in the fast-twitch than in the slow-twitch fibres and was accompanied by a reduction in the expression of SNAT2 and LAT2 at the protein level. Again, the decrease in the expression of the amino acid transporters was greater in the fast-twitch than in the slow-twitch fibres. In contrast, ageing had no effect on SNAT2 and LAT2 expressions at the mRNA level. Treating the muscle fibre bundles with physiological concentrations (~2 nM) of DHT for 1 h completely reversed the effects of ageing on protein synthesis and the expression of SNAT2 and LAT2 protein in both fibre types. From the observations that ageing is accompanied by a reduction in protein synthesis and transporter expression and that these effects are reversed by DHT treatment, we conclude that sarcopenia arises from an age-dependent reduction in protein synthesis caused, in part, by the lack of or by the low bioavailability of the male sex steroid, DHT.

Light-regulation of enzyme activity in anacystis nidulans (Richt.).

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Duggan, J X; Anderson, L E

1975-01-01

The effect of light on the levels of activity of six enzymes which are light-modulated in higher plants was examined in the photosynthetic procaryot Anacystis nidulans. Ribulose-5-phosphate kinase (EC 2.7.1.19) was found to be light-activated in vivo and dithiothreitol-activated in vitro while glucose-6-phosphate dehydrogenase (EC 1.1.1.49) was light-inactivated and dithiothreitol-inactivated. The enzymes fructose-1,6-diphosphate phosphatase (EC 3.1.3.11), sedoheptulose-1,7-diphosphate phosphatase, NAD- and NADP-linked glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12; EC 1.2.1.13) were not affected by light treatment of the intact algae, but sedoheptulose-diphosphate phosphatase and the glyceraldehyde-3-phosphate dehydrogenases were dithiothreitol-activated in crude extracts. Light apparently controls the activity of the reductive and oxidative pentose phosphate pathway in this photosynthetic procaryot as in higher plants, through a process which probably involves reductive modulation of enzyme activity.

Sources and oxidative potential of water-soluble humic-like substances (HULISWS) in fine particulate matter (PM2.5) in Beijing

Science.gov (United States)

Ma, Yiqiu; Cheng, Yubo; Qiu, Xinghua; Cao, Gang; Fang, Yanhua; Wang, Junxia; Zhu, Tong; Yu, Jianzhen; Hu, Di

2018-04-01

Water-soluble humic-like substances (HULISWS) are a major redox-active component of ambient fine particulate matter (PM2.5); however, information on their sources and associated redox activity is limited. In this study, HULISWS mass concentration, various HULISWS species, and dithiothreitol (DTT) activity of HULISWS were quantified in PM2.5 samples collected during a 1-year period in Beijing. Strong correlation was observed between HULISWS and DTT activity; both exhibited higher levels during the heating season than during the nonheating season. Positive matrix factorization analysis of both HULISWS and DTT activity was performed. Four combustion-related sources, namely coal combustion, biomass burning, waste incineration, and vehicle exhausts, and one secondary factor were resolved. In particular, waste incineration was identified as a source of HULISWS for the first time. Biomass burning and secondary aerosol formation were the major contributors ( > 59 %) to both HULISWS and associated DTT activity throughout the year. During the nonheating season, secondary aerosol formation was the most important source, whereas during the heating season, the predominant contributor was biomass burning. The four combustion-related sources accounted for > 70 % of HULISWS and DTT activity, implying that future reduction in PM2.5 emissions from combustion activities can substantially reduce the HULISWS burden and their potential health impact in Beijing.

Decrease of Na, K-ATPase Electrogenic Contribution and Resting Membrane Potential of Rat Soleus after 3 Days of Hindlimb Unloading

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Krivoi, I. I.; Kravtsova, V. V.; Drabkina, T. M.; Prokofiev, A. V.; Nikolsky, E. E.; Shenkman, B. S.

2008-06-01

The Na,K-ATPase activity is critically important for excitability, electrogenesis and contractility of skeletal muscle expressing ? and ? isoforms of the enzyme [6, 9]. It is well known that disuse induced by hindlimb unloading (HU) leads to progressive atrophy of skeletal muscle; the muscle undergoes a number of dramatic remodeling events. In particular, changes in ion channel expression in response to muscle unweighting were observed [1, 8]. Decrease of resting membrane potential (RMP), electrogenic contribution of Na,K-ATPase and membrane resistance during 7-28 days of HU was shown [8, 10]. The intrinsic mechanisms involved in the process have not been revealed until present. At the same time, the understanding of these mechanisms could be crucial for the disclosing the mechanisms underlying the resting Ca2+ accumulation in the cytoplasm of the unloaded muscle [3, 7]. In the present study, the effect of early (3 days) HU-induced disuse of slow-twitch soleus muscle on membrane electrogenesis as well as on electrogenic contribution of Na,K-ATPase isoforms was investigated.

Phantom Limbs, Neuroprosthetics, and the Developmental Origins of Embodiment.

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Blumberg, Mark S; Dooley, James C

2017-10-01

Amputees who wish to rid themselves of a phantom limb must weaken the neural representation of the absent limb. Conversely, amputees who wish to replace a lost limb must assimilate a neuroprosthetic with the existing neural representation. Whether we wish to remove a phantom limb or assimilate a synthetic one, we will benefit from knowing more about the developmental process that enables embodiment. A potentially critical contributor to that process is the spontaneous activity - in the form of limb twitches - that occurs exclusively and abundantly during active (REM) sleep, a particularly prominent state in early development. The sensorimotor circuits activated by twitching limbs, and the developmental context in which activation occurs, could provide a roadmap for creating neuroprosthetics that feel as if they are part of the body. Copyright © 2017 Elsevier Ltd. All rights reserved.

Visualization of the diaphragm muscle with ultrasound improves diagnostic accuracy of phrenic nerve conduction studies.

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Johnson, Nicholas E; Utz, Michael; Patrick, Erica; Rheinwald, Nicole; Downs, Marlene; Dilek, Nuran; Dogra, Vikram; Logigian, Eric L

2014-05-01

Evaluation of phrenic neuropathy (PN) with phrenic nerve conduction studies (PNCS) is associated with false negatives. Visualization of diaphragmatic muscle twitch with diaphragm ultrasound (DUS) when performing PNCS may help to solve this problem. We performed bilateral, simultaneous DUS-PNCS in 10 healthy adults and 12 patients with PN. The amplitude of the diaphragm compound muscle action potential (CMAP) (on PNCS) and twitch (on DUS) was calculated. Control subjects had phrenic CMAP (on PCNS). In the 12 patients with PN, 12 phrenic neuropathies were detected. Three of these patients had either significant side-to-side asymmetry or absolute reduction in diaphragm movement that was not detected with PNCS. There were no cases in which the PNCS showed an abnormality but the DUS did not. The addition of DUS to PNCS enhances diagnostic accuracy in PN. Copyright © 2013 Wiley Periodicals, Inc.

Beyond dreams: do sleep-related movements contribute to brain development?

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Mark S Blumberg

2010-11-01

Full Text Available Conventional wisdom has long held that the twitches of sleeping infants and adults are by-products of a dreaming brain. With the discovery of active (or REM sleep in the 1950s and the recognition soon thereafter that active sleep is characterized by inhibition of motor outflow, researchers elaborated on conventional wisdom and concluded that sleep-related twitches are epiphenomena that result from incomplete blockade of dream-related cortical activity. This view persists despite the fact that twitching is unaffected in infant and adults when the cortex is disconnected from the brainstem. In 1966, Roffwarg and colleagues introduced the ontogenetic hypothesis, which addressed the preponderance of active sleep in early infancy. This hypothesis posited that the brainstem mechanisms that produce active sleep provide direct ascending stimulation to the forebrain and descending stimulation to the musculature, thereby promoting brain and neuromuscular development. However, this hypothesis and the subsequent work that tested it did not directly address the developmental significance of twitching or sensory feedback as a contributor to activity-dependent development. Here I review recent findings that have inspired an elaboration of the ontogenetic hypothesis. Specifically, in addition to direct brainstem activation of cortex during active sleep, sensory feedback arising from limb twitches produces discrete and substantial activation of somatosensory cortex and, beyond that, of hippocampus. Delineating how twitching during active sleep contributes to the establishment, refinement, and maintenance of neural circuits may aid our understanding of the early developmental events that make sensorimotor integration possible. In addition, twitches may prove to be sensitive and powerful tools for assessing somatosensory function in humans across the lifespan as well as functional recovery in individuals with injuries or conditions that affect sensorimotor function.

Effect of Age and Sex on Histomorphometrical Characteristics of Two Muscles of Laticauda Lambs

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Salvatore Velotto

2010-01-01

Full Text Available The aim of the present experiment was to determine the effect of sex and age on histochemical and morphometric characteristics of muscle fibres (myocytes in lambs born by single, twin, triplet and quadruplet birth. Thirty lambs were slaughtered at 60 days of age; thirty were weaned at 60 days and fed until 120 days with flakes (60% and food supplements, and then slaughtered. Muscle tissues were obtained from two muscles, namely m. semitendinosus and m. longissimus dorsi of all lambs. For each fibre type, area perimeter and diameter (maximum and minimum were measured and slow-twitch oxidative fibres, fast-twitch glycolytic fibres, fast-twitch oxidative-glycolytic fibres were histochemically differentiated. The muscles were stained for myosin ATPase, and succinic dehydrogenase. At 60 days, females had fibres larger than males, whereas the opposite was observed at 120 days. Besides, at 60 days, the lambs born by single birth had fibres larger than those born by multiple birth, whereas the opposite was observed at 120 days. Single lambs were heavier than twin lambs and multiple lambs. Fast-twitch glycolytic fibres had the largest size, followed by slow-twitch oxidative and fast-twitch oxidative glycolytic fibres. The dimensions of fibre types in m. longissimus dorsi were larger than in m. semitendinosus (P < 0.001.These muscle fibre characteristics are thought to be important factors influencing meat quality, which is often related to metabolic and contractile properties as determined by the muscle fibre type distribution.

Oxidative potential of particulate matter 2.5 as predictive indicator of cellular stress

International Nuclear Information System (INIS)

Crobeddu, Bélinda; Aragao-Santiago, Leticia; Bui, Linh-Chi; Boland, Sonja; Baeza Squiban, Armelle

2017-01-01

Particulate air pollution being recognized to be responsible for short and long term health effects, regulations for particulate matter with an aerodynamic diameter less than 2.5 (PM 2.5 ) are more and more restrictive. PM 2.5 regulation is based on mass without taking into account PM 2.5 composition that drives toxicity. Measurement of the oxidative potential (OP) of PM could be an additional PM indicator that would encompass the PM components involved in oxidative stress, the main mechanism of PM toxicity. We compared different methods to evaluate the intrinsic oxidative potential of PM 2.5 sampled in Paris and their ability to reflect the oxidative and inflammatory response in bronchial epithelial cells used as relevant target organ cells. The dithiothreitol depletion assay, the antioxidant (ascorbic acid and glutathione) depletion assay (OP AO ), the plasmid scission assay and the dichlorofluorescein (DCFH) oxidation assay used to characterize the OP of PM 2.5 (10–100 μg/mL) provided positive results of different magnitude with all the PM 2.5 samples used with significant correlation with different metals such as Cu and Zn as well as total polyaromatic hydrocarbons and the soluble organic fraction. The OP AO assay showed the best correlation with the production of intracellular reactive oxygen species by NCI-H292 cell line assessed by DCFH oxidation and with the expression of anti-oxidant genes (superoxide dismutase 2, heme-oxygenase-1) as well as the proinflammatory response (Interleukin 6) when exposed from 1 to 10 μg/cm 2 . The OP AO assay appears as the most prone to predict the biological effect driven by PM 2.5 and related to oxidative stress. - Highlights: • 5 Acellular assays were used to compare the intrinsic oxidative potential (OP) of PM. • The amount of ROS generation in bronchial cells is particle dependent. • Particles induce the expression of anti-oxidant and proinflammatory genes. • Biological effects correlates with OP assay

The effects of ramp stretches on active contractions in intact mammalian fast and slow muscle fibres.

Science.gov (United States)

Mutungi, G; Ranatunga, K W

2001-01-01

The effects of a ramp stretch (amplitude muscle fibre length (L0), speed twitch tension and twitch tension re-development were examined in intact mammalian (rat) fast and slow muscle fibre bundles. The experiments were done in vitro at 20 degrees C and at an initial sarcomere length of 2.68 microm. In both fibre types, a stretch applied during the rising phase of the twitch response (including the time of stimulation) increased the re-developed twitch tension (15-35%). A stretch applied before the stimulus had little or no effect on the twitch myogram in fast muscle fibres, but it increased the twitch tension (approximately 5%) in slow muscle fibres. A similar stretch had little or no effect on tetanic tension in either muscle fibre type. In general, the results indicate that the contractile-activation mechanism may be stretch sensitive and this is particularly pronounced in slow muscle fibres. Recorded at a high sampling rate and examined at an appropriate time scale, the transitory tension response to a stretch rose in at least two phases; an initial rapid tension rise to a break (break point tension, P1a) followed by a slower tension rise (apparent P2a) to a peak reached at the end of the stretch. Plotted against stretch velocity, P1a tension increased in direct proportion to stretch velocity (viscous-like) whereas, P2a tension (calculated as peak tension minus P1a tension) increased with stretch velocity to a plateau (visco-elastic). Examined at the peak of a twitch, P1a tension had a slope (viscosity coefficient) of 1.8 kN m(-2) per L0 s(-1) in fast fibres and 4.7 kN m(-2) per L0 s(-1) in slow muscle fibres. In the same preparations, P2a tension had a relaxation time of 8 ms in the fast muscle fibres and 25 ms in the slow muscle fibres. The amplitudes of both tension components scaled with the instantaneous twitch tension in qualitatively the same way as the instantaneous fibre stiffness. These fast/slow fibre type differences probably reflect differences in

Antagonism of botulinum toxin-induced muscle weakness by aminopyridines in rat phrenic nerve-hemidiaphragm preparations

Energy Technology Data Exchange (ETDEWEB)

Adler, M.; Scovill, J.; Deshpande, S.S.

1993-05-13

The effects of the potassium channel inhibitor and putative botulinum toxin antagonists 4-aminopyridine (4-AP) and 3,4-diaminopyridine (3,4-DAP) were investigated in vitro on the contractile and electrophysiological properties of rat diaphragm muscle. In the presence of 300 pM botulinum toxin A (BoTx A), twitches elicited by supramaximal nerve stimulation (0. 1 Hz) were reduced by over 80% in 3 hr. The time to block decreased with increases in temperature, toxin concentration and stimulation frequency. Addition of 4-AP or 3,4-DAP led to a prompt reversal of the BoTx A-induced depression of twitch tension. This reversal was concentration-dependent such that, in the presence of 1 mM 4-AP, reversal of the BoTx A-induced blockade was complete in 6.7 min. The beneficial effect of the APs were well maintained and persisted for up to 6 hr after addition. Application of 1 microns M neostigmine 1 hr after 3,4-DAP produced a further potentiation of twitch tensions, but this action lasted for < 5 min and led to the appearance of tetanic fade during repetitive stimulation. It is concluded that the APs are of benefit in antagonizing the muscle paralysis following exposure to botulinum toxin. Co-application of neostigmine, however, appears to confer no additional benefit.

Dihydrotestosterone activates the MAPK pathway and modulates maximum isometric force through the EGF receptor in isolated intact mouse skeletal muscle fibres.

Science.gov (United States)

Hamdi, M M; Mutungi, G

2010-02-01

It is generally believed that steroid hormones have both genomic and non-genomic (rapid) actions. Although the latter form an important component of the physiological response of these hormones, little is known about the cellular signalling pathway(s) mediating these effects and their physiological functions in adult mammalian skeletal muscle fibres. Therefore, the primary aim of this study was to investigate the non-genomic actions of dihydrotestosterone (DHT) and their physiological role in isolated intact mammalian skeletal muscle fibre bundles. Our results show that treating the fibre bundles with physiological concentrations of DHT increases both twitch and tetanic contractions in fast twitch fibres. However, it decreases them in slow twitch fibres. These changes in force are accompanied by an increase in the phosphorylation of MAPK/ERK1/2 in both fibre types and that of regulatory myosin light chains in fast twitch fibres. Both effects were insensitive to inhibitors of Src kinase, androgen receptor, insulin-like growth factor 1 receptor and platelet-derived growth factor receptor. However, they were abolished by the MAPK/ERK1/2 kinase inhibitor PD98059 and the epidermal growth factor (EGF) receptor inhibitor tyrphostin AG 1478. In contrast, testosterone had no effect on force and increased the phosphorylation of ERK1/2 in slow twitch fibres only. From these results we conclude that sex steroids have non-genomic actions in isolated intact mammalian skeletal muscle fibres. These are mediated through the EGF receptor and one of their main physiological functions is the enhancement of force production in fast twitch skeletal muscle fibres.

Mechanisms limiting glycogen storage in muscle during prolonged insulin stimulation

DEFF Research Database (Denmark)

Richter, Erik; Hansen, S A; Hansen, B F

1988-01-01

increased muscle glycogen concentrations to maximal values 2, 3, and 3.5 times above normal fed levels in fast-twitch white, slow-twitch red, and fast-twitch red fibers, respectively. Glucose uptake decreased (mean +/- SE) from 34.9 +/- 1.2 mumol.g-1.h-1 at 0 h to 7.5 +/- 0.7 after 7 h of perfusion. During...... compared with initial values. Total muscle water concentration decreased during glycogen loading of the muscles. Mechanisms limiting glycogen storage under maximal insulin stimulation include impaired insulin-stimulated membrane transport of glucose as well as impaired intracellular glucose disposal....

The augmenting action of banana tree juice on skeletal muscle contraction.

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Singh, Y N; Dryden, W F

1990-01-01

An extract obtained from juice expressed from the stem of the plantain banana tree (Musa sapientum L., var. paradisiaca) induces twitch augmentation in skeletal muscles. The mechanism of this action was investigated in the mouse hemi-diaphragm preparation. Directly evoked twitches and potassium induced (K+) contractures were both augmented by the extract. Twitch augmentation was partly dependent on extracellular Ca2+. The action on K(+)-contractures was unaffected by tetrodotoxin, but the rate of relaxation was enhanced in the absence of extracellular calcium (0[Ca2+]o). Muscle contracture induced by high concentrations of extract was also augmented in 0[Ca2+]o and in the presence of the Ca2(+)-channel blocking agent, nifedipine. The time course of the contracture was shortened in 0[Ca2+]o, but not by nifedipine. Nifedipine enhanced the augmenting effect of the extract on twitches but shortened the time-course of this action. In addition, a muscle contracture was superimposed on the twitching muscle at higher concentrations of nifedipine. Manganese, on the other hand, reduced or abolished the augmenting action of the extract. The results are consistent with an action of banana tree juice on the molecule responsible for excitation-contraction coupling in skeletal muscle, resulting in a labilization of intracellular Ca2+.

Single muscle fiber adaptations with marathon training.

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Trappe, Scott; Harber, Matthew; Creer, Andrew; Gallagher, Philip; Slivka, Dustin; Minchev, Kiril; Whitsett, David

2006-09-01

The purpose of this investigation was to characterize the effects of marathon training on single muscle fiber contractile function in a group of recreational runners. Muscle biopsies were obtained from the gastrocnemius muscle of seven individuals (22 +/- 1 yr, 177 +/- 3 cm, and 68 +/- 2 kg) before, after 13 wk of run training, and after 3 wk of taper. Slow-twitch myosin heavy chain [(MHC) I] and fast-twitch (MHC IIa) muscle fibers were analyzed for size, strength (P(o)), speed (V(o)), and power. The run training program led to the successful completion of a marathon (range 3 h 56 min to 5 h 35 min). Oxygen uptake during submaximal running and citrate synthase activity were improved (P training program. Muscle fiber size declined (P training. P(o) was maintained in both fiber types with training and increased (P 60% increase (P training and was unchanged in MHC IIa fibers. Peak power increased (P training with a further increase (P marathon training decreased slow-twitch and fast-twitch muscle fiber size but that it maintained or improved the functional profile of these fibers. A taper period before the marathon further improved the functional profile of the muscle, which was targeted to the fast-twitch muscle fibers.

Chemical characterization and oxidative potential of particles emitted from open burning of cereal straws and rice husk under flaming and smoldering conditions

Science.gov (United States)

Fushimi, Akihiro; Saitoh, Katsumi; Hayashi, Kentaro; Ono, Keisuke; Fujitani, Yuji; Villalobos, Ana M.; Shelton, Brandon R.; Takami, Akinori; Tanabe, Kiyoshi; Schauer, James J.

2017-08-01

Open burning of crop residue is a major source of atmospheric fine particle emissions. We burned crop residues (rice straws, barley straws, wheat straws, and rice husks produced in Japan) in an outdoor chamber and measured particle mass, composition (elemental carbon: EC, organic carbon: OC, ions, elements, and organic species), and oxidative potential in the exhausts. The fine particulate emission factors from the literature were within the range of our values for rice straws but were 1.4-1.9 and 0.34-0.44 times higher than our measured values for barley straw and wheat straw, respectively. For rice husks and wheat straws, which typically lead to combustion conditions that are relatively mild, the EC content of the particles was less than 5%. Levoglucosan seems more suitable as a biomass burning marker than K+, since levoglucosan/OC ratios were more stable than K+/particulate mass ratios among crop species. Stigmasterol and β-sitosterol could also be used as markers of biomass burning with levoglucosan or instead of levoglucosan. Correlation analysis between chemical composition and combustion condition suggests that hot or flaming combustions enhance EC, K+, Cl- and polycyclic aromatic hydrocarbons emissions, while low-temperature or smoldering combustions enhance levoglucosan and water-soluble organic carbon emissions. Oxidative potential, measured with macrophage-based reactive oxygen species (ROS) assay and dithiothreitol (DTT) assay, of open burning fine particles per particulate mass as well as fine particulate emission factors were the highest for wheat straws and second highest for rice husks and rice straws. Oxidative potential per particulate mass was in the lower range of vehicle exhaust and atmosphere. These results suggest that the contribution of open burning is relatively small to the oxidative potential of atmospheric particles. In addition, oxidative potential (both ROS and DTT activities) correlated well with water-insoluble organic species

Optimization of methodology to analyze ascorbic and dehydroascorbic acid in vegetables

Directory of Open Access Journals (Sweden)

Flávia Milagres Campos

2009-01-01

Full Text Available In this study, different solutions to extract vitamin C were tested. High-performance liquid chromatography was chosen and the conditions were based on isocratic elution in reverse phase column. Dehydroascorbic acid was determined indirectly after its reduction using dithiothreitol. The use of metaphosphoric acid to stabilize the vitamin C was shown to be required and it was necessary to neutralize the pH of the extract to apply dithiothreitol. The average recovery was 90% in collard and tomato samples. The presence of oil did not interfere in extraction and the methodology can be used to analyze stir fried vegetables.

CONTRACTION CHARACTERISTICS AND MYOSIN HEAVY-CHAIN COMPOSITION OF RABBIT MASSETER MOTOR UNITS

NARCIS (Netherlands)

KWA, SHS; WEIJS, WA; JUCH, PJW

1. We studied isometric twitch peak force (TPF) and twitch contraction time (TCT) of 249 motor units of the masseter muscle in 41 rabbits after extracellular electrical stimulation of single trigeminal motoneurons in the brain stem. In 41 of these units we determined the amount of tension decrease

Sources and oxidative potential of water-soluble humic-like substances (HULISWS in fine particulate matter (PM2.5 in Beijing

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Y. Ma

2018-04-01

Full Text Available Water-soluble humic-like substances (HULISWS are a major redox-active component of ambient fine particulate matter (PM2.5; however, information on their sources and associated redox activity is limited. In this study, HULISWS mass concentration, various HULISWS species, and dithiothreitol (DTT activity of HULISWS were quantified in PM2.5 samples collected during a 1-year period in Beijing. Strong correlation was observed between HULISWS and DTT activity; both exhibited higher levels during the heating season than during the nonheating season. Positive matrix factorization analysis of both HULISWS and DTT activity was performed. Four combustion-related sources, namely coal combustion, biomass burning, waste incineration, and vehicle exhausts, and one secondary factor were resolved. In particular, waste incineration was identified as a source of HULISWS for the first time. Biomass burning and secondary aerosol formation were the major contributors ( >  59 % to both HULISWS and associated DTT activity throughout the year. During the nonheating season, secondary aerosol formation was the most important source, whereas during the heating season, the predominant contributor was biomass burning. The four combustion-related sources accounted for  >  70 % of HULISWS and DTT activity, implying that future reduction in PM2.5 emissions from combustion activities can substantially reduce the HULISWS burden and their potential health impact in Beijing.

Skeletal Myocyte Types and Vascularity in the Black Sicilian Pig

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S. Velotto

2007-01-01

Full Text Available The objective of this study was to verify the presence of giant fibres in the Black Sicilian pig skeletal muscle and to evaluate the effect of sex on histochemical and morphometric characteristics of the myocytes (myofibres as well as vascularity of the muscle. Twenty Black Sicilian pigs (10 males, 10 females from a farm in Sicily (Italy were slaughtered at two years of age. Muscle tissues were obtained from three muscles: psoas major, longissimus dorsi, and trapezius. Myofibres were stained for myosin ATPase, succinic dehydrogenase, and α-amylase-PAS. For all fibre types, area and perimeter were measured. Slow-twitch oxidative fibres, fast-twitch glycolytic fibres and fast-twitch oxidative-glycolytic fibres were histochemically differentiated; an image-analyzing system was used. The results showed no differences between males and females in percentage of the fibre types, but there were significant differences between sexes in size of all the three fibre types. Psoas major muscle had a high percentage of slow-twitch oxidative fibres and contained more capillaries per fibre and per mm2 than trapezius and longissimus dorsi, in which fast-twitch glycolytic fibres dominated. The cross-sectional area of all fibres types was larger in longissimus dorsi than in trapezius and psoas major muscles; the giant fibres were absent in all the muscles studied. Fibre type composition may contribute to the variation of meat quality.

Effects of capsaicin in the motor nerve.

Science.gov (United States)

Pettorossi, V E; Bortolami, R; Della Torre, G; Brunetti, O

1994-08-01

The injection of capsaicin into the lateral gastrocnemius (LG) muscle of the rat induced an immediate and sustained reduction in the A delta and C components of the compound action potential (CAP) of the LG motor nerve. Conversely, the drug did not immediately affect the CAP wave belonging to fast-conducting fibers or the motor responses to LG nerve stimulation. It seems that capsaicin only affects the group III and IV afferents of LG nerve. However, a week after the injection the capsaicin also altered the motor responses, as shown by the threshold enhancement and amplitude reduction of the muscle twitch and by the decrease of the A alpha-beta CAP components. This late motor impairment was attributed to a central depression following a reduction of capsaicin-sensitive neuron input into the CNS. However, this motor effect was transient since the LG nerve regained the preinjection excitability level in a week and the muscle twitch amplitude reached the control value in a month.

Relationship between chemical composition and oxidative potential of secondary organic aerosol from polycyclic aromatic hydrocarbons

Science.gov (United States)

Wang, Shunyao; Ye, Jianhuai; Soong, Ronald; Wu, Bing; Yu, Legeng; Simpson, André J.; Chan, Arthur W. H.

2018-03-01

Owing to the complex nature and dynamic behaviors of secondary organic aerosol (SOA), its ability to cause oxidative stress (known as oxidative potential, or OP) and adverse health outcomes remains poorly understood. In this work, we probed the linkages between the chemical composition of SOA and its OP, and investigated impacts from various SOA evolution pathways, including atmospheric oligomerization, heterogeneous oxidation, and mixing with metal. SOA formed from photooxidation of the two most common polycyclic aromatic hydrocarbons (naphthalene and phenanthrene) were studied as model systems. OP was evaluated using the dithiothreitol (DTT) assay. The oligomer-rich fraction separated by liquid chromatography dominates DTT activity in both SOA systems (52 ± 10 % for naphthalene SOA (NSOA), and 56 ± 5 % for phenanthrene SOA (PSOA)). Heterogeneous ozonolysis of NSOA was found to enhance its OP, which is consistent with the trend observed in selected individual oxidation products. DTT activities from redox-active organic compounds and metals were found to be not additive. When mixing with highly redox-active metal (Cu), OP of the mixture decreased significantly for 1,2-naphthoquinone (42 ± 7 %), 2,3-dihydroxynaphthalene (35 ± 1 %), NSOA (50 ± 6 %), and PSOA (43 ± 4 %). Evidence from proton nuclear magnetic resonance (1H NMR) spectroscopy illustrates that such OP reduction upon mixing can be ascribed to metal-organic binding interactions. Our results highlight the role of aerosol chemical composition under atmospheric aging processes in determining the OP of SOA, which is needed for more accurate and explicit prediction of the toxicological impacts from particulate matter.

Antidepressant-like effects of a water-soluble extract from the culture medium of Ganoderma lucidum mycelia in rats.

Science.gov (United States)

Matsuzaki, Hirokazu; Shimizu, Yuta; Iwata, Naohiro; Kamiuchi, Shinya; Suzuki, Fumiko; Iizuka, Hiroshi; Hibino, Yasuhide; Okazaki, Mari

2013-12-26

Ganoderma lucidum is a popular medicinal mushroom used for promoting health and longevity in Asian countries. Previously, we reported that a water-soluble extract from a culture medium of Ganoderma lucidum mycelia (MAK) exerts antioxidative and cerebroprotective effects against ischemia-reperfusion injury in vivo. Here, we evaluated the antidepressant and anxiolytic activities of MAK in rats. MAK (0.3 or 1 g/kg, p.o.) was administered in the experimental animals 60 min before the forced swimming, open-field, elevated plus-maze, contextual fear-conditioning, and head twitch tests. Additionally, the mechanisms involved in the antidepressant-like action of MAK were investigated by the serotonin precursor 5-hydroxy-L-tryptophan (5-HTP)- or 5-HT2A agonist (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI)-induced head twitch responses. Treatment with MAK (1 g/kg) exhibited antidepressant-like effects in the forced swimming test, attenuated freezing behavior in the contextual fear-conditioning test, and decreased the number of head twitches induced by DOI, but not with 5-HTP. No significant response was observed in locomotion or anxiety-like behavior, when the animals were evaluated in the open-field or elevated plus-maze test, respectively. These data suggest that MAK has antidepressant-like potential, which is most likely due to the antagonism of 5-HT2A receptors, and possesses anxiolytic-like effects toward memory-dependent and/or stress-induced anxiety in rats.

Left Phrenic Nerve Stimulation Due to Breakage of the Endocardial Right Ventricular Lead at the Costoclavicular Ligament

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Mariko Fujimori, MD

2007-01-01

Full Text Available A 78-year-old man with a permanent pacemaker (PM implanted in his left prepectoral area reported twitches in his left lateral abdominal region. Chest X-rays revealed a broken right atrial (RA lead and a fracture of the right ventricular (RV lead at the left costoclavicular ligament. The electrocardiogram (ECG and the Holler ECG revealed atrial fibrillation (AF and an improperly functioning PM. We observed that the twitching seemed to correspond with each pacing beat and that it did not appear with his own beat. We suspected that the twitching was due to electric current leakage from the broken RV lead. We performed a PM re-implantation with a screw-in RV lead using the extrathoracic approach. After re-implantation the twitching disappeared. Costoclavicular ligament related electrode lead fractures are not uncommon and electric current leaks can be a source of problems in cardiac pacing. In this case, the electric current leak from the broken RV lead at the costoclavicular ligament stimulated the left phrenic nerve.

Catalase-positive microperoxisomes in rat soleus and extensor digitorum longus muscle fiber types

Science.gov (United States)

Riley, Danny A.; Bain, James L. W.; Ellis, Stanley

1988-01-01

The size, distribution, and content of catalase-reactive microperoxisomes were investigated cytochemically in three types of muscle fibers from the soleus and the extensor digitorum longus (EDL) of male rats. Muscle fibers were classified on the basis of the mitochondrial content and distribution, the Z-band widths, and the size and shape of myofibrils as the slow-twitch oxidative (SO), the fast-twitch oxidative glycolytic (FOG), and the fast-twitch glycolytic (FG) fibers. It was found that both the EDL and soleus SO fibers possessed the largest microperoxisomes. A comparison of microperoxisome number per muscle fiber area or the microperoxisome area per fiber area revealed following ranking, starting from the largest number and the area-ratio values: soleus SO, EDL SO, EDL FOG, and EDL FG.

Electrophoretic and chemical studies on the X-ray damage of malate synthase

International Nuclear Information System (INIS)

Durchschlag, H.; Zipper, P.

1981-01-01

1) Both X-irradiation and treatment with H 2 O 2 caused a decrease of total and an increase of available sulfhydryl groups of the enzyme and led to a loss of enzymic activity. The presence of dithiothreitol turned out to be able to protect the enzyme against X-ray or H 2 O 2 induced inactivation. Moreover, addition of dithiothreitol after X-irradiation or H 2 O 2 treatment allowed a considerable repair of enzymic activity. 2) Polyacrylamide gel disc electrophoreses of X-irradiated enzyme solutions, performed in the presence of sodium dodecyl sulfate, showed the occurrence of covalently cross-linked subunits (preferably dimers and trimers) and of various definite fragments. Electrophoreses in the absence of the denaturant indicated the occurrence of enzyme aggregation. The effects were more pronounced with increasing X-ray doses. The electrophoreses also clearly reflected a radioprotection by dithiothreitol against cross-linking, but not against fragmentation. Addition of excess of 2-mercaptoethanol or of dithiothreitol to the X-irradiated enzyme clearly demonstrated that part of the covalent cross-links were disulfide bridges; the aggregates themselves, however, were held together primarily by non-covalent bonds. Blocking of exposed enzyme sulfhydryls by means of Ellman's reagent prevented both covalent cross-linking and enzyme aggregation. 3) Similar electrophoretic patterns as found for the X-irradiated enzyme were obtained for the unirradiated enzyme after treatment with H 2 O 2 . The similarity of the electropherograms, as well as the reversible diminution of enzymic activity and the loss of sulfhydryls in the presence of H 2 O 2 , suggest an involvement of H 2 O 2 in the radiation damage of the enzyme. It seems plausible that oxidation reactions are responsible for the effects caused by X-irradiation or H 2 O 2 treatment. (orig./AJ)

Changes in the cholinergic system of rat sciatic nerve and skeletal muscle following suspension induced disuse

Science.gov (United States)

Gupta, R. C.; Misulis, K. E.; Dettbarn, W. D.

1984-01-01

Muscle disused induced changes in the cholinergic system of sciatic nerve, slow twitch soleus (SOL) and fast twitch extensor digitorum longus (EDL) muscle were studied in rats. Rats with hindlimbs suspended for 2 to 3 weeks showed marked elevation in the activity of choline acetyltransferase (ChAT) in sciatic nerve (38%), in SOL (108%) and in EDL (67%). Acetylcholinesterase (AChE) activity in SOL increased by 163% without changing the molecular forms pattern of 4S, 10S, 12S, and 16S. No significant changes in activity and molecular forms pattern of AChE were seen in EDL or in AChE activity of sciatic nerve. Nicotinic receptor binding of 3H-acetylcholine was increased in both muscles. When measured after 3 weeks of hindlimb suspension the normal distribution of type 1 fibers in SOL was reduced and a corresponding increase in type IIa and IIb fibers is seen. In EDL no significant change in fiber proportion is observed. Muscle activity, such as loadbearing, appears to have a greater controlling influence on the characteristics of the slow twitch SOL muscle than upon the fast twitch EDL muscle.

S-glutathionylation of troponin I (fast) increases contractile apparatus Ca2+ sensitivity in fast-twitch muscle fibres of rats and humans.

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Mollica, J P; Dutka, T L; Merry, T L; Lamboley, C R; McConell, G K; McKenna, M J; Murphy, R M; Lamb, G D

2012-03-15

Oxidation can decrease or increase the Ca2+ sensitivity of the contractile apparatus in rodent fast-twitch (type II) skeletal muscle fibres, but the reactions and molecular targets involved are unknown. This study examined whether increased Ca2+ sensitivity is due to S-glutathionylation of particular cysteine residues. Skinned muscle fibres were directly activated in heavily buffered Ca2+ solutions to assess contractile apparatus Ca2+ sensitivity. Rat type II fibres were subjected to S-glutathionylation by successive treatments with 2,2′-dithiodipyridine (DTDP) and glutathione (GSH), and displayed a maximal increase in pCa50 (−log10 [Ca2+] at half-maximal force) of ∼0.24 pCa units, with little or no effect on maximum force or Hill coefficient. Partial similar effect was produced by exposure to oxidized gluthathione (GSSG, 10 mM) for 10 min at pH 7.1, and near-maximal effect by GSSG treatment at pH 8.5. None of these treatments significantly altered Ca2+ sensitivity in rat type I fibres. Western blotting showed that both the DTDP–GSH and GSSG–pH 8.5 treatments caused marked S-glutathionylation of the fast troponin I isoform (TnI(f)) present in type II fibres, but not of troponin C (TnC) or myosin light chain 2. Both the increased Ca2+ sensitivity and glutathionylation of TnI(f) were blocked by N-ethylmaleimide (NEM). S-nitrosoglutathione (GSNO) also increased Ca2+ sensitivity, but only in conditions where it caused S-glutathionylation of TnI(f). In human type II fibres from vastus lateralis muscle, DTDP–GSH treatment also caused similar increased Ca2+ sensitivity and S-glutathionylation of TnI(f). When the slow isoform of TnI in type I fibres of rat was partially substituted (∼30%) with TnI(f), DTDP–GSH treatment caused a significant increase in Ca2+ sensitivity (∼0.08 pCa units). TnIf in type II fibres from toad and chicken muscle lack Cys133 present in mammalian TnIf, and such fibres showed no change in Ca2+ sensitivity with DTDP–GSH nor any S

New Insights into Muscle Fibre Types in Casertana Pig

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Salvatore Velotto

2010-01-01

Full Text Available Little is known about the Casertana pig. The aim of this study was to evaluate the effect of sex on histochemical and morphometrical characteristics of muscle fibres (myocytes in this pure breed and to verify the presence of giant fibres as well as vascularity of the muscle. Finally, maximum shortening velocity and isometric tension were measured in single muscle fibres. Sixteen Casertana pigs (8 males, 8 females from a farm in Campania (Italy were slaughtered at one year of age. Muscle tissues were obtained from psoas minor, rhomboideus and longissimus dorsi. Myofibres were stained for myosin adenosine triphosphatase, succinic dehydrogenase, and α-amylase-periodic acid schiff. For all fibre types, the area and perimeter were measured. Slowtwitch oxidative fibres, fast-twitch glycolytic fibres and fast-twitch oxidative-glycolytic fibres were histochemically differentiated; an image-analyzing system was used. The results showed significant differences between the sexes in the size of all three fibre types. The psoas minor muscle had a high percentage of slow-twitch oxidative fibres and contained more capillaries per fibre and per mm2 than rhomboideus and longissimus dorsi, in which fast-twitch glycolytic fibres dominated. The cross-sectional area of all fibre types was larger in longissimus dorsi than in rhomboideus and psoas minor muscles; the giant fibres were present in the longissimus dorsi muscle only. Besides, isometric tension values were higher in fast-twitch glycolytic fibres than in the other ones. Variations in fibre type composition may contribute to meat quality.

Cardiac troponin T and fast skeletal muscle denervation in ageing.

Science.gov (United States)

Xu, Zherong; Feng, Xin; Dong, Juan; Wang, Zhong-Min; Lee, Jingyun; Furdui, Cristina; Files, Daniel Clark; Beavers, Kristen M; Kritchevsky, Stephen; Milligan, Carolanne; Jin, Jian-Ping; Delbono, Osvaldo; Zhang, Tan

2017-10-01

Ageing skeletal muscle undergoes chronic denervation, and the neuromuscular junction (NMJ), the key structure that connects motor neuron nerves with muscle cells, shows increased defects with ageing. Previous studies in various species have shown that with ageing, type II fast-twitch skeletal muscle fibres show more atrophy and NMJ deterioration than type I slow-twitch fibres. However, how this process is regulated is largely unknown. A better understanding of the mechanisms regulating skeletal muscle fibre-type specific denervation at the NMJ could be critical to identifying novel treatments for sarcopenia. Cardiac troponin T (cTnT), the heart muscle-specific isoform of TnT, is a key component of the mechanisms of muscle contraction. It is expressed in skeletal muscle during early development, after acute sciatic nerve denervation, in various neuromuscular diseases and possibly in ageing muscle. Yet the subcellular localization and function of cTnT in skeletal muscle is largely unknown. Studies were carried out on isolated skeletal muscles from mice, vervet monkeys, and humans. Immunoblotting, immunoprecipitation, and mass spectrometry were used to analyse protein expression, real-time reverse transcription polymerase chain reaction was used to measure gene expression, immunofluorescence staining was performed for subcellular distribution assay of proteins, and electromyographic recording was used to analyse neurotransmission at the NMJ. Levels of cTnT expression in skeletal muscle increased with ageing in mice. In addition, cTnT was highly enriched at the NMJ region-but mainly in the fast-twitch, not the slow-twitch, muscle of old mice. We further found that the protein kinase A (PKA) RIα subunit was largely removed from, while PKA RIIα and RIIβ are enriched at, the NMJ-again, preferentially in fast-twitch but not slow-twitch muscle in old mice. Knocking down cTnT in fast skeletal muscle of old mice: (i) increased PKA RIα and reduced PKA RIIα at the NMJ; (ii

GLUT11, but not GLUT8 or GLUT12, is expressed in human skeletal muscle in a fibre type-specific pattern

DEFF Research Database (Denmark)

Gaster, M; Handberg, A; Schürmann, A

2004-01-01

or amyotrophic lateral sclerosis (ALS) were studied. GLUT8 and 12 immunoreactivity was below detection level in both developing and adult muscle fibres. GLUT11 immunoreactivity, however, was present in slow-twitch muscle fibres, but not in fast twitch fibres. Since, in contrast, GLUT4 was expressed in all...... exclusively in slow-twitch muscle fibres and is unaffected by physiological and pathophysiological conditions except in primary myopathy. GLUT8 and GLUT12 do not appear to be of importance in human muscle under physiological and pathophysiological conditions....... to induce GLUT8 or -12 expression. Likewise, the fibre type-dependent pattern of GLUT11 immunoreactivity was unaltered. However, some slow muscle fibres lose their GLUT11 immunoreactivity under regeneration. Our results indicate that GLUT11 immunoreactivity, in contrast to that of GLUT4, is expressed...

Mitochondrial oxidative enzyme activity in individual fibre types in hypo- and hyperthyroid rat skeletal muscles.

Science.gov (United States)

Johnson, M A; Turnbull, D M

1984-04-01

Quantitative cytochemical and biochemical techniques have been used in combination to study the response of mitochondrial oxidative enzymes in individual muscle fibre types to hypo- and hyperthyroidism. Hypothyroidism resulted in decreased activity of succinate dehydrogenase (SDH), L-glycerol-3-phosphate dehydrogenase (L-GPDH), and D-3-hydroxybutyrate dehydrogenase (D-HBDH) in all fibre types of both slow-twitch soleus and fast-twitch extensor digitorum longus (e.d.l.) muscles. In hyperthyroidism, only L-GPDH activity increased in e.d.l. but more marked increases were seen in soleus muscles, which also showed increased SDH activity. In addition to these alterations in the enzyme activity in individual fibre types the metabolic profile of the muscle is further modified by the hormone-induced interconversion of slow- to fast-twitch fibres and vice versa.

Neuromuscular activity of Bothrops neuwiedi pauloensis snake venom in mouse nerve-muscle preparations

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A. M. Durigon

2005-03-01

Full Text Available The pharmacological effects of Bothrops neuwiedi pauloensis venom on mouse phrenic nerve-diaphragm (PND preparations were studied. Venom (20 mug/ml irreversibly inhibited indirectly evoked twitches in PND preparations (60 ± 10% inhibition, mean ± SEM; p<0.05; n=6. At 50 mug/ml, the venom blocked indirectly and directly (curarized preparations evoked twitches in mouse hemidiaphragms. In the absence of Ca2+, venom (50 mug/ml, produced partial blockade only after an 80 min incubation, which reached 40.3 ± 7.8% (p<0.05; n=3 after 120 min. Venom (20 mug/ml increased (25 ± 2%, p< 0.05 the frequency of giant miniature end-plate potentials in 9 of 10 end-plates after 30 min and the number of miniature end-plate potentials which was maximum (562 ± 3%, p<0.05 after 120 min. During the same period, the resting membrane potential decreased from - 81 ± 1.4 mV to - 41.3 ± 3.6 mV 24 fibers; p<0.01; n=4 in the end-plate region and from - 77.4 ± 1.4 to -44.6 ± 3.9 mV (24 fibers; p<0.01; n=4 in regions distant from the end-plate. These results indicate that B. n. pauloensis venom acts primarily at presynaptic sites. They also suggest that enzymatic activity may be involved in this pharmacological action.

In vitro, in vivo, and clinical studies of tedizolid to assess the potential for peripheral or central monoamine oxidase interactions.

Science.gov (United States)

Flanagan, S; Bartizal, K; Minassian, S L; Fang, E; Prokocimer, P

2013-07-01

Tedizolid phosphate is a novel oxazolidinone prodrug whose active moiety, tedizolid, has improved potency against Gram-positive pathogens and pharmacokinetics, allowing once-daily administration. Given linezolid warnings for drug-drug and drug-food interactions mediated by monoamine oxidase (MAO) inhibition, including sporadic serotonergic toxicity, these studies evaluated tedizolid for potential MAO interactions. In vitro, tedizolid and linezolid were reversible inhibitors of human MAO-A and MAO-B; the 50% inhibitory concentration (IC50) for tedizolid was 8.7 μM for MAO-A and 5.7 μM for MAO-B and 46.0 and 2.1 μM, respectively, with linezolid. Tedizolid phosphate was negative in the mouse head twitch model of serotonergic activity. Two randomized placebo-controlled crossover clinical studies assessed the potential of 200 mg/day tedizolid phosphate (at steady state) to enhance pressor responses to coadministered oral tyramine or pseudoephedrine. Sensitivity to tyramine was determined by comparing the concentration of tyramine required to elicit a ≥ 30-mmHg increase in systolic blood pressure (TYR30) when administered with placebo versus tedizolid phosphate. The geometric mean tyramine sensitivity ratio (placebo TYR30/tedizolid phosphate TYR30) was 1.33; a ratio of ≥ 2 is considered clinically relevant. In the pseudoephedrine study, mean maximum systolic blood pressure was not significantly different when pseudoephedrine was coadministered with tedizolid phosphate versus placebo. In summary, tedizolid is a weak, reversible inhibitor of MAO-A and MAO-B in vitro. Provocative testing in humans and animal models failed to uncover significant signals that would suggest potential for hypertensive or serotonergic adverse consequences at the therapeutic dose of tedizolid phosphate. Clinical studies are registered at www.clinicaltrials.gov as NCT01539473 (tyramine interaction study conducted at Covance Clinical Research Center, Evansville, IN) and NCT01577459

Transcranial magnetic stimulation and the human brain

Science.gov (United States)

Hallett, Mark

2000-07-01

Transcranial magnetic stimulation (TMS) is rapidly developing as a powerful, non-invasive tool for studying the human brain. A pulsed magnetic field creates current flow in the brain and can temporarily excite or inhibit specific areas. TMS of motor cortex can produce a muscle twitch or block movement; TMS of occipital cortex can produce visual phosphenes or scotomas. TMS can also alter the functioning of the brain beyond the time of stimulation, offering potential for therapy.

[Case of anti VGKC-complex antibody associated disorder presenting with severe pain and fasciculations predominant in unilateral upper extremity].

Science.gov (United States)

Hara, Kenju; Watanabe, Osamu; Shibano, Ken; Ishiguro, Hideaki

2012-01-01

A 21-year-old man complained of severe pain and muscle twitching localized in his right arm. Neurological examination showed muscle fasciculations in his right forearm but no myokymia or myotonia. Needle electromyography revealed fibrillation potentials in his biceps brachii muscle and extensor carpi radialis muscle at rest but no myokymic discharges. His serum anti-voltage-gated potassium channel (VGKC)-complex antibody level was significantly high (194.2pM; controls VGKC-complex antibody associated disorder.

A study of fatigue in rabbit skeletal muscle by in vivo 31P MRS

International Nuclear Information System (INIS)

Koga, Keiko; Miura, Iwao

1989-01-01

Energy metabolism during exercise and recovery process of rabbit skeletal muscle was obserbed by in vivo 31 P MRS. The small value of the ratio of the intensities between inorganic phosphate and phosphocreatine at rest indicated that the observed moiety of muscle had high fast-twitch fiber content. More than half of ATP and almost all of phosphocreatine were depleted by electric stimulation at 4Hz. The extreme intracellular pH was 5.9. The recovery from this metabolic state was very slow, and only a small amount of ATP was resynthesized after 40 minutes of recovery. These phenomena show the characteristic features of the energy metabolism in the fatigue of fast-twitch muscle. The metabolic state as indicated by the intensity of phosphocreatine and intracellular pH during exercise was not always parallel to contraction power measured by straingauge. Two inorganic phosphate peaks were observed, which are regarded as the signals from fast-twitch fiber and slow-twitch fiber from their pH values. The ratios of these two peaks were different between 1Hz, 2Hz, and 4Hz electric stimulation. We conclude that we are observing the different recruitment of fiber types at different exercise level in vivo. (author)

Department of Clinical Investigation Annual Research Progress Report, Fiscal Year 1993. Volume 1

Science.gov (United States)

1993-10-01

Compare the Therapeutic Imlant 5FU -e TI (5003) to its Placebo Vehicle when Administered xviii Project Page Number to Patients with External Condylomata...results: Objective(s): 1) To determine the ED" of vecuronium bromide for train-of-four twitch height depression in the nonthermally injured patient. 2) To...vocal cord position and patient response when monitoring for 95% T1 twitch height depression at the adductor pollicis versus the orbicularis oculi

Factors in Maximal Power Production and in Exercise Endurance Relative to Maximal Power

Science.gov (United States)

1988-10-13

Mechanical efficiency of fast -and slow - twitch muscle fibers in mnan during cycling. J. ADLi Physiol.:Reespirat. Environ. Exercise Physiol. 47: 263- 267...R.S. Hikida, and F.C. Hagerman. Myofibrillar ATPase activity in hu-man muscle fast - twitch subtypes. Histochem. 78: 405-408, 1983. 31. Suzuki, Y...capacity and muscle fibre composition in mnan. J. Physiol (London) 354: 73P, 1984. 21. Margaria, R., P. Aghemo, and E. Rovelli. Measurement of muscular

Coordination Mechanism in Fast Human Movements - Experimental and Modelling Studies. Volume 2.

Science.gov (United States)

1982-02-01

the involved muscle . Knapik and Ramos (33) have proposed that fast twitch muscle fiber may be responsible for a maximun speed move- ment, while slow ...repetition with submaximun weight (low intensity), Wolcott (56) attempted to selectively fatigue the fast and slow twitch muscle fiber. Both intensities of...B. "Enzyme Activity and Fibre Composition in Skeletal Muscle of Trained and Untrained Men." Journal of Applied Physiology 33: 312-319, 1972. 40

Fnip1 regulates skeletal muscle fiber type specification, fatigue resistance, and susceptibility to muscular dystrophy

Science.gov (United States)

Reyes, Nicholas L.; Banks, Glen B.; Tsang, Mark; Margineantu, Daciana; Gu, Haiwei; Djukovic, Danijel; Chan, Jacky; Torres, Michelle; Liggitt, H. Denny; Hirenallur-S, Dinesh K.; Hockenbery, David M.; Raftery, Daniel; Iritani, Brian M.

2015-01-01

Mammalian skeletal muscle is broadly characterized by the presence of two distinct categories of muscle fibers called type I “red” slow twitch and type II “white” fast twitch, which display marked differences in contraction strength, metabolic strategies, and susceptibility to fatigue. The relative representation of each fiber type can have major influences on susceptibility to obesity, diabetes, and muscular dystrophies. However, the molecular factors controlling fiber type specification remain incompletely defined. In this study, we describe the control of fiber type specification and susceptibility to metabolic disease by folliculin interacting protein-1 (Fnip1). Using Fnip1 null mice, we found that loss of Fnip1 increased the representation of type I fibers characterized by increased myoglobin, slow twitch markers [myosin heavy chain 7 (MyH7), succinate dehydrogenase, troponin I 1, troponin C1, troponin T1], capillary density, and mitochondria number. Cultured Fnip1-null muscle fibers had higher oxidative capacity, and isolated Fnip1-null skeletal muscles were more resistant to postcontraction fatigue relative to WT skeletal muscles. Biochemical analyses revealed increased activation of the metabolic sensor AMP kinase (AMPK), and increased expression of the AMPK-target and transcriptional coactivator PGC1α in Fnip1 null skeletal muscle. Genetic disruption of PGC1α rescued normal levels of type I fiber markers MyH7 and myoglobin in Fnip1-null mice. Remarkably, loss of Fnip1 profoundly mitigated muscle damage in a murine model of Duchenne muscular dystrophy. These results indicate that Fnip1 controls skeletal muscle fiber type specification and warrant further study to determine whether inhibition of Fnip1 has therapeutic potential in muscular dystrophy diseases. PMID:25548157

Synthesis, Characterization and DNA Cleavage of Copper(II ...

African Journals Online (AJOL)

Keywords: DNA shearing, Copper(II) complex, Dithiothreitol, Attenuated total reflectance-Fourier transform .... confirm the fragmentation of DNA by the newly .... sperm. Biochem Biophys Acta 1986; 884: 124-134. 7. Cornell NW, Crivaro KE.

Slow and fast fatigable frog muscle fibres: electrophysiological and histochemical characteristics.

Science.gov (United States)

Vydevska-Chichova, M; Mileva, K; Todorova, R; Dimitrova, M; Radicheva, N

2005-12-01

Continuous activity of isolated frog gastrocnemius muscle fibres provoked by repetitive stimulation of 5 Hz was used as an experimental model for fatigue development in different fibre types. Parameter changes of the elicited intracellular action potentials and mechanical twitches during the period of uninterrupted activity were used as criteria for fatigue evaluation. Slow fatigable muscle fibre (SMF) and fast fatigable muscle fibre (FMF) types were distinguished depending on the duration of their uninterrupted activity, which was significantly longer in SMFs than in FMFs. The normalized changes of action potential amplitude and duration were significantly smaller in FMFs than in SMFs. The average twitch force and velocity of contraction and relaxation were significantly higher in FMFs than in SMFs. Myosin ATPase (mATPase) and succinate dehydrogenase activity were studied by histochemical assessment in order to validate the fibre type classification based on their electrophysiological characteristics. Based on the relative mATPase reactivity, the fibres of the studied muscle were classified as one of five different types (1-2, 2, 2-3, 3 and tonic). Smaller sized fibres (tonic and type 3) expressed higher succinate dehydrogenase activity than larger sized fibres (type 1-2, 2), which is related to the fatigue resistance. The differences between fatigue development in SMFs and FMFs during continuous activity were associated with fibre-type specific mATPase and succinate dehydrogenase activity.

Muscle sarcomere lesions and thrombosis after spaceflight and suspension unloading

Energy Technology Data Exchange (ETDEWEB)

Riley, D.A.; Ellis, S.; Giometti, C.S.; Hoh, J.F.Y.; Ilyina-Kakueva, E.I.; Oganov, V.S.; Slocum, G.R.; Bain, J.L.W.; Sedlak, F.R. (Argonne National Lab., IL (United States))

1992-08-01

Extended exposure of humans to spaceflight produces a progressive loss of skeletal muscle strength. This process must be understood to design effective countermeasures. The present investigation examined hindlimb muscles from flight rats killed as close to landing as possible. Spaceflight and tail suspension-hindlimb unloading (unloaded) produced significant decreases in fiber cross-sectional areas of the adductor longus (AL), a slow-twitch antigravity muscle. However, the mean wet weight of the flight AL muscles was near normal, whereas that of the suspension unloaded AL muscles was significantly reduced. Interstitial edema within the flight AL, but not in the unloaded AL, appeared to account for this apparent disagreement.In both conditions, the slow-twitch oxidative fibers atrophied more than the fast-twitch oxidative-glycolytic fibers. Microcirculation was also compromised by spaceflight, such that there was increased formation of thrombi in the postcapillary venules and capillaries.

Whole-body hypothermia has central and peripheral influences on elbow flexor performance.

Science.gov (United States)

Cahill, Farrell; Kalmar, Jayne M; Pretorius, Thea; Gardiner, Phillip F; Giesbrecht, Gordon G

2011-05-01

The superimposed twitch technique was used to study the effect of whole-body hypothermia on maximal voluntary activation of elbow flexors. Seven subjects [26.4 ± 4 years old (mean ± SD)] were exposed to 60 min of either immersion in 8°C water (hypothermia) or sitting in 22°C air (control). Voluntary activation was assessed during brief (3 s) maximal voluntary contractions (MVCs) and then during a 2 min fatiguing sustained MVC. Hypothermia (core temperature 34.8 ± 0.9°C) decreased maximal voluntary torque from 98.2 ± 1.0 to 82.8 ± 5.8% MVC (P < 0.001) and increased central conduction time from 7.9 ± 0.4 to 9.1 ± 0.7 ms (P < 0.05). Hypothermia also decreased maximal resting twitch amplitude from 17.6 ± 4.0 to 10.0 ± 1.7% MVC (P < 0.005) and increased the time-to-peak twitch tension from 55.4 ± 4.0 to 79.0 ± 11.7 ms (P < 0.001). During the 2 min contraction, hypothermia decreased initial torque (P < 0.01) but attenuated the subsequent rate of torque decline (control from 95.5 ± 4 to 29.4 ± 8% MVC; and hypothermia from 85.3 ± 8 to 37.3 ± 5% MVC; P < 0.01). Cortical superimposed twitches increased as fatigue developed but were always lower in the hypothermic conditions. Cortical superimposed twitches increased from a value of 0.4 ± 0.3% MVC prefatigue to 3.9 ± 1.4% MVC postfatigue (P < 0.001) in the hypothermic conditions and from 1.7 ± 0.9 to 5.5 ± 2.3% MVC in control conditions. Our results suggest that hypothermia decreases MVCs primarily via peripheral mechanisms and attenuates the rate of fatigue development by reducing central fatigue.

Effect of ageing on the myosin heavy chain composition of the human sternocleidomastoid muscle.

Science.gov (United States)

Meznaric, M; Eržen, I; Karen, P; Cvetko, E

2018-03-01

The myosin heavy chain (MyHC) composition of ageing limb muscles is transformed into a slower phenotype and expresses fast-twitch fibre type atrophy, presumably due to age-related motor unit remodelling and a change in the patterns of physical activity. It is not known if ageing affects the sternocleidomastoid muscle (SCM) in a similar way. The goal of the study was to analyze the MyHC composition and the size of muscle fibres in the ageing SCM by immunohistochemical methods and quantitative analysis and stereology using our own software for morphometry. We hypothesize that with ageing the MyHC composition of SCM transforms similarly as in ageing limb muscles, but the size of the muscle fibres is less effected as in limb muscles. The study was performed on the autopsy samples of the SCM in 12 older males. The results were compared with those published in our previous study on 15 young adult males. An ageing SCM transforms into a slower MyHC profile: the percentage of slow-twitch fibres is enhanced (numerical proportion 44.6 vs. 31.5%, Pfibres is diminished (numerical proportion 14.1 vs. 26.8%, Pfast-twitch fibres expressing MyHC-2a and 2x is smaller (50.6 vs. 63.5%, Pfibres expressing the fastest myosin isoform MyHC-2x is smaller too (19.0 vs. 34.5%, Pfibres expressing the fastest MyHC-2x provide circumstantial evidence for: (i) more fast-twitch than slow-twitch motor units being lost; and (ii) reinnervation by the surviving motor units. There appears to be no significant influence on muscle fibre size, which is congruent with relatively unchanged SCM activity during life. Copyright © 2017 Elsevier GmbH. All rights reserved.

Differential dependence of apple ( Malus domestica Borkh.) cultivars ...

African Journals Online (AJOL)

Golden Delicious' and 'Topred' apple (Malus domestica Borkh.) peel on the xanthophyll cycle for photoprotection was studied under laboratory conditions. Mature fruit peel was either treated with 1 mm dithiothreitol (DTT) to inhibit the ...

Calsequestrin content and SERCA determine normal and maximal Ca2+ storage levels in sarcoplasmic reticulum of fast- and slow-twitch fibres of rat.

Science.gov (United States)

Murphy, Robyn M; Larkins, Noni T; Mollica, Janelle P; Beard, Nicole A; Lamb, Graham D

2009-01-15

Whilst calsequestrin (CSQ) is widely recognized as the primary Ca2+ buffer in the sarcoplasmic reticulum (SR) in skeletal muscle fibres, its total buffering capacity and importance have come into question. This study quantified the absolute amount of CSQ isoform 1 (CSQ1, the primary isoform) present in rat extensor digitorum longus (EDL) and soleus fibres, and related this to their endogenous and maximal SR Ca2+ content. Using Western blotting, the entire constituents of minute samples of muscle homogenates or segments of individual muscle fibres were compared with known amounts of purified CSQ1. The fidelity of the analysis was proven by examining the relative signal intensity when mixing muscle samples and purified CSQ1. The CSQ1 contents of EDL fibres, almost exclusively type II fibres, and soleus type I fibres [SOL (I)] were, respectively, 36 +/- 2 and 10 +/- 1 micromol (l fibre volume)(-1), quantitatively accounting for the maximal SR Ca2+ content of each. Soleus type II [SOL (II)] fibres (approximately 20% of soleus fibres) had an intermediate amount of CSQ1. Every SOL (I) fibre examined also contained some CSQ isoform 2 (CSQ2), which was absent in every EDL and other type II fibre except for trace amounts in one case. Every EDL and other type II fibre had a high density of SERCA1, the fast-twitch muscle sarco(endo)plasmic reticulum Ca2+-ATPase isoform, whereas there was virtually no SERCA1 in any SOL (I) fibre. Maximal SR Ca2+ content measured in skinned fibres increased with CSQ1 content, and the ratio of endogenous to maximal Ca2+ content was inversely correlated with CSQ1 content. The relative SR Ca2+ content that could be maintained in resting cytoplasmic conditions was found to be much lower in EDL fibres than in SOL (I) fibres (approximately 20 versus >60%). Leakage of Ca2+ from the SR in EDL fibres could be substantially reduced with a SR Ca2+ pump blocker and increased by adding creatine to buffer cytoplasmic [ADP] at a higher level, both results

Calcium currents in a fast-twitch skeletal muscle of the rat.

Science.gov (United States)

Donaldson, P L; Beam, K G

1983-10-01

Slow ionic currents were measured in the rat omohyoid muscle with the three-microelectrode voltage-clamp technique. Sodium and delayed rectifier potassium currents were blocked pharmacologically. Under these conditions, depolarizing test pulses elicited an early outward current, followed by a transient slow inward current, followed in turn by a late outward current. The early outward current appeared to be a residual delayed rectifier current. The slow inward current was identified as a calcium current on the basis that (a) its magnitude depended on extracellular calcium concentration, (b) it was blocked by the addition of the divalent cations cadmium or nickel, and reduced in magnitude by the addition of manganese or cobalt, and (c) barium was able to replace calcium as an inward current carrier. The threshold potential for inward calcium current was around -20 mV in 10mM extracellular calcium and about -35 mV in 2 mM calcium. Currents were net inward over part of their time course for potentials up to at least +30 mV. At temperatures of 20-26 degrees C, the peak inward current (at approximately 0 mV) was 139 +/- 14 microA/cm2 (mean +/- SD), increasing to 226 +/- 28 microA/cm2 at temperatures of 27-37 degrees C. The late outward current exhibited considerable fiber-to-fiber variability. In some fibers it was primarily a time-independent, nonlinear leakage current. In other fibers it was primarily a time-independent, nonlinear leakage current. In other fibers it appeared to be the sum of both leak and a slowly activated outward current. The rate of activation of inward calcium current was strongly temperature dependent. For example, in a representative fiber, the time-to-peak inward current for a +10-mV test pulse decreased from approximately 250 ms at 20 degrees C to 100 ms at 30 degrees C. At 37 degrees C, the time-to-peak current was typically approximately 25 ms. The earliest phase of activation was difficult to quantify because the ionic current was partially

Biochemical and physical characterisation of urinary nanovesicles following CHAPS treatment.

Science.gov (United States)

Musante, Luca; Saraswat, Mayank; Duriez, Elodie; Byrne, Barry; Ravidà, Alessandra; Domon, Bruno; Holthofer, Harry

2012-01-01

Urinary exosomes represent a precious source of potential biomarkers for disease biology. Currently, the methods for vesicle isolation are severely restricted by the tendency of vesicle entrapment, e.g. by the abundant Tamm-Horsfall protein (THP) polymers. Treatment by reducing agents such as dithiothreitol (DTT) releases entrapped vesicles, thus increasing the final yield. However, this harsh treatment can cause remodelling of all those proteins which feature extra-vesicular domains stabilized by internal disulfide bridges and have detrimental effects on their biological activity. In order to optimize exosomal yield, we explore two vesicle treatment protocols - dithiothreitol (DTT) and 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic (CHAPS) - applied to the differential centrifugation protocol for exosomal vesicle isolation. The results show that CHAPS treatment does not affect vesicle morphology or exosomal marker distribution, thus eliminating most of THP interference. Moreover, the recovery and preservation of catalytic activity of two trans-membrane proteases, dipeptidyl peptidase IV and nephrilysin, was examined and found to be clearly superior after CHAPS treatment compared to DTT. Finally, proteomic profiling by mass spectrometry (MS) revealed that 76.2% of proteins recovered by CHAPS are common to those seen for DTT treatment, which illustrates underlining similarities between the two approaches. In conclusion, we provide a major improvement to currently-utilized urinary vesicle isolation strategies to allow recovery of urinary vesicles without the deleterious interference of abundant urinary proteins, while preserving typical protein folding and, consequently, the precious biological activity of urinary proteins which serve as valuable biomarkers.

Lactate Accumulation in Muscle and Blood during Submaximal Exercise

Science.gov (United States)

1981-09-21

exercise, fast and slow twitch fibers Short title: Lactate in muscle and blood P.A. Tesch, W.L. Daniels and D.S. Sharp Exercise Physiology Division, U.S...KIRBY, R.L. & BELCASTRO, A.N. 1978. Relationship between slow - twitch muscle fibres and lactic acid removal. Can J Appl Sports Sci 3:160-162. BRODAL, P...oxygen uptake (Karlsson 1971, Knuttgen & Saltin 1972). It is generally agreed that the main muscle fiber type to be recruited below this level is the slow

β-Nicotinamide adenine dinucleotide acts at prejunctional adenosine A1 receptors to suppress inhibitory musculomotor neurotransmission in guinea pig colon and human jejunum

Science.gov (United States)

Wang, Guo-Du; Wang, Xi-Yu; Liu, Sumei; Xia, Yun; Zou, Fei; Qu, Meihua; Needleman, Bradley J.; Mikami, Dean J.

2015-01-01

Intracellular microelectrodes were used to record neurogenic inhibitory junction potentials in the intestinal circular muscle coat. Electrical field stimulation was used to stimulate intramural neurons and evoke contraction of the smooth musculature. Exposure to β-nicotinamide adenine dinucleotide (β-NAD) did not alter smooth muscle membrane potential in guinea pig colon or human jejunum. ATP, ADP, β-NAD, and adenosine, as well as the purinergic P2Y1 receptor antagonists MRS 2179 and MRS 2500 and the adenosine A1 receptor agonist 2-chloro-N6-cyclopentyladenosine, each suppressed inhibitory junction potentials in guinea pig and human preparations. β-NAD suppressed contractile force of twitch-like contractions evoked by electrical field stimulation in guinea pig and human preparations. P2Y1 receptor antagonists did not reverse this action. Stimulation of adenosine A1 receptors with 2-chloro-N6-cyclopentyladenosine suppressed the force of twitch contractions evoked by electrical field stimulation in like manner to the action of β-NAD. Blockade of adenosine A1 receptors with 8-cyclopentyl-1,3-dipropylxanthine suppressed the inhibitory action of β-NAD on the force of electrically evoked contractions. The results do not support an inhibitory neurotransmitter role for β-NAD at intestinal neuromuscular junctions. The data suggest that β-NAD is a ligand for the adenosine A1 receptor subtype expressed by neurons in the enteric nervous system. The influence of β-NAD on intestinal motility emerges from adenosine A1 receptor-mediated suppression of neurotransmitter release at inhibitory neuromuscular junctions. PMID:25813057

The effects of inorganic phosphate and arsenate on both passive muscle visco-elasticity and maximum Ca2+ activated tension in chemically skinned rat fast and slow twitch muscle fibres.

Science.gov (United States)

Mutungi, Gabriel

2003-01-01

The effects of adding either 25 mM inorganic phosphate (Pi) or its structural analogue arsenate (ASi) on both the maximum Ca2+ activated tension (Po) and passive muscle visco-elasticity (P2 tension) were investigated at 10 degrees C, using segments of single, chemically skinned rat muscle fibres. Whilst the results confirmed some previous findings on the effects of Pi on Po, they also showed that the addition of 25 mM ASi led to a large (approximately 50%) but completely reversible depression of Po in both the fast and slow twitch rat muscle fibres. Moreover, the depression of Po by ASi was greater at low than at high pH values. Examined in the presence of Dextran T-500, the passive tension and sarcomere length responses to a ramp stretch were found to be qualitatively and quantitatively similar to those previously reported in intact rat muscle fibres. Thus, the tension response to a ramp stretch, in the presence and absence of either 25 mM Pi or ASi, consisted of a viscous (P1), a visco-elastic (P2) and an elastic (P3) tension. However, the addition of either 25 mM Pi or ASi led to approximately 15-18% increase in the amplitude of the visco-elastic (P2) tension but had little or no effect on the amplitudes of the other two tension components (viscous, P1 and elastic, P3 tensions). Furthermore, neither compound significantly altered the relaxation rate of the passive muscle visco-elasticity (P2 tension). These results show that Po (arising from cycling cross-bridges) and passive muscle visco-elasticity (P2 tension) are affected differently by both Pi and ASi and suggest that they may not share a common structural basis. The possibility that passive muscle visco-elasticity (P2 tension) arises from the gap-(titin) filament (as suggested previously by Mutungi and Ranatunga, 1996b J Physiol 496: 827-837) and that Pi and ASi increase its amplitude by interacting with the PEVK region of the filament are discussed.

Spatial distribution of motor units recruited during electrical stimulation of the quadriceps muscle versus the femoral nerve.

Science.gov (United States)

Rodriguez-Falces, Javier; Maffiuletti, Nicola A; Place, Nicolas

2013-11-01

In this study we investigated differences in the spatial recruitment of motor units (MUs) in the quadriceps when electrical stimulation is applied over the quadriceps belly versus the femoral nerve. M-waves and mechanical twitches were evoked using over-the-quadriceps and femoral nerve stimulation of gradually increasing intensity from 22 young, healthy subjects. Spatial recruitment was investigated using recruitment curves of M-waves recorded from the vastus medialis (VM) and vastus lateralis (VL) and of twitches recorded from the quadriceps. At maximal stimulation intensity (Imax), no differences were found between nerve and over-the-quadriceps stimulation. At submaximal intensities, VL M-wave amplitude was higher for over-the-quadriceps stimulation at 40% Imax, and peak twitch force was greater for nerve stimulation at 60% and 80% Imax. For the VM, MU spatial recruitment during nerve and over-the-quadriceps stimulation of increasing intensity occurred in a similar manner, whereas significant differences were observed for the VL. Copyright © 2013 Wiley Periodicals, Inc.

Interleaved neuromuscular electrical stimulation: Motor unit recruitment overlap.

Science.gov (United States)

Wiest, Matheus J; Bergquist, Austin J; Schimidt, Helen L; Jones, Kelvin E; Collins, David F

2017-04-01

In this study, we quantified the "overlap" between motor units recruited by single pulses of neuromuscular electrical stimulation (NMES) delivered over the tibialis anterior muscle (mNMES) and the common peroneal nerve (nNMES). We then quantified the torque produced when pulses were alternated between the mNMES and nNMES sites at 40 Hz ("interleaved" NMES; iNMES). Overlap was assessed by comparing torque produced by twitches evoked by mNMES, nNMES, and both delivered together, over a range of stimulus intensities. Trains of iNMES were delivered at the intensity that produced the lowest overlap. Overlap was lowest (5%) when twitches evoked by both mNMES and nNMES produced 10% peak twitch torque. iNMES delivered at this intensity generated 25% of maximal voluntary dorsiflexion torque (11 Nm). Low intensity iNMES leads to low overlap and produces torque that is functionally relevant to evoke dorsiflexion during walking. Muscle Nerve 55: 490-499, 2017. © 2016 Wiley Periodicals, Inc.

Low-intensity electromagnetic irradiation of 70.6 and 73 GHz frequencies enhances the effects of disulfide bonds reducer on Escherichia coli growth and affects the bacterial surface oxidation-reduction state

International Nuclear Information System (INIS)

Torgomyan, Heghine; Trchounian, Armen

2011-01-01

Highlights: → Low intensity 70.6 and 73 GHz electromagnetic irradiation (EMI) strongly suppressed Escherichia coli growth at 73 GHz and pH 7.3. → Reducer DL-dithiothreitol had bactericidal effect and disturbed the SH-groups number. → EMI enhanced E. coli sensitivity toward dithiothreitol. → EMI decreased the SH-groups number of membrane disturbed by ATP and N,N'-dicyclohexycarbodiimide. → The changed membrane oxidation-reduction state could be the primary mechanisms in EMI effects. -- Abstract: Low-intensity electromagnetic irradiation (EMI) of 70.6 and 73 GHz frequencies (flux capacity - 0.06 mW cm -2 ) had bactericidal effects on Escherichia coli. This EMI (1 h) exposure suppressed the growth of E. coli K-12(λ). The pH value (6.0-8.0) did not significantly affect the growth. The lag-phase duration was prolonged, and the growth specific rate was inhibited, and these effects were more noticeable after 73 GHz irradiation. These effects were enhanced by the addition of DL-dithiothreitol (DTT), a strong reducer of disulfide bonds in surface membrane proteins, which in its turn also has bactericidal effect. Further, the number of accessible SH-groups in membrane vesicles was markedly decreased by EMI that was augmented by N,N'-dicyclohexycarbodiimide and DTT. These results indicate a change in the oxidation-reduction state of bacterial cell membrane proteins that could be the primary membranous mechanism in the bactericidal effects of low-intensity EMI of the 70.6 and 73 GHz frequencies.

Primary structure of the light-dependent regulatory site of corn NADP-malate dehydrogenase

Energy Technology Data Exchange (ETDEWEB)

Decottignies, P.; Schmitter, J.M.; Miginiac-Maslow, M.; Le Marechal, P.; Jacquot, J.P.; Gadal, P.

1988-08-25

The light-activated NADP-malate dehydrogenase (NADP-MDH) catalyzes the reduction of oxaloacetate to malate in higher plant chloroplasts. This enzyme is regulated in vivo by the ferredoxin-thioredoxin system through redox reactions. NADP-MDH has been photoactivated in vitro in a chloroplast system reconstituted from the pure protein components and thylakoid membranes. Photoactivation was accompanied by the appearance of new thiol groups (followed by (14C)iodoacetate incorporation). 14C-Carboxymethylated NADP-MDH has been purified from the incubation mixture and its amino-terminal sequence analyzed. Two (14C)carboxymethylcysteines were identified at positions 10 and 15 after light activation, while they were not detected in the dark-treated protein. In addition, the analysis of the tryptic digest of light-activated (14C)carboxymethylated NADP-MDH revealed that the radioactive label was mostly incorporated in Cys10 and Cys15, indicating that these 2 residues play a major role in the light activation mechanism. Moreover, an activation model, in which photoreduced thio-redoxin was replaced by the dithiol reductant dithio-threitol, has been developed. When NADP-MDH was activated in this way, the same sulfhydryls were found to be labeled, and alternatively, they did not incorporate any radioactivity when dithiothreitol reduction was performed after carboxymethylation in denaturating conditions. These results indicate that activation (by light or by dithiothreitol) proceeds on each subunit by reduction of a disulfide bridge located at the amino terminus of the enzyme between Cys10 and Cys15.

Location of the redox-active thiols of ribonucleotide reductase: sequences similarity between the Escherichia coli and Lactobacillus leichmannii enzymes

International Nuclear Information System (INIS)

Lin, A.N.I.; Ashley, G.W.; Stubbe, J.

1987-01-01

The redox-active thiols of Escherichia coli ribonucleoside diphosphate reductase and of Lactobacillus leichmannii ribonucleoside triphosphate reductase have been located by a procedure involving (1) prereduction of enzyme with dithiothreitol, (2) specific oxidation of the redox-active thiols by treatment with substrate in the absence of exogenous reductant, (3) alkylation of other thiols with iodoacetamide, and (4) reduction of the disulfides with dithiothreitol and alkylation with [1- 14 C]iodoacetamide. The dithiothreitol-reduce E. coli B1 subunit is able to convert 3 equiv of CDP to dCDP and is labeled with 5.4 equiv of 14 C. Sequencing of tryptic peptides shows that 2.8 equiv of 14 C is on cysteines-752 and -757 at the C-terminus of B1, while 1.0-1.5 equiv of 14 C is on cysteines-222 and -227. It thus appears that two sets of redox-active dithiols are involved in substrate reduction. The L. leichmannii reductase is able to convert 1.1 equiv of CTP to dCTP and is labeled with 2.1 equiv of 14 C. Sequencing of tryptic peptides shows that 1.4 equiv of 14 C is located on the two cysteines of C-E-G-G-A-C-P-I-K. This peptide shows remarkable and unexpected similarity to the thiol-containing region of the C-terminal peptide of E. coli B1, C-E-S-G-A-C-K-I

Primary structure of the light-dependent regulatory site of corn NADP-malate dehydrogenase

International Nuclear Information System (INIS)

Decottignies, P.; Schmitter, J.M.; Miginiac-Maslow, M.; Le Marechal, P.; Jacquot, J.P.; Gadal, P.

1988-01-01

The light-activated NADP-malate dehydrogenase (NADP-MDH) catalyzes the reduction of oxaloacetate to malate in higher plant chloroplasts. This enzyme is regulated in vivo by the ferredoxin-thioredoxin system through redox reactions. NADP-MDH has been photoactivated in vitro in a chloroplast system reconstituted from the pure protein components and thylakoid membranes. Photoactivation was accompanied by the appearance of new thiol groups (followed by [14C]iodoacetate incorporation). 14C-Carboxymethylated NADP-MDH has been purified from the incubation mixture and its amino-terminal sequence analyzed. Two [14C]carboxymethylcysteines were identified at positions 10 and 15 after light activation, while they were not detected in the dark-treated protein. In addition, the analysis of the tryptic digest of light-activated [14C]carboxymethylated NADP-MDH revealed that the radioactive label was mostly incorporated in Cys10 and Cys15, indicating that these 2 residues play a major role in the light activation mechanism. Moreover, an activation model, in which photoreduced thio-redoxin was replaced by the dithiol reductant dithio-threitol, has been developed. When NADP-MDH was activated in this way, the same sulfhydryls were found to be labeled, and alternatively, they did not incorporate any radioactivity when dithiothreitol reduction was performed after carboxymethylation in denaturating conditions. These results indicate that activation (by light or by dithiothreitol) proceeds on each subunit by reduction of a disulfide bridge located at the amino terminus of the enzyme between Cys10 and Cys15

Cleome gynandra L.

African Journals Online (AJOL)

STORAGESEVER

2009-09-15

Sep 15, 2009 ... INTRODUCTION. Spiderplant is an herbaceous .... 0.5% SDS; 0.1% dithiothreitol (DTT)] and a pinch (about 10 - 20 mg) of polyvinyl ... Operon Technologies, Alameda Calif., USA and ABgene, Epsom,. UK. Based on results of ...

Brain-derived neurotrophic factor in asthmatic children.

African Journals Online (AJOL)

Ehab

of airway smooth muscle contractility remains to be defined2. ... and sputum) were significantly elevated in asthmatic patients, whether studied as one group or .... by adding equal volumes of 0.1% of dithiothreitol. (sputa-lysine 10%, Gibco BRL ...

Agreement of central site measurements and land use regression modeled oxidative potential of PM{sub 2.5} with personal exposure

Energy Technology Data Exchange (ETDEWEB)

Yang, Aileen, E-mail: Yang@uu.nl [National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720BA Bilthoven (Netherlands); Institute for Risk Assessment Sciences, Division Environmental Epidemiology, Utrecht University, P.O. Box 80.178, 3508TD Utrecht (Netherlands); Hoek, Gerard; Montagne, Denise [Institute for Risk Assessment Sciences, Division Environmental Epidemiology, Utrecht University, P.O. Box 80.178, 3508TD Utrecht (Netherlands); Leseman, Daan L.A.C. [National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720BA Bilthoven (Netherlands); Hellack, Bryan [Air Quality & Sustainable Nanotechnology, Institute of Energy and Environmental Technology (IUTA), e.V., Blierheimer Str. 58-60, 47229 Duisburg (Germany); Kuhlbusch, Thomas A.J. [Air Quality & Sustainable Nanotechnology, Institute of Energy and Environmental Technology (IUTA), e.V., Blierheimer Str. 58-60, 47229 Duisburg (Germany); Center for Nanointegration Duisburg-Essen (CENIDE), University Duisburg-Essen, Carl-Benz-Straße 199, 47057 Duisburg (Germany); Cassee, Flemming R. [National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720BA Bilthoven (Netherlands); Institute for Risk Assessment Sciences, Division Environmental Epidemiology, Utrecht University, P.O. Box 80.178, 3508TD Utrecht (Netherlands); Brunekreef, Bert [Institute for Risk Assessment Sciences, Division Environmental Epidemiology, Utrecht University, P.O. Box 80.178, 3508TD Utrecht (Netherlands); Julius Center for Health Sciences and Primary Care, University Medical Center Utrecht, Utrecht (Netherlands); Janssen, Nicole A.H. [National Institute for Public Health and the Environment (RIVM), P.O. Box 1, 3720BA Bilthoven (Netherlands)

2015-07-15

Oxidative potential (OP) of ambient particulate matter (PM) has been suggested as a health-relevant exposure metric. In order to use OP for exposure assessment, information is needed about how well central site OP measurements and modeled average OP at the home address reflect temporal and spatial variation of personal OP. We collected 96-hour personal, home outdoor and indoor PM{sub 2.5} samples from 15 volunteers living either at traffic, urban or regional background locations in Utrecht, the Netherlands. OP was also measured at one central reference site to account for temporal variations. OP was assessed using electron spin resonance (OP{sup ESR}) and dithiothreitol (OP{sup DTT}). Spatial variation of average OP at the home address was modeled using land use regression (LUR) models. For both OP{sup ESR} and OP{sup DTT}, temporal correlations of central site measurements with home outdoor measurements were high (R>0.75), and moderate to high (R=0.49–0.70) with personal measurements. The LUR model predictions for OP correlated significantly with the home outdoor concentrations for OP{sup DTT} and OP{sup ESR} (R=0.65 and 0.62, respectively). LUR model predictions were moderately correlated with personal OP{sup DTT} measurements (R=0.50). Adjustment for indoor sources, such as vacuum cleaning and absence of fume-hood, improved the temporal and spatial agreement with measured personal exposure for OP{sup ESR}. OP{sup DTT} was not associated with any indoor sources. Our study results support the use of central site OP for exposure assessment of epidemiological studies focusing on short-term health effects. - Highlights: • Oxidative potential (OP) of PM was proposed as a health-relevant exposure metric. • We evaluated the relationship between measured and modeled outdoor and personal OP. • Temporal correlations of central site with personal OP are moderate to high. • Adjusting for indoor sources improved the agreement with personal OP. • Our results

The Gross Morphology and Histochemistry of Respiratory Muscles in Bottlenose Dolphins, Tursiops truncatus

Science.gov (United States)

Cotten, Pamela B.; Piscitelli, Marina A.; McLellan, William A.; Rommel, Sentiel A.; Dearolf, Jennifer L.; Pabst, D. Ann

2011-01-01

Most mammals possess stamina because their locomotor and respiratory (i.e., ventilatory) systems are mechanically coupled. These systems are decoupled, however, in bottlenose dolphins (Tursiops truncatus) as they swim on a breath-hold. Locomotion and ventilation are coupled only during their brief surfacing event, when they respire explosively (up to 90% of total lung volume in approximately 0.3s) (Ridgway et al., 1969). The predominantly slow-twitch fiber profile of their diaphragm (Dearolf, 2003) suggests that this muscle does not likely power their rapid ventilatory event. Based upon Bramble's (1989) biomechanical model of locomotor-respiratory coupling in galloping mammals, it was hypothesized that locomotor muscles function to power ventilation in bottlenose dolphins. It was further hypothesized that these muscles would be composed predominantly of fast-twitch fibers to facilitate the bottlenose dolphin's rapid ventilation. The gross morphology of cranio-cervical (scalenus, sternocephalicus, sternohyoid), thoracic (intercostals, transverse thoracis), and lumbo-pelvic (hypaxialis, rectus abdominis, abdominal obliques) muscles (n=7) and the fiber-type profiles (n=6) of selected muscles (scalenus, sternocephalicus, sternohyoid, rectus abdominis) of bottlenose dolphins were investigated. Physical manipulations of excised thoracic units were carried out to investigate potential actions of these muscles. Results suggest that the cranio-cervical muscles act to draw the sternum and associated ribs cranio-dorsally, which flares the ribs laterally, and increases the thoracic cavity volume required for inspiration. The lumbo-pelvic muscles act to draw the sternum and caudal ribs caudally, which decreases the volumes of the thoracic and abdominal cavities required for expiration. All muscles investigated were composed predominantly of fast-twitch fibers (range 61-88% by area) and appear histochemically poised for rapid contraction. These combined results suggest that

On the Mechanism(s of Membrane Permeability Transition in Liver Mitochondria of Lamprey, Lampetra fluviatilis L.: Insights from Cadmium

Directory of Open Access Journals (Sweden)

Elena A. Belyaeva

2014-01-01

Full Text Available Previously we have shown that opening of the mitochondrial permeability transition pore in its low conductance state is the case in hepatocytes of the Baltic lamprey (Lampetra fluviatilis L. during reversible metabolic depression taking place in the period of its prespawning migration when the exogenous feeding is switched off. The depression is observed in the last year of the lamprey life cycle and is conditioned by reversible mitochondrial dysfunction (mitochondrial uncoupling in winter and coupling in spring. To further elucidate the mechanism(s of induction of the mitochondrial permeability transition pore in the lamprey liver, we used Cd2+ and Ca2+ plus Pi as the pore inducers. We found that Ca2+ plus Pi induced the high-amplitude swelling of the isolated “winter” mitochondria both in isotonic sucrose and ammonium nitrate medium while both low and high Cd2+ did not produce the mitochondrial swelling in these media. Low Cd2+ enhanced the inhibition of basal respiration rate of the “winter” mitochondria energized by NAD-dependent substrates whereas the same concentrations of the heavy metal evoked its partial stimulation on FAD-dependent substrates. The above changes produced by Cd2+ or Ca2+ plus Pi in the “winter” mitochondria were only weakly (if so sensitive to cyclosporine A (a potent pharmacological desensitizer of the nonselective pore added alone and they were not sensitive to dithiothreitol (a dithiol reducing agent. Under monitoring of the transmembrane potential of the “spring” lamprey liver mitochondria, we revealed that Cd2+ produced its decrease on both types of the respiratory substrates used that was strongly hampered by cyclosporine A, and the membrane potential was partially restored by dithiothreitol. The effects of different membrane permeability modulators on the lamprey liver mitochondria function and the seasonal changes in their action are discussed.

Aggregation of bovine serum albumin upon cleavage of its disulfide bonds, studied by the time-resolved small-angle X-ray scattering technique with synchrotron radiation

Energy Technology Data Exchange (ETDEWEB)

Ueki, Tatzuo; Inoko, Yoji; Hiragi, Yuzuru; Kataoka, Mikio; Amemiya, Yoshiyuki; Izumi, Yoshinobu; Tagawa, Hiroyuki; Muroga, Yoshio

1985-11-01

A rapid mixing system of the stopped-flow type, used with small-angle X-ray scattering equipment using synchrotron radiation, is described. The process of aggregation of bovine serum albumin was traced with a time interval of 50s, initiated upon cleavage of its disulfide bonds by reduction with dithiothreitol. The results indicate that a 218-fold molar excess of dithiothreitol over the number of moles of disulfide bonds in bovine serum albumin is sufficient to initiate the reaction immediately after mixing, which reaches equilibrium in about 15 min. On the other hand, half this amount is not sufficient to initiate the reaction, so that the reaction is delayed by about 150s. Such a single-shot time-resolved experiment showed that experiments with a time interval of 100 ms are possible with repeated multi-shot runs. 26 refs.; 8 figs.

Aggregation of bovine serum albumin upon cleavage of its disulfide bonds, studied by the time-resolved small-angle X-ray scattering technique with synchrotron radiation

International Nuclear Information System (INIS)

Ueki, Tatzuo; Inoko, Yoji; Izumi, Yoshinobu; Tagawa, Hiroyuki; Muroga, Yoshio

1985-01-01

A rapid mixing system of the stopped-flow type, used with small-angle X-ray scattering equipment using synchrotron radiation, is described. The process of aggregation of bovine serum albumin was traced with a time interval of 50 s, initiated upon cleavage of its disulfide bonds by reduction with dithiothreitol. The results indicate that a 218-fold molar excess of dithiothreitol over the number of moles of disulfide bonds in bovine serum albumin is sufficient to initiate the reaction immediately after mixing, which reaches equilibrium in about 15 min. On the other hand, half this amount is not sufficient to initiate the reaction, so that the reaction is delayed by about 150 s. Such a single-shot time-resolved experiment showed that experiments with a time interval of 100 ms are possible with repeated multi-shot runs. (Auth.)

Proteomic detection of oxidized and reduced thiol proteins in cultured cells.

Science.gov (United States)

Cuddihy, Sarah L; Baty, James W; Brown, Kristin K; Winterbourn, Christine C; Hampton, Mark B

2009-01-01

The oxidation and reduction of cysteine residues is emerging as an important post-translational control of protein function. We describe a method for fluorescent labelling of either reduced or oxidized thiols in combination with two-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (2DE) to detect changes in the redox proteome of cultured cells. Reduced thiols are labelled with the fluorescent compound 5-iodoacetamidofluorescein. To monitor oxidized thiols, the reduced thiols are first blocked with N-ethyl-maleimide, then the oxidized thiols reduced with dithiothreitol and labelled with 5-iodoacetamidofluorescein. The method is illustrated by treating Jurkat T-lymphoma cells with hydrogen peroxide and monitoring increased labelling of oxidized thiol proteins. A decrease in labelling can also be detected, and this is attributed to the formation of higher oxidation states of cysteine that are not reduced by dithiothreitol.

Advanced Development of an Active Neuromusculature Response to Mechanical Stress.

Science.gov (United States)

1984-10-31

muscle (Hatze, 1981, p. 62) For slow motor units (n-0) m • 3-72 and for fast motor units (n-1) m » 18.5. Again an average value is used for...skeletal muscle fibre arrangement. z-disk I-band Fig. 2 Molecular substructure of mammalian skeletal muscle . 11 • • • • ••_» ’J. -J.-J... twitch ) motor units and Type II ( fast twitch ) motor units (Close, 1972). Type I motor units have slower contraction times, tend to be more aerobic and

Electrical Stimulation of Artificial Heart Muscle: a look into the electrophysiological and genetic implications

Science.gov (United States)

Mohamed, Mohamed A; Islas, Jose F; Schwartz, Robert J; Birla, Ravi K

2016-01-01

Development of tissue-engineered hearts for treatment of myocardial infarction or biological pacemakers has been hindered by the production of mostly arrhythmic or in-synergistic constructs. Electrical stimulation (ES) of these constructs has been shown to produce tissues with greater twitch force and better adrenergic response. In order to further our understanding of the mechanisms underlying the effect of ES, we fabricated a bioreactor capable of delivering continuous or intermittent waveforms of various types to multiple constructs simultaneously. In this study, we examined the effect of an intermittent biphasic square wave on our artificial heart muscle (AHM) composed of neonatal rat cardiac cells and fibrin gel. Twitch forces, spontaneous contraction rates, biopotentials, gene expression profiles, and histological observations were examined for the ES protocol over a 12 day culture period. We demonstrate improved consistency between samples for twitch force and contraction rate, and higher normalized twitch force amplitudes for electrically stimulated AHM. Improvements in electrophysiology within the AHM was noted by higher conduction velocities and lower latency in electrical response for electrically stimulated AHM. Genes expressing key electrophysiological and structural markers peaked at days 6 and 8 of culture, only a few days after the initiation of ES. These results may be used for optimization strategies to establish protocols for producing AHM capable of replacing damaged heart tissue in either a contractile or electrophysiological capacity. Optimized AHM can lead to alternative treatments to heart failure and alleviate the limited donor supply crisis. PMID:28459744

Electrical Stimulation of Artificial Heart Muscle: A Look Into the Electrophysiologic and Genetic Implications.

Science.gov (United States)

Mohamed, Mohamed A; Islas, Jose F; Schwartz, Robert J; Birla, Ravi K

Development of tissue-engineered hearts for treatment of myocardial infarction or biologic pacemakers has been hindered by the production of mostly arrhythmic or in-synergistic constructs. Electrical stimulation (ES) of these constructs has been shown to produce tissues with greater twitch force and better adrenergic response. To further our understanding of the mechanisms underlying the effect of ES, we fabricated a bioreactor capable of delivering continuous or intermittent waveforms of various types to multiple constructs simultaneously. In this study, we examined the effect of an intermittent biphasic square wave on our artificial heart muscle (AHM) composed of neonatal rat cardiac cells and fibrin gel. Twitch forces, spontaneous contraction rates, biopotentials, gene expression profiles, and histologic observations were examined for the ES protocol over a 12 day culture period. We demonstrate improved consistency between samples for twitch force and contraction rate, and higher normalized twitch force amplitudes for electrically stimulated AHMs. Improvements in electrophysiology within the AHM were noted by higher conduction velocities and lower latency in electrical response for electrically stimulated AHMs. Genes expressing key electrophysiologic and structural markers peaked at days 6 and 8 of culture, only a few days after the initiation of ES. These results may be used for optimization strategies to establish protocols for producing AHMs capable of replacing damaged heart tissue in either a contractile or electrophysiologic capacity. Optimized AHMs can lead to alternative treatments to heart failure and alleviate the limited donor supply crisis.

A potential human hepatocellular carcinoma inhibitor from Bauhinia purpurea L. seeds: from purification to mechanism exploration.

Science.gov (United States)

Fang, Evandro Fei; Bah, Clara Shui Fern; Wong, Jack Ho; Pan, Wen Liang; Chan, Yau Sang; Ye, Xiu Juan; Ng, Tzi Bun

2012-02-01

A 20-kDa Kunitz-type trypsin-chymotrypsin inhibitor, Bauhinia purpurea trypsin inhibitor (BPLTI), has been isolated from the seeds of B. purpurea L. by using liquid chromatography procedures that involved ion exchange chromatography on Sp-Sepharose and Mono S and gel filtration on Superdex 75. BPLTI demonstrated protease inhibitory activities of 7226 BAEE units/mg and 65 BTEE units/mg toward trypsin and α-chymotrypsin, respectively. BPLTI was relatively thermal (0-60°C) and pH (3-10) stable and its activity could be decreased by dithiothreitol treatment. BPLTI exhibited a wide spectrum of anti-proliferative and pro-apoptotic activities especially on human hepatocellular carcinoma Hep G2 cells. However, it was devoid of a significant antiproliferative effect on immortal human hepatic WRL 68 cells. We show here that BPLTI stimulates apoptosis in Hep G2 cells, including (1) evoking DNA damage including the production of chromatin condensation and apoptotic bodies; (2) induction of cell apoptosis/necrosis; (3) mitochondrial membrane depolarization; and (4) increasing the production of cytokines. Taken together, our findings show for the first time that purified protease inhibitor from B. purpurea L. seeds is a promising candidate for the treatment of human hepatocellular carcinoma.

Neuromuscular activity of Bothrops fonsecai snake venom in vertebrate preparations

Science.gov (United States)

Fernandes, Carla T; Giaretta, Vânia MA; Prudêncio, Luiz S; Toledo, Edvana O; da Silva, Igor RF; Collaço, Rita CO; Barbosa, Ana M; Hyslop, Stephen; Rodrigues-Simioni, Léa; Cogo, José C

2014-01-01

The neuromuscular activity of venom from Bothrops fonsecai, a lancehead endemic to southeastern Brazil, was investigated. Chick biventer cervicis (CBC) and mouse phrenic nerve-diaphragm (PND) preparations were used for myographic recordings and mouse diaphragm muscle was used for membrane resting potential (RP) and miniature end-plate potential (MEPP) recordings. Creatine kinase release and muscle damage were also assessed. In CBC, venom (40, 80 and 160μg/ml) produced concentration- and time-dependent neuromuscular blockade (50% blockade in 85±9 min and 73±8 min with 80 and 160μg/ml, respectively) and attenuated the contractures to 110μM ACh (78–100% inhibition) and 40mM KCl (45–90% inhibition). The venom-induced decrease in twitch-tension in curarized, directly-stimulated preparations was similar to that in indirectly stimulated preparations. Venom (100 and 200μg/ml) also caused blockade in PND preparations (50% blockade in 94±13 min and 49±8 min with 100 and 200μg/ml, respectively) but did not alter the RP or MEPP amplitude. In CBC, venom caused creatine kinase release and myonecrosis. The venom-induced decrease in twitch-tension and in the contractures to ACh and K+ were abolished by preincubating venom with commercial antivenom. These findings indicate that Bothrops fonsecai venom interferes with neuromuscular transmission essentially through postsynaptic muscle damage that affects responses to ACh and KCl. These actions are effectively prevented by commercial antivenom. PMID:25028603

Simple protein structure-sensitive chronopotentiometric analysis with dithiothreitol-modified Hg electrodes

Czech Academy of Sciences Publication Activity Database

Ostatná, Veronika; Černocká, Hana; Paleček, Emil

2012-01-01

Roč. 87, SI (2012), s. 84-88 ISSN 1567-5394 R&D Projects: GA AV ČR(CZ) KJB100040901; GA ČR(CZ) GAP301/11/2055; GA MŠk(CZ) LC06035 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : protein electroanalysis * DTT-modified electrodes * electrocatalysis Subject RIV: BO - Biophysics Impact factor: 3.947, year: 2012

Isokinetic and isometric muscle strength combined with transcutaneous electrical muscle stimulation in primary fibromyalgia syndrome

DEFF Research Database (Denmark)

Jacobsen, Søren; Wildschiødtz, Gordon; Danneskiold-Samsøe, B

1991-01-01

Twenty women with primary fibromyalgia syndrome and 20 age matched healthy women were investigated. The subjects performed maximum voluntary isokinetic contractions of the right quadriceps in an isokinetic dynamometer. Maximum voluntary isometric contractions of the right quadriceps were performed...... of superimposed twitches was 65% in the patient group and 15% in the control group (p = 0.003). Patients with primary fibromyalgia have a lower maximum voluntary muscle strength than expected. The increased presence of superimposed electrically elicited twitches during maximum voluntary contraction indicates...... submaximal force application in primary fibromyalgia syndrome....

Calcium and the role of motoneuronal doublets in skeletal muscle control

DEFF Research Database (Denmark)

Nielsen, Bjørn Gilbert

2009-01-01

+ resources and the dynamics of calcium transport is proposed. The model correctly accounts for catch-like effects in slow and fast-twitch fibers during long-train stimulations and force-frequency relations in different muscle types. Results obtained from the model compare favorably to experiments showing...... in the central nervous system. This is a potentially very useful property directly mediated by the catch-like process modeled here. One further prediction of the model is that the slope of the frequency-tension profile of a given muscle is highly sensitive to changes in the efficiency and temporal...

Effects of caffeine at different temperatures on contractile properties of slow-twitch and fast-twitch rat muscles

Czech Academy of Sciences Publication Activity Database

Wondmikun, Y.; Soukup, Tomáš; Asmussen, G.

2006-01-01

Roč. 55, č. 6 (2006), s. 641-652 ISSN 0862-8408 R&D Projects: GA ČR(CZ) GA304/05/0327 Grant - others:Schwerpunkt Muskelforschung(DE) As 74/1-2; MYORES(XE) 511978 Institutional research plan: CEZ:AV0Z50110509 Keywords : MyHC mRNA transcripts and protein isoforms * altered thyroid status * rat muscles Subject RIV: ED - Physiology Impact factor: 2.093, year: 2006

Influence of Saharan dust outbreaks and carbon content on oxidative potential of water-soluble fractions of PM2.5 and PM10

Science.gov (United States)

Chirizzi, Daniela; Cesari, Daniela; Guascito, Maria Rachele; Dinoi, Adelaide; Giotta, Livia; Donateo, Antonio; Contini, Daniele

2017-08-01

Exposure to atmospheric particulate matter (PM) leads to adverse health effects although the exact mechanisms of toxicity are still poorly understood. Several studies suggested that a large number of PM health effects could be due to the oxidative potential (OP) of ambient particles leading to high concentrations of reactive oxygen species (ROS). The contribution to OP of specific anthropogenic sources like road traffic, biomass burning, and industrial emissions has been investigated in several sites. However, information about the OP of natural sources are scarce and no data is available regarding the OP during Saharan dust outbreaks (SDO) in Mediterranean regions. This work uses the a-cellular DTT (dithiothreitol) assay to evaluate OP of the water-soluble fraction of PM2.5 and PM10 collected at an urban background site in Southern Italy. OP values in three groups of samples were compared: standard characterised by concentrations similar to the yearly averages; high carbon samples associated to combustion sources (mainly road traffic and biomass burning) and SDO events. DTT activity normalised by sampled air volume (DTTV), representative of personal exposure, and normalised by collected aerosol mass (DTTM), representing source-specific characteristics, were investigated. The DTTV is larger for high PM concentrations. DTTV is well correlated with secondary organic carbon concentration. An increased DTTV response was found for PM2.5 compared to the coarse fraction PM2.5-10. DTTV is larger for high carbon content samples but during SDO events is statistically comparable with that of standard samples. DTTM is larger for PM2.5 compared to PM10 and the relative difference between the two size fractions is maximised during SDO events. This indicates that Saharan dust advection is a natural source of particles having a lower specific OP with respect to the other sources acting on the area (for water-soluble fraction). OP should be taken into account in epidemiological

Fiber specific changes in sphingolipid metabolism in skeletal muscles of hyperthyroid rats.

Science.gov (United States)

Chabowski, A; Zendzian-Piotrowska, M; Mikłosz, A; Łukaszuk, B; Kurek, K; Górski, J

2013-07-01

Thyroid hormones (T3, T4) are well known modulators of different cellular signals including the sphingomyelin pathway. However, studies regarding downstream effects of T3 on sphingolipid metabolism in skeletal muscle are scarce. In the present work we sought to investigate the effects of hyperthyroidism on the activity of the key enzymes of ceramide metabolism as well as the content of fundamental sphingolipids. Based on fiber/metabolic differences, we chose three different skeletal muscles, with diverse fiber compositions: soleus (slow-twitch oxidative), red (fast-twitch oxidative-glycolytic) and white (fast-twitch glycolytic) section of gastrocnemius. We demonstrated that T3 induced accumulation of sphinganine, ceramide, sphingosine, as well as sphingomyelin, mostly in soleus and in red, but not white section of gastrocnemius. Concomitantly, the activity of serine palmitoyltransferase and acid/neutral ceramidase was increased in more oxidative muscles. In conclusion, hyperthyroidism induced fiber specific changes in the content of sphingolipids that were relatively more related to de novo synthesis of ceramide rather than to its generation via hydrolysis of sphingomyelin.

Low-intensity electromagnetic irradiation of 70.6 and 73 GHz frequencies enhances the effects of disulfide bonds reducer on Escherichia coli growth and affects the bacterial surface oxidation-reduction state

Energy Technology Data Exchange (ETDEWEB)

Torgomyan, Heghine [Department of Biophysics of Biology Faculty, Yerevan State University, Yerevan 0025 (Armenia); Trchounian, Armen, E-mail: Trchounian@ysu.am [Department of Biophysics of Biology Faculty, Yerevan State University, Yerevan 0025 (Armenia)

2011-10-14

Highlights: {yields} Low intensity 70.6 and 73 GHz electromagnetic irradiation (EMI) strongly suppressed Escherichia coli growth at 73 GHz and pH 7.3. {yields} Reducer DL-dithiothreitol had bactericidal effect and disturbed the SH-groups number. {yields} EMI enhanced E. coli sensitivity toward dithiothreitol. {yields} EMI decreased the SH-groups number of membrane disturbed by ATP and N,N'-dicyclohexycarbodiimide. {yields} The changed membrane oxidation-reduction state could be the primary mechanisms in EMI effects. -- Abstract: Low-intensity electromagnetic irradiation (EMI) of 70.6 and 73 GHz frequencies (flux capacity - 0.06 mW cm{sup -2}) had bactericidal effects on Escherichia coli. This EMI (1 h) exposure suppressed the growth of E. coli K-12({lambda}). The pH value (6.0-8.0) did not significantly affect the growth. The lag-phase duration was prolonged, and the growth specific rate was inhibited, and these effects were more noticeable after 73 GHz irradiation. These effects were enhanced by the addition of DL-dithiothreitol (DTT), a strong reducer of disulfide bonds in surface membrane proteins, which in its turn also has bactericidal effect. Further, the number of accessible SH-groups in membrane vesicles was markedly decreased by EMI that was augmented by N,N'-dicyclohexycarbodiimide and DTT. These results indicate a change in the oxidation-reduction state of bacterial cell membrane proteins that could be the primary membranous mechanism in the bactericidal effects of low-intensity EMI of the 70.6 and 73 GHz frequencies.

The effect of acute hypoxia on short-circuit current and epithelial resistivity in biopsies from human colon.

Science.gov (United States)

Carra, Graciela E; Ibáñez, Jorge E; Saraví, Fernando D

2013-09-01

In isolated colonic mucosa, decreases in short-circuit current (ISC) and transepithelial resistivity (RTE) occur when hypoxia is either induced at both sides or only at the serosal side of the epithelium. We assessed in human colon biopsies the sensitivity to serosal-only hypoxia and mucosal-only hypoxia and whether Na, K-ATPase blockade with ouabain interacts with hypoxia. Biopsy material from patients undergoing colonoscopy was mounted in an Ussing chamber for small samples (1-mm2 window). In a series of experiments we assessed viability and the electrical response to the mucolytic, dithiothreitol (1 mmol/l). In a second series, we explored the effect of hypoxia without and with ouabain. In a third series, we evaluated the response to a cycle of hypoxia and reoxygenation induced at the serosal or mucosal side while keeping the oxygenation of the opposite side. 1st series: Dithiothreitol significantly decreased the unstirred layer and ISC but increased RTE. 2nd series: Both hypoxia and ouabain decreased ISC, but ouabain increased RTE and this effect on RTE prevailed even during hypoxia. 3rd series: Mucosal hypoxia caused lesser decreases of ISC and RTE than serosal hypoxia; in the former, but not in the latter, recovery was complete upon reoxygenation. In mucolytic concentration, dithiothreitol modifies ISC and RTE. Oxygen supply from the serosal side is more important to sustain ISC and RTE in biopsy samples. The different effect of hypoxia and Na, K-ATPase blockade on RTE suggests that their depressing effect on ISC involves different mechanisms.

Biochemical and physical characterisation of urinary nanovesicles following CHAPS treatment.

Directory of Open Access Journals (Sweden)

Luca Musante

Full Text Available Urinary exosomes represent a precious source of potential biomarkers for disease biology. Currently, the methods for vesicle isolation are severely restricted by the tendency of vesicle entrapment, e.g. by the abundant Tamm-Horsfall protein (THP polymers. Treatment by reducing agents such as dithiothreitol (DTT releases entrapped vesicles, thus increasing the final yield. However, this harsh treatment can cause remodelling of all those proteins which feature extra-vesicular domains stabilized by internal disulfide bridges and have detrimental effects on their biological activity. In order to optimize exosomal yield, we explore two vesicle treatment protocols - dithiothreitol (DTT and 3-[(3-cholamidopropyldimethylammonio]-1-propanesulfonic (CHAPS - applied to the differential centrifugation protocol for exosomal vesicle isolation. The results show that CHAPS treatment does not affect vesicle morphology or exosomal marker distribution, thus eliminating most of THP interference. Moreover, the recovery and preservation of catalytic activity of two trans-membrane proteases, dipeptidyl peptidase IV and nephrilysin, was examined and found to be clearly superior after CHAPS treatment compared to DTT. Finally, proteomic profiling by mass spectrometry (MS revealed that 76.2% of proteins recovered by CHAPS are common to those seen for DTT treatment, which illustrates underlining similarities between the two approaches. In conclusion, we provide a major improvement to currently-utilized urinary vesicle isolation strategies to allow recovery of urinary vesicles without the deleterious interference of abundant urinary proteins, while preserving typical protein folding and, consequently, the precious biological activity of urinary proteins which serve as valuable biomarkers.

Topological analysis of Chlamydia trachomatis L2 outer membrane protein 2

DEFF Research Database (Denmark)

Mygind, P; Christiansen, Gunna; Birkelund, Svend

1998-01-01

Using monospecific polyclonal antisera to different parts of Chlamydia trachomatis L2 outer membrane protein 2 (Omp2), we show that the protein is localized at the inner surface of the outer membrane. Omp2 becomes immunoaccessible when Chlamydia elementary bodies are treated with dithiothreitol...

Effects of dantrolene and its derivatives on Ca2+ release from the sarcoplasmic reticulum of mouse skeletal muscle fibres

Science.gov (United States)

Ikemoto, Takaaki; Hosoya, Takamitsu; Aoyama, Hiroshi; Kihara, Yasutaka; Suzuki, Masaaki; Endo, Makoto

2001-01-01

We analysed the effect of dantrolene (Dan) and five newly synthesized derivatives (GIFs) on Ca2+ release from the sarcoplasmic reticulum (SR) of mouse skeletal muscle.In intact muscles, GIF-0185 reduced the size of twitch contraction induced by electrical stimulation to the same extent as Dan. GIF-0082, an azido-functionalized Dan derivative, also inhibited twitch contraction, although the extent of inhibition was less than that of Dan and of GIF-0185.In skinned fibres, Dan inhibited Ca2+-induced Ca2+ release (CICR) under Mg2+-free conditions at room temperature. In contrast, GIF-0082 and GIF-0185 showed no inhibitory effect on CICR under the same conditions.Dan-induced inhibition of CICR was not affected by the presence of GIF-0082, whereas it was diminished in the presence of GIF-0185.GIF-0082 and GIF-0185 significantly inhibited clofibric acid (Clof)-induced Ca2+ release, as did Dan.Several Dan derivatives other than GIF-0082 and GIF-0185 showed an inhibitory effect on twitch tension but not on the CICR mechanism. All of these derivatives inhibited Clof-induced Ca2+ release.The magnitudes of inhibition of Clof-induced Ca2+ release by all Dan derivatives were well correlated with those of twitch inhibition. This supports the notion that the mode of Clof-induced opening of the RyR-Ca2+ release channel may be similar to that of physiological Ca2+ release (PCR).These results indicate that the difference in opening modes of the RyR-Ca2+ release channel is recognized by certain Dan derivatives. PMID:11606312

Torque decrease during submaximal evoked contractions of the quadriceps muscle is linked not only to muscle fatigue.

Science.gov (United States)

Matkowski, Boris; Lepers, Romuald; Martin, Alain

2015-05-01

The aim of this study was to analyze the neuromuscular mechanisms involved in the torque decrease induced by submaximal electromyostimulation (EMS) of the quadriceps muscle. It was hypothesized that torque decrease after EMS would reflect the fatigability of the activated motor units (MUs), but also a reduction in the number of MUs recruited as a result of changes in axonal excitability threshold. Two experiments were performed on 20 men to analyze 1) the supramaximal twitch superimposed and evoked at rest during EMS (Experiment 1, n = 9) and 2) the twitch response and torque-frequency relation of the MUs activated by EMS (Experiment 2, n = 11). Torque loss was assessed by 15 EMS-evoked contractions (50 Hz; 6 s on/6 s off), elicited at a constant intensity that evoked 20% of the maximal voluntary contraction (MVC) torque. The same stimulation intensity delivered over the muscles was used to induce the torque-frequency relation and the single electrical pulse evoked after each EMS contraction (Experiment 2). In Experiment 1, supramaximal twitch was induced by femoral nerve stimulation. Torque decreased by ~60% during EMS-evoked contractions and by only ~18% during MVCs. This was accompanied by a rightward shift of the torque-frequency relation of MUs activated and an increase of the ratio between the superimposed and posttetanic maximal twitch evoked during EMS contraction. These findings suggest that the torque decrease observed during submaximal EMS-evoked contractions involved muscular mechanisms but also a reduction in the number of MUs recruited due to changes in axonal excitability. Copyright © 2015 the American Physiological Society.

Serum levo-carnitine levels and skeletal muscle functions in type 2 diabetes mellitus in rodents

International Nuclear Information System (INIS)

Aleem, S.B.; Hussain, M.M.; Farooq, Y.

2013-01-01

Objective: To study serum levo-carnitine (l-carnitine) levels and isometric contraction, force frequency relationship and fatigue of rodent skeletal muscles in type 2 diabetes mellitus. Study Design: Randomized controlled trial. Place and Duration of Study: Physiology Department, Army Medical College, Rawalpindi, from January 2009 to January 2010. Methodology: Sixty Sprague-Dawley rats were randomly divided into two groups; group I (control), fed on normal diet ad libitum and Group II (diabetic), fed on high fat diet and administered streptozocin to induce type 2 diabetes mellitus (T2DM). At 21st day, plasma glucose and TG/HDL ratio were measured to confirm the development of T2DM in group II. At 28th day, blood was drawn by intracardiac puncture to estimate serum levo-carnitine levels. Contractile functions of skeletal muscles were assessed by using iWorx AHK/214 physiological data acquisition unit. Simple muscle twitches, maximum isometric twitch tension (MITT), time-to-peak twitch tension (TPTT) and time-to-relax to 50% of the peak twitch tension (1/2RT) of extensor digitorum muscles were recorded. Muscles were stimulated at higher frequencies to determine maximum fused tetanic tension (MFTT), maximum fused tetanic tension after fatigue protocol (TTFP) and recovery from fatigue (RF). Results: Serum levo-carnitine level decreased significantly in the diabetic group. Both groups had similar MITT, TPTT and 1/2RT but decline in MFTT, TTFP and RF was significant in the diabetic rats. Conclusion: T2DM adversely affected serum levo-carnitine levels and the contractile functions of rodent skeletal muscle at high frequency stimulation. (author)

A nerve stimulation method to selectively recruit smaller motor-units in rat skeletal muscle.

Science.gov (United States)

van Bolhuis, A I; Holsheimer, J; Savelberg, H H

2001-05-30

Electrical stimulation of peripheral nerve results in a motor-unit recruitment order opposite to that attained by natural neural control, i.e. from large, fast-fatiguing to progressively smaller, fatigue-resistant motor-units. Yet animal studies involving physiological exercise protocols of low intensity and long duration require minimal fatigue. The present study sought to apply a nerve stimulation method to selectively recruit smaller motor-units in rat skeletal muscle. Two pulse generators were used, independently supplying short supramaximal cathodal stimulating pulses (0.5 ms) and long subthreshold cathodal inactivating pulses (1.5 s) to the sciatic nerve. Propagation of action potentials was selectively blocked in nerve fibres of different diameter by adjusting the strength of the inactivating current. A tensile-testing machine was used to gauge isometric muscle force of the plantaris and both heads of the gastrocnemius muscle. The order of motor-unit recruitment was estimated from twitch characteristics, i.e. peak force and relaxation time. The results showed prolonged relaxation at lower twitch peak forces as the intensity of the inactivating current increased, indicating a reduction of the number of large motor-units to force production. It is shown that the nerve stimulation method described is effective in mimicking physiological muscle control.

Sodium bicarbonate supplementation delays neuromuscular fatigue without changes in performance outcomes during a basketball match simulation protocol.

Science.gov (United States)

Ansdell, Paul; Dekerle, Jeanne

2017-10-10

To investigate the development of neuromuscular fatigue during a basketball game simulation and ascertain whether sodium bicarbonate (NaHCO3) supplementation attenuates any neuromuscular fatigue that persists. Ten participants ingested 0.2 g.kg of NaHCO3 (or an equimolar placebo dosage of sodium chloride [NaCl]) 90 and 60 minutes prior to commencing a basketball game simulation (ALK-T vs PLA-T). Isometric maximal voluntary contractions of the knee extensors (MVIC) and potentiated high (100 Hz) and low (10 Hz) frequency doublet twitches were recorded before and after each match quarter for both trials. In addition, 15 m sprint times and layup completion (%) were recorded during each quarter. MVIC, 100 and 10 Hz twitch forces declined progressively in both trials (P0.05). A basketball simulation protocol induces a substantial amount of neuromuscular (reduction in knee extensor MVICs) and peripheral fatigue with a concomitant increase in 15 m sprint time over the protocol. NaHCO3 supplementation attenuated the rate of fatigue development by protecting contractile elements of the muscle fibres. This study provides coaches with information about the magnitude of fatigue induced by a simulated basketball game, and provides evidence of the efficacy of NaHCO3 in attenuating fatigue.

Upper motor neurone modulation of the structure of the terminal cisternae in rat skeletal muscle fibres.

Science.gov (United States)

Dulhunty, A F; Gage, P W; Valois, A A

1981-12-23

There are fewer indentations on the flat surfaces of terminal cisternae in soleus (slow-twitch) than in extensor digitorum longus (EDL, fast-twitch) muscle fibres of rats. Following mid-thoracic spinal cord transection, there is an increase in the number of indentations in soleus fibres but no change in EDL fibres. The increase in the numbers of indentations after spinal cord transections is correlated with changes in the contractile and charge movement properties of the soleus fibres so that they resemble normal EDL fibres. The indentations appear to have an important role in excitation-contraction coupling.

Heterogeneous recruitment of quadriceps muscle portions and fibre types during moderate intensity knee-extensor exercise: effect of thigh occlusion

DEFF Research Database (Denmark)

Krustrup, Peter; Söderlund, Karin; Relu, Mihai U.

2009-01-01

temperature increase (DeltaT(m)) in RF was 0.52+/-0.09 degrees C, which was 57% and 73% higher (Pmuscle CP in slow twitch (ST) and fast......The involvement of quadriceps femoris muscle portions and fibre type recruitment was studied during submaximal knee-extensor exercise without and with thigh occlusion (OCC) and compared with responses during intense exercise. Six healthy male subjects performed 90-s of moderate exercise without...... twitch (FT) fibres was 81% and 91% of resting levels, respectively, with lower (Pfibres had CP levels below mean-1 SD, respectively, with corresponding values for FT fibres being 41...

Easy design of colorimetric logic gates based on nonnatural base pairing and controlled assembly of gold nanoparticles.

Science.gov (United States)

Zhang, Li; Wang, Zhong-Xia; Liang, Ru-Ping; Qiu, Jian-Ding

2013-07-16

Utilizing the principles of metal-ion-mediated base pairs (C-Ag-C and T-Hg-T), the pH-sensitive conformational transition of C-rich DNA strand, and the ligand-exchange process triggered by DL-dithiothreitol (DTT), a system of colorimetric logic gates (YES, AND, INHIBIT, and XOR) can be rationally constructed based on the aggregation of the DNA-modified Au NPs. The proposed logic operation system is simple, which consists of only T-/C-rich DNA-modified Au NPs, and it is unnecessary to exquisitely design and alter the DNA sequence for different multiple molecular logic operations. The nonnatural base pairing combined with unique optical properties of Au NPs promises great potential in multiplexed ion sensing, molecular-scale computers, and other computational logic devices.

p-Toluenethiol as an Initiator of Autolysis in Bakers' Yeast

Science.gov (United States)

Arnold, Wilfred N.

1972-01-01

Cysteine or dithiothreitol enhances the rate of autolysis in toluene-treated yeast. p-Toluenethiol alone is even more effective and is recommended for the isolation of β-fructofuranosidase. This suggests a more general application of p-toluenethiol in the isolation of enzymes from yeast. PMID:5058457

Evidence of significant central fatigue in patients with cancer-related fatigue during repetitive elbow flexions till perceived exhaustion.

Directory of Open Access Journals (Sweden)

Bin Cai

Full Text Available To investigate whether fatigue induced by an intermittent motor task in patients with cancer-related fatigue (CRF is more central or peripheral.Ten patients with CRF who were off chemo and radiation therapies and 14 age-matched healthy controls were enrolled. Participants completed a Brief Fatigue Inventory (BFI and performed a fatigue task consisting of intermittent elbow-flexion contractions at submaximal (40% maximal voluntary contraction intensity till self-perceived exhaustion. Twitch force was elicited by an electrical stimulation applied to the biceps brachii muscle. The relative degree of peripheral (muscle vs. central contribution to fatigue induced by the intermittent motor task (IMT was assessed using twitch force ratio (TF ratio defined as post IMT twitch force to pre IMT twitch force. The total number of trials (intermittent contractions and total duration of all trials performed by each subject were also quantified.BFI scores were higher (p < 0.001 in CRF than controls, indicating greater feeling of fatigue in CRF patients than controls. A significantly smaller number of trials and shorter total duration of the trials (p < 0.05 were observed in CRF than control participants. The TF ratio (0.81 ± 0.05 in CRF was higher (p < 0.05 compared with that of controls (0.62 ± 0.05, suggesting CRF patients experienced a significantly lower degree of muscle (peripheral fatigue at the time of perceived exhaustion.Consistent with prior findings for fatigue under submaximal sustained contraction, our results indicate that motor fatigue in CRF is more of central than peripheral origin during IMT. Significant central fatigue in CRF patients limits their ability to prolong motor performance.

Quantitative differences among EMG activities of muscles innervated by subpopulations of hypoglossal and upper spinal motoneurons during non-REM sleep - REM sleep transitions: a window on neural processes in the sleeping brain.

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Rukhadze, I; Kamani, H; Kubin, L

2011-12-01

In the rat, a species widely used to study the neural mechanisms of sleep and motor control, lingual electromyographic activity (EMG) is minimal during non-rapid eye movement (non-REM) sleep and then phasic twitches gradually increase after the onset of REM sleep. To better characterize the central neural processes underlying this pattern, we quantified EMG of muscles innervated by distinct subpopulations of hypoglossal motoneurons and nuchal (N) EMG during transitions from non-REM sleep to REM sleep. In 8 chronically instrumented rats, we recorded cortical EEG, EMG at sites near the base of the tongue where genioglossal and intrinsic muscle fibers predominate (GG-I), EMG of the geniohyoid (GH) muscle, and N EMG. Sleep-wake states were identified and EMGs quantified relative to their mean levels in wakefulness in successive 10 s epochs. During non-REM sleep, the average EMG levels differed among the three muscles, with the order being N>GH>GG-I. During REM sleep, due to different magnitudes of phasic twitches, the order was reversed to GG-I>GH>N. GG-I and GH exhibited a gradual increase of twitching that peaked at 70-120 s after the onset of REM sleep and then declined if the REM sleep episode lasted longer. We propose that a common phasic excitatory generator impinges on motoneuron pools that innervate different muscles, but twitching magnitudes are different due to different levels of tonic motoneuronal hyperpolarization. We also propose that REM sleep episodes of average durations are terminated by intense activity of the central generator of phasic events, whereas long REM sleep episodes end as a result of a gradual waning of the tonic disfacilitatory and inhibitory processes.

The role of the sarcoplasmic reticulum in the generation of high heart rates and blood pressures in reptiles.

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Galli, Gina L J; Gesser, Hans; Taylor, Edwin W; Shiels, Holly A; Wang, Tobias

2006-05-01

The functional significance of the sarcoplasmic reticulum (SR) in the generation of high heart rates and blood pressures was investigated in four species of reptile; the turtle, Trachemys scripta; the python, Python regius, the tegu lizard, Tupinanvis merianae, and the varanid lizard, Varanus exanthematicus. Force-frequency trials and imposed pauses were performed on ventricular and atrial tissue from each species with and without the SR inhibitor ryanodine, and in the absence and presence of adrenaline. In all species, an imposed pause of 1 or 5 min caused a post-rest decay of force, and a negative force-frequency response was observed in all species within their in vivo frequency range of heart rates. These relationships were not affected by either ryanodine or adrenaline. In ventricular strips from varanid lizards and pythons, ryanodine caused significant reductions in twitch force within their physiologically relevant frequency range. In atrial tissue from the tegu and varanid lizards, SR inhibition reduced twitch force across the whole of their physiological frequency range. In contrast, in the more sedentary species, the turtle and the python, SR inhibition only decreased twitch force at stimulation frequencies above maximal in vivo heart rates. Adrenaline caused an increase in twitch force in all species studied. In ventricular tissue, this positive inotropic effect was sufficient to overcome the negative effects of ryanodine. In atrial tissue however, adrenaline could only ameliorate the negative effects of ryanodine at the lower pacing frequencies. Our results indicate that reptiles recruit Ca2+ from the SR for force development in a frequency and tissue dependent manner. This is discussed in the context of the development of high reptilian heart rates and blood pressures.

A membrane glucocorticoid receptor mediates the rapid/non-genomic actions of glucocorticoids in mammalian skeletal muscle fibres.

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Pérez, María Hernández-Alcalá; Cormack, Jonathan; Mallinson, David; Mutungi, Gabriel

2013-10-15

Glucocorticoids (GCs) are steroid hormones released from the adrenal gland in response to stress. They are also some of the most potent anti-inflammatory and immunosuppressive drugs currently in clinical use. They exert most of their physiological and pharmacological actions through the classical/genomic pathway. However, they also have rapid/non-genomic actions whose physiological and pharmacological functions are still poorly understood. Therefore, the primary aim of this study was to investigate the rapid/non-genomic effects of two widely prescribed glucocorticoids, beclomethasone dipropionate (BDP) and prednisolone acetate (PDNA), on force production in isolated, intact, mouse skeletal muscle fibre bundles. The results show that the effects of both GCs on maximum isometric force (Po) were fibre-type dependent. Thus, they increased Po in the slow-twitch fibre bundles without significantly affecting that of the fast-twitch fibre bundles. The increase in Po occurred within 10 min and was insensitive to the transcriptional inhibitor actinomycin D. Also, it was maximal at ∼250 nM and was blocked by the glucocorticoid receptor (GCR) inhibitor RU486 and a monoclonal anti-GCR, suggesting that it was mediated by a membrane (m) GCR. Both muscle fibre types expressed a cytosolic GCR. However, a mGCR was present only in the slow-twitch fibres. The receptor was more abundant in oxidative than in glycolytic fibres and was confined mainly to the periphery of the fibres where it co-localised with laminin. From these findings we conclude that the rapid/non-genomic actions of GCs are mediated by a mGCR and that they are physiologically/therapeutically beneficial, especially in slow-twitch muscle fibres.

Time-resolved x-ray diffraction from frog skeletal muscle during shortening against an inertial load and a quick release

International Nuclear Information System (INIS)

Amemiya, Yoshiyuki; Hashizume, Hiroo; Tameyasu, Tsukasa; Tanaka, Hidehiro; Sugi, Haruo.

1980-01-01

A group of Japanese researchers conducted, for the first time in this field, experiments on time-resolved x-ray diffraction of frog (bullfrog, Rana catesbeiana) skeletal muscle in conditions where both the force and the muscle length change with time. During an isotonic twitch under a load of about 0.3 P 0 , the intensity ratio started falling on stimulation and reached a minimum value of 0.5 - 0.6 at the early shortening phase, which was maintained until the beginning of relaxation. Except that the minimum value was not retained until the start of relaxation, the same was observed during a twitch against an inertial load whereby the peak force exerted by the muscle was about 0.4 P 0 . The results may be taken to indicate that the change in the intensity ratio reflects not the time course of shortening but that of force generation. When a quick release (3 - 4% of muscle length) was applied during the rising phase of an isometric twitch, the intensity ratio showed no distinct change. Judging from tentative calculation results, however, the foregoing result is subject to further experiments with a much improved time resolution of the measurements. (Kitajima, A.)

Mechanisms limiting glycogen storage in muscle during prolonged insulin stimulation

International Nuclear Information System (INIS)

Richter, E.A.; Hansen, S.A.; Hansen, B.F.

1988-01-01

The extent to which muscle glycogen concentrations can be increased during exposure to maximal insulin concentrations and abundant glucose was investigated in the isolated perfused rat hindquarter preparation. Perfusion for 7 h in the presence of 20,000 μU/ml insulin and 11-13 mM glucose increased muscle glycogen concentrations to maximal values 2, 3, and 3.5 times above normal fed levels in fast-twitch white, slow-twitch red, and fast-twitch red fibers, respectively. Glucose uptake decreased from 34.9 μmol·g -1 ·h -1 at 0 h to 7.5 after 7 h of perfusion. During the perfusion muscle glycogen synthase activity decreased and free intracellular glucose and glucose 6-phosphate increased indicating that glucose disposal was impaired. However, glucose transport as measured by the uptake of 3-O-[ 14 C]methyl-D-glucose was also markedly decreased after 5 and 7 h of perfusion compared with initial values. Total muscle water concentration decreased during glycogen loading of the muscles. Mechanisms limiting glycogen storage under maximal insulin stimulation include impaired insulin-stimulated membrane transport of glucose as well as impaired intracellular glucose disposal

The non-competitive acetylcholinesterase inhibitor APS12-2 is a potent antagonist of skeletal muscle nicotinic acetylcholine receptors

International Nuclear Information System (INIS)

Grandič, Marjana; Aráoz, Romulo; Molgó, Jordi; Turk, Tom; Sepčić, Kristina; Benoit, Evelyne; Frangež, Robert

2012-01-01

APS12-2, a non-competitive acetylcholinesterase inhibitor, is one of the synthetic analogs of polymeric alkylpyridinium salts (poly-APS) isolated from the marine sponge Reniera sarai. In the present work the effects of APS12-2 were studied on isolated mouse phrenic nerve–hemidiaphragm muscle preparations, using twitch tension measurements and electrophysiological recordings. APS12-2 in a concentration-dependent manner blocked nerve-evoked isometric muscle contraction (IC 50 = 0.74 μM), without affecting directly-elicited twitch tension up to 2.72 μM. The compound (0.007–3.40 μM) decreased the amplitude of miniature endplate potentials until a complete block by concentrations higher than 0.68 μM, without affecting their frequency. Full size endplate potentials, recorded after blocking voltage-gated muscle sodium channels, were inhibited by APS12-2 in a concentration-dependent manner (IC 50 = 0.36 μM) without significant change in the resting membrane potential of the muscle fibers up to 3.40 μM. The compound also blocked acetylcholine-evoked inward currents in Xenopus oocytes in which Torpedo (α1 2 β1γδ) muscle-type nicotinic acetylcholine receptors (nAChRs) have been incorporated (IC 50 = 0.0005 μM), indicating a higher affinity of the compound for Torpedo (α1 2 β1γδ) than for the mouse (α1 2 β1γε) nAChR. Our data show for the first time that APS12-2 blocks neuromuscular transmission by a non-depolarizing mechanism through an action on postsynaptic nAChRs of the skeletal neuromuscular junction. -- Highlights: ► APS12-2 produces concentration-dependent inhibition of nerve-evoked muscle contraction in vitro. ► APS12-2 blocks MEPPs and EPPs at the neuromuscular junction. APS12-2 blocks ACh-activated current in Xenopus oocytes incorporated with Torpedo nAChRs.

Oxidative potential of ambient PM2.5 in the coastal cities of the Bohai Sea, northern China: Seasonal variation and source apportionment.

Science.gov (United States)

Liu, WeiJian; Xu, YunSong; Liu, WenXin; Liu, QingYang; Yu, ShuangYu; Liu, Yang; Wang, Xin; Tao, Shu

2018-05-01

Emissions of air pollutants from primary and secondary sources in China are considerably higher than those in developed countries, and exposure to air pollution is main risk of public health. Identifying specific particulate matter (PM) compositions and sources are essential for policy makers to propose effective control measures for pollutant emissions. Ambient PM 2.5 samples covered a whole year were collected from three coastal cities of the Bohai Sea. Oxidative potential (OP) was selected as the indicator to characterize associated PM compositions and sources most responsible for adverse impacts on human health. Positive matrix factorization (PMF) and multiple linear regression (MLR) were employed to estimate correlations of PM 2.5 sources with OP. The volume- and mass-based dithiothreitol (DTT v and DTT m ) activities of PM 2.5 were significantly higher in local winter or autumn (p  0.700, p water-soluble organic carbon (WSOC) and some transition metals. Using PMF, source fractions of PM 2.5 were resolved as secondary source, traffic source, biomass burning, sea spray and urban dust, industry, coal combustion, and mineral dust. Further quantified by MLR, coal combustion, biomass burning, secondary sources, industry, and traffic source were dominant contributors to the water-soluble DTT v activity. Our results also suggested large differences in seasonal contributions of different sources to DTT v variability. A higher contribution of DTT v was derived from coal combustion during the local heating period. Secondary sources exhibited a greater fraction of DTT v in summer, when there was stronger solar radiation. Traffic sources exhibited a prevailing contribution in summer, and industry contributed larger proportions in spring and winter. Future abatement priority of air pollution should reduce the sources contributing to OP of PM 2.5 . Copyright © 2018 Elsevier Ltd. All rights reserved.

Antimicrobial Peptides with Differential Bacterial Binding Characteristics

Science.gov (United States)

2013-03-01

organisms [5]. AMPs exhibit broad- spectrum antimicrobial activity against Gram-positive and Gram-negative bacteria, viruses, and fungi [6]. Hundreds of...polymerase chain reaction PE: PBS with 1mM EDTA PED: PBS with 1mM EDTA and 0.1µM dithiothreitol PEG: polyethylene glycol PL: pleurocidin RP-HPLC

Hierarchical self-assembly of heparin-PEG end-capped porous silica as a redox sensitive nanocarrier for doxorubicin delivery

Energy Technology Data Exchange (ETDEWEB)

Nguyen Thi, Thu Thao; Tran, Tuong Vi; Tran, Ngoc Quyen [Institute of Research and Development, Duy Tan University, Da Nang City 550000 (Viet Nam); Institute of Applied Materials Science, Vietnam Academy of Science and Technology, Ho Chi Minh City 70000 (Viet Nam); Nguyen, Cuu Khoa [Institute of Applied Materials Science, Vietnam Academy of Science and Technology, Ho Chi Minh City 70000 (Viet Nam); Nguyen, Dai Hai, E-mail: nguyendaihai0511@gmail.com [Institute of Applied Materials Science, Vietnam Academy of Science and Technology, Ho Chi Minh City 70000 (Viet Nam)

2017-01-01

Porous nanosilica (PNS) has been attracting a great attention in fabrication carriers for drug delivery system (DDS). However, unmodified PNS-based carriers exhibited the initial burst release of loaded bioactive molecules, which may limit their potential clinical application. In this study, the surface of PNS was conjugated with adamantylamine (A) via disulfide bonds (PNS-SS-A) which was functionalized with cyclodextrin-heparin-polyethylene glycol (CD-HPEG) for redox triggered doxorubicin (DOX) delivery. The modified PNS was successfully formed with spherical shape and diameter around 50 nm determined by transmission electron microscopy (TEM). DOX was efficiently trapped in the PNS-SS-A@CD-HPEG and slowly released in phosphate buffered saline (PBS) without any initial burst effect. Importantly, the release of DOX was triggered due to the cleavage of the disulfide bonds in the presence of dithiothreitol (DTT). In addition, the MTT assay data showed that PNS-SS-A@CD-HPEG was a biocompatible nanocarrier and reduced the toxicity of DOX. These results demonstrated that PNS-SS-A@CD-HPEG has great potential as a novel nanocarrier for anticancer drug in cancer therapy. - Graphic abstract: Hierarchical self-assembly of heparin-PEG end-capped mesoporous silica through host-guest interaction for trapping doxorubicin. The copolymer attached on PNS via disulfide bond which is rapidly cleaved in redox environment, and as a result a huge amount of doxorubicin will release. - Highlights: • Novel redox-responsive nanocarriers based on surface-modified porous nanosilica (PNS) were developed. • Spherical-shaped PNS nanoparticles with diameter around 50 nm were obtained. • Doxorubicin (DOX) was effectively loaded and released in a controlled manner without any initial burst release by surface modification of PNS. • The redox-responsive properties of the modified PNS were demonstrated due to reductive cleavage of disulfide bonds in dithiothreitol (DTT). • The

When climate twitches, evolution takes great leaps

International Nuclear Information System (INIS)

Kerr, R.A.

1992-01-01

Earth's climate system has ridden a slow roller coaster for the past 60 million years. It warmed gradually until about 53 million years ago, reaching a peak that brought crocodiles to northern Canada; then it entered a long, undulating downgrade toward the ice-age world of the past few million years. Because the same span of time brought major evolutionary changes, including the rise of modern mammals, paleontologists have long assumed that these slow temperature changes helped spur the processes of evolution. But recent findings have suggested that another, more potent mechanisms was also at work: Abrupt climate excursions, superimposed on these long-term trends, have now been linked to rapid periods of mammal evolution. The evidence for short, sharp evolutionary shocks comes from studies of a pair of mirror-image climate shifts at opposite ends of the Eocene epoch, 55 million and 33.5 million years ago. In both cases, researchers studying the record of climate preserved in sea-floor sediments have found that a gradual climate change-warming in the first case, cooling in the second - suddenly steepened into a short-lived pulse of extreme warming or cooling. The short time scale of these events and their clear coincidence with turning points in the evolution of mammals are providing the tightest links yet between global climate change and evolution on land

Epiberberine, a natural protoberberine alkaloid, inhibits urease of Helicobacter pylori and jack bean: Susceptibility and mechanism.

Science.gov (United States)

Tan, Lihua; Li, Cailan; Chen, Hanbin; Mo, Zhizhun; Zhou, Jiangtao; Liu, Yuhong; Ma, Zhilin; Xu, Yuyao; Yang, Xiaobo; Xie, Jianhui; Su, Ziren

2017-12-15

In our previous study, Rhizoma Coptidis extract was found to exert more potent inhibitory effect than its major component berberine towards urease from Helicobacter pylori (HPU) and jack bean (JBU). In continuation of our work, the present study was designed to further comparatively investigate the urease inhibitory activities of five major protoberberine alkaloids in Rhizoma Coptidis, namely berberine, palmatine, coptisine, epiberberine, jateorhizine to identify the bioactive constituent, and illuminate the potential mechanism of action. Results indicated that the five protoberberine alkaloids acted as concentration-dependent inactivators of urease with IC 50 values ranging between 3.0 and 5087μM for HPU and 2.3->10,000μM for JBU, respectively. Notably, epiberberine (EB) was found to be the most potent inhibitor against both ureases with IC 50 values of 3.0±0.01μM for HPU and 2.3±0.01μM for JBU, which was more effective than the standard urease inhibitor, acetohydroxamic acid (83±0.01μM for HPU and 22±0.01μM for JBU, respectively). Further kinetic analysis revealed that the type of EB inhibition against HPU was slow-binding and uncompetitive, with K i of 10.6±0.01μM, while slow-binding and competitive against JBU with K i of 4.6±0.01μM. Addition of thiol reagents, such as l-cysteine, glutathione and dithiothreitol, significantly abolished the inhibition, while Ni 2+ competitive inhibitors, boric acid and sodium fluoride, synergetically inhibited urease with EB, indicating the obligatory role of the active site sulfhydryl group for the inhibition. In addition, binding of EB with the urease proved to be reversible, as about 65% and 90% enzymatic activity of HPU and JBU, respectively, could be restored by dithiothreitol application. These findings highlighted the potential role of Rhizoma Coptidis protoberberine alkaloids, especially EB, as a lead urease inhibitor in the treatment of diseases associated with ureolytic bacteria. Thus, EB had good

78 FR 6824 - Considerations Regarding Food and Drug Administration Review and Regulation of Drugs for the...

Science.gov (United States)

2013-01-31

..., the muscles gradually weaken, waste away (atrophy), and twitch (fasciculations). Eventually, the... providers, the pharmaceutical industry, and other interested parties on their experience with, concerns...

Human faces are slower than chimpanzee faces.

Directory of Open Access Journals (Sweden)

Anne M Burrows

Full Text Available While humans (like other primates communicate with facial expressions, the evolution of speech added a new function to the facial muscles (facial expression muscles. The evolution of speech required the development of a coordinated action between visual (movement of the lips and auditory signals in a rhythmic fashion to produce "visemes" (visual movements of the lips that correspond to specific sounds. Visemes depend upon facial muscles to regulate shape of the lips, which themselves act as speech articulators. This movement necessitates a more controlled, sustained muscle contraction than that produced during spontaneous facial expressions which occur rapidly and last only a short period of time. Recently, it was found that human tongue musculature contains a higher proportion of slow-twitch myosin fibers than in rhesus macaques, which is related to the slower, more controlled movements of the human tongue in the production of speech. Are there similar unique, evolutionary physiologic biases found in human facial musculature related to the evolution of speech?Using myosin immunohistochemistry, we tested the hypothesis that human facial musculature has a higher percentage of slow-twitch myosin fibers relative to chimpanzees (Pan troglodytes and rhesus macaques (Macaca mulatta. We sampled the orbicularis oris and zygomaticus major muscles from three cadavers of each species and compared proportions of fiber-types. Results confirmed our hypothesis: humans had the highest proportion of slow-twitch myosin fibers while chimpanzees had the highest proportion of fast-twitch fibers.These findings demonstrate that the human face is slower than that of rhesus macaques and our closest living relative, the chimpanzee. They also support the assertion that human facial musculature and speech co-evolved. Further, these results suggest a unique set of evolutionary selective pressures on human facial musculature to slow down while the function of this muscle

Changes in force and calcium sensitivity in the developing avian heart.

Science.gov (United States)

Godt, R E; Fogaça, R T; Nosek, T M

1991-11-01

The aim of this study was to characterize the development of the contractile properties of intact and chemically skinned muscle from chicken heart and to compare these characteristics with those of developing mammalian heart reported by others. Small trabeculae were dissected from left ventricles of Arbor Acre chickens between embryonic day 7 and young adulthood (7 weeks post-hatching). At all ages, increasing extracellular calcium (0.45-3.6 mM) progressively increased twitch force of electrically stimulated trabeculae. Twitch force at 1.8 mM extracellular calcium, normalized to cross-sectional area, increased to a maximum at 1 day post-hatching, remained constant through 3 weeks post-hatching, but then decreased at 7 weeks post-hatching. The maximal calcium-activated force of trabeculae chemically skinned with Triton X-100 detergent increased to a maximum 2 days before the time of hatching and was not significantly changed up to 7 weeks post-hatching. Over the ages studied, average twitch force in 1.8 mM calcium was between 26 and 66% of maximal calcium-activated force after skinning, suggesting that the contractile apparatus is not fully activated during the twitch in normal Ringer. In skinned trabeculae, the calcium sensitivity of the contractile apparatus was higher in the embryo than in the young adult. These age-dependent changes in calcium sensitivity are correlated with isoform switching in troponin T. A decrease in pH from 7.0 to 6.5 decreased the calcium sensitivity of the contractile apparatus to a greater degree in skinned trabeculae from young adult hearts than in those from embryonic hearts. This change in susceptibility to acidosis is temporally associated with isoform switching in troponin I.(ABSTRACT TRUNCATED AT 250 WORDS)

Differentiation and fiber type-specific activity of a muscle creatine kinase intronic enhancer

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Tai Phillip WL

2011-07-01

Full Text Available Abstract Background Hundreds of genes, including muscle creatine kinase (MCK, are differentially expressed in fast- and slow-twitch muscle fibers, but the fiber type-specific regulatory mechanisms are not well understood. Results Modulatory region 1 (MR1 is a 1-kb regulatory region within MCK intron 1 that is highly active in terminally differentiating skeletal myocytes in vitro. A MCK small intronic enhancer (MCK-SIE containing a paired E-box/myocyte enhancer factor 2 (MEF2 regulatory motif resides within MR1. The SIE's transcriptional activity equals that of the extensively characterized 206-bp MCK 5'-enhancer, but the MCK-SIE is flanked by regions that can repress its activity via the individual and combined effects of about 15 different but highly conserved 9- to 24-bp sequences. ChIP and ChIP-Seq analyses indicate that the SIE and the MCK 5'-enhancer are occupied by MyoD, myogenin and MEF2. Many other E-boxes located within or immediately adjacent to intron 1 are not occupied by MyoD or myogenin. Transgenic analysis of a 6.5-kb MCK genomic fragment containing the 5'-enhancer and proximal promoter plus the 3.2-kb intron 1, with and without MR1, indicates that MR1 is critical for MCK expression in slow- and intermediate-twitch muscle fibers (types I and IIa, respectively, but is not required for expression in fast-twitch muscle fibers (types IIb and IId. Conclusions In this study, we discovered that MR1 is critical for MCK expression in slow- and intermediate-twitch muscle fibers and that MR1's positive transcriptional activity depends on a paired E-box MEF2 site motif within a SIE. This is the first study to delineate the DNA controls for MCK expression in different skeletal muscle fiber types.

Collective defense of Aphis nerii and Uroleucon hypochoeridis (Homoptera, Aphididae against natural enemies.

Directory of Open Access Journals (Sweden)

Manfred Hartbauer

2010-04-01

Full Text Available The prevalent way aphids accomplish colony defense against natural enemies is a mutualistic relationship with ants or the occurrence of a specialised soldier caste typical for eusocial aphids, or even both. Despite a group-living life style of those aphid species lacking these defense lines, communal defense against natural predators has not yet been observed there. Individuals of Aphis nerii (Oleander aphid and Uroleucon hypochoeridis, an aphid species feeding on Hypochoeris radicata (hairy cat's ear, show a behavioral response to visual stimulation in the form of spinning or twitching, which is often accompanied by coordinated kicks executed with hind legs. Interestingly, this behaviour is highly synchronized among members of a colony and repetitive visual stimulation caused strong habituation. Observations of natural aphid colonies revealed that a collective twitching and kicking response (CTKR was frequently evoked during oviposition attempts of the parasitoid wasp Aphidius colemani and during attacks of aphidophagous larvae. CTKR effectively interrupted oviposition attempts of this parasitoid wasp and even repelled this parasitoid from colonies after evoking consecutive CTKRs. In contrast, solitary feeding A. nerii individuals were not able to successfully repel this parasitoid wasp. In addition, CTKR was also evoked through gentle substrate vibrations. Laser vibrometry of the substrate revealed twitching-associated vibrations that form a train of sharp acceleration peaks in the course of a CTKR. This suggests that visual signals in combination with twitching-related substrate vibrations may play an important role in synchronising defense among members of a colony. In both aphid species collective defense in encounters with different natural enemies was executed in a stereotypical way and was similar to CTKR evoked through visual stimulation. This cooperative defense behavior provides an example of a surprising sociality that can be found

Application of 31P-NMR spectroscopy to the study of striated muscle metabolism

International Nuclear Information System (INIS)

Meyer, R.A.; Kushmerick, M.J.; Brown, T.R.

1982-01-01

This review presents the principles and limitations of phosphorus nuclear magnetic resonance ( 31 P-NMR) spectroscopy as applied to the study of striated muscle metabolism. Application of the techniques discussed include noninvasive measurement of high-energy phosphate, intracellular pH, intracellular free Mg 2+ , and metabolite compartmentation. In perfused cat biceps (fast-twitch) muscles, but not in soleus (slow-twitch), NMR spectra indicate a substantially lower (1 mM) free inorganic phosphate level than when measured chemically (6 mM). In addition, saturation and inversion spin-transfer methods that enable direct measurement of the unidirectional fluxes through creatine kinase are described. In perfused cat biceps muscle, results suggest that this enzyme and its substrates are in simple chemical equilibrium

Motor unit recruitment strategies and muscle properties determine the influence of synaptic noise on force steadiness

Science.gov (United States)

Dideriksen, Jakob L.; Negro, Francesco; Enoka, Roger M.

2012-01-01

Motoneurons receive synaptic inputs from tens of thousands of connections that cause membrane potential to fluctuate continuously (synaptic noise), which introduces variability in discharge times of action potentials. We hypothesized that the influence of synaptic noise on force steadiness during voluntary contractions is limited to low muscle forces. The hypothesis was examined with an analytical description of transduction of motor unit spike trains into muscle force, a computational model of motor unit recruitment and rate coding, and experimental analysis of interspike interval variability during steady contractions with the abductor digiti minimi muscle. Simulations varied contraction force, level of synaptic noise, size of motor unit population, recruitment range, twitch contraction times, and level of motor unit short-term synchronization. Consistent with the analytical derivations, simulations and experimental data showed that force variability at target forces above a threshold was primarily due to low-frequency oscillations in neural drive, whereas the influence of synaptic noise was almost completely attenuated by two low-pass filters, one related to convolution of motoneuron spike trains with motor unit twitches (temporal summation) and the other attributable to summation of single motor unit forces (spatial summation). The threshold force above which synaptic noise ceased to influence force steadiness depended on recruitment range, size of motor unit population, and muscle contractile properties. This threshold was low (motor unit recruitment and muscle properties of a typical muscle are tuned to limit the influence of synaptic noise on force steadiness to low forces and that the inability to produce a constant force during stronger contractions is mainly attributable to the common low-frequency oscillations in motoneuron discharge rates. PMID:22423000

Neuromuscular Activity of Micrurus laticollaris (Squamata: Elapidae Venom in Vitro

Directory of Open Access Journals (Sweden)

Alejandro Carbajal-Saucedo

2014-01-01

Full Text Available In this work, we have examined the neuromuscular activity of Micrurus laticollaris (Mexican coral snake venom (MLV in vertebrate isolated nerve-muscle preparations. In chick biventer cervicis preparations, the MLV induced an irreversible concentration- and time-dependent (1–30 µg/mL neuromuscular blockade, with 50% blockade occurring between 8 and 30 min. Muscle contractures evoked by exogenous acetylcholine were completely abolished by MLV, whereas those of KCl were also significantly altered (86% ± 11%, 53% ± 11%, 89% ± 5% and 89% ± 7% for one, three, 10 and 30 µg of venom/mL, respectively; n = 4; p < 0.05. In mouse phrenic nerve-diaphragm preparations, MLV (1–10 µg/mL promoted a slight increase in the amplitude of twitch-tension (3 µg/mL, followed by neuromuscular blockade (n = 4; the highest concentration caused complete inhibition of the twitches (time for 50% blockade = 26 ± 3 min, without exhibiting a previous neuromuscular facilitation. The venom (3 µg/mL induced a biphasic modulation in the frequency of miniature end-plate potentials (MEPPs/min, causing a significant increase after 15 min, followed by a decrease after 60 min (from 17 ± 1.4 (basal to 28 ± 2.5 (t15 and 12 ± 2 (t60. The membrane resting potential of mouse diaphragm preparations pre-exposed or not to d-tubocurarine (5 µg/mL was also significantly less negative with MLV (10 µg/mL. Together, these results indicate that M. laticollaris venom induces neuromuscular blockade by a combination of pre- and post-synaptic activities.

Thiol-modified gold nanoparticles deposited on silica support using dip coating

International Nuclear Information System (INIS)

Magura, Jozef; Zeleňáková, Adriana; Zeleňák, Vladimír; Kaňuchová, Maria

2014-01-01

Graphical abstract: - Highlights: • Thin layers of gold were deposited on glass substrate. • Layers were modified by two different ligands, 1,4-dithiothreitol and L-glutathione. • Red shift of SPR band was observed in spectra after modification of Au by thiols. • Charge transfer between Au and S atoms leads to ferromagnetic behaviour of samples. - Abstract: In our work, we have prepared thin layers of gold nanoparticles deposited via dip coating technique on silica glass substrate. The prepared thin layers were modified by two different ligands, namely 1,4-dithiothreitol (sample Au-DTT NPs) and L-glutathione (sample Au-GSH NPs). The spectral, structural and magnetic properties of the prepared samples were investigated. The modification of Au nanoparticles with thiol ligands leads to change of their plasmon resonance fields, as indicated by UV–vis spectra. The magnetic measurements showed that the magnetization of the samples is composed from two magnetic contributions: diamagnetic contribution and low field ferromagnetic contribution. Our experimental results show that the charge transfer between Au and S atoms gives rise to the ferromagnetic behaviour of prepared thin layers

Cortical tremor: a variant of cortical reflex myoclonus.

Science.gov (United States)

Ikeda, A; Kakigi, R; Funai, N; Neshige, R; Kuroda, Y; Shibasaki, H

1990-10-01

Two patients with action tremor that was thought to originate in the cerebral cortex showed fine shivering-like finger twitching provoked mainly by action and posture. Surface EMG showed relatively rhythmic discharge at a rate of about 9 Hz, which resembled essential tremor. However, electrophysiologic studies revealed giant somatosensory evoked potentials (SEPs) with enhanced long-loop reflex and premovement cortical spike by the jerk-locked averaging method. Treatment with beta-blocker showed no effect, but anticonvulsants such as clonazepam, valproate, and primidone were effective to suppress the tremor and the amplitude of SEPs. We call this involuntary movement "cortical tremor," which is in fact a variant of cortical reflex myoclonus.

Characterization of components released by alkali disruption of simian virus 40

DEFF Research Database (Denmark)

Christiansen, Gunna; Landers, T; Griffith, J

1977-01-01

Treatment of simian virus 40 (SV40) particles at pH 9.8 in the presence of 1 mM dithiothreitol for 5 min at 37 degrees C disrupted the virions into a 60S DNA-protein complex and DNA-free 7S protein particles. The DNA-protein complex contained approximately equal amounts of DNA and protein, and ap...

Oxidative potential of ambient fine aerosol over a semi-urban site in the Indo-Gangetic Plain

Science.gov (United States)

Patel, Anil; Rastogi, Neeraj

2018-02-01

Indo-Gangetic Plain (IGP) receives emissions from variety of pollutant sources such as post-harvest crop residue burning, vehicles, industries, power plants, and bio-fuel burning. Several studies have documented physical, chemical and optical properties of aerosol over the IGP; however, their oxidative potential (OP) has not yet documented. Present study reports the OP (measured through dithiothreitol (DTT) assay) of soluble particulate matter smaller than 2.5 μm aerodynamic diameter (PM2.5) over Patiala (30.3°N, 76.4°E, 249 m amsl), a semi-urban site located in the IGP, during winter 2014. Volume-normalized OP (range: 1.3-7.2 nmol DTT min-1 m-3, average: 3.8 ± 1.4, 1σ) is found to be ∼3 to 20 times higher, and mass-normalized OP (range: 13-50 pmol DTT min-1 μg-1, average: 27 ± 8, 1σ) is found to be similar or higher than those documented in literature. Further, observed OP is found to depend more on PM2.5 composition rather than mass concentration. Mass fractions of organic carbon (OC), elemental carbon (EC) and water-soluble organic carbon (WSOC) correlate positively whereas that of secondary inorganic aerosol (SIA, sum of the concentrations of SO42-, NO3- and NH4+) correlate negatively with OP μg-1 at considerable significance level (p < 0.05). Negative correlation of SIA with OP μg-1 has been assessed in laboratory experiment and attributed to their DTT inactive nature. It is suggested to use WSOC/SIA ratio as a measure of DTT activity of secondary particles over the study region. Further, biomass burning derived species are observed to be more DTT active than those derived from fossil fuel burning. It was also observed that the slope of OP μg-1 and WSOC/SIA ratio linear relationship enhances significantly in samples collected during days following foggy nights in comparison to that in samples collected during non-foggy period, which may be due to the production of redox-active species by fog processing. Such studies have implications in assessing

A novel noninvasive method for measuring fatigability of the quadriceps muscle in noncooperating healthy subjects

DEFF Research Database (Denmark)

Poulsen, Jesper Brøndum; Rose, Martin Høyer; Møller, Kirsten

2015-01-01

days, nonvoluntary isometric contractions (twitch and tetanic) of the quadriceps femoris muscle evoked by transcutaneous electrical muscle stimulation were recorded in twelve healthy adults. For tetanic contractions, the Fatigue Index (ratio of peak torque values) and the slope of the regression line...... of peak torque values were primary outcome measures. For twitch contractions, maximum peak torque and rise time were calculated. Relative (intraclass correlation, ICC3.1) and absolute (standard error of measurement, SEM) reliability were assessed and minimum detectable change was calculated using a 95...... fatigability of the quadriceps muscle produces reliable results in healthy subjects and may provide valuable data on quantitative changes in muscle working capacity and treatment effects in patients who are incapable of producing voluntary muscle contractions....

Serodiagnostic potential of immuno-PCR using a cocktail of mycobacterial antigen 85B, ESAT-6 and cord factor in tuberculosis patients.

Science.gov (United States)

Singh, Netrapal; Sreenivas, Vishnubhatla; Sheoran, Abhishek; Sharma, Suman; Gupta, Krishna B; Khuller, Gopal K; Mehta, Promod K

2016-01-01

A novel indirect immuno-polymerase chain reaction (I-PCR) assay was developed for the detection of circulating anti-Ag85B (antigen 85B, Rv1886c), anti-ESAT-6 (early secretory antigenic target-6, Rv3875) and anti-cord factor (trehalose 6,6'-dimycolate) antibodies from the sera samples of pulmonary tuberculosis (PTB) and extrapulmonary tuberculosis (EPTB) patients and the results were compared with an analogous enzyme-linked immunosorbent assay (ELISA). We covalently attached the amino-modified reporter DNA to the dithiothreitol (DTT)-reduced anti-human IgG antibody through a chemical linker succinimidyl 4-[N-maleimidomethyl]-cyclohexane-1-carboxylate (SMCC). The detection of cocktail of anti-Ag85B, anti-ESAT-6 and anti-cord factor antibodies was found to be superior to the detection of individual antibodies. The sensitivities of 89.5% and 77.5% with I-PCR and 70.8% and 65% with ELISA were observed in smear-positive and smear-negative PTB cases, respectively with high specificity (90.9%). On the other hand, a sensitivity of 77.5% with I-PCR and 65% with ELISA was observed in EBTB cases. The detection of cocktail of antibodies by I-PCR is likely to improve the utility of existing algorithms for TB diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

Neuromuscular rate of force development deficit in Parkinson disease.

Science.gov (United States)

Hammond, Kelley G; Pfeiffer, Ronald F; LeDoux, Mark S; Schilling, Brian K

2017-06-01

Bradykinesia and reduced neuromuscular force exist in Parkinson disease. The interpolated twitch technique has been used to evaluate central versus peripheral manifestations of neuromuscular strength in healthy, aging, and athletic populations, as well as moderate to advanced Parkinson disease, but this method has not been used in mild Parkinson disease. This study aimed to evaluate quadriceps femoris rate of force development and quantify potential central and peripheral activation deficits in individuals with Parkinson disease. Nine persons with mild Parkinson Disease (Hoehn & Yahr≤2, Unified Parkinson Disease Rating Scale total score=mean 19.1 (SD 5.0)) and eight age-matched controls were recruited in a cross-sectional investigation. Quadriceps femoris voluntary and stimulated maximal force and rate of force development were evaluated using the interpolated twitch technique. Thirteen participants satisfactorily completed the protocol. Individuals with early Parkinson disease (n=7) had significantly slower voluntary rate of force development (p=0.008; d=1.97) and rate of force development ratio (p=0.004; d=2.18) than controls (n=6). No significant differences were found between groups for all other variables. Persons with mild-to-moderate Parkinson disease display disparities in rate of force development, even without deficits in maximal force. The inability to produce force at a rate comparable to controls is likely a downstream effect of central dysfunction of the motor pathway in Parkinson disease. Copyright © 2017. Published by Elsevier Ltd.

The effects of local forearm muscle cooling on motor unit properties.

Science.gov (United States)

Mallette, Matthew M; Green, Lara A; Gabriel, David A; Cheung, Stephen S

2018-02-01

Muscle cooling impairs maximal force. Using needle electromyography (EMG) to assess motor unit properties during muscle cooling, is limited and equivocal. Therefore, we aimed to determine the impact of local muscle cooling on motor unit firing properties using surface EMG decomposition. Twenty participants (12 M, 8 F) completed maximal, evoked, and trapezoidal contractions during thermoneutral and cold muscle conditions. Forearm muscle temperature was manipulated using 10-min neutral (~ 32 °C) or 20-min cold (~ 3 °C) water baths. Twitches and maximal voluntary contractions were performed prior to, and after, forearm immersion in neutral or cold water. Motor unit properties were assessed during trapezoidal contractions to 50% baseline force using surface EMG decomposition. Impaired contractile properties from muscle cooling were evident in the twitch amplitude, duration, and rate of force development indicating that the muscle was successfully cooled from the cold water bath (all d ≥ 0.5, P motor units (d = 0.7, P = 0.01) and motor unit action potential (MUAP) duration (d = 0.6, P motor unit firing rates (d = 0.1, P = 0.843) nor recruitment threshold (d = 0.1, P = 0.746) changed; however, the relationship between the recruitment threshold and motor unit firing rate was steeper (d = 1.0, P motor units, and small but coupled changes in motor unit firing rates and recruitment threshold to produce the same force.

Controlled atmosphere stunning of broiler chickens. I. Effects on behaviour, physiology and meat quality in a pilot scale system at a processing plant.

Science.gov (United States)

Abeyesinghe, S M; McKeegan, D E F; McLeman, M A; Lowe, J C; Demmers, T G M; White, R P; Kranen, R W; van Bemmel, H; Lankhaar, J A C; Wathes, C M

2007-08-01

1. The effects of controlled atmosphere stunning on the behaviour, physiology and carcase and meat quality of broiler chickens were studied experimentally in a pilot scale plant. 2. Gas mixtures tested were: single phase anoxic mixture (90% Ar in air, posture occurred soonest with hypercapnic anoxia with the earliest and most twitching and wing flapping in individuals and earliest leg paddling. Biphasic birds were most alert, exhibited most respiratory disruption and mandibulation, and had the latest loss of posture and fewest, but longest bouts of wing flapping and least leg paddling and twitching. 4. Significant and sudden bradycardia and arrhythmia were evident with all gas mixtures and were not related solely to anoxia or hypercapnia. Birds stunned by Ar anoxia showed a slightly more gradual decline from baseline rates, compared with hypercapnic mixtures. 5. Few differences were found between gas mixes in terms of carcase and meat quality. Initial bleeding rate was slowest in biphasic-stunned birds, but total blood loss was not affected. Acceleration of post-mortem metabolism in anoxic-stunned birds was not sufficient to allow de-boning within 5 h without the risk of tough meat. 6. On welfare grounds and taking into account other laboratory and field studies, a biphasic method (using consecutive phases of anaesthesia and euthanasia) of controlled atmosphere stunning of broilers is potentially more humane than anoxic or hypercapnic anoxic methods using argon or nitrogen.

Electrofishing and Its Harmful Effects on Fish

National Research Council Canada - National Science Library

Snyder, Darrel

2003-01-01

... to populations of endangered species. Spinal injuries and associated hemorrhages, although often not externally obvious or fatal, can occur anywhere in the electrofishing field at or above the intensity threshold for twitch...

Chemical Protection Against Ionizing Radiation.

Science.gov (United States)

1984-08-01

cystathionine [3763. Exogen- ous sources of cysteine such as - acetylcysteine and L-2-oxothiazolldine-4 carboxylate promote glutathione synthesis. Whereas the...tubular epithelium and catalyzes the oxidation of extracellular thfols only. Substrates include GSH, cysteine, N- acetylcysteine and dithiothreitol [328,329...extracellular C&2+ . These studies have demonstrated the depression of Ca 2+ sequestration in liver microsomes and mitochondrta after treatment with

Inhibitors for Androgen Receptor Activation Surfaces

Science.gov (United States)

2007-09-01

times and the electron-rich iodine groups of Triac representing particularly good markers. Control soaks with solvent ( DMSO ) reveal no similar...electron-rich iodine groups of Triac represent particu- larly good markers. Control soaks with solvent ( DMSO ) reveal no similar effects on coregulator...3-(dibutylamino)-1-(4-hexylphenyl)propan-1-one DMSO , dimethylsulfoxide DTT, dithiothreitol ER, estrogen receptor GST, glutathione S-transferase

Properties of ribulose diphosphate carboxylase immobilized on porous glass

Science.gov (United States)

Shapira, J.; Hanson, C. L.; Lyding, J. M.; Reilly, P. J.

1974-01-01

Ribulose-1,5-diphosphate carboxylase from spinach has been bound to arylamine porous glass with a diazo linkage and to alklamine porous glass with glutaraldehyde. Stability at elevated temperatures and responses to changes of pH and ribulose-1,5-diphosphate, Mg(2+), and dithiothreitol concentrations were not significantly different from the soluble enzyme, though stability at 4 C was somewhat improved.

Sulphur depletion altered somatic embryogenesis in Theobroma ...

African Journals Online (AJOL)

USER

2010-08-30

Aug 30, 2010 ... embryo) occur in response to alteration of genes expression patterns ... the proteins synthesis and the rate with which all amino ... thiamine-HCl, 1 mgml-1 nicotinic acid and 2 mgml-1 glycine), 20 gL-1 ... Amino acids and soluble sugar extraction .... 0.1% (v/v) Triton X-100, 0.1 % (w/v) dithiothreitol, and 0.2%.

Effects of magnesium chloride on rocuronium-induced neuromuscular blockade and sugammadex reversal in an isolated rat phrenic nerve-hemidiaphragm preparation: An in-vitro study.

Science.gov (United States)

Sung, Tae-Yun; You, Hwang-Ju; Cho, Choon-Kyu; Choi, Hey Ran; Kim, Yong Beom; Shin, Yong Sup; Yang, Hong Seuk

2018-03-01

Magnesium potentiates the effects of nondepolarising muscle relaxants. However, few studies have used magnesium chloride (MgCl2). Sugammadex reverses neuromuscular block by steroidal nondepolarising muscle relaxants. To assess the effects of MgCl2 on rocuronium-induced neuromuscular blockade and its reversal by sugammadex. In-vitro experimental study. Animal laboratory, Asan Medical Center, Seoul, South Korea, from 20 March 2016 to 3 April 2016. Forty male Sprague Dawley rats. Left phrenic nerve-hemidiaphragms from 40 Sprague Dawley rats were allocated randomly to four groups (1, 2, 3 and 4 mmol l MgCl2 group, n = 10 each). Rocuronium was administered cumulatively until the first twitch of train-of-four (TOF) disappeared completely. Then, equimolar sugammadex was administered. The effective concentration (EC) of rocuronium was obtained in each group. After administering sugammadex, recovery of the first twitch height and the TOF ratio were measured for 30 min. EC50, EC90 and EC95 significantly decreased as the concentration of MgCl2 increased (all P ≤ 0.001), except the comparison between the 3 and 4 mmol l MgCl2 groups. After administration of sugammadex, the maximal TOF ratio (%) was lower in the 4 mmol l MgCl2 group than the 1 mmol l MgCl2 group [median 91.7 interquartile range (83.4 to 95.8) vs. 98.3 interquartile range (92.2 to 103.4), P = 0.049]. The mean time (s) from sugammadex injection to achieving maximal first twitch was significantly prolonged in the 4 mmol l MgCl2 group vs. the 1 mmol l MgCl2 and 2 mmol l MgCl2 groups [1483.9 (± 237.0) vs. 1039.0 (± 351.8) and 926.0 (± 278.1), P = 0.022 and 0.002, respectively]. Increases in MgCl2 concentration reduce the ECs of rocuronium. In addition, administering sugammadex equimolar to the administered rocuronium shows limited efficacy as MgCl2 concentration is increased. The in-vitro study was not registered in a database.

Cloning of a neonatal calcium atpase isoform (SERCA 1B) from extraocular muscle of adult blue marlin (Makaira nigricans).

Science.gov (United States)

Londraville, R L; Cramer, T D; Franck, J P; Tullis, A; Block, B A

2000-10-01

Complete cDNAs for the fast-twitch Ca2+ -ATPase isoform (SERCA 1) were cloned and sequenced from blue marlin (Makaira nigricans) extraocular muscle (EOM). Complete cDNAs for SERCA 1 were also cloned from fast-twitch skeletal muscle of the same species. The two sequences are identical over the coding region except for the last five codons on the carboxyl end; EOM SERCA 1 cDNA codes for 996 amino acids and the fast-twitch cDNAs code for 991 aa. Phylogenetic analysis revealed that EOM SERCA 1 clusters with an isoform of Ca2+ -ATPase normally expressed in early development of mammals (SERCA 1B). This is the first report of SERCA 1B in an adult vertebrate. RNA hybridization assays indicate that 1B expression is limited to extraocular muscles. Because EOM gives rise to the thermogenic heater organ in marlin, we investigated whether SERCA 1B may play a role in heat generation, or if 1B expression is common in EOM among vertebrates. Chicken also expresses SERCA 1B in EOM, but rat expresses SERCA 1A; because SERCA 1B is not specific to heater tissue we conclude it is unlikely that it plays a specific role in intracellular heat production. Comparative sequence analysis does reveal, however, several sites that may be the source of functional differences between fish and mammalian SERCAs.

Expression of Dihydropyridine and Ryanodine Receptors in Type IIA Fibers of Rat Skeletal Muscle

International Nuclear Information System (INIS)

Anttila, Katja; Mänttäri, Satu; Järvilehto, Matti

2007-01-01

In this study, the fiber type specificity of dihydropyridine receptors (DHPRs) and ryanodine receptors (RyRs) in different rat limb muscles was investigated. Western blot and histochemical analyses provided for the first time evidence that the expression of both receptors correlates to a specific myosin heavy chain (MHC) composition. We observed a significant (p=0.01) correlation between DHP as well as Ry receptor density and the expression of MHC IIa (correlation factor r=0.674 and r=0.645, respectively) in one slow-twitch, postural muscle (m. soleus), one mixed, fast-twitch muscle (m. gastrocnemius) and two fast-twitch muscles (m. rectus femoris, m. extensor digitorum longus). The highest DHP and Ry receptor density was found in the white part of m. rectus femoris (0.058±0.0060 and 0.057±0.0158 ODu, respectively). As expected, the highest relative percentage of MHC IIa was also found in the white part of m. rectus femoris (70.0±7.77%). Furthermore, histochemical experiments revealed that the IIA fibers stained most strongly for the fluorophore-conjugated receptor blockers. Our data clearly suggest that the expression of DHPRs and RyRs follows a fiber type-specific pattern, indicating an important role for these proteins in the maintenance of an effective Ca 2+ cycle in the fast contracting fiber type IIA

Anatomy and histochemistry of hindlimb flight posture in birds. I. The extended hindlimb posture of shorebirds.

Science.gov (United States)

McFarland, Joshua C; Meyers, Ron A

2008-08-01

Birds utilize one of two hindlimb postures during flight: an extended posture (with the hip and knee joints flexed, while the ankle joint is extended caudally) or a flexed posture (with the hip, knee, and ankle joints flexed beneath the body). American Avocets (Recurvirostra americana) and Black-necked Stilts (Himantopus mexicanus) extend their legs caudally during flight and support them for extended periods. Slow tonic and slow twitch muscle fibers are typically found in muscles functioning in postural support due to the fatigue resistance of these fibers. We hypothesized that a set of small muscles composed of high percentages of slow fibers and thus dedicated to postural support would function in securing the legs in the extended posture during flight. This study examined the anatomy and histochemical profile of eleven hindlimb muscles to gain insight into their functional roles during flight. Contrary to our hypothesis, all muscles possessed both fast twitch and slow twitch or slow tonic fibers. We believe this finding is due to the versatility of dynamic and postural functions the leg muscles must facilitate, including standing, walking, running, swimming, and hindlimb support during flight. Whether birds use an extended or flexed hindlimb flight posture may be related to the aerodynamic effect of leg position or may reflect evolutionary history. (c) 2008 Wiley-Liss, Inc.

Oxidative Potential of ambient particulate matter in Athens, Greece.

Science.gov (United States)

Paraskevopoulou, Despina; Bougiatioti, Aikaterini; Fang, Ting; Liakakou, Eleni; Weber, Rodney; Nenes, Athanasios; Mihalopoulos, Nikolaos

2017-04-01

Exposure of populations to airborne particulate matter (PM) is a leading cause of premature death worldwide. Oxidative stress resulting from exposure of chemical species present in PM is a mechanism thought to cause adverse health effects. Apart from radicals present in aerosol, species that can catalytically deplete the antioxidant buffering capacity of cells, called Oxidative Potential (OP), are thought to be particularly toxic. The variability of OP over location, particle age, source and environmental conditions is virtually unknown for most populated regions of the world. Motivated by this, we have built and deployed one of the first operational measurements of OP in Europe at the National Observatory of Athens site in downtown Athens, Greece. OP for fine and coarse mode is measured using a semi-automated dithiothreitol (DTT) assay developed at the Georgia Institute of Technology; the assay measures the oxidation rate of DTT by water-soluble aerosol constituents, and simulates the rate at which the same compounds would deplete antioxidants in-vivo. The DTT oxidation rate per unit volume of air (water-soluble "DTT activity") and aerosol size class (fine, coarse) are used as a measure of aerosol toxicity. We present continuous (24hr average) OP measurements in downtown Athens from July 2016 to January 2017, conducted through quartz fiber filter analysis. The dataset covers a broad range of aerosol sources (pollution from Europe, regional and local biomass burning, dust, marine aerosol, biogenic aerosol) and meteorological conditions. The daily water-soluble DTT activity ranges between 0.02-0.81 nmolmin-1 m-3 (averaging at 0.24 nmolmin-1 m-3) for fine aerosol and between 0.01-0.52 nmolmin-1 m-3 (averaging at 0.08 nmolmin-1 m-3) for coarse particulate matter, indicating that water-soluble fine mode aerosol components possess a significant fraction of the OP. The seasonal variability demonstrates a higher DTT activity during the coldest period of the year for both

Prediction of Recovery from Coma After CPR

Science.gov (United States)

... to pain. There is good evidence* that myoclonus status epilepticus within the first day after CPR accurately predicts poor recovery from coma. Myoclonus status epilepticus is a constant twitching of muscles, including the ...

A survey on some biochemical and pharmacological activities of venom from two Colombian colubrid snakes, Erythrolamprus bizona (Double-banded coral snake mimic) and Pseudoboa neuwiedii (Neuwied's false boa).

Science.gov (United States)

Torres-Bonilla, Kristian A; Floriano, Rafael S; Schezaro-Ramos, Raphael; Rodrigues-Simioni, Léa; da Cruz-Höfling, Maria Alice

2017-06-01

Colombian colubrid snake venoms have been poorly studied. They represent a great resource of biological, ecological, toxinological and pharmacological research. We assessed some enzymatic properties and neuromuscular effects of Erythrolamprus bizona and Pseudoboa neuwiedii venoms from Colombia. Proteolytic, amidolytic and phospholipase A 2 (PLA 2 ) activities were analyzed using colorimetric assays and the neuromuscular activity was analyzed in chick biventer cervicis (BC) preparations. The venom of both species showed very low PLA 2 and amidolytic activities; however, both exhibited high proteolytic activity, which in E. bizona venom surpassed that of P. neuwiedii venom. E. bizona and P. neuwiedii venoms provoked partial neuromuscular blockade, which was more prominent in P. neuwiedii venom. E. bizona venom (30 μg/ml) induced a significant potentiation of the contracture response to exogenous ACh (110 μM), which was not accompanied by twitch height alteration, whereas the highest venom concentration (100 μg/ml) inhibited contracture responses to both ACh and KCl (40 mM). In contrast, P. neuwiedii venom (30 and 100 μg/ml) caused significant reduction in the contracture responses to exogenous ACh and KCl. The morphological analyses showed high myotoxic effects in the muscle fibers of BC incubated with either venoms; however, they are more prominent in the P. neuwiedii venom. Our results suggest that the myotoxicity of the venom of the two Colombian species can be ascribed to their high proteolytic activity. An interesting data was the potentiation of the ACh-induced contracture, but not the twitch height, caused by E. bizona venom, at a concentration that is harmless to muscle fibers integrity. This phenomenon remains to be further elucidated, and suggest that a possible involvement of post-synaptic receptors cannot be discarded. This work is a contribution to expand the knowledge on colubrid venoms; it allows envisaging that the two venoms offer

The non-competitive acetylcholinesterase inhibitor APS12-2 is a potent antagonist of skeletal muscle nicotinic acetylcholine receptors

Energy Technology Data Exchange (ETDEWEB)

Grandič, Marjana [Institute of Physiology, Pharmacology and Toxicology, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, SI-1000 Ljubljana (Slovenia); Aráoz, Romulo; Molgó, Jordi [CNRS, Institut de Neurobiologie Alfred Fessard, FRC 2118, Laboratoire de Neurobiologie et Développement, UPR 3294, F-91198 Gif-sur-Yvette Cedex (France); Turk, Tom; Sepčić, Kristina [Department of Biology, Biotechnical Faculty, University of Ljubljana, Večna pot 111, SI-1000 Ljubljana (Slovenia); Benoit, Evelyne [CNRS, Institut de Neurobiologie Alfred Fessard, FRC 2118, Laboratoire de Neurobiologie et Développement, UPR 3294, F-91198 Gif-sur-Yvette Cedex (France); Frangež, Robert, E-mail: robert.frangez@vf.uni-lj.si [Institute of Physiology, Pharmacology and Toxicology, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, SI-1000 Ljubljana (Slovenia)

2012-12-01

APS12-2, a non-competitive acetylcholinesterase inhibitor, is one of the synthetic analogs of polymeric alkylpyridinium salts (poly-APS) isolated from the marine sponge Reniera sarai. In the present work the effects of APS12-2 were studied on isolated mouse phrenic nerve–hemidiaphragm muscle preparations, using twitch tension measurements and electrophysiological recordings. APS12-2 in a concentration-dependent manner blocked nerve-evoked isometric muscle contraction (IC{sub 50} = 0.74 μM), without affecting directly-elicited twitch tension up to 2.72 μM. The compound (0.007–3.40 μM) decreased the amplitude of miniature endplate potentials until a complete block by concentrations higher than 0.68 μM, without affecting their frequency. Full size endplate potentials, recorded after blocking voltage-gated muscle sodium channels, were inhibited by APS12-2 in a concentration-dependent manner (IC{sub 50} = 0.36 μM) without significant change in the resting membrane potential of the muscle fibers up to 3.40 μM. The compound also blocked acetylcholine-evoked inward currents in Xenopus oocytes in which Torpedo (α1{sub 2}β1γδ) muscle-type nicotinic acetylcholine receptors (nAChRs) have been incorporated (IC{sub 50} = 0.0005 μM), indicating a higher affinity of the compound for Torpedo (α1{sub 2}β1γδ) than for the mouse (α1{sub 2}β1γε) nAChR. Our data show for the first time that APS12-2 blocks neuromuscular transmission by a non-depolarizing mechanism through an action on postsynaptic nAChRs of the skeletal neuromuscular junction. -- Highlights: ► APS12-2 produces concentration-dependent inhibition of nerve-evoked muscle contraction in vitro. ► APS12-2 blocks MEPPs and EPPs at the neuromuscular junction. APS12-2 blocks ACh-activated current in Xenopus oocytes incorporated with Torpedo nAChRs.

Human Engineered Cardiac Tissues Created Using Induced Pluripotent Stem Cells Reveal Functional Characteristics of BRAF-Mediated Hypertrophic Cardiomyopathy.

Directory of Open Access Journals (Sweden)

Timothy J Cashman

Full Text Available Hypertrophic cardiomyopathy (HCM is a leading cause of sudden cardiac death that often goes undetected in the general population. HCM is also prevalent in patients with cardio-facio-cutaneous syndrome (CFCS, which is a genetic disorder characterized by aberrant signaling in the RAS/MAPK signaling cascade. Understanding the mechanisms of HCM development in such RASopathies may lead to novel therapeutic strategies, but relevant experimental models of the human condition are lacking. Therefore, the objective of this study was to develop the first 3D human engineered cardiac tissue (hECT model of HCM. The hECTs were created using human cardiomyocytes obtained by directed differentiation of induced pluripotent stem cells derived from a patient with CFCS due to an activating BRAF mutation. The mutant myocytes were directly conjugated at a 3:1 ratio with a stromal cell population to create a tissue of defined composition. Compared to healthy patient control hECTs, BRAF-hECTs displayed a hypertrophic phenotype by culture day 6, with significantly increased tissue size, twitch force, and atrial natriuretic peptide (ANP gene expression. Twitch characteristics reflected increased contraction and relaxation rates and shorter twitch duration in BRAF-hECTs, which also had a significantly higher maximum capture rate and lower excitation threshold during electrical pacing, consistent with a more arrhythmogenic substrate. By culture day 11, twitch force was no longer different between BRAF and wild-type hECTs, revealing a temporal aspect of disease modeling with tissue engineering. Principal component analysis identified diastolic force as a key factor that changed from day 6 to day 11, supported by a higher passive stiffness in day 11 BRAF-hECTs. In summary, human engineered cardiac tissues created from BRAF mutant cells recapitulated, for the first time, key aspects of the HCM phenotype, offering a new in vitro model for studying intrinsic mechanisms and

Onset and effectiveness of rocuronium for rapid onset of paralysis in patients with major burns: priming or large bolus

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Han, T.-H.; Martyn, J. A. J.

2009-01-01

Background Burn injury leads to resistance to the effects of non-depolarizing muscle relaxants. We tested the hypothesis that a larger bolus dose is as effective as priming for rapid onset of paralysis after burns. Methods Ninety adults, aged 18–59 yr with 40 (2)% [mean (se)] burn and 30 (2) days after injury, received rocuronium as a priming dose followed by bolus (0.06+0.94 mg kg−1), or single bolus of either 1.0 or 1.5 mg kg−1. Sixty-one non-burned, receiving 1.0 mg kg−1 as a primed (0.06+0.94 mg kg−1) or full bolus dose, served as controls. Acceleromyography measured the onset times. Results Priming when compared with 1.0 mg kg−1 bolus in burned patients shortened the time to first appearance of twitch depression (30 vs 45 s, P<0.05) and time to maximum twitch inhibition (135 vs 210 s, P<0.05). The onset times between priming and higher bolus dose (1.5 mg kg−1) were not different (30 vs 30 s for first twitch depression and 135 vs 135 s for maximal depression, respectively). The onset times in controls, however, were significantly (P<0.05) faster than burns both for priming and for full bolus (15 and 15 s, respectively, for first twitch depression and 75 and 75 s for maximal depression). Priming caused respiratory distress in 10% of patients in both groups. Intubating conditions in burns were significantly better with 1.5 mg kg−1 than with priming or full 1.0 mg kg−1 bolus. Conclusions A dose of 1.5 mg kg−1 not only produces an initial onset of paralysis as early as 30 s, which we speculate could be a reasonable onset time for relief of laryngospasm, but also has an onset as fast as priming with superior intubating conditions and no respiratory side-effects. PMID:19029093

Objectivity of two methods of differentiating fibre types and repeatability of measurements by application of the TEMA image analysis system.

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Henckel, P; Ducro, B; Oksbjerg, N; Hassing, L

1998-01-01

The objectivity of two of the most widely used methods for differentiation of fibre types, i.e. 1) the myosin ATP-ase method (Brooke and Kaiser, 1970a,b) and 2) the combined method, by which the myosin ATP-ase reaction is used to differentiate between fast and slow twitch fibres and NADH-tetrazolium reductase activity is used to identify the subgroups of fast twitch fibres (Ashmore and Doerr, 1970, Peter et al., 1972), was assessed in muscle samples from horses, calves and pigs. We also assessed the objectivity of the alpha-amylase-PAS preparation for the visualisation of capillaries (Andersen, 1975) in these species. For the purpose of reducing the time costs of histochemical analysis of muscle samples, we have developed an interactive image analysis system which is described. All analyses are performed on this system. In accordance with several other investigations, differences between the two methods of differentiating fibre types were found only for the relative distribution of the fast-twitch fibre subgroups (p 87%), the impact of differences in pre-requisites (varied degrees of overlap between the fibre types) for performing the differentiation by the combined method raises a question of the reliability of this method. Apparently, no general rules for comparison of results of distribution of the two subgroups of fast twitch fibres by the two methods are applicable. The alpha-amylase-PAS method was found to be a fairly objective method to identify capillaries in muscles from horses, calves and pigs. However, as capillarity described in combination with other traits to give an indication of diffusion characteristics is significantly influenced by person, it is recommended that the same person perform all the analysis of a project. In addition to the methodological results in this study, we have shown that by application of the TEMA image analysis system, which is more rapid compared with the time-consuming traditional method for evaluation of histochemical

Neuromuscular Disorders

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... lead to twitching, cramps, aches and pains, and joint and movement problems. Sometimes it also affects heart function and your ability to breathe. Examples of neuromuscular disorders include Amyotrophic lateral sclerosis Multiple sclerosis Myasthenia ...

Genetics Home Reference: benign essential blepharospasm

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... eyes, and eye irritation that is aggravated by wind, air pollution, sunlight, and other irritants. These symptoms ... find a genetics professional in my area? Other Names for This Condition essential blepharospasm eyelid twitching primary ...

Dolasetron

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... stiff or twitching muscles seizures coma (loss of consciousness) Dolasetron may cause other side effects. Call your ... for more information if you do not have access to a take-back program.It is important ...

Granisetron

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... stiff or twitching muscles seizures coma (loss of consciousness) Granisetron may cause other side effects. Call your ... for more information if you do not have access to a take-back program.It is important ...

Morphine Rectal

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... of lung function) or other breathing problems; prostatic hypertrophy (enlargement of a male reproductive gland); urethral stricture ( ... exist), fever, sweating, confusion, fast heartbeat, shivering, severe muscle stiffness or twitching, loss of coordination, nausea, vomiting, ...

Reduced Acetylcholine Receptor Density, Morphological Remodeling, and Butyrylcholinesterase Activity Can Sustain Muscle Function in Acetylcholinesterase Knockout Mice

National Research Council Canada - National Science Library

Adler, Michael; Manley, Heather A; Deshpande, Sharad S; Hamilton, Tracey A; Kan, Robert K; Oyler, George; Lockridge, Oksana; Duysen, Ellen G; Sheridan, Robert E

2004-01-01

...) and can survive into adulthood. Twitch tensions elicited in diaphragms of AChE-/- mice by single supramaximal stimuli had larger amplitudes and slower rise and decay times than did those in wild-type animals...

Genetics Home Reference: hereditary hyperekplexia

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... hyperekplexia can include muscle twitches when falling asleep (hypnagogic myoclonus) and movements of the arms or legs ... fall down. They may also continue to have hypnagogic myoclonus or movements during sleep. As they get ...

Free radicals in alcoholic myopathy: indices of damage and preventive studies.

Science.gov (United States)

Preedy, Victor R; Adachi, Junko; Asano, Migiwa; Koll, Michael; Mantle, David; Niemela, Onni; Parkkila, Seppo; Paice, Alistair G; Peters, Timothy; Rajendram, Rajkumar; Seitz, Helmut; Ueno, Yasuhiro; Worrall, Simon

2002-04-15

Chronic alcoholic myopathy affects up to two-thirds of all alcohol misusers and is characterized by selective atrophy of Type II (glycolytic, fast-twitch, anaerobic) fibers. In contrast, the Type I fibers (oxidative, slow-twitch, aerobic) are relatively protected. Alcohol increases the concentration of cholesterol hydroperoxides and malondialdehyde-protein adducts, though protein-carbonyl concentration levels do not appear to be overtly increased and may actually decrease in some studies. In alcoholics, plasma concentrations of alpha-tocopherol may be reduced in myopathic patients. However, alpha-tocopherol supplementation has failed to prevent either the loss of skeletal muscle protein or the reductions in protein synthesis in alcohol-dosed animals. The evidence for increased oxidative stress in alcohol-exposed skeletal muscle is thus inconsistent. Further work into the role of ROS in alcoholic myopathy is clearly warranted.

Alpha and beta adrenergic effects on metabolism in contracting, perfused muscle

DEFF Research Database (Denmark)

Richter, Erik; Ruderman, N B; Galbo, H

1982-01-01

The role of alpha- and beta-adrenergic receptor stimulation for the effect of epinephrine on muscle glycogenolysis, glucose- and oxygen uptake and muscle performance was studied in the perfused rat hindquarter at rest and during electrical stimulation (60 contractions/min). Adrenergic stimulation...... was obtained by epinephrine in a physiological concentration (2.4 X 10(-8) M) and alpha- and beta-adrenergic blockade by 10(-5) M phentolamine and propranolol, respectively. Epinephrine enhanced net glycogenolysis during contractions most markedly in slow-twitch red fibers. In these fibers the effect...... was mediated by alpha- as well as by beta-adrenergic stimulation, the latter involving production of cAMP, phosphorylase activation and synthase inactivation. In contrast, in fast-twitch fibers only beta-adrenergic mechanisms were involved in the glycogenolytic effect of epinephrine. Moreover, inactivation...

Muscle and liver glycogen, protein, and triglyceride in the rat

DEFF Research Database (Denmark)

Richter, Erik; Sonne, Bente; Joensen Mikines, Kari

1984-01-01

in skeletal muscle was accompanied by increased breakdown of triglyceride and/or protein. Thus, the effect of exhausting swimming and of running on concentrations of glycogen, protein, and triglyceride in skeletal muscle and liver were studied in rats with and without deficiencies of the sympatho......-adrenal system. In control rats, both swimming and running decreased the concentration of glycogen in fast-twitch red and slow-twitch red muscle whereas concentrations of protein and triglyceride did not decrease. In the liver, swimming depleted glycogen stores but protein and triglyceride concentrations did...... not decrease. In exercising rats, muscle glycogen breakdown was impaired by adrenodemedullation and restored by infusion of epinephrine. However, impaired glycogen breakdown during exercise was not accompanied by a significant net breakdown of protein or triglyceride. Surgical sympathectomy of the muscles did...

Genetics Home Reference: dystonia 6

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... neck, causing problems with speaking (dysarthria) and eating (dysphagia). Eyelid twitching (blepharospasm) may also occur. Involvement of ... dystonia, DYT6 type The Bachmann-Strauss Dystonia and Parkinson Foundation: What Is Dystonia? Patient Support and Advocacy ...

Effects of Bright Light Therapy of Sleep, Cognition, Brain Function, and Neurochemistry in Mild Traumatic Brain Injury

Science.gov (United States)

2012-01-01

DTI ) 128 killgore@mclean.harvard.edu Table of Contents...the Less than Once or twice Three or more past month_____ once a week_____ a week_____ times a week_____ c) Legs twitching or jerking while you

Characterizing SHP2 as a Novel Therapeutic Target in Breast Cancer

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2014-04-01

collected and stored in ‐80oC with 10mM  dithiothreitol. The  Bradford   assay  (Biorad, City, State) was used to quantify the product.  In vitro...activity assay ............................................................................. 1   5.3.4) Site-directed mutagenesis, subcloning, and...independent growth assay ........................................................................   5.3.8) Molecular docking

Analysis of the protein profiles of the antibiotic- resistant Salmonella ...

African Journals Online (AJOL)

Owner

2005-05-18

May 18, 2005 ... ice-cold 100% acetone and air-dried. The dried whole cell proteins and other samples (flagillin, CFUS) for 2DE were digested (100oC,. 5 min) in 4 µl of 10% SDS and dissolved in 100 µl of urea sample buffer containing 8 M urea, 4% Triton X-100, 20 mM dithiothreitol,. 2% ampholyte (pH 3.5~10) and traces ...

Bell's Palsy: Treatment with Steroids and Antiviral Drugs

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... Drooping of a corner of the mouth • Difficulty smiling, frowning, or making other facial expressions • Twitching or ... no definite added improvement. If there is any benefit to adding an antiviral to steroid treatment, it ...

Active site mutations in yeast protein disulfide isomerase cause dithiothreitol sensitivity and a reduced rate of protein folding in the endoplasmic reticulum

DEFF Research Database (Denmark)

Holst, B; Tachibana, C; Winther, Jakob R.

1997-01-01

Aspects of protein disulfide isomerase (PDI) function have been studied in yeast in vivo. PDI contains two thioredoxin-like domains, a and a', each of which contains an active-site CXXC motif. The relative importance of the two domains was analyzed by rendering each one inactive by mutation to SGAS....... Such mutations had no significant effect on growth. The domains however, were not equivalent since the rate of folding of carboxypeptidase Y (CPY) in vivo was reduced by inactivation of the a domain but not the a' domain. To investigate the relevance of PDI redox potential, the G and H positions of each CGHC......-deleted strains overexpressing the yeast PDI homologue EUG1 are viable. Exchanging the wild-type Eug1p C(L/I)HS active site sequences for C(L/I)HC increased the growth rate significantly, however, further highlighting the importance of the oxidizing function for optimal growth....

Flavonoids from Rhizophora conjugata fruit extract blocks virulence factors of Pseudomonas aeruginosa

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Naik, D.; Tilvi, S.; DeSouza, L.

: chloroform (1:1) extracts of 7 mangrove plants on P. aeruginosa ATCC 27853 motilities (swarming, swimming and twitching). Amongst the 22 extracts tested, methanolic extract of Rhizophora conjugata fruit showed maximum inhibition. The butanol (RcBu) fraction...

Tics

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... Staying Safe Videos for Educators Search English Español Tics KidsHealth / For Teens / Tics What's in this article? ... tic, such as a nose twitch. What's a Tic? A tic is an uncontrolled sudden, repetitive movement ...

Effect of lung-protective ventilation-induced respiratory acidosis on the duration of neuromuscular blockade by rocuronium.

Science.gov (United States)

Taguchi, Shinya; Ono, Kazumi; Hidaka, Hidekuni; Koyama, Yusuke

2016-12-01

The purpose of this study was to elucidate whether lung-protective ventilation-induced respiratory acidosis increased the duration of neuromuscular blockade by rocuronium. A total of 72 patients were enrolled. After the induction of general anesthesia, rocuronium 0.6 mg/kg real body weight was administered. Tidal volume and positive end-expiratory pressure were randomly assigned as either 10 ml/kg predicted body weight and 0 cmH 2 O (group S) or 6 ml/kg and 5 cmH 2 O (group L), respectively. Respiratory rate was started at 10/min. Neuromuscular blockade was monitored by acceleromyography at the adductor pollicis with train-of-four stimulation. The time from the initial bolus injection of rocuronium to first recovery of the first twitch was defined as DUR1. Immediately, rocuronium 0.15 mg/kg was administered. The time from first recovery of the first twitch to second recovery of the first twitch was defined as DUR2. We also measured arterial pH (pH1 and pH2, respectively). Data from 66 patients (33 each in groups L and S) were eventually available. pH1 and pH2 were significantly lower in group L compared with group S [pH1: 7.308 (7.288-7.334) vs. 7.439 (7.423-7.466); p respiratory acidosis increased the duration of neuromuscular blockade by rocuronium.

Bion 11 Spaceflight Project: Effect of Weightlessness on Single Muscle Fiber Function in Rhesus Monkeys

Science.gov (United States)

Fitts, Robert H.; Romatowski, Janell G.; Widrick, Jeffrey J.; DeLaCruz, Lourdes

1999-01-01

Although it is well known that microgravity induces considerable limb muscle atrophy, little is known about how weightlessness alters cell function. In this study, we investigated how weightlessness altered the functional properties of single fast and slow striated muscle fibers. Physiological studies were carried out to test the hypothesis that microgravity causes fiber atrophy, a decreased peak force (Newtons), tension (Newtons/cross-sectional area) and power, an elevated peak rate of tension development (dp/dt), and an increased maximal shortening velocity (V(sub o)) in the slow type I fiber, while changes in the fast-twitch fiber are restricted to atrophy and a reduced peak force. For each fiber, we determined the peak force (P(sub o)), V(sub o), dp/dt, the force-velocity relationship, peak power, the power-force relationship, the force-pCa relationship, and fiber stiffness. Biochemical studies were carried out to assess the effects of weightlessness on the enzyme and substrate profile of the fast- and slow-twitch fibers. We predicted that microgravity would increase resting muscle glycogen and glycolytic metabolism in the slow fiber type, while the fast-twitch fiber enzyme profile would be unaltered. The increased muscle glycogen would in part result from an elevated hexokinase and glycogen synthase. The enzymes selected for study represent markers for mitochondrial function (citrate synthase and 0-hydroxyacyl-CoA dehydrogenase), glycolysis (Phosphofructokinase and lactate dehydrogenase), and fatty acid transport (Carnitine acetyl transferase). The substrates analyzed will include glycogen, lactate, adenosine triphosphate, and phosphocreatine.

Effect of endurance training on glucose transport capacity and glucose transporter expression in rat skeletal muscle

International Nuclear Information System (INIS)

Ploug, T.; Stallknecht, B.M.; Pedersen, O.; Kahn, B.B.; Ohkuwa, T.; Vinten, J.; Galbo, H.

1990-01-01

The effect of 10 wk endurance swim training on 3-O-methylglucose (3-MG) uptake (at 40 mM 3-MG) in skeletal muscle was studied in the perfused rat hindquarter. Training resulted in an increase of approximately 33% for maximum insulin-stimulated 3-MG transport in fast-twitch red fibers and an increase of approximately 33% for contraction-stimulated transport in slow-twitch red fibers compared with nonexercised sedentary muscle. A fully additive effect of insulin and contractions was observed both in trained and untrained muscle. Compared with transport in control rats subjected to an almost exhaustive single exercise session the day before experiment both maximum insulin- and contraction-stimulated transport rates were increased in all muscle types in trained rats. Accordingly, the increased glucose transport capacity in trained muscle was not due to a residual effect of the last training session. Half-times for reversal of contraction-induced glucose transport were similar in trained and untrained muscles. The concentrations of mRNA for GLUT-1 (the erythrocyte-brain-Hep G2 glucose transporter) and GLUT-4 (the adipocyte-muscle glucose transporter) were increased approximately twofold by training in fast-twitch red muscle fibers. In parallel to this, Western blot demonstrated a approximately 47% increase in GLUT-1 protein and a approximately 31% increase in GLUT-4 protein. This indicates that the increases in maximum velocity for 3-MG transport in trained muscle is due to an increased number of glucose transporters

De novo synthesis of adenine nucleotides in different skeletal muscle fiber types

International Nuclear Information System (INIS)

Tullson, P.C.; John-Alder, H.B.; Hood, D.A.; Terjung, R.L.

1988-01-01

Management of adenine nucleotide catabolism differs among skeletal muscle fiber types. This study evaluated whether there are corresponding differences in the rates of de novo synthesis of adenine nucleotide among fiber type sections of skeletal muscle using an isolated perfused rat hindquarter preparation. Label incorporation into adenine nucleotides from the [1-14C]glycine precursor was determined and used to calculate synthesis rates based on the intracellular glycine specific radioactivity. Results show that intracellular glycine is closely related to the direct precursor pool. Rates of de novo synthesis were highest in fast-twitch red muscle (57.0 +/- 4.0, 58.2 +/- 4.4 nmol.h-1.g-1; deep red gastrocnemius and vastus lateralis), relatively high in slow-twitch red muscle (47.0 +/- 3.1; soleus), and low in fast-twitch white muscle (26.1 +/- 2.0 and 21.6 +/- 2.3; superficial white gastrocnemius and vastus lateralis). Rates for four mixed muscles were intermediate, ranging between 32.3 and 37.3. Specific de novo synthesis rates exhibited a strong correlation (r = 0.986) with muscle section citrate synthase activity. Turnover rates (de novo synthesis rate/adenine nucleotide pool size) were highest in high oxidative muscle (0.82-1.06%/h), lowest in low oxidative muscle (0.30-0.35%/h), and intermediate in mixed muscle (0.44-0.55%/h). Our results demonstrate that differences in adenine nucleotide management among fiber types extends to the process of de novo adenine nucleotide synthesis

Reappraisal of VAChT-Cre: Preference in slow motor neurons innervating type I or IIa muscle fibers.

Science.gov (United States)

Misawa, Hidemi; Inomata, Daijiro; Kikuchi, Miseri; Maruyama, Sae; Moriwaki, Yasuhiro; Okuda, Takashi; Nukina, Nobuyuki; Yamanaka, Tomoyuki

2016-11-01

VAChT-Cre.Fast and VAChT-Cre.Slow mice selectively express Cre recombinase in approximately one half of postnatal somatic motor neurons. The mouse lines have been used in various studies with selective genetic modifications in adult motor neurons. In the present study, we crossed VAChT-Cre lines with a reporter line, CAG-Syp/tdTomato, in which synaptophysin-tdTomato fusion proteins are efficiently sorted to axon terminals, making it possible to label both cell bodies and axon terminals of motor neurons. In the mice, Syp/tdTomato fluorescence preferentially co-localized with osteopontin, a recently discovered motor neuron marker for slow-twitch fatigue-resistant (S) and fast-twitch fatigue-resistant (FR) types. The fluorescence did not preferentially co-localize with matrix metalloproteinase-9, a marker for fast-twitch fatigable (FF) motor neurons. In the neuromuscular junctions, Syp/tdTomato fluorescence was detected mainly in motor nerve terminals that innervate type I or IIa muscle fibers. These results suggest that the VAChT-Cre lines are Cre-drivers that have selectivity in S and FR motor neurons. In order to avoid confusion, we have changed the mouse line names from VAChT-Cre.Fast and VAChT-Cre.Slow to VAChT-Cre.Early and VAChT-Cre.Late, respectively. The mouse lines will be useful tools to study slow-type motor neurons, in relation to physiology and pathology. © 2016 Wiley Periodicals, Inc.

Predicting Effects of Tropomyosin Mutations on Cardiac Muscle Contraction through Myofilament Modeling

Directory of Open Access Journals (Sweden)

Lorenzo Rakesh Sewanan

2016-10-01

Full Text Available Point mutations to the human gene TPM1 have been implicated in the development of both hypertrophic and dilated cardiomyopathies. Such observations have led to studies investigating the link between single residue changes and the biophysical behavior of the tropomyosin molecule. However, the degree to which these molecular perturbations explain the performance of intact sarcomeres containing mutant tropomyosin remains uncertain. Here, we present a modeling approach that integrates various aspects of tropomyosin’s molecular properties into a cohesive paradigm representing their impact on muscle function. In particular, we considered the effects of tropomyosin mutations on (1 persistence length, (2 equilibrium between thin filament blocked and closed regulatory states, and (3 the crossbridge duty cycle. After demonstrating the ability of the new model to capture Ca-dependent myofilament responses during both dynamic and steady-state activation, we used it to capture the effects of hypertrophic cardiomyopathy (HCM related E180G and D175N mutations on skinned myofiber mechanics. Our analysis indicates that the fiber-level effects of the two mutations can be accurately described by a combination of changes to the three tropomyosin properties represented in the model. Subsequently, we used the model to predict mutation effects on muscle twitch. Both mutations led to increased twitch contractility as a consequence of diminished cooperative inhibition between thin filament regulatory units. Overall, simulations suggest that a common twitch phenotype for HCM-linked tropomyosin mutations includes both increased contractility and elevated diastolic tension.

A Ca2+-calmodulin-eEF2K-eEF2 signalling cascade, but not AMPK, contributes to the suppression of skeletal muscle protein synthesis during contractions

DEFF Research Database (Denmark)

Rose, Adam John; Alsted, Thomas Junker; Jensen, Thomas Elbenhardt

2009-01-01

Skeletal muscle protein synthesis rate decreases during contractions but the underlying regulatory mechanisms are poorly understood. It was hypothesised that there would be a coordinated regulation of eukaryotic elongation factor 2 (eEF2) and eukaryotic initiation factor 4E-binding protein 1 (4EBP1......) phosphorylation by signalling cascades downstream of rises in intracellular [Ca(2+)] and decreased energy charge via AMP activated protein kinase (AMPK) in contracting skeletal muscle. When fast-twitch skeletal muscles were contracted ex vivo using different protocols, the suppression of protein synthesis...... correlated more closely with changes in eEF2 rather than 4EBP1 phosphorylation. Using a combination of Ca(2+) release agents and ATPase inhibitors it was shown that the 60-70% suppression of fast-twitch skeletal muscle protein synthesis during contraction was equally distributed between Ca(2+) and energy...

[Acupuncture therapy for regaining consciousness in terms of acupoint location, needle insertion and needle manipulation].

Science.gov (United States)

Meng, Xianggang; Gu, Wenlong; Ma, Fen; Du, Yuzheng; Zhao, Qi

2015-03-01

Acupuncture therapy for regaining consciousness activates soreness, numbness, distention, heaviness, radiating and moving, electric shock and ant climbing sensations at the specific acupoints in the stroke patients. Radiating and moving sensations are the summary of needling sensations such as soreness, numbness and twitching presenting during lifting and thrusting manipulation. These sensations are the essential factors of the therapeutic effect of regaining consciousness. Radiating sensation refers to the conduction along meridians and radiation of soreness and numbness. Moving sensation refers to the local muscular twitching at acupoints and the involuntary movement of limbs, joints and the distal. Acupuncture at the specific acupoints achieves radiating and moving sensations for promoting the circulation in meridians, regulating qi and mind and balancing yin and yang in stroke patients. This therapy was introduced in the paper in view of acupoint location, needle insertion and manipulation.

GLUT4 expression in human muscle fibres is not correlated with intracellular triglyceride (TG) content. Is TG a maker or a marker of insulin resistance?

DEFF Research Database (Denmark)

Gaster, M; Ottosen, P D; Vach, W

2003-01-01

diabetic subjects, and young lean controls. TG density was significantly higher in slow compared to fast fibres in all studied subjects (pslow twitch fibres of obese diabetic subjects compared to obese (p...We have recently reported a progressive decline in the expression of glucose transporter isoform 4 (GLUT4) from control subjects through obese non-diabetics to obese type 2 diabetic subjects, indicating that the reduced GLUT4 in slow twitch fibres could be secondary to obesity. In this study we...... densities in slow and fast fibres did not correlate with the corresponding GLUT4 density in the same fibres in our study groups (p>0.05). Plasma TG and FFA did not correlate with GLUT4 expression in slow or fast fibres (p>0.05). In conclusion, TG content was increased in diabetic slow fibres with a reduced...

Limited effects of exogenous glucose during severe hypoxia and a lack of hypoxia-stimulated glucose uptake in isolated rainbow trout cardiac muscle

DEFF Research Database (Denmark)

Becker, Tracy A; Della Valle, Brian William; Gesser, Hans

2013-01-01

We examined whether exogenous glucose affects contractile performance of electrically paced ventricle strips from rainbow trout under conditions known to alter cardiomyocyte performance, ion regulation and energy demands. Physiological levels of d-glucose did not influence twitch force developmen...

GH/IGF-I Transgene Expression on Muscle Homeostasis

Science.gov (United States)

Schwartz, Robert J.

1999-01-01

We propose to test the hypothesis that the growth hormone/ insulin like growth factor-I axis through autocrine/paracrine mechanisms may provide long term muscle homeostasis under conditions of prolonged weightlessness. As a key alternative to hormone replacement therapy, ectopic production of hGH, growth hormone releasing hormone (GHRH), and IGF-I will be studied for its potential on muscle mass impact in transgenic mice under simulated microgravity. Expression of either hGH or IGF-I would provide a chronic source of a growth-promoting protein whose biosynthesis or secretion is shut down in space. Muscle expression of the IGF-I transgene has demonstrated about a 20% increase in hind limb muscle mass over control nontransgenic litter mates. These recent experiments, also establish the utility of hind-limb suspension in mice as a workable model to study atrophy in weight bearing muscles. Thus, transgenic mice will be used in hind-limb suspension models to determine the role of GH/IGF-I on maintenance of muscle mass and whether concentric exercises might act in synergy with hormone treatment. As a means to engineer and ensure long-term protein production that would be workable in humans, gene therapy technology will be used by to monitor muscle mass preservation during hind-limb suspension, after direct intramuscular injection of a genetically engineered muscle-specific vector expressing GHRH. Effects of this gene-based therapy will be assessed in both fast twitch (medial gastrocnemius) and slow twitch muscle (soleus). End-points include muscle size, ultrastructure, fiber type, and contractile function, in normal animals, hind limb suspension, and reambutation.

Voluntary resistance running with short distance enhances spatial memory related to hippocampal BDNF signaling.

Science.gov (United States)

Lee, Min Chul; Okamoto, Masahiro; Liu, Yu Fan; Inoue, Koshiro; Matsui, Takashi; Nogami, Haruo; Soya, Hideaki

2012-10-15

Although voluntary running has beneficial effects on hippocampal cognitive functions if done abundantly, it is still uncertain whether resistance running would be the same. For this purpose, voluntary resistance wheel running (RWR) with a load is a suitable model, since it allows increased work levels and resultant muscular adaptation in fast-twitch muscle. Here, we examined whether RWR would have potential effects on hippocampal cognitive functions with enhanced hippocampal brain-derived neurotrophic factor (BDNF), as does wheel running without a load (WR). Ten-week-old male Wistar rats were assigned randomly to sedentary (Sed), WR, and RWR (to a maximum load of 30% of body weight) groups for 4 wk. We found that in RWR, work levels increased with load, but running distance decreased by about half, which elicited muscular adaptation for fast-twitch plantaris muscle without causing any negative stress effects. Both RWR and WR led to improved spatial learning and memory as well as gene expressions of hippocampal BDNF signaling-related molecules. RWR increased hippocampal BDNF, tyrosine-related kinase B (TrkB), and cAMP response element-binding (CREB) protein levels, whereas WR increased only BDNF. With both exercise groups, there were correlations between spatial memory and BDNF protein (r = 0.41), p-CREB protein (r = 0.44), and work levels (r = 0.77). These results suggest that RWR plays a beneficial role in hippocampus-related cognitive functions associated with hippocampal BDNF signaling, even with short distances, and that work levels rather than running distance are more determinant of exercise-induced beneficial effects in wheel running with and without a load.

Multidrug-resistant Salmonella enterica serovar Typhimurium isolates are resistant to antibiotics that influence their swimming and swarming motility

Science.gov (United States)

Motile bacteria utilize one or more strategies for movement, such as darting, gliding, sliding, swarming, swimming, and twitching. The ability to move is considered a virulence factor in many pathogenic bacteria, including Salmonella. Multidrug-resistant (MDR) Salmonella encodes acquired factors t...

Crystallization and preliminary X-ray analysis of a decameric form of cytosolic thioredoxin peroxidase 1 (Tsa1), C47S mutant, from Saccharomyces cerevisiae

Energy Technology Data Exchange (ETDEWEB)

Oliveira, Marcos Antonio de, E-mail: scaff@lnls.br; Genu, Victor; Discola, Karen Fulan; Alves, Simone Vidigal; Netto, Luis Eduardo Soares [Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, 05508-900 São Paulo-SP (Brazil); Guimarães, Beatriz Gomes, E-mail: scaff@lnls.br [Centro de Biologia Molecular Estrutural, Laboratório Nacional de Luz Síncrotron, 13084-971 Campinas-SP (Brazil); Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo, 05508-900 São Paulo-SP (Brazil)

2007-08-01

A recombinant mutant (C47S) of cytosolic thioredoxin peroxidase 1 from S. cerevisiae was expressed, purified and crystallized by the hanging-drop vapour-diffusion method from protein previously treated with 1,4-dithiothreitol. The crystals belong to the monoclinic space group C2 and diffraction data were collected to 2.8 Å resolution using a synchrotron-radiation source. Saccharomyces cerevisiae cytosolic thioredoxin peroxidase 1 (cTPxI or Tsa1) is a bifunctional enzyme with protective roles in cellular defence against oxidative and thermal stress that exhibits both peroxidase and chaperone activities. Protein overoxidation and/or high temperatures induce great changes in its quaternary structure and lead to its assembly into large complexes that possess chaperone activity. A recombinant mutant of Tsa1 from S. cerevisiae, with Cys47 substituted by serine, was overexpressed in Escherichia coli as a His{sub 6}-tagged fusion protein and purified by nickel-affinity chromatography. Crystals were obtained from protein previously treated with 1,4-dithiothreitol by the hanging-drop vapour-diffusion method using PEG 3000 as precipitant and sodium fluoride as an additive. Diffraction data were collected to 2.8 Å resolution using a synchrotron-radiation source. The crystal structure was solved by molecular-replacement methods and structure refinement is currently in progress.

Crystal structures of the F and pSLT plasmid TraJ N-terminal regions reveal similar homodimeric PAS folds with functional interchangeability

Energy Technology Data Exchange (ETDEWEB)

Lu, Jun; Wu, Ruiying; Adkins, Joshua N.; Joachimiak, Andrzej; Glover, Mark

2014-09-16

In the F-family of conjugative plasmids, TraJ is an essential transcriptional activator of the tra operon that encodes most of the proteins required for conjugation. Here we report for the first time the X-ray crystal structures of the TraJ N-terminal regions from the prototypic F plasmid (TraJF11-130) and from the Salmonella virulence plasmid pSLT (TraJpSLT 1-128). Both proteins form similar homodimeric Per-ARNT-Sim (PAS) fold structures. Mutational analysis reveals that the observed dimeric interface is critical for TraJF transcriptional activation, indicating that dimerization of TraJ is required for its in vivo function. An artificial ligand (oxidized dithiothreitol) occupies a cavity in the TraJF dimer interface, while a smaller cavity in corresponding region of the TraJpSLT structure lacks a ligand. Gas chromatography/mass spectrometry-electron ionization analysis of dithiothreitol-free TraJF suggests indole may be the natural TraJ ligand; however, disruption of the indole biosynthetic pathway does not affect TraJF function. Heterologous PAS domains from pSLT and R100 TraJ can functionally replace the TraJF PAS domain, suggesting that TraJ allelic specificity is mediated by the region C-terminal to the PAS domain.

Gastrin receptor characterization: affinity cross-linking of the gastrin receptor on canine gastric parietal cells

International Nuclear Information System (INIS)

Matsumoto, M.; Park, J.; Yamada, T.

1987-01-01

The authors applied affinity cross-linking methods to label the gastrin receptor on isolated canine gastric parietal cells in order to elucidate the nature of its chemical structure. 125 I-labeled Leu 15 -gastrin and 125 I-labeled gastrin/sub 2-17/ bound to intact parietal cells and their membranes with equal affinity, and half-maximal inhibition of binding was obtained at an incubation concentration of 3.2 x 10 -10 M unlabeled gastrin. 125 I-gastrin/sub 2-17/ was cross-linked to plasma membranes or intact parietal cells by incubation in disuccinimidyl suberate. The membrane pellets were solubilized with or without dithiothreitol and applied to electrophoresis on 7.5% sodium dodecyl sulfate polyacrylamide gels. Autoradiograms revealed a band of labeling at M/sub r/ 76,000 and labeling of this band was inhibited in a dose-dependent fashion by addition of unlabeled gastrin to the incubation mixture. Dithiothreitol in concentrations as high as 100 mM did not later the electrophoretic mobility of the labeled band. After taking into account the molecular weight of 125 I-gastrin/sub 2-17/, the results suggest that the gastrin receptor on parietal cells is a single protein of M/sub r/ 74,000 without disulfide-linked subunits

Skeletal muscle, but not cardiovascular function, is altered in a mouse model of autosomal recessive hypophosphatemic rickets

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Michael J. Wacker

2016-05-01

Full Text Available Autosomal recessive hypophosphatemic rickets (ARHR is a heritable disorder characterized by hypophosphatemia, osteomalacia, and poor bone development. ARHR results from inactivating mutations in the DMP1 gene with the human phenotype being recapitulated in the Dmp1 null mouse model which displays elevated plasma fibroblast growth factor 23. While the bone phenotype has been well characterized, it is not known what effects ARHR may also have on skeletal, cardiac, or vascular smooth muscle function, which is critical to understand to treat patients suffering from this condition. In this study, the extensor digitorum longus (EDL- fast-twitch muscle, soleus (SOL- slow-twitch muscle, heart, and aorta were removed from Dmp1 null mice and ex-vivo functional tests were simultaneously performed in collaboration by three different laboratories. Dmp1 null EDL and SOL muscles produced less force than wildtype muscles after normalization for physiological cross sectional area of the muscles. Both EDL and SOL muscles from Dmp1 null mice also produced less force after the addition of caffeine (which releases calcium from the sarcoplasmic reticulum which may indicate problems in excitation contraction coupling in these mice. While the body weights of the Dmp1 null were smaller than wildtype, the heart weight to body weight ratio was higher. However, there were no differences in pathological hypertrophic gene expression compared to wildtype and maximal force of contraction was not different indicating that there may not be cardiac pathology under the tested conditions. We did observe a decrease in the rate of force development generated by cardiac muscle in the Dmp1 null which may be related to some of the deficits observed in skeletal muscle. There were no differences observed in aortic contractions induced by PGF2a or 5-HT or in endothelium-mediated acetylcholine-induced relaxations or endothelium-independent sodium nitroprusside-induced relaxations. In

High glycogen levels enhance glycogen breakdown in isolated contracting skeletal muscle

DEFF Research Database (Denmark)

Richter, Erik; Galbo, H

1986-01-01

and after 15 min of intermittent electrical muscle stimulation. Before stimulation, glycogen was higher in rats that swam on the preceding day (supercompensated rats) compared with controls. During muscle contractions, glycogen breakdown in fast-twitch red and white fibers was larger in supercompensated...

Genetics Home Reference: spinocerebellar ataxia type 3

Science.gov (United States)

... time, individuals with SCA3 may develop loss of sensation and weakness in the limbs (peripheral neuropathy), muscle cramps, muscle twitches (fasciculations), and swallowing difficulties. Individuals with SCA3 may have problems with memory, planning, and problem solving. Signs and symptoms of ...

Does interscalene catheter placement with stimulating catheters improve postoperative pain or functional outcome after shoulder surgery? A prospective, randomized and double-blinded trial

NARCIS (Netherlands)

Stevens, Markus F.; Werdehausen, Robert; Golla, Elisabeth; Braun, Sebastian; Hermanns, Henning; Ilg, Ansgar; Willers, Reinhardt; Lipfert, Peter

2007-01-01

BACKGROUND: In this prospective, randomized, double-blind trial we investigated the use of stimulating catheters in patients during and after shoulder surgery; functional improvement being the primary outcome measurement. METHODS: After eliciting an adequate muscular twitch at

Immobilization of free-ranging maned wolf (Chrysocyon brachyurus) with tiletamine and zolazepam in central Brazil.

Science.gov (United States)

Furtado, Mariana Malzoni; Kashivakura, Cynthia Kayo; Ferro, Claudia; de Almeida, Jácomo Anah Tereza; Silveira, Leandro; Astete, Samuel

2006-03-01

A tiletamine hydrochloride-zolazepam hydrochloride combination was used successfully to immobilize 27 free-ranging maned wolves (Chrysocyon brachyurus) at a mean dose of 2.77+/-0.56 (mean+/-SD) mg/kg. The induction time ranged from 3-15 min. Animals remained immobilized for periods of 48.56 +/-12.65 min. Compulsive licking, excessive salivation, muscle twitching, muscle tremors, tachypnea, and bradycardia were observed associated with the induction of the anesthesia in 13 of 27 maned wolves. Muscle twitching, pedal withdrawal reflex, muscle tremors, and ataxia were observed during recovery in three (11%) maned wolves. There were no significant differences in heart rates (P = 0.44), respiratory rates (P = 0.82), and rectal temperatures (P = 0.54) recorded at 5, 15, and 25 min after induction at these dosages. The tiletamine hydrochloride-zolazepam hydrochloride combination was shown to be an effective and safe immobilizing agent for free-ranging maned wolves.

Non-stationarity and power spectral shifts in EMG activity reflect motor unit recruitment in rat diaphragm muscle.

Science.gov (United States)

Seven, Yasin B; Mantilla, Carlos B; Zhan, Wen-Zhi; Sieck, Gary C

2013-01-15

We hypothesized that a shift in diaphragm muscle (DIAm) EMG power spectral density (PSD) to higher frequencies reflects recruitment of more fatigable fast-twitch motor units and motor unit recruitment is reflected by EMG non-stationarity. DIAm EMG was recorded in anesthetized rats during eupnea, hypoxia-hypercapnia (10% O(2)-5% CO(2)), airway occlusion, and sneezing (maximal DIAm force). Although power in all frequency bands increased progressively across motor behaviors, PSD centroid frequency increased only during sneezing (pmotor units were recruited during different motor behaviors. Motor units augmented their discharge frequencies progressively beyond the non-stationary period; yet, EMG signal became stationary. In conclusion, non-stationarity of DIAm EMG reflects the period of motor unit recruitment, while a shift in the PSD towards higher frequencies reflects recruitment of more fatigable fast-twitch motor units. Copyright © 2012 Elsevier B.V. All rights reserved.

The Functional Role of Calcineurin in Hypertrophy, Regeneration, and Disorders of Skeletal Muscle

Directory of Open Access Journals (Sweden)

Kunihiro Sakuma

2010-01-01

Full Text Available Skeletal muscle uses calcium as a second messenger to respond and adapt to environmental stimuli. Elevations in intracellular calcium levels activate calcineurin, a serine/threonine phosphatase, resulting in the expression of a set of genes involved in the maintenance, growth, and remodeling of skeletal muscle. In this review, we discuss the effects of calcineurin activity on hypertrophy, regeneration, and disorders of skeletal muscle. Calcineurin is a potent regulator of muscle remodeling, enhancing the differentiation through upregulation of myogenin or MEF2A and downregulation of the Id1 family and myostatin. Foxo may also be a downstream candidate for a calcineurin signaling molecule during muscle regeneration. The strategy of controlling the amount of calcineurin may be effective for the treatment of muscular disorders such as DMD, UCMD, and LGMD. Activation of calcineurin produces muscular hypertrophy of the slow-twitch soleus muscle but not fast-twitch muscles.

Toward a comprehensive hybrid physical-virtual reality simulator of peripheral anesthesia with ultrasound and neurostimulator guidance.

Science.gov (United States)

Samosky, Joseph T; Allen, Pete; Boronyak, Steve; Branstetter, Barton; Hein, Steven; Juhas, Mark; Nelson, Douglas A; Orebaugh, Steven; Pinto, Rohan; Smelko, Adam; Thompson, Mitch; Weaver, Robert A

2011-01-01

We are developing a simulator of peripheral nerve block utilizing a mixed-reality approach: the combination of a physical model, an MRI-derived virtual model, mechatronics and spatial tracking. Our design uses tangible (physical) interfaces to simulate surface anatomy, haptic feedback during needle insertion, mechatronic display of muscle twitch corresponding to the specific nerve stimulated, and visual and haptic feedback for the injection syringe. The twitch response is calculated incorporating the sensed output of a real neurostimulator. The virtual model is isomorphic with the physical model and is derived from segmented MRI data. This model provides the subsurface anatomy and, combined with electromagnetic tracking of a sham ultrasound probe and a standard nerve block needle, supports simulated ultrasound display and measurement of needle location and proximity to nerves and vessels. The needle tracking and virtual model also support objective performance metrics of needle targeting technique.

Nox4 Is Dispensable for Exercise Induced Muscle Fibre Switch.

Directory of Open Access Journals (Sweden)

Juri Vogel

Full Text Available By producing H2O2, the NADPH oxidase Nox4 is involved in differentiation of mesenchymal cells. Exercise alters the composition of slow and fast twitch fibres in skeletal. Here we hypothesized that Nox4 contributes to exercise-induced adaptation such as changes in muscle metabolism or muscle fibre specification and studied this in wildtype and Nox4-/- mice.Exercise, as induced by voluntary running in a running wheel or forced running on a treadmill induced a switch from fast twitch to intermediate fibres. However the induced muscle fibre switch was similar between Nox4-/- and wildtype mice. The same held true for exercise-induced expression of PGC1α or AMPK activation. Both are increased in response to exercise, but with no difference was observed between wildtype and Nox4-/- mice.Thus, exercise-induced muscle fibre switch is Nox4-independent.

L-acetylcarnitine enhances functional muscle re-innervation.

Science.gov (United States)

Pettorossi, V E; Brunetti, O; Carobi, C; Della Torre, G; Grassi, S

1991-01-01

The efficacy of L-acetylcarnitine and L-carnitine treatment on motor re-innervation was analyzed by evaluating different muscular parameters describing functional muscle recovery after denervation and re-innervation. The results show that L-acetylcarnitine markedly enhances functional muscle re-innervation, which on the contrary is unaffected by L-carnitine. The medial gastrocnemius muscle was denervated by cutting the nerve at the muscle entry point. After 20 days the sectioned nerve was resutured into the medial gastrocnemius muscle, and the extent of re-innervation was monitored 45 days later. L-acetylcarnitine-treated animals show significantly higher twitch and tetanic tensions of re-innervated muscle. Furthermore the results, obtained by analysing the twitch time to peak and tetanic contraction-relaxation times, suggest that L-acetylcarnitine mostly affects the functional re-innervation of slow motor units. The possible mechanisms by which L-acetylcarnitine facilitates such motor and nerve recovery are discussed.

Comprehensive glycan analysis of recombinant Aspergillus niger endo-polygalacturonase C.

Science.gov (United States)

Woosley, Bryan; Xie, Min; Wells, Lance; Orlando, Ron; Garrison, Derek; King, Daniel; Bergmann, Carl

2006-07-01

The enzyme PGC is produced by the fungus Aspergillus niger during invasion of plant cell walls. The enzyme has been homologously overexpressed to provide sufficient quantities of purified enzyme for biological studies. We have characterized this enzyme in terms of its posttranslational modifications (PTMs) and found it to be both N- and O-glycosylated. The glycosyl moieties have also been characterized. This has involved a combination of matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), liquid chromatography (LC)-ion trap, and LC-electrospray ionization (ESI) mass spectrometries in conjunction with trypsin degradation and beta-elimination, followed by Michael addition with dithiothreitol (BEMAD). This is the first demonstration of the ability of BEMAD to map glycosylation sites other than O-GlcNAc sites. The complete characterization of all PTMs on PGC allows us to model them on the peptide backbone, revealing potential roles played by the glycans in modulating the interaction of the enzyme with other macromolecules.

Novel Peptidase Kunitz Inhibitor from Platypodium elegans Seeds Is Active against Spodoptera frugiperda Larvae.

Science.gov (United States)

Ramalho, Suellen Rodrigues; Bezerra, Cézar da Silva; Lourenço de Oliveira, Daniella Gorete; Souza Lima, Letícia; Maria Neto, Simone; Ramalho de Oliveira, Caio Fernando; Valério Verbisck, Newton; Rodrigues Macedo, Maria Lígia

2018-02-14

A novel Kunitz-type inhibitor from Platypodium elegans seeds (PeTI) was purified and characterized. The mass spectrometry analyses of PeTI indicated an intact mass of 19 701 Da and a partial sequence homologous to Kunitz inhibitors. PeTI was purified by ion exchange and affinity chromatographies. A complex with a 1:1 ratio was obtained only for bovine trypsin, showing a K i = 0.16 nM. Stability studies showed that PeTI was stable over a wide range of temperature (37-80 °C) and pH (2-10). The inhibitory activity of PeTI was affected by dithiothreitol (DTT). Bioassays of PeTI on Spodoptera frugiperda showed negative effects on larval development and weight gain, besides extending the insect life cycle. The activities of digestive enzymes, trypsin and chymotrypsin, were reduced by feeding larvae with 0.2% PeTI in an artificial diet. In summary, we describe a novel Kunitz inhibitor with promising biotechnological potential for pest control.

Dehalogenation of lindane by a variety of porphyrins and corrins.

OpenAIRE

Marks, T S; Allpress, J D; Maule, A

1989-01-01

The dehalogenation of lindane by a range of hemoproteins, porphyrins, and corrins has been tested under reducing conditions in the presence of dithiothreitol. In addition, a series of porphyrin-metal ion complexes have been prepared and have also been screened for the capacity to dehalogenate lindane. Hemoglobin, hemin, hematin, and chlorophyll alpha all catalyzed the dehalogenation of lindane, as did all of the corrins tested. The porphyrins which did not contain metal centers--coproporphyri...

Effects of heavy metals on the neuromuscular transmission of the mouse diaphragm

Energy Technology Data Exchange (ETDEWEB)

Fu, W M; Shiau, S Y.L.

1978-04-01

Effects of heavy metals including Mn, Co, Ni, Cd, Zn, Cu, Sr, Ba, and UO/sub 2//sup +/ ions on the neuromuscular transmission of the mouse diaphragm were studied and compared. From the dose-inhibition curves, the concentrations (mM) required to inhibit 50% of the contraction (ID/sub 50/) for Cd, Mn, Co, Ni, Zn and Sr are 0.03, 0.8, 0.75, 0.82, 1.2 and >20 respectively. In addition to the potent neuromuscular blocking action, both Cd and Zn induce a contracture of the mouse diaphram. Among the cations tested, Cu is the most potent in inducing the contracture. Mn does not cause a contracture, while Co and Ni induce a contracture only after a prolonged incubation for 3 hours. The neuromuscular blocking action of most of the cations tested can be completely or partially reversed by either high Ca or cysteine except the irreversible action of Zn and Cu. These findings suggest that most divalent cations block the neuromuscular transmission by binding to the -SH group of the cell membrane and inhibiting Ca influx. On the other hand, both Ba and UO/sub 2/ at low concentration increase but at high concentration inhibit the twitch response. Sine Ba increases the twitch response of the mouse diaphragm stimulated directly in the presence of d-tubocurarine as well as that stimulated indirectly, Ba/sup + +/ acts mainly directly on the muscle. In contrast, UO/sub 2//sup +/ ions at low concentration increases the twitch response possibly by releasing acetylcholine from the nerve endings.

Phrenic motor unit recruitment during ventilatory and non-ventilatory behaviors.

Science.gov (United States)

Mantilla, Carlos B; Sieck, Gary C

2011-10-15

Phrenic motoneurons are located in the cervical spinal cord and innervate the diaphragm muscle, the main inspiratory muscle in mammals. Similar to other skeletal muscles, phrenic motoneurons and diaphragm muscle fibers form motor units which are the final element of neuromotor control. In addition to their role in sustaining ventilation, phrenic motor units are active in other non-ventilatory behaviors important for airway clearance such as coughing or sneezing. Diaphragm muscle fibers comprise all fiber types and are commonly classified based on expression of contractile proteins including myosin heavy chain isoforms. Although there are differences in contractile and fatigue properties across motor units, there is a matching of properties for the motor neuron and muscle fibers within a motor unit. Motor units are generally recruited in order such that fatigue-resistant motor units are recruited earlier and more often than more fatigable motor units. Thus, in sustaining ventilation, fatigue-resistant motor units are likely required. Based on a series of studies in cats, hamsters and rats, an orderly model of motor unit recruitment was proposed that takes into consideration the maximum forces generated by single type-identified diaphragm muscle fibers as well as the proportion of the different motor unit types. Using this model, eupnea can be accomplished by activation of only slow-twitch diaphragm motor units and only a subset of fast-twitch, fatigue-resistant units. Activation of fast-twitch fatigable motor units only becomes necessary when accomplishing tasks that require greater force generation by the diaphragm muscle, e.g., sneezing and coughing. Copyright © 2011 Elsevier B.V. All rights reserved.

Effects of intra-arterial epinephrine on energy metabolism in exercising rabbit gastrocnemius muscle, studied by in vivo phosphorus nuclear magnetic resonance.

Science.gov (United States)

Argov, Z; Nioka, S; Eleff, S; Chance, B

1991-10-01

Epinephrine has an inotropic effect on skeletal muscle, especially on glycolytic type 2 fibers. The mechanism of this effect is not completely clear and its association with a change in oxidative metabolism or glycolytic activation was not fully investigated. Epinephrine's effects on muscle bioenergetics were studied by in vivo 31P nuclear magnetic resonance to find if mitochondrial metabolism is changed during the inotropic action and if the known glycolytic activation by epinephrine is operative during muscle twitch. The study was also used as a model for the application of in vivo 31P nuclear magnetic resonance in the evaluation of short-term acting drugs. When injected intra-arterially, epinephrine (1 micrograms/kg) augmented the twitch tension of indirectly stimulated, continuously working rabbit gastrocnemius muscle by 15.4 + 6.5%. This increase in work was associated with reduction of phosphocreatine to inorganic phosphate ratio (PCr/Pi) from 3.4 to 2.1 without change in ATP levels. Intracellular pH was reduced from 6.9 to 6.75, but no accumulation of glycolytic intermediates could be observed. The increase in work was not associated with a rise in ADP. All these changes occurred for a few minutes only. The findings suggest that epinephrine's inotropic action is not mediated by a change in mitochondrial metabolism. Glycolytic activation by epinephrine occurs even during twitch and contributes partly to the energy demands of the augmented force. Epinephrine's inotropic effect is, however, not primarily due to changes in bioenergetic kinetics, but to effects on force generating mechanisms, with secondary reduction in energy state.

The expression of NFATc1 in adult rat skeletal muscle fibres.

Science.gov (United States)

Mutungi, Gabriel

2008-03-01

Although numerous studies have recently implicated the calcineurin-nuclear factor of activated T-cells (Cn-NFAT) signalling pathway in the regulation of activity-dependent fibre type switching in adult mammalian skeletal muscles, little is known about the endogenous expression of NFAT proteins in the various fibre types present in these muscles. In this study, the immunolocalization of NFATc1 (also known as NFATc or NFAT2) in the extensor digitorum longus (EDL; a mainly fast-twitch muscle) and the soleus (a predominantly slow-twitch muscle) muscles of adult ( approximately 90-day-old) Wistar rats was investigated. The results show that NFATc1 is expressed only in oxidative fibres (i.e. type I and type IIA fibres) that stain intensely for succinate dehydrogenase activity irrespective of whether they are from the fast- or slow-twitch muscle. Thus, 99 +/- 4% (n = 7 rats) of the muscle fibres in the soleus and 42 +/- 2% (n = 7 rats) of those in the EDL expressed NFATc1. In the soleus muscle fibres, NFATc1 was localized mainly in the fibre nuclei, whereas in the EDL fibres it was localized in both the cytoplasm and the nuclei. However, no difference in its localization was observed between type I and type IIA fibres in both muscles. Western blot experiments showed that the soleus expressed more NFATc1 proteins than the EDL. From these results, we suggest that NFATc1 controls the number and distribution of both type I and type IIA fibres, as well as the oxidative capacity of adult mammalian skeletal muscles.

Rocuronium duration of action under sevoflurane, desflurane or propofol anaesthesia.

Science.gov (United States)

Maidatsi, P G; Zaralidou, A Th; Gorgias, N K; Amaniti, E N; Karakoulas, K A; Giala, M M

2004-10-01

We conducted a prospective randomized study to evaluate whether the duration of action of a single bolus dose of rocuronium is influenced by maintenance of anaesthesia with sevoflurane, desflurane or propofol infusion. Fifty-seven ASA I-II patients undergoing elective abdominal surgery were enrolled in this study. Anaesthesia was induced with thiopental 3-5 mg kg(-1) or propofol 2.5 mg kg(-1) and fentanyl 5 microg kg(-1) and tracheal intubation was facilitated with rocuronium 0.9 mg kg(-1). Thereafter patients were randomly allocated to three different groups to receive sevoflurane, desflurane or propofol for maintenance of anaesthesia. Recovery of neuromuscular function was monitored by single twitch stimulation of the ulnar nerve and by recording the adductor pollicis response using accelerometry. Intergroup recovery times to 5% of control value of single twitch were analysed using analysis of variance with Bonferroni correction. The mean (95% confidence interval) recovery time to 5% of control value of single twitch during desflurane anaesthesia was 90.18 (86.11-94.25) min. Significantly shorter recovery times were observed during sevoflurane or propofol anaesthesia, 58.86 (54.73-62.99) min and 51.11 (45.47-56.74) min, respectively (P < 0.001). There were also significant differences in the recovery time between groups receiving desflurane vs. sevoflurane (P < 0.001) and desflurane vs. propofol (P < 0.001). Desflurane anaesthesia significantly prolongs the duration of action of rocuronium at 0.9 mg kg(-1) single bolus dose, compared to sevoflurane or propofol anaesthesia maintenance regimens.

Thoracoscopic patch insulation to correct phrenic nerve stimulation secondary to cardiac resynchronization therapy.

Science.gov (United States)

Mediratta, Neeraj; Barker, Diane; McKevith, James; Davies, Peter; Belchambers, Sandra; Rao, Archana

2012-07-01

Cardiac resynchronization therapy is an established therapy for heart failure, improving quality of life and prognosis. Despite advances in technique, available leads and delivery systems, trans-venous left ventricular (LV) lead positioning remains dependent on the patient's underlying venous anatomy. The left phrenic nerve courses over the surface of the pericardium laterally and may be stimulated by the LV pacing lead, causing uncomfortable diaphragmatic twitch. This paper describes a video-assisted thoracoscopic (VATS) procedure to correct phrenic nerve stimulation secondary to cardiac resynchronization therapy. Most current ways of avoiding phrenic stimulation involve either electronic reprogramming to distance the phrenic nerve from the stimulation circuit or repositioning the lead. We describe a case where the phrenic nerve was surgically insulated from the stimulating current by insinuating a patch of bovine pericardium between the epicardium and native pericardium of the heart thus completely resolving previously intolerable and incessant diaphragmatic twitch. The procedure was performed under general anaesthesia with single-lung ventilation and minimal use of neuromuscular blocking agents. Surgical patch insulation of the phrenic nerve was performed using minimally invasive VATS surgery, as a short-stay procedure, with no complications. No diaphragmatic twitch occurred post-surgery and the patient continued to gain symptomatic benefit from cardiac synchronization therapy (New York Heart Association Class III to II), enabling return to work. In cases where the trans-venous position of a LV lead is limited by troublesome phrenic nerve stimulation, thoracoscopic surgical patch insulation of the phrenic nerve could be considered to allow beneficial cardiac resynchronization therapy.

Rice Seed Priming with Picomolar Rutin Enhances Rhizospheric Bacillus subtilis CIM Colonization and Plant Growth.

Directory of Open Access Journals (Sweden)

Akanksha Singh

Full Text Available The effect of rutin, a bioflavonoid on the growth and biofilm formation of Bacillus subtilis strain CIM was investigated. In addition to swimming, swarming, and twitching potentials of B. subtilis CIM (BS, one picomolar (1 pM of rutin was also observed to boost the biofilm forming ability of the bacterium. Bio-priming of rice seeds with BS and rutin not only augmented root and shoot lengths but also the photosynthetic pigments like chlorophyll and carotenoid. Similarly, high accumulation of phenolic and flavonoid contents was observed in the leaves. Fluorescent microscopic images revealed that BS plus rutin enhanced callose deposition in the leaves. It was also established that the least formation of reactive oxygen species in BS plus rutin treated rice plants was due to higher free radicals scavenging activity and total antioxidant potential. The results highlight chemo attractant nature of BS towards rutin, which by enhancing biofilm formation and root colonization indirectly strengthened the plants' defensive state.

Anti-quorum sensing activity of some medicinal plants | Al-Haidari ...

African Journals Online (AJOL)

On the other hand, Psidium guajava and Mentha longifolia extracts showed lower QSI activity. These extracts exhibited significant elimination of pyocyanin formation and biofilm development of Pseudomonas aeruginosa PA14. In addition, they significantly inhibited twitching and swimming motilities of P. aeruginosa PA14.

Myopathy

Science.gov (United States)

... alternating episodes of twitching and stiffness; and stiff-man syndrome: characterized by episodes of rigidity and reflex spasms common muscle cramps and stiffness, and tetany: characterized by prolonged spasms of the arms and legs × Definition The myopathies are neuromuscular disorders in which the ...

Rhamnolipid but not motility is associated with the initiation of biofilm ...

Indian Academy of Sciences (India)

2013-01-10

Jan 10, 2013 ... In this study, confocal scanning laser microscope combined with .... Images were obtained by ... plates were inverted and incubated at 37°C for 24 h. .... and twitching motility of PAO1; d, e and f represent the motility of PA17.

The effect of antibiotics on swimming and swarming motility of multidrug-resistant Salmonella enterica serovar Typhimurium

Science.gov (United States)

Motile bacteria can employ one or more different strategies to move, including swimming, swarming, twitching, gliding, sliding, and darting. Swimming is considered the movement of individual bacteria through a liquid or semi-solid medium, while swarming is the concerted movement of a group of bacte...

The final common pathway in postural control - Developmental perspective

NARCIS (Netherlands)

Kernell, D

A brief review is given concerning postural specialisations among mammalian muscle fibres and motor units. Most skeletal muscles contain a mixture of fibres with different characteristics, and their slow-twitch (S) units are well-known to possess properties suitable for postural tasks: they are

Differences in motor evoked potentials induced in rats by transcranial magnetic stimulation under two separate anesthetics: implications for plasticity studies

Directory of Open Access Journals (Sweden)

Matthew Sykes

2016-10-01

Full Text Available Repetitive transcranial magnetic stimulation (rTMS is primarily used in humans to change the state of corticospinal excitability. To assess the efficacy of different rTMS stimulation protocols, motor evoked potentials (MEPs are used as a readout due to their non-invasive nature. Stimulation of the motor cortex produces a response in a targeted muscle, and the amplitude of this twitch provides an indirect measure of the current state of the cortex. When applied to the motor cortex, rTMS can alter MEP amplitude, however results are variable between participants and across studies. In addition, the mechanisms underlying any change and its locus are poorly understood. In order to better understand these effects, MEPs have been investigated in vivo in animal models, primarily in rats. One major difference in protocols between rats and humans is the use of general anesthesia in animal experiments. Anesthetics are known to affect plasticity-like mechanisms and so may contaminate the effects of an rTMS protocol. In the present study, we explored the effect of anesthetic on MEP amplitude, recorded before and after intermittent theta burst stimulation (iTBS, a patterned rTMS protocol with reported facilitatory effects. MEPs were assessed in the brachioradialis muscle of the upper forelimb under two anesthetics: a xylazine/zoletil combination and urethane. We found MEPs could be induced under both anesthetics, with no differences in the resting motor threshold or the average baseline amplitudes. However, MEPs were highly variable between animals under both anesthetics, with the xylazine/zoletil combination showing higher variability and most prominently a rise in amplitude across the baseline recording period. Interestingly, application of iTBS did not facilitate MEP amplitude under either anesthetic condition. Although it is important to underpin human application of TMS with mechanistic examination of effects in animals, caution must be taken when

Differences in Motor Evoked Potentials Induced in Rats by Transcranial Magnetic Stimulation under Two Separate Anesthetics: Implications for Plasticity Studies.

Science.gov (United States)

Sykes, Matthew; Matheson, Natalie A; Brownjohn, Philip W; Tang, Alexander D; Rodger, Jennifer; Shemmell, Jonathan B H; Reynolds, John N J

2016-01-01

Repetitive transcranial magnetic stimulation (rTMS) is primarily used in humans to change the state of corticospinal excitability. To assess the efficacy of different rTMS stimulation protocols, motor evoked potentials (MEPs) are used as a readout due to their non-invasive nature. Stimulation of the motor cortex produces a response in a targeted muscle, and the amplitude of this twitch provides an indirect measure of the current state of the cortex. When applied to the motor cortex, rTMS can alter MEP amplitude, however, results are variable between participants and across studies. In addition, the mechanisms underlying any change and its locus are poorly understood. In order to better understand these effects, MEPs have been investigated in vivo in animal models, primarily in rats. One major difference in protocols between rats and humans is the use of general anesthesia in animal experiments. Anesthetics are known to affect plasticity-like mechanisms and so may contaminate the effects of an rTMS protocol. In the present study, we explored the effect of anesthetic on MEP amplitude, recorded before and after intermittent theta burst stimulation (iTBS), a patterned rTMS protocol with reported facilitatory effects. MEPs were assessed in the brachioradialis muscle of the upper forelimb under two anesthetics: a xylazine/zoletil combination and urethane. We found MEPs could be induced under both anesthetics, with no differences in the resting motor threshold or the average baseline amplitudes. However, MEPs were highly variable between animals under both anesthetics, with the xylazine/zoletil combination showing higher variability and most prominently a rise in amplitude across the baseline recording period. Interestingly, application of iTBS did not facilitate MEP amplitude under either anesthetic condition. Although it is important to underpin human application of TMS with mechanistic examination of effects in animals, caution must be taken when selecting an

Glycogen depletion and resynthesis during 14 days of chronic low-frequency stimulation of rabbit muscle

DEFF Research Database (Denmark)

Prats, C; Bernal, C; Cadefau, J A

2002-01-01

Electro-stimulation alters muscle metabolism and the extent of this change depends on application intensity and duration. The effect of 14 days of chronic electro-stimulation on glycogen turnover and on the regulation of glycogen synthase in fast-twitch muscle was studied. The results showed that...

Persistent effects of chronic clozapine on the cellular and behavioral responses to LSD in mice

Science.gov (United States)

Moreno, José L.; Holloway, Terrell; Umali, Adrienne; Rayannavar, Vinayak; Sealfon, Stuart C.

2013-01-01

Rationale In schizophrenia patients, optimal treatment with antipsychotics requires weeks to months of sustained drug therapy. However, single administration of antipsychotic drugs can reverse schizophrenia-like behavioral alterations in rodent models of psychosis. This raises questions about the physiological relevance of such antipsychotic-like activity. Objective This study evaluates the effects of chronic treatment with clozapine on the cellular and behavioral responses induced by the hallucinogenic serotonin 5-HT2A receptor agonist lysergic acid diethylamide (LSD) as a mouse model of psychosis. Method Mice were treated chronically (21 days) with 25 mg/kg/day clozapine. Experiments were conducted 1, 7, 14, and 21 days after the last clozapine administration. [3H]Ketanserin binding and 5-HT2A mRNA expression were determined in mouse somatosensory cortex. Head-twitch behavior, expression of c-fos, which is induced by all 5-HT2A agonists, and expression of egr-1 and egr-2, which are LSD-like specific, were assayed. Results Head-twitch response was decreased and [3H]ketanserin binding was downregulated in 1, 7, and 14 days after chronic clozapine. 5-HT2A mRNA was reduced 1 day after chronic clozapine. Induction of c-fos, but not egr-1 and egr-2, was rescued 7 days after chronic clozapine. These effects were not observed after short treatment (2 days) with clozapine or chronic haloperidol (1 mg/kg/day). Conclusion Our findings provide a murine model of chronic atypical antipsychotic drug action and suggest downregulation of the 5-HT2A receptor as a potential mechanism involved in these persistent therapeutic-like effects. PMID:22842765

Contractile properties of motor units and expression of myosin heavy chain isoforms in rat fast-type muscle after volitional weight-lifting training.

Science.gov (United States)

Łochyński, Dawid; Kaczmarek, Dominik; Mrówczyński, Włodzimierz; Warchoł, Wojciech; Majerczak, Joanna; Karasiński, Janusz; Korostyński, Michał; Zoladz, Jerzy A; Celichowski, Jan

2016-10-01

Dynamic resistance training increases the force and speed of muscle contraction, but little is known about modifications to the contractile properties of the main physiological types of motor units (MUs) that contribute to these muscle adaptations. Although the contractile profile of MU muscle fibers is tightly coupled to myosin heavy chain (MyHC) protein expression, it is not well understood if MyHC transition is a prerequisite for modifications to the contractile characteristics of MUs. In this study, we examined MU contractile properties, the mRNA expression of MyHC, parvalbumin, and sarcoendoplasmic reticulum Ca 2+ pump isoforms, as well as the MyHC protein content after 5 wk of volitional progressive weight-lifting training in the medial gastrocnemius muscle in rats. The training had no effect on MyHC profiling or Ca 2+ -handling protein gene expression. Maximum force increased in slow (by 49%) and fast (by 21%) MUs. Within fast MUs, the maximum force increased in most fatigue-resistant and intermediate but not most fatigable MUs. Twitch contraction time was shortened in slow and fast fatigue-resistant MUs. Twitch half-relaxation was shortened in fast most fatigue-resistant and intermediate MUs. The force-frequency curve shifted rightward in fast fatigue-resistant MUs. Fast fatigable MUs fatigued less within the initial 15 s while fast fatigue-resistant units increased the ability to potentiate the force within the first minute of the standard fatigue test. In conclusion, at the early stage of resistance training, modifications to the contractile characteristics of MUs appear in the absence of MyHC transition and the upregulation of Ca 2+ -handling genes. Copyright © 2016 the American Physiological Society.

Histochemical and functional parameters in Nordic combination athletes.

Science.gov (United States)

Matolín, S; Vaverka, F; Lunák, J; Novák, J; Horák, V; Krejcí, P

1994-01-01

Bioptic samples from the vastus lateralis muscle were analyzed in a group of Czechoslovak representatives in the Nordic combination (ski-jumping and 15 km cross-country skiing). The distribution of individual muscle fibre types (FG, FOG and SO) was detected and correlated with values obtained by motor and functional performance tests. Histochemical analysis of the bioptic samples revealed a considerably heterogeneous distribution of muscle fibre types in the group studied. No typical profilation for this sport discipline was found. Weak correlation between the proportion of fast muscle fibres and explosive strength parameters was ascertained. The correlation between the proportion of slow muscle fibres and the capacity of O2 utilization (VO2max) was statistically significant. Strong correlation between the proportion of fast twitch fibres and relative maximal strength of knee extensors (N/kg) was disclosed. A non-linear relation between the area of fast twitch fibres and vigour of take-off was found.

EFFECTS OF PSYCHOTROPIC DRUGS AS BACTERIAL EFFLUX PUMP INHIBITORS ON QUORUM SENSING REGULATED BEHAVIORS

Directory of Open Access Journals (Sweden)

Aynur Aybey

2014-10-01

Full Text Available Psychotropic drugs are known to have antimicrobial activity against several groups of microorganisms. The antidepressant agents such as duloxetine, paroxetine, hydroxyzine and venlafaxine are shown to act as efflux pump inhibitors in bacterial cells. In order to the investigation of the effects of psychotropic drugs were determined for clinically significant pathogens by using standart broth microdillusion method. The anti-quorum sensing (anti-QS activity of psychotropic drugs was tested against four test pathogens using the agar well diffusion method. All drugs showed strong inhibitory effect on the growth of S. typhimurium. Additionally, quorum sensing-regulated behaviors of Pseudomonas aeruginosa, including swarming, swimming and twitching motility and alkaline protease production were investigated. Most effective drugs on swarming, swimming and twitching motility and alkaline protease production, respectively, were paroxetine and duloxetine; duloxetine; hydroxyzine and venlafaxine; paroxetine and venlafaxine; venlafaxine. Accordingly, psychotropic drugs were shown strongly anti-QS activity by acting as bacterial efflux pump inhibitors and effection on motility and alkaline protease production of P. aeruginosa.

Tbx15 controls skeletal muscle fibre-type determination and muscle metabolism

Science.gov (United States)

Lee, Kevin Y.; Singh, Manvendra K.; Ussar, Siegfried; Wetzel, Petra; Hirshman, Michael F.; Goodyear, Laurie J.; Kispert, Andreas; Kahn, C. Ronald

2015-01-01

Skeletal muscle is composed of both slow-twitch oxidative myofibers and fast-twitch glycolytic myofibers that differentially impact muscle metabolism, function and eventually whole-body physiology. Here we show that the mesodermal transcription factor T-box 15 (Tbx15) is highly and specifically expressed in glycolytic myofibers. Ablation of Tbx15 in vivo leads to a decrease in muscle size due to a decrease in the number of glycolytic fibres, associated with a small increase in the number of oxidative fibres. This shift in fibre composition results in muscles with slower myofiber contraction and relaxation, and also decreases whole-body oxygen consumption, reduces spontaneous activity, increases adiposity and glucose intolerance. Mechanistically, ablation of Tbx15 leads to activation of AMPK signalling and a decrease in Igf2 expression. Thus, Tbx15 is one of a limited number of transcription factors to be identified with a critical role in regulating glycolytic fibre identity and muscle metabolism. PMID:26299309

Fish protein intake induces fast-muscle hypertrophy and reduces liver lipids and serum glucose levels in rats.

Science.gov (United States)

Kawabata, Fuminori; Mizushige, Takafumi; Uozumi, Keisuke; Hayamizu, Kohsuke; Han, Li; Tsuji, Tomoko; Kishida, Taro

2015-01-01

In our previous study, fish protein was proven to reduce serum lipids and body fat accumulation by skeletal muscle hypertrophy and enhancing basal energy expenditure in rats. In the present study, we examined the precise effects of fish protein intake on different skeletal muscle fiber types and metabolic gene expression of the muscle. Fish protein increased fast-twitch muscle weight, reduced liver triglycerides and serum glucose levels, compared with the casein diet after 6 or 8 weeks of feeding. Furthermore, fish protein upregulated the gene expressions of a fast-twitch muscle-type marker and a glucose transporter in the muscle. These results suggest that fish protein induces fast-muscle hypertrophy, and the enhancement of basal energy expenditure by muscle hypertrophy and the increase in muscle glucose uptake reduced liver lipids and serum glucose levels. The present results also imply that fish protein intake causes a slow-to-fast shift in muscle fiber type.

Muscle glycogen depletion and lactate concentration during downhill skiing.

Science.gov (United States)

Tesch, P; Larsson, L; Eriksson, A; Karlsson, J

1978-01-01

Skilled and unskilled skiers were studied during downhill skiing. Muscle glycogen and muscle lactate concentrations in the vastus lateralis muscle were determined following different skiing conditions. Heavy glycogen utilization was found in the groups studied during a day of skiing. The skilled and unskilled skiers differed with respect to selective glycogen depletion pattern and the skilled subjects demonstrated greater depletion of slow twitch fibers than the unskilled subjects. Lactate concentrations ranged from approximately 5-26 mmoles x kg-1 wet muscle after approximately one minute of maximal skiing. This wide range was not found to be related to the level of skiing proficiency. However, skiing with varyingly angled boots, resulting in different knee angles, did affect lactate concentration. Lactate concentration was positively correlated to individual muscle fiber composition expressed as a percent of fast twitch fibers. The results suggest more pronounced involvement of aerobic energy metabolism in skilled skiers than in unskilled skiers.

Manual of Hemorrhagic Fever with Renal Syndrome

Science.gov (United States)

1989-04-01

added to form 2,000 ml solution. (3) Fresh guinea pig serum (GPS), which may be stored at -80’ C. (4) Dithiothreitol solution (DTT), 3 mg/ml, which...named Seoul virus, has been serol gically identified in Africa, North and South America, India, te Ihighlands of New Guinea , 3 Y/ j5 HFRS several...kalium and protein free and essentially natrium-free, is recommended. If parenteral alimentation is necessary, the usual treatment is to infuse

Markers of Ovarian Cancer Using a Glycoprotein/Antibody Array

Science.gov (United States)

2014-05-01

Article dx.doi.org/10.1021/pr400169n | J. Proteome Res. 2013, 12, 3342−33523350 osteopontin fragments for ovarian cancer in urine . Clin. Cancer Res. 2006...absorbent under urea /dithiothreitol denatured environment. Anal. Biochem. 2010, 398 (1), 34−44. (26) Chiu, P. C.; Chung, M. K.; Koistinen, R.; Koistinen... Synthesis and functional analyses of nuclear clusterin, a cell death protein. J. Biol. Chem. 2003, 278 (13), 11590−600. (60) Hassan, M. K.; Watari, H

Assay of vitamin B12

International Nuclear Information System (INIS)

Tovey, K.C.; Carrick, D.T.

1982-01-01

A radioassay is described for vitamin B12 which involves denaturing serum protein binding proteins with alkali. In the denaturation step a dithiopolyol and cyanide are used and in the intrinsic factor assay step a vitamin B12 analogue such as cobinamide is used to bind with any remaining serum proteins. The invention also includes a kit in which the dithiopolyol is provided in admixture with the alkali. The dithiopolyol may be dithiothreitol or dithioerythritol. (author)

Respiratory Responses to Stimulation of Branchial Vagus Nerve Ganglia of a Teleost Fish

NARCIS (Netherlands)

Ballintijn, C.M.; Luiten, P.G.M.

1983-01-01

The effects of electrical stimulation of epibranchial vagus ganglia upon respiration of the carp were investigated. Single shocks evoked fast twitch responses in a number of respiratory muscles with latencies around 18 msec to the beginning and 30-35 msec to the peak of activity. Shocks given during

Sepsis Strengthens Antagonistic Actions of Neostigmine on Rocuronium in a Rat Model of Cecal Ligation and Puncture

Directory of Open Access Journals (Sweden)

Jin Wu

2016-01-01

Conclusions: Sepsis strengthened the antagonistic actions of neostigmine on Roc-depressed twitch tension of the diaphragm by inhibiting the activity of AChE in the NMJ. The reduced content of AChE might be one of the possible causes of the decreased AChE activity in the NMJ.

(TNNC1) gene in goat

African Journals Online (AJOL)

Yomi

2012-02-23

Feb 23, 2012 ... (soleus), but was not expressed in fast skeletal muscle (longissimus muscle, gluteus maximus) and brain, kidney, lung ... Muscle fibre can be classified according to their ... muscle and TNNC1 express in slow skeletal muscle and cardiac ..... and expression of the human slow twitch skeletal muscle/cardiac.

Intraneural stimulation using wire-microelectrode arrays: analysis of force steps in recruitment curves

NARCIS (Netherlands)

Smit, J.P.A.; Rutten, Wim; Boom, H.B.K.

1996-01-01

In acute experiments on six Wistar rats, a wire-microelectrode array was inserted into the common peroneal nerve. A 5-channel array and a 24-channel array were available. Each electrode in the array was used to generate a twitch contraction force recruitment curve for the extensor digitorum longus

Studying somatosensory function in Parkinson’s disease using Magnetoencephalography

DEFF Research Database (Denmark)

Sridharan, Kousik Sarathy; Johnsen, Erik Lisbjerg; Beniczky, Sándor

-stimulus interval of 350 ms. Current was setat the level of an observable twitch of the thumb. MEGdata were filtered using MaxFilter, a software that implements the tSSS(Temporal signal space separation) algorithm to remove DBS artefacts. The datais processed using the Fieldtrip toolbox. After preprocessing steps...

Electrical stimulation to the trigeminal proprioceptive fibres that innervate the mechanoreceptors in Müller's muscle induces involuntary reflex contraction of the frontalis muscles.

Science.gov (United States)

Matsuo, Kiyoshi; Osada, Yoshiro; Ban, Ryokuya

2013-02-01

The levator and frontalis muscles lack interior muscle spindles, despite consisting of slow-twitch fibres that involuntarily sustain eyelid-opening and eyebrow-raising against gravity. To compensate for this anatomical defect, this study hypothetically proposes that initial voluntary contraction of the levator fast-twitch muscle fibres stretches the mechanoreceptors in Müller's muscle and evokes proprioception, which continuously induces reflex contraction of slow-twitch fibres of the levator and frontalis muscles. This study sought to determine whether unilateral transcutaneous electrical stimulation to the trigeminal proprioceptive fibres that innervate the mechanoreceptors in Müller's muscle could induce electromyographic responses in the frontalis muscles, with monitoring responses in the orbicularis oculi muscles. The study population included 27 normal subjects and 23 subjects with aponeurotic blepharoptosis, who displayed persistently raised eyebrows on primary gaze and light eyelid closure. The stimulation induced a short-latency response in the ipsilateral frontalis muscle of all subjects and long-latency responses in the bilateral frontalis muscles of normal subjects. However, it did not induce long-latency responses in the bilateral frontalis muscles of subjects with aponeurotic blepharoptosis. The orbicularis oculi muscles showed R1 and/or R2 responses. The stimulation might reach not only the proprioceptive fibres, but also other sensory fibres related to the blink or corneal reflex. The experimental system can provoke a monosynaptic short-latency response in the ipsilateral frontalis muscle, probably through the mesencephalic trigeminal proprioceptive neuron and the frontalis motor neuron, and polysynaptic long-latency responses in the bilateral frontalis muscles through an unknown pathway. The latter neural circuit appeared to be engaged by the circumstances of aponeurotic blepharoptosis.

Plasticity of the transverse tubules following denervation and subsequent reinnervation in rat slow and fast muscle fibres.

Science.gov (United States)

Takekura, Hiroaki; Tamaki, Hiroyuki; Nishizawa, Tomie; Kasuga, Norikatsu

2003-01-01

We have studied the effects of short term denervation followed by reinnervation on the ultrastructure of the membrane systems and on the content of and distribution of key proteins involved in calcium regulation of fast-twitch (FT) extensor digitorum longus (EDL) and slow-twitch (ST) soleus (SOL) muscle fibres. Ischiadic nerve freezing resulted in total lack of neuromuscular transmission for 3 days followed by a slow recovery, but no decline in twitch force elicited by direct stimulation. The latter measurements indicate no significant atrophy within this time frame. The membrane systems of skeletal muscle fibres were visualized using Ca92+)-K3Fe(CN)6-OsO4 techniques and observed using a high voltage electron microscope. [3H]nitrendipine binding was used to detect levels of dihydropyridine receptor (DHPR) expression. The Ca2+ pumping free sarcoplasmic reticulum domains were not affected by the denervation, but the Ca2+ release domains were dramatically increased, particularly in the FT-EDL muscle fibres. The increase is evidenced by a doubling up of the areas of contacts between SR and transverse (t-) tubules, so that in place of the normal triadic arrangement, pentadic and heptadic junctions, formed by multiple interacting layers of ST and t-tubules are seen. Frequency of pentads and heptads increases and declines in parallel to the denervation and reinnervation but with a delay. Immunofluorecence and electron microscopy observations show presence of DHPR and ryanodine receptor clusters at pentads and heptads junctions. A significant (P muscle fibres indicating that overexpression of DHPRs accompanies the build up extra junctional contacts. The results indicate that denervation reversibly affects the domains of the membrane systems involved in excitation-contraction coupling.

Changes in contractile activation characteristics of rat fast and slow skeletal muscle fibres during regeneration.

Science.gov (United States)

Gregorevic, Paul; Plant, David R; Stupka, Nicole; Lynch, Gordon S

2004-07-15

Damaged skeletal muscle fibres are replaced with new contractile units via muscle regeneration. Regenerating muscle fibres synthesize functionally distinct isoforms of contractile and regulatory proteins but little is known of their functional properties during the regeneration process. An advantage of utilizing single muscle fibre preparations is that assessment of their function is based on the overall characteristics of the contractile apparatus and regulatory system and as such, these preparations are sensitive in revealing not only coarse, but also subtle functional differences between muscle fibres. We examined the Ca(2+)- and Sr(2+)-activated contractile characteristics of permeabilized fibres from rat fast-twitch (extensor digitorum longus) and slow-twitch (soleus) muscles at 7, 14 and 21 days following myotoxic injury, to test the hypothesis that fibres from regenerating fast and slow muscles have different functional characteristics to fibres from uninjured muscles. Regenerating muscle fibres had approximately 10% of the maximal force producing capacity (P(o)) of control (uninjured) fibres, and an altered sensitivity to Ca(2+) and Sr(2+) at 7 days post-injury. Increased force production and a shift in Ca(2+) sensitivity consistent with fibre maturation were observed during regeneration such that P(o) was restored to 36-45% of that in control fibres by 21 days, and sensitivity to Ca(2+) and Sr(2+) was similar to that of control (uninjured) fibres. The findings support the hypothesis that regenerating muscle fibres have different contractile activation characteristics compared with mature fibres, and that they adopt properties of mature fast- or slow-twitch muscle fibres in a progressive manner as the regeneration process is completed.

Impact of titin strain on the cardiac slow force response.

Science.gov (United States)

Ait-Mou, Younss; Zhang, Mengjie; Martin, Jody L; Greaser, Marion L; de Tombe, Pieter P

2017-11-01

Stretch of myocardium, such as occurs upon increased filling of the cardiac chamber, induces two distinct responses: an immediate increase in twitch force followed by a slower increase in twitch force that develops over the course of several minutes. The immediate response is due, in part, to modulation of myofilament Ca 2+ sensitivity by sarcomere length (SL). The slowly developing force response, termed the Slow Force Response (SFR), is caused by a slowly developing increase in intracellular Ca 2+ upon sustained stretch. A blunted immediate force response was recently reported for myocardium isolated from homozygous giant titin mutant rats (HM) compared to muscle from wild-type littermates (WT). Here, we examined the impact of titin isoform on the SFR. Right ventricular trabeculae were isolated and mounted in an experimental chamber. SL was measured by laser diffraction. The SFR was recorded in response to a 0.2 μm SL stretch in the presence of [Ca 2+ ] o  = 0.4 mM, a bathing concentration reflecting ∼50% of maximum twitch force development at 25 °C. Presence of the giant titin isoform (HM) was associated with a significant reduction in diastolic passive force upon stretch, and ∼50% reduction of the magnitude of the SFR; the rate of SFR development was unaffected. The sustained SL stretch was identical in both muscle groups. Therefore, our data suggest that cytoskeletal strain may underlie directly the cellular mechanisms that lead to the increased intracellular [Ca 2+ ] i that causes the SFR, possibly by involving cardiac myocyte integrin signaling pathways. Copyright © 2017 Elsevier Ltd. All rights reserved.

Altered myoplasmic Ca(2+) handling in rat fast-twitch skeletal muscle fibres during disuse atrophy.

Science.gov (United States)

Weiss, Norbert; Andrianjafiniony, Tina; Dupré-Aucouturier, Sylvie; Pouvreau, Sandrine; Desplanches, Dominique; Jacquemond, Vincent

2010-03-01

Calcium-dependent signalling pathways are believed to play an important role in skeletal muscle atrophy, but whether intracellular Ca(2+) homeostasis is affected in that situation remains obscure. We show here that there is a 20% atrophy of the fast-type flexor digitorum brevis (FDB) muscle in rats hind limb unloaded (HU) for 2 weeks, with no change in fibre type distribution. In voltage-clamp experiments, the amplitude of the slow Ca(2+) current was found similar in fibres from control and HU animals. In fibres loaded with the Ca(2+) dye indo-1, the value for the rate of [Ca(2+)] decay after the end of 5-100-ms-long voltage-clamp depolarisations from -80 to +10 mV was found to be 30-50% lower in fibres from HU animals. This effect was consistent with a reduced contribution of both saturable and non-saturable components of myoplasmic Ca(2+) removal. However, there was no change in the relative amount of parvalbumin, and type 1 sarco-endoplasmic reticulum Ca(2+)-ATPase was increased by a factor of three in the atrophied muscles. Confocal imaging of mitochondrial membrane potential showed that atrophied FDB fibres had significantly depolarized mitochondria as compared to control fibres. Depolarization of mitochondria in control fibres with carbonyl cyanide-p-trifluoromethoxyphenylhydrazone induced a slowing of the decay of [Ca(2+)] transients accompanied by an increase in resting [Ca(2+)] and a reduction of the peak amplitude of the transients. Overall results provide the first functional evidence for severely altered intracellular Ca(2+) removal capabilities in atrophied fast-type muscle fibres and highlight the possible contribution of reduced mitochondrial polarisation.

Evaluation of the effect of cooling and of the addition of collagenase on llama sperm DNA using toluidine blue.

Science.gov (United States)

Carretero, M I; Giuliano, S M; Casaretto, C I; Gambarotta, M C; Neild, D M

2012-05-01

The effect cryopreservation has on sperm chromatin condensation has been studied in many species but not in South American camelids. The objectives of this study were to evaluate with toluidine blue (TB) the effects of cooling and of adding collagenase on llama sperm DNA condensation. The optimum incubation time (30 s, 1.5 and 3 min) with a reducing agent (dithiothreitol) was also determined. When comparing cooled samples with the raw ejaculate, a significant increase in sperm showing a high degree of decondensation (TB positive) was observed (P = 0.005). A positive correlation was observed, both in raw and cooled semen, between sperm head morphological abnormalities observed in TB-stained cells and TB-positive sperm (highly decondensed DNA), but not with TB-intermediate spermatozoa (moderately decondensed DNA). No significant differences (P > 0.05) were observed in samples incubated with or without 0.1% collagenase. In cooled semen, but not in raw, a significant increase (P = 0.000) in reacted sperm (TB positive) was observed using 3-min incubation with 1% dithiothreitol (DTT). To conclude, cooling would seem to produce an increase in llama sperm chromatin decondensation. Also, 0.1% collagenase in H-TALP-BSA could be added to raw semen to aid its manipulation as it would not seem to increase DNA decondensation. © 2011 Blackwell Verlag GmbH.

Functional characterization of the role of the PilG in Xylella fastidiosa

Science.gov (United States)

Type IV pili of Xylella fastidiosa are regulated by pilG, a chemotaxis regulator in the Pil-Chp operon involving signal transduction pathways. To elucidate the role of pilG in twitching motility and pathogenicity of X. fastidiosa, phenotypes of wild type, a pilG-mutant, and a complementary strain we...

Studies of association of the CASQ1 rs2275703 polymorphism in relation to type 2 diabetes and related quantitative metabolic traits among 7,088 Danish whites

DEFF Research Database (Denmark)

Sparsø, Thomas; Hussain, Meena Shaheen; Borch-Johnsen, Knut

2007-01-01

Calsequestrin 1 (CASQ1) is a calcium storage protein of fast-twitch skeletal muscle cells. In previous human association studies the results have been contradictory regarding the association between a CASQ1 rs2275703 polymorphism and type 2 diabetes. In the present study of the CASQ1 rs2275703 po...

Task-dependent inhibition of slow-twitch soleus and excitation of fast-twitch gastrocnemius do not require high movement speed and velocity-dependent sensory feedback

Directory of Open Access Journals (Sweden)

Ricky eMehta

2014-10-01

Full Text Available Although individual heads of triceps surae, soleus (SO and medial gastrocnemius (MG muscles, are often considered close functional synergists, previous studies have shown distinct activity patterns between them in some motor behaviors. The goal of this study was to test two hypotheses explaining inhibition of slow SO with respect to fast MG: (1 inhibition occurs at high movement velocities and mediated by velocity-dependent sensory feedback and (2 inhibition depends on the ankle-knee joint moment combination and does not require high movement velocities. The hypotheses were tested by comparing the SO EMG/MG EMG ratio during fast and slow motor behaviors (cat paw shake responses vs. back, straight leg load lifting in humans, which had the same ankle extension-knee flexion moment combination; and during fast and slow behaviors with the ankle extension-knee extension moment combination (human vertical jumping and stance phase of walking in cats and leg load lifting in humans. In addition, SO EMG/MG EMG ratio was determined during cat paw shake responses and walking before and after removal of stretch velocity-dependent sensory feedback by self-reinnervating SO and/or gastrocnemius. We found the ratio SO EMG/MG EMG below 1 (p<0.05 during fast paw shake responses and slow back load lifting, requiring the ankle extension-knee flexion moment combination; whereas the ratio SO EMG/MG EMG was above 1 (p<0.05 during fast vertical jumping and slow tasks of walking and leg load lifting, requiring ankle extension-knee extension moments. Removal of velocity-dependent sensory feedback did not affect the SO EMG/MG EMG ratio in cats. We concluded that the relative inhibition of SO does not require high muscle velocities, depends on ankle-knee moment combinations, and is mechanically advantageous for allowing a greater MG contribution to ankle extension and knee flexion moments.

Assessing the cytotoxicity of ambient particulate matter (PM) using Chinese hamster ovary (CHO) cells and its relationship with the PM chemical composition and oxidative potential

Science.gov (United States)

Wang, Yixiang; Plewa, Michael J.; Mukherjee, Ujjal Kumar; Verma, Vishal

2018-04-01

We assessed mammalian cell cytotoxicity of ambient PM2.5 and investigated its association with the oxidative potential (OP) and chemical composition of the particles. Sixteen PM samples spanning in various seasons (fall, winter, spring and summer) were collected from an urban site in central Illinois. Cytotoxicity (LC50) in terms of the volume of air that kills 50% of the cells were calculated, which varied from 4.3 to 7.2 m3 of air. The OP was measured by two assays - the dithiothreitol (DTT) and the surrogate lung fluid (SLF) assay. In DTT assay, we measured two endpoints - hydroxyl radicals (•OH) generation and DTT consumption (the conventionally measured endpoint), while only •OH generation was measured in the SLF assay. Although, all three endpoints in the OP assays correlated significantly (P ≤ 0.05) with LC50, the correlation of reactive oxygen species (ROS) generation in DTT and SLF assays was much higher (r > 0.80 for •OH generation versus LC50) than the DTT consumption (r = 0.58). To further understand the components in PM that drive cytotoxicity and OP, concentration of water-soluble metals (Fe, Cu, Co, Cr, Mn, Ni, Pb, V, Hg, and Zn), organic carbon (OC), water soluble organic carbon (WSOC), and elemental carbon (EC) were measured. Among all the chemical components, Fe, Cu and WSOC correlated most (r > 0.70; P ≤ 0.01) with the cytotoxicity. DTT consumption correlated only with OC and WSOC (r > 0.80; P ≤ 0.01), while •OH generation in DTT and SLF assay correlated with both WSOC (r > 0.70; P ≤ 0.01) and metals (i.e. Fe and Cu; r > 0.75; P ≤ 0.01). Our results suggest a strong link between the PM2.5 OP and its cytotoxicity. Furthermore, the synergistic interactions among the organic compounds (i.e. WSOC) and metals (Fe and Cu) to enhance the ROS generation, which are more effectively captured in •OH generation endpoints in DTT and SLF assay than the DTT consumption, appear to be largely responsible for the observed mammalian cell

Time course of action and endotracheal intubating conditions of Org 9487, a new short-acting steroidal muscle relaxant; a comparison with succinylcholine

NARCIS (Netherlands)

Wierda, JMKH; van den Broek, L; Proost, JH; Verbaan, BW; Hennis, PJ

In a randomized study, we evaluated lag time (time from the end of injection of muscle relaxant until the first depression of the train-of-four response [TOF]), onset time (time from the end of injection of muscle relaxant until the maximum depression of the first twitch of the TOF [T1]),

Excessive Fragmentary Myoclonus: What Do We Know?

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Jiří Nepožitek

2017-03-01

Full Text Available Excessive fragmentary myoclonus (EFM is a polysomnographic finding registered by the surface electromyography (EMG and characterized as a result of the muscle activity consisting of sudden, isolated, arrhythmic, asynchronous and asymmetric brief twitches. The EMG potentials are defined by the exact criteria in The International Classification of the Sleep Disorders, 3rd edition and they appear with high intensity in all sleep stages. Clinical significance of EFM is unclear. It was observed in combination with other diseases and features such as obstructive and central sleep apnea, narcolepsy, periodic limb movements, insomnia, neurodegenerative disorders and peripheral nerve dysfunction. Relation to such wide range of diseases supports the opinion that EFM is nor a specific sleep disorder nor a specific polysomnographic sign. The option that EFM is a normal variant has also not been ruled out so far.

Research on acute toxicity and the behavioral effects of methanolic extract from psilocybin mushrooms and psilocin in mice.

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Zhuk, Olga; Jasicka-Misiak, Izabela; Poliwoda, Anna; Kazakova, Anastasia; Godovan, Vladlena V; Halama, Marek; Wieczorek, Piotr P

2015-03-27

The pharmacological activities and acute toxicity of the psilocin (PC) and dried residues of the crude extracts of psychotropic mushrooms were investigated in mice. The hallucinogenic substances were effectively isolated, by using methanol, from the species of Psilocybe semilanceata and Pholiotina cyanopus, that were collected in the north-east region of Poland. The chemical analysis of these extracts, which was performed by liquid chromatography with mass spectrometry detection (LC-MS), indicated the presence of psilocin and other hallucinogenic substances, including indolealkylamines and their phosphorylated analogues. When the pure psilocin or fungal extracts were used, slight differences in determined LD50 values were observed. However, the application of PC evoked the highest level of toxicity (293.07 mg/kg) compared to the activity of extracts from Ph. cyanopus and P. semilanceata, where the level of LD50 was 316.87 mg/kg and 324.37 mg/kg, respectively. Furthermore, the behavioral test, which considered the head-twitching response (HTR), was used to assess the effects of the studied psychotropic factors on the serotonergic system. Both, the fungal extracts and psilocin evoked characteristic serotoninergic effects depending on the dose administered to mice, acting as an agonist/partial agonist on the serotonergic system. A dose of 200 mg/kg 5-hydroxytryptophan (5-HTP) induced spontaneous head-twitching in mice (100% effect), as a result of the formation of 5-hydroxytryptamine (5-HT) in the brain. Compared to the activity of 5-HTP, the intraperitoneal administration of 1mg/kg of psilocin or hallucinogenic extracts of studied mushrooms (Ph. cyanopus and P. semilanceata) reduced the number of head-twitch responses of about 46% and 30%, respectively. In contrast, the administration of PC exhibited a reduction of about 60% in HTR numbers.

Research on Acute Toxicity and the Behavioral Effects of Methanolic Extract from Psilocybin Mushrooms and Psilocin in Mice

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Olga Zhuk

2015-03-01

Full Text Available The pharmacological activities and acute toxicity of the psilocin (PC and dried residues of the crude extracts of psychotropic mushrooms were investigated in mice. The hallucinogenic substances were effectively isolated, by using methanol, from the species of Psilocybe semilanceata and Pholiotina cyanopus, that were collected in the north-east region of Poland. The chemical analysis of these extracts, which was performed by liquid chromatography with mass spectrometry detection (LC-MS, indicated the presence of psilocin and other hallucinogenic substances, including indolealkylamines and their phosphorylated analogues. When the pure psilocin or fungal extracts were used, slight differences in determined LD50 values were observed. However, the application of PC evoked the highest level of toxicity (293.07 mg/kg compared to the activity of extracts from Ph. cyanopus and P. semilanceata, where the level of LD50 was 316.87 mg/kg and 324.37 mg/kg, respectively. Furthermore, the behavioral test, which considered the head-twitching response (HTR, was used to assess the effects of the studied psychotropic factors on the serotonergic system. Both, the fungal extracts and psilocin evoked characteristic serotoninergic effects depending on the dose administered to mice, acting as an agonist/partial agonist on the serotonergic system. A dose of 200 mg/kg 5-hydroxytryptophan (5-HTP induced spontaneous head-twitching in mice (100% effect, as a result of the formation of 5-hydroxytryptamine (5-HT in the brain. Compared to the activity of 5-HTP, the intraperitoneal administration of 1mg/kg of psilocin or hallucinogenic extracts of studied mushrooms (Ph. cyanopus and P. semilanceata reduced the number of head-twitch responses of about 46% and 30%, respectively. In contrast, the administration of PC exhibited a reduction of about 60% in HTR numbers.

Weekly Time Course of Neuro-Muscular Adaptation to Intensive Strength Training.

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Brown, Niklas; Bubeck, Dieter; Haeufle, Daniel F B; Weickenmeier, Johannes; Kuhl, Ellen; Alt, Wilfried; Schmitt, Syn

2017-01-01

Detailed description of the time course of muscular adaptation is rarely found in literature. Thus, models of muscular adaptation are difficult to validate since no detailed data of adaptation are available. In this article, as an initial step toward a detailed description and analysis of muscular adaptation, we provide a case report of 8 weeks of intense strength training with two active, male participants. Muscular adaptations were analyzed on a morphological level with MRI scans of the right quadriceps muscle and the calculation of muscle volume, on a voluntary strength level by isometric voluntary contractions with doublet stimulation (interpolated twitch technique) and on a non-voluntary level by resting twitch torques. Further, training volume and isokinetic power were closely monitored during the training phase. Data were analyzed weekly for 1 week prior to training, pre-training, 8 weeks of training and 2 weeks of detraining (no strength training). Results show a very individual adaptation to the intense strength training protocol. While training volume and isokinetic power increased linearly during the training phase, resting twitch parameters decreased for both participants after the first week of training and stayed below baseline until de-training. Voluntary activation level showed an increase in the first 4 weeks of training, while maximum voluntary contraction showed only little increase compared to baseline. Muscle volume increased for both subjects. Especially training status seemed to influence the acute reaction to intense strength training. Fatigue had a major influence on performance and could only be overcome by one participant. The results give a first detailed insight into muscular adaptation to intense strength training on various levels, providing a basis of data for a validation of muscle fatigue and adaptation models.

Single muscle fiber gene expression with run taper.

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Kevin Murach

Full Text Available This study evaluated gene expression changes in gastrocnemius slow-twitch myosin heavy chain I (MHC I and fast-twitch (MHC IIa muscle fibers of collegiate cross-country runners (n = 6, 20±1 y, VO₂max = 70±1 ml•kg-1•min-1 during two distinct training phases. In a controlled environment, runners performed identical 8 kilometer runs (30:18±0:30 min:s, 89±1% HRmax while in heavy training (∼72 km/wk and following a 3 wk taper. Training volume during the taper leading into peak competition was reduced ∼50% which resulted in improved race times and greater cross-section and improved function of MHC IIa fibers. Single muscle fibers were isolated from pre and 4 hour post run biopsies in heavily trained and tapered states to examine the dynamic acute exercise response of the growth-related genes Fibroblast growth factor-inducible 14 (FN14, Myostatin (MSTN, Heat shock protein 72 (HSP72, Muscle ring-finger protein-1 (MURF1, Myogenic factor 6 (MRF4, and Insulin-like growth factor 1 (IGF1 via qPCR. FN14 increased 4.3-fold in MHC IIa fibers with exercise in the tapered state (P<0.05. MSTN was suppressed with exercise in both fiber types and training states (P<0.05 while MURF1 and HSP72 responded to running in MHC IIa and I fibers, respectively, regardless of training state (P<0.05. Robust induction of FN14 (previously shown to strongly correlate with hypertrophy and greater overall transcriptional flexibility with exercise in the tapered state provides an initial molecular basis for fast-twitch muscle fiber performance gains previously observed after taper in competitive endurance athletes.

Qualitative alteration of peripheral motor system begins prior to appearance of typical sarcopenia syndrome in middle-aged rats

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Tetsuro eTamaki

2014-10-01

Full Text Available Qualitative changes in the peripheral motor system were examined using Young, Adult, Middle-aged and Old-aged rats in order to assess before and after the appearance of sarcopenia symptoms. Significant loss of muscle mass and strength, and slow-type fiber grouping with a loss of innervated nerve fibers were used as typical markers of sarcopenia. Dynamic twitch and tetanus tension and evoked electromyogram (EEMG were measured via electrical stimulation through the sciatic nerve under anesthesia using our force-distance transducer system before and after sciatectomy. Digital and analogue data sampling was performed and shortening and relaxing velocity of serial twitches was calculated with tension force. Muscle tenderness in passive stretching was also measured as stretch absorption ability, associated with histological quantitation of muscle connective tissues. The results indicated the validity of the present model, in which Old-aged rats clearly showed the typical signs of sarcopenia, specifically in the fast-type plantaris muscles, while the slow-type soleus showed relatively mild syndromes. These observations suggest the following qualitative alterations as the pathophysiological mechanism of sarcopenia: 1 reduction of shortening and relaxing velocity of twitch; 2 decline of muscle tenderness following an increase in the connective tissue component; 3 impaired recruitment of motor units (sudden depression of tetanic force and EEMG in higher stimulation frequencies over 50-60 Hz; and 4 easy fatigability in the neuromuscular junctions. These findings are likely to be closely related to significant losses in fast-type motor units, muscle strength and contraction velocity, which could be a causative factor in falls in the elderly. Importantly, some of these symptoms began in Middle-aged rats that showed no other signs of sarcopenia. Thus, prevention should be started in middle age that could be retained relatively higher movement ability.

ATP sensitive potassium channels in the skeletal muscle functions : involvement of the KCNJ11(Kir6.2 gene in the determination of Warner Bratzer shear force

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Domenico eTricarico

2016-05-01

Full Text Available The ATP-sensitive K+-channels (KATP are distributed in the tissues coupling metabolism with K+ ions efflux. KATP subunits are encoded by KCNJ8 (Kir6.1, KCNJ11 (Kir6.2, ABCC8 (SUR1 and ABCC9 (SUR2 genes, alternative RNA splicing give rise to SUR variants that confer distinct physiological properties on the channel. An high expression/activity of the sarco-KATP channel is observed in various rat fast-twitch muscles, characterized by elevated muscle strength, while a low expression/activity is observed in the slow-twitch muscles characterized by reduced strength and frailty. Down-regulation of the KATP subunits of fast-twitch fibres is found in conditions characterized by weakness and frailty. KCNJ11 gene knockout mice have reduced glycogen, lean phenotype, lower body fat, and weakness. KATP channel is also a sensor of muscle atrophy. The KCNJ11 gene is located on BTA15, close to a QTL for meat tenderness, it has also a role in glycogen storage, a key mechanism of the postmortem transformation of muscle into meat. The role of KCNJ11 gene in muscle function may underlie an effect of KCNJ11 genotypes on meat tenderness, as recently reported. The fiber phenotype and genotype are important in livestock production science. Quantitative traits including meat production and quality are influenced both by environment and genes. Molecular markers can play an important role in the genetic improvement of animals through breeding strategies. Many factors influence the muscle Warner-Bratzler shear force including breed, age, feeding, the biochemical and functional parameters. The role of KCNJ11gene and related genes on muscle tenderness will be discussed in the present review.

Dihydrotestosterone stimulates amino acid uptake and the expression of LAT2 in mouse skeletal muscle fibres through an ERK1/2-dependent mechanism

Science.gov (United States)

Hamdi, M M; Mutungi, G

2011-01-01

Abstract Dihydrotestosterone (DHT) has acute/non-genomic actions in adult mammalian skeletal muscles whose physiological functions are still poorly understood. Therefore, the primary aim of this study was to investigate the acute/non-genomic effects of DHT on amino acid uptake as well as the cellular signal transduction events underlying these actions in mouse fast- and slow-twitch skeletal muscle fibre bundles. 14C-Labelled amino acids were used to investigate the effects of DHT and testosterone (T) on amino acid uptake and pharmacological interventions were used to determine the cellular signal transduction events mediating these actions. While T had no effect on the uptake of isoleucine (Ile) and α-methylaminoisobutyric acid (MeAIB) in both fibre types, DHT increased their uptake in the fast-twitch fibre bundles. This effect was reversed by inhibitors of protein translation, the epidermal growth factor receptor (EGFR), system A, system L, mTOR and MEK. However, it was relatively insensitive to inhibitors of transcription, androgen receptors and PI3K/Akt. Additionally, DHT treatment increased the expression of LAT2 and the phosphorylation of the EGFR in the fast-twitch fibre bundles and that of ERK1/2, RSK1/2 and ATF2 in both fibre types. Also, it decreased the phosphorylation of eEF2 and increased the incorporation of Ile into proteins in both fibre types. Most of these effects were reversed by EGFR and MEK inhibitors. From these findings we suggest that another physiological function of the acute/non-genomic actions of DHT in isolated mammalian skeletal muscle fibres is to stimulate amino acid uptake. This effect is mediated through the EGFR and involves the activation of the MAPK pathway and an increase in LAT2 expression. PMID:21606113

Carbon Dioxide Fixation in Isolated Kalanchoe Chloroplasts 1

Science.gov (United States)

Levi, Carolyn; Gibbs, Martin

1975-01-01

Chloroplasts isolated from Kalanchoe diagremontiana leaves were capable of photosynthesizing at a rate of 5.4 μmoles of CO2 per milligram of chlorophyll per hour. The dark rate of fixation was about 1% of the light rate. A high photosynthetic rate was associated with low starch content of the leaves. Ribose 5-phosphate, fructose 1,6-diphosphate, and dithiothreitol stimulated fixation, whereas phosphoenolpyruvate and azide were inhibitors. The products of CO2 fixation were primarily those of the photosynthetic carbon reduction cycle. PMID:16659249

Effective potential for non-convex potentials

International Nuclear Information System (INIS)

Fujimoto, Y.; O'Raifeartaigh, L.; Parravicini, G.

1983-01-01

It is shown that the well-known relationship between the effective potential GAMMA and the vacuum graphs μ of scalar QFT follows directly from the translational invariance of the measure, and that it holds for all values of the fields phi if, and only if, the classical potential is convex. In the non-convex case μ appears to become complex for some values of phi, but it is shown that the complexity is only apparent and is due to the failure of the loop expansion. The effective potential actually remains real and well-defined for all phi, and reduces to μ in the neighbourhood of the classical minima. A number of examples are considered, notably potentials which are spontaneously broken. In particular the mechanism by which a spontaneous breakdown may be generated by radiative corrections is re-investigated and some new insights obtained. Finally, it is shown that the renormalization group equations for the parameters may be obtained by inspection from the effective potential, and among the examples considered are SU(n) fields and supermultiplets. In particular, it is shown that for supermultiplets the effective potential is not only real but positive. (orig.)

Disulfiram is a potent modulator of multidrug transporter Cdr1p of Candida albicans

International Nuclear Information System (INIS)

Shukla, Suneet; Sauna, Zuben E.; Prasad, Rajendra; Ambudkar, Suresh V.

2004-01-01

To find novel drugs for effective antifungal therapy in candidiasis, we examined disulfiram, a drug used for the treatment of alcoholism, for its role as a potential modulator of Candida multidrug transporter Cdr1p. We show that disulfiram inhibits the oligomycin-sensitive ATPase activity of Cdr1p and 2.5 mM dithiothreitol reverses this inhibition. Disulfiram inhibited the binding of photoaffinity analogs of both ATP ([α- 32 P]8-azidoATP; IC 50 = 0.76 μM) and drug-substrates ([ 3 H]azidopine and [ 125 I]iodoarylazidoprazosin; IC 50 ∼ 12 μM) to Cdr1p in a concentration-dependent manner, suggesting that it can interact with both ATP and substrate-binding site(s) of Cdr1p. Furthermore, a non-toxic concentration of disulfiram (1 μM) increased the sensitivity of Cdr1p expressing Saccharomyces cerevisiae cells to antifungal agents (fluconazole, miconazole, nystatin, and cycloheximide). Collectively these results demonstrate that disulfiram reverses Cdr1p-mediated drug resistance by interaction with both ATP and substrate-binding sites of the transporter and may be useful for antifungal therapy

The Control of Human Immunosystem by Using Paeony Root Drug

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Hideo Tsuboi

2010-10-01

Full Text Available Paeoniflorin (PF, isolated from paeony root, has been used as a herbal medicine for more than 1200 years in China, Korea and Japan for its anti-allergic, anti-inflamatory and immunoregulatory effects. In this study, we found that PF induces apoptosis in both murine T-lineage cells and human T-cell leukemia Jurkat cells. This apoptosis was mediated through the reduction of mitochondrial membrane potential, activation of caspase and fragmentation of DNA. Interestingly, PF induced generation of reactive oxygen species (ROS and a reducing agent, dithiothreitol (DTT, and a ROS scavenger, N-acetyl cysteine (NAC, successfully attenuated the PF-induced apoptosis. Additionally, PF induced the phosphorylation of three mitogen-activated protein (MAP family kinases, extracellular signal-regulated kinase, c-Jun amino-terminal kinase (JNK and p38 MAP kinase. Curcumin, an anti-oxidant and JNK inhibitor, inhibited PF-induced apoptosis, suggesting the possible involvement of curcumin-sensitive JNK or other redox-sensitive elements in PF-induced apoptosis. These results partially explain the action mechanism of PF-containing paeony root as a herbal medicine.

Skeletal muscle fiber type composition and performance during repeated bouts of maximal, concentric contractions

Science.gov (United States)

Colliander, E. B.; Dudley, G. A.; Tesch, P. A.

1988-01-01

Force output and fatigue and recovery patterns were studied during intermittent short-term exercise. 27 men performed three bouts of 30 maximal unilateral knee extensions on 2 different occasions. Blood flow was maintained or occluded during recovery periods (60 s). Blood flow was restricted by inflating a pneumatic cuff placed around the proximal thigh. Muscle biopsies from vastus lateralis were analyzed for identification of fast twitch (FT) and slow twitch (ST) fibers and relative FT area. Peak torque decreased during each bout of exercise and more when blood flow was restricted during recovery. Initial peak torque (IPT) and average peak torque (APT) decreased over the three exercise bouts. This response was 3 fold greater without than with blood flow during recovery. IPT and APT decreased more in individuals with mainly FT fibers than in those with mainly ST fibers. It is suggested that performance during repeated bouts of maximal concentric contractions differs between individuals with different fiber type composition. Specifically, in high intensity, intermittent exercise with emphasis on anaerobic energy release a high FT composition may not necessarily be advantageous for performance.

Heat production during contraction in skeletal muscle of hypothyroid mice

International Nuclear Information System (INIS)

Leijendekker, W.J.; van Hardeveld, C.; Elzinga, G.

1987-01-01

The effect of hypothyroidism on tension-independent and -dependent heat produced during a twitch and a tetanic contraction of extensor digitorum longus (EDL) and soleus muscle of mice was examined. The amount of heat produced during a twitch and the rate of heat development during a tetanus of EDL and soleus were measured at and above optimal length. The effect of hypothyroidism on force production was 2+ cycling, these findings suggest that ATP splitting due to the Ca 2+ cycling is reduced in hypothyroid mice. This conclusion was strengthened by the observation that the oxalate-supported 45 Ca 2+ -uptake activity and 45 Ca 2+ -loading capacity of muscle homogenates from hypothyroid mice were reduced, respectively, to 51 and to 65% in soleus and to 63 and 73% in EDL muscle as compared with euthyroid mice. The tension-dependent rate of heat development during a tetanus was also decreased in soleus muscle of hypothyroid mice. This suggests a lower rate of ATP hydrolysis related to cross-bridge cycling in this muscle due to the hypothyroid state

Roles of nitric oxide, nitrite and myoglobin on myocardial efficiency in trout (Oncorthynchus mykiss) and goldfish (Carassius auratus): implications for hypoxia tolerance

DEFF Research Database (Denmark)

Pedersen, Claus Lunde; Faggiano, Serena; Helbo, Signe

2010-01-01

The roles of nitric oxide synthase activity (NOS), nitrite and myoglobin (Mb) in the regulation of myocardial function during hypoxia were examined in trout and goldfish, a hypoxia-intolerant and hypoxia-tolerant species, respectively. We measured the effect of NOS inhibition, adrenaline and nitr......The roles of nitric oxide synthase activity (NOS), nitrite and myoglobin (Mb) in the regulation of myocardial function during hypoxia were examined in trout and goldfish, a hypoxia-intolerant and hypoxia-tolerant species, respectively. We measured the effect of NOS inhibition, adrenaline...... in both trout and goldfish myocardium, with trout showing a significant increase in the O2 utilization efficiency, i.e. the ratio of twitch force to O2 consumption, suggesting an increased anaerobic metabolism. NOS inhibition enhanced myocardial O2 consumption and decreased efficiency, indicating...... that mitochondrial respiration is under a tone of NOS-produced NO. When trout myocardial twitch force and O2 consumption are enhanced by adrenaline, this NO tone disappears. Consistent with its conversion to NO, nitrite reduced O2 consumption and increased myocardial efficiency in trout but not in goldfish...

Pseudo potentials and model potentials in atomic collisions

International Nuclear Information System (INIS)

Reyes, O.; Jouin, H.; Fuentealba, P.

1988-01-01

In this work, it is discussed the main differences between the use of pseudo-potentials and model potentials in collision problems . It is shown the potential energy curves for distinct systems obtained with both kinds of potentials. (A.C.A.S.) [pt

Epileptic palatal myoclonus

International Nuclear Information System (INIS)

Tatum, W.O.; Sperling, M.R.; Jacobstein, J.G.

1991-01-01

Palatal myoclonus (PM) is usually caused by lesions of the brainstem. The authors report a case of PM of focal cortical origin in a patient with epilepsia partialis continua. The PM sometimes occurred in isolation, and at other times was accompanied by unilateral face, neck, and arm twitching. This was documented by both EEG and SPECT

Epileptic palatal myoclonus

Energy Technology Data Exchange (ETDEWEB)

Tatum, W.O.; Sperling, M.R.; Jacobstein, J.G. (Graduate Hospital, Philadelphia, PA (USA))

1991-08-01

Palatal myoclonus (PM) is usually caused by lesions of the brainstem. The authors report a case of PM of focal cortical origin in a patient with epilepsia partialis continua. The PM sometimes occurred in isolation, and at other times was accompanied by unilateral face, neck, and arm twitching. This was documented by both EEG and SPECT.

The Impact of Muscle Disuse on Muscle Atrophy in Severely Burned Rats

Science.gov (United States)

2010-12-01

fascia around the opened wound were retracted to make a reservoir to hold warm mineral oil to maintain the temperature between 36.5°C to 37.5°C monitored...EDL) as the representative fast-twitch muscle. The EDL is a dorsi flexor, while the PL is a planter flexor. It is possible, as with HLU, plantar

Localization of the 5-phospho-alpha-D-ribosyl-1-pyrophosphate binding site of human hypoxanthine-guanine phosphoribosyltransferase.

Science.gov (United States)

Keough, D T; Emmerson, B T; de Jersey, J

1991-02-22

Human erythrocyte hypoxanthine-guanine phosphoribosyltransferase (HPRT) is inactivated by iodoacetate in the absence, but not in the presence, of the substrate, 5-phospho-alpha-D-ribosyl-1-pyrophosphate (PRib-PP). Treatment of HPRT with [14C]iodoacetate followed by tryptic digestion, peptide separation and sequencing has shown that Cys-22 reacts with iodoacetate only in the absence of PRib-PP; this strongly suggests that Cys-22 is in or near the PRib-PP binding site. In contrast, Cys-105 reacts with [14C]iodoacetate both in the presence and absence of PRib-PP. Carboxymethylation of Cys-22 resulted in an increase in the Km for PRib-PP, but no change in Vmax. Storage of HPRT also resulted in an increase in the Km for PRib-PP and a decrease in its susceptibility to inactivation by iodoacetate. Dialysis of stored enzyme against 1 mM dithiothreitol resulted in a marked decrease in Km for PRib-PP. The stoichiometry of the reaction of [14C]iodoacetate with Cys-22 in HPRT leading to inactivation (approx. 1 residue modified per tetramer) showed that, in this preparation of HPRT purified from erythrocytes, only about 25% of the Cys-22 side chains were present as free and accessible thiols. Titration of thiol groups [with 5,5'-dithiobis(2-nitrobenzoic acid)] and the effect of dithiothreitol on Km for PRib-PP indicate that oxidation of thiol groups occurs on storage of HPRT, even in the presence of 1 mM beta-mercaptoethanol.

Influence of some radioprotective and radiosensitizing compounds on the replicative and repair induced DNA synthesis of rats spleen cells in vitro

International Nuclear Information System (INIS)

Goette, A.

1982-01-01

The effect of cysteine, dithiothreitol, N-ethylmaleimide, cytosinearabinoside, ethidiumbromide, bleomycine and diethyldithiocarbamate on the replicative and repair induced DNA synthesis in vitro was tested by using rats spleen cells. Besides the incorporation of a labeled DNA precursor (TdR- 3 H) the sedimentation of DNA in sucrose gradients was inquired. With respect to the DNA synthesis an uniform mechanism of action for the radioprotective substances can't be seen. Thymocytes and spleen cells seem to possess different systems of repair; this may be an explanation for their different sensibility against ionizing radiation. (orig./MG) [de

High-performance liquid chromatography of rat and mouse islet polypeptides

DEFF Research Database (Denmark)

Linde, S; Hansen, B; Welinder, B S

1990-01-01

After preparative high-performance liquid chromatography of mouse islet culture medium, concentrated on disposable C18 cartridges (Sep-Pak), an unexpected insulin immunoreactive peak eluting earlier than mouse insulin I and II was detected. Molecular mass determination by mass spectrometry...... on the buffer, the organic modifier and the procedure. In particular the use of methanol-trifluoroacetic acid resulted in extensive oxidation. The oxidation could be minimized by adding 2 mM dithiothreitol to the buffer and by degassing and/or nitrogen-bubbling of the buffer. Minimal formation of Met...

Carbon dioxide fixation in isolated Kalanchoe chloroplasts

Energy Technology Data Exchange (ETDEWEB)

Levi, C.; Gibbs, M.

1975-07-01

Chloroplasts isolated from Kalanchoe diagremontiana leaves were capable of photosynthesizing at a rate of 5.4 ..mu..moles of CO/sub 2/ per milligram of chlorophyll per hour. The dark rate of fixation was about 1 percent of the light rate. A high photosynthetic rate was associated with low starch content of the leaves. Ribose 5-phosphate, fructose 1, 6-diphosphate, and dithiothreitol stimulated fixation, whereas phosphoenolpyruvate and azide were inhibitors. The products of CO/sub 2/ fixation were primarily those of the photosynthetic carbon reduction cycle. (auth)

A General Mathematical Algorithm for Predicting the Course of Unfused Tetanic Contractions of Motor Units in Rat Muscle.

Directory of Open Access Journals (Sweden)

Rositsa Raikova

Full Text Available An unfused tetanus of a motor unit (MU evoked by a train of pulses at variable interpulse intervals is the sum of non-equal twitch-like responses to these stimuli. A tool for a precise prediction of these successive contractions for MUs of different physiological types with different contractile properties is crucial for modeling the whole muscle behavior during various types of activity. The aim of this paper is to develop such a general mathematical algorithm for the MUs of the medial gastrocnemius muscle of rats. For this purpose, tetanic curves recorded for 30 MUs (10 slow, 10 fast fatigue-resistant and 10 fast fatigable were mathematically decomposed into twitch-like contractions. Each contraction was modeled by the previously proposed 6-parameter analytical function, and the analysis of these six parameters allowed us to develop a prediction algorithm based on the following input data: parameters of the initial twitch, the maximum force of a MU and the series of pulses. Linear relationship was found between the normalized amplitudes of the successive contractions and the remainder between the actual force levels at which the contraction started and the maximum tetanic force. The normalization was made according to the amplitude of the first decomposed twitch. However, the respective approximation lines had different specific angles with respect to the ordinate. These angles had different and non-overlapping ranges for slow and fast MUs. A sensitivity analysis concerning this slope was performed and the dependence between the angles and the maximal fused tetanic force normalized to the amplitude of the first contraction was approximated by a power function. The normalized MU contraction and half-relaxation times were approximated by linear functions depending on the normalized actual force levels at which each contraction starts. The normalization was made according to the contraction time of the first contraction. The actual force levels

Maternal creatine supplementation during pregnancy prevents acute and long-term deficits in skeletal muscle after birth asphyxia: a study of structure and function of hind limb muscle in the spiny mouse.

Science.gov (United States)

LaRosa, Domenic A; Ellery, Stacey J; Snow, Rod J; Walker, David W; Dickinson, Hayley

2016-12-01

Maternal antenatal creatine supplementation protects the brain, kidney, and diaphragm against the effects of birth asphyxia in the spiny mouse. In this study, we examined creatine's potential to prevent damage to axial skeletal muscles. Pregnant spiny mice were fed a control or creatine-supplemented diet from mid-pregnancy, and 1 d before term (39 d), fetuses were delivered by c-section with or without 7.5 min of birth asphyxia. At 24 h or 33 ± 2 d after birth, gastrocnemius muscles were obtained for ex-vivo study of twitch-tension, muscle fatigue, and structural and histochemical analysis. Birth asphyxia significantly reduced cross-sectional area of all muscle fiber types (P creatine treatment prevented all asphyxia-induced changes in the gastrocnemius, improved motor performance. This study demonstrates that creatine loading before birth protects the muscle from asphyxia-induced damage at birth.

SLEEP phenomena as an early biomarker for Parkinsonism

DEFF Research Database (Denmark)

Kempfner, Jacob; Jennum, Poul; Nikolic, Miki

2013-01-01

Idiopathic Rapid-Eye-Movement (REM) sleep Behavior Disorder (iRBD) is one of the most potential biomarkers for Parkinson's Disease (PD) and some atypical PD (AP). It is characterized by REM sleep with abnormal high surface EMG (sEMG) activity. Some twitching during REM sleep is normal, but no one...... has defined what normal is, and no well-defined methodology for measuring muscle activity in REM sleep exists. The purpose of this study is to investigate the possibility of detecting abnormal high muscle activity during REM sleep in subjects diagnosed with iRBD. This has been achieved by considering...... the abnormal high muscle activity during REM sleep in iRBD subjects as an outlier detection problem, while exploiting that iRBD muscle activity is more grouped. It was possible to correctly discriminate all iRBD subjects from healthy elderly control subjects and subjects diagnosed with periodic limb movement...

Know Thy Learner: User Characteristics Underlying Effective Videogame-Based Training

Science.gov (United States)

Orvis, Karin A.; Horn, Daniel B.; Belanich, James

2008-01-01

Some proponents of training games argue that younger adults (Soldiers) are part of the "digital"or "twitch" generation, having grown up uing computers and playing videogames (e.g.,Prensky,2001). The Entertainment Software Association (ESA) reports that 69%of American heads of households play computer and/or videogames. "65% of college students reported being regular or occasional game players" (Jones, 2003).

A new view of “dream enactment” in REM sleep behavior disorder

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Blumberg, Mark S.; Plumeau, Alan M.

2015-01-01

SUMMARY REM sleep behavior disorder (RBD) is a disorder in which patients exhibit increased muscle tone and exaggerated myoclonic twitching during REM sleep. In addition, violent movements of the limbs, and complex behaviors that can sometimes appear to involve the enactment of dreams, are associated with RBD. These behaviors are widely thought to result from a dysfunction involving atonia-producing neural circuitry in the brainstem, thereby unmasking cortically generated dreams. Here we scrutinize the assumptions that led to this interpretation of RBD. In particular, we challenge the assumption that motor cortex produces twitches during REM sleep, thus calling into question the related assumption that motor cortex is primarily responsible for all of the pathological movements of RBD. Moreover, motor cortex is not even necessary to produce complex behavior; for example, stimulation of some brainstem structures can produce defensive and aggressive behaviors in rats and monkeys that are striking similar to those reported in human patients with RBD. Accordingly, we suggest an interpretation of RBD that focuses increased attention on the brainstem as a source of the pathological movements and that considers sensory feedback from moving limbs as an important influence on the content of dream mentation. PMID:26802823

Discharge properties of motor units during steady isometric contractions performed with the dorsiflexor muscles.

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Jesunathadas, Mark; Klass, Malgorzata; Duchateau, Jacques; Enoka, Roger M

2012-06-01

The purpose of this study was to record the discharge characteristics of tibialis anterior motor units over a range of target forces and to import these data, along with previously reported observations, into a computational model to compare experimental and simulated measures of torque variability during isometric contractions with the dorsiflexor muscles. The discharge characteristics of 44 motor units were quantified during brief isometric contractions at torques that ranged from recruitment threshold to an average of 22 ± 14.4% maximal voluntary contraction (MVC) torque above recruitment threshold. The minimal [range: 5.8-19.8 pulses per second (pps)] and peak (range: 8.6-37.5 pps) discharge rates of motor units were positively related to the recruitment threshold torque (R(2) ≥ 0.266; P recruitment was positively associated with recruitment threshold torque (R(2) = 0.443; P recruitment threshold torque. The variability in the simulated torque did not differ from the experimental values once the recruitment range was set to ∼85% MVC torque, and the association between motor twitch contraction times and peak twitch torque was defined as a weak linear association (R(2) = 0.096; P motor units in the tibialis anterior.

Exploring the Role of PGC-1α in Defining Nuclear Organisation in Skeletal Muscle Fibres.

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Ross, Jacob A; Pearson, Adam; Levy, Yotam; Cardel, Bettina; Handschin, Christoph; Ochala, Julien

2017-06-01

Muscle fibres are multinucleated cells, with each nucleus controlling the protein synthesis in a finite volume of cytoplasm termed the myonuclear domain (MND). What determines MND size remains unclear. In the present study, we aimed to test the hypothesis that the level of expression of the transcriptional coactivator PGC-1α and subsequent activation of the mitochondrial biogenesis are major contributors. Hence, we used two transgenic mouse models with varying expression of PGC-1α in skeletal muscles. We isolated myofibres from the fast twitch extensor digitorum longus (EDL) and slow twitch diaphragm muscles. We then membrane-permeabilised them and analysed the 3D spatial arrangements of myonuclei. In EDL muscles, when PGC-1α is over-expressed, MND volume decreases; whereas, when PGC-1α is lacking, no change occurs. In the diaphragm, no clear difference was noted. This indicates that PGC-1α and the related mitochondrial biogenesis programme are determinants of MND size. PGC-1α may facilitate the addition of new myonuclei in order to reach MND volumes that can support an increased mitochondrial density. J. Cell. Physiol. 232: 1270-1274, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Electrophysiological, histochemical, and hormonal adaptation of rat muscle after prolonged hindlimb suspension

Science.gov (United States)

Kourtidou-Papadeli, Chrysoula; Kyparos, Antonios; Albani, Maria; Frossinis, Athanasios; Papadelis, Christos L.; Bamidis, Panagiotis; Vivas, Ana; Guiba-Tziampiri, Olympia

2004-05-01

The perspective of long-duration flights for future exploration, imply more research in the field of human adaptation. Previous studies in rat muscles hindlimb suspension (HLS), indicated muscle atrophy and a change of fibre composition from slow-to-fast twitch types. However, the contractile responses to long-term unloading is still unclear. Fifteen adult Wistar rats were studied in 45 and 70 days of muscle unweighting and soleus (SOL) muscle as well as extensor digitorum longus (EDL) were prepared for electrophysiological recordings (single, twitch, tetanic contraction and fatigue) and histochemical stainings. The loss of muscle mass observed was greater in the soleus muscle. The analysis of electrophysiological properties of both EDL and SOL showed significant main effects of group, of number of unweighting days and fatigue properties. Single contraction for soleus muscle remained unchanged but there was statistically significant difference for tetanic contraction and fatigue. Fatigue index showed a decrease for the control rats, but increase for the HLS rats. According to the histochemical findings there was a shift from oxidative to glycolytic metabolism during HLS. The data suggested that muscles atrophied, but they presented an adaptation pattern, while their endurance in fatigue was decreased.

The pharmacological effect of Bothrops neuwiedii pauloensis (jararaca-pintada snake venom on avian neuromuscular transmission

Directory of Open Access Journals (Sweden)

C.R. Borja-Oliveira

2003-05-01

Full Text Available The neuromuscular effects of Bothrops neuwiedii pauloensis (jararaca-pintada venom were studied on isolated chick biventer cervicis nerve-muscle preparations. Venom concentrations of 5-50 µg/ml produced an initial inhibition and a secondary increase of indirectly evoked twitches followed by a progressive concentration-dependent and irreversible neuromuscular blockade. At venom concentrations of 1-20 µg/ml, the responses to 13.4 mM KCl were inhibited whereas those to 110 µM acetylcholine alone and cumulative concentrations of 1 µM to 10 mM were unaffected. At venom concentrations higher than 50 µg/ml, there was pronounced muscle contracture with inhibition of the responses to acetylcholine, KCl and direct stimulation. At 20-24ºC, the venom (50 µg/ml produced only partial neuromuscular blockade (30.7 ± 8.0%, N = 3 after 120 min and the initial inhibition and the secondary increase of the twitch responses caused by the venom were prolonged and pronounced and the response to KCl was unchanged. These results indicate that B.n. pauloensis venom is neurotoxic, acting primarily at presynaptic sites, and that enzyme activity may be involved in this pharmacological action.

Inhibition of micturition reflex by activation of somatic afferents in posterior femoral cutaneous nerve.

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Tai, Changfeng; Shen, Bing; Mally, Abhijith D; Zhang, Fan; Zhao, Shouguo; Wang, Jicheng; Roppolo, James R; de Groat, William C

2012-10-01

This study determined if activation of somatic afferents in posterior femoral cutaneous nerve (PFCN) could modulate the micturition reflex recorded under isovolumetric conditions in α-chloralose anaesthetized cats. PFCN stimulation inhibited reflex bladder activity and significantly (P acid (AA). The optimal frequency for PFCN stimulation-induced bladder inhibition was between 3 and 10 Hz, and a minimal stimulation intensity of half of the threshold for inducing anal twitching was required. Bilateral pudendal nerve transection eliminated PFCN stimulation-induced anal twitching but did not change the stimulation-induced bladder inhibition, excluding the involvement of pudendal afferent or efferent axons in PFCN afferent inhibition.Mechanical or electrical stimulation on the skin surface in the PFCN dermatome also inhibited bladder activity. Prolonged (2 × 30 min) PFCN stimulation induced a post-stimulation inhibition that persists for at least 2 h. This study revealed a new cutaneous-bladder reflex activated by PFCN afferents. Although the mechanisms and physiological functions of this cutaneous-bladder reflex need to be further studied, our data raise the possibility that stimulation of PFCN afferents might be useful clinically for the treatment of overactive bladder symptoms.

Composition of Muscle Fiber Types in Rat Rotator Cuff Muscles.

Science.gov (United States)

Rui, Yongjun; Pan, Feng; Mi, Jingyi

2016-10-01

The rat is a suitable model to study human rotator cuff pathology owing to the similarities in morphological anatomy structure. However, few studies have reported the composition muscle fiber types of rotator cuff muscles in the rat. In this study, the myosin heavy chain (MyHC) isoforms were stained by immunofluorescence to show the muscle fiber types composition and distribution in rotator cuff muscles of the rat. It was found that rotator cuff muscles in the rat were of mixed fiber type composition. The majority of rotator cuff fibers labeled positively for MyHCII. Moreover, the rat rotator cuff muscles contained hybrid fibers. So, compared with human rotator cuff muscles composed partly of slow-twitch fibers, the majority of fast-twitch fibers in rat rotator cuff muscles should be considered when the rat model study focus on the pathological process of rotator cuff muscles after injury. Gaining greater insight into muscle fiber types in rotator cuff muscles of the rat may contribute to elucidate the mechanism of pathological change in rotator cuff muscles-related diseases. Anat Rec, 299:1397-1401, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

PV potential and potential PV rent in European regions

DEFF Research Database (Denmark)

Hansen, Anders Chr.; Thorn, Paul

2013-01-01

The paper provides a GIS based model for assessing the potentials of photovoltaic electricity in Europe by NUTS 2 regions. The location specific energy potential per PV-­‐panel area is estimated based on observations of solar irradiation, conversion efficiency, levelised costs and the social value...... of PV-­‐electricity. Combined with the potential density of PV-­‐panel area based on land cover and environental restrictions, the PV energy potential and the potential PV ressource rent is calculated. These calculations enbable the model to estimate the regional patterns at NUTS 2 level...

Structure and Properties of Hydrophobic Aggregation Hydrogel with Chemical Sensitive Switch

Directory of Open Access Journals (Sweden)

Jiufang Duan

2017-01-01

Full Text Available Hydrogels with chemical sensitive switch have control release properties in special environments. A series of polyacrylamide-octadecyl methacrylate hydrogels crosslinked by N,N′-bis (acryloyl cystamine were synthesized as potential chemical sensitive system. When this hydrogel encounters dithiothreitol it can change its quality. The properties of the hydrogels were characterized by infrared spectroscopy, contact angle, and scanning electron microscopy. The water absorption of the hydrogel has the maximum value of 475%, when the content of octadecyl methacrylate is 5 wt%. The amount of weight loss was changed from 34.6% to 17.2%, as the content of octadecyl methacrylate increased from 3 wt% to 9.4 wt%. At the same time, the stress of the hydrogel decreased from 67.01% to 47.61%; the strength of the hydrogel reaches to the maximum 0.367 Mpa at 7 wt% octadecyl methacrylate. The increasing content of octadecyl methacrylate from 3 wt% to 9.4 wt% can enhance the hydrophobicity of the hydrogel; the contact angle of water to hydrogel changed from 14.10° to 19.62°. This hydrogel has the porous structure which permits loading of oils into the gel matrix. The functionalities of the hydrogel make it have more widely potential applications in chemical sensitive response materials.

Characterization of Transfluthrin Emissions Over Time in an Enclosed Space Over a Range of Discreet Temperatures

Science.gov (United States)

2014-05-14

MEDICINE AND BIOMETRICS Thesis Advisor L~Ph.D. !""𔃻hfr’f NAVAL MEDICAL RESEARCH CENTER-ASIA Committee Member Eleanor S. i\\lie~calf, Ph D. Dean ii...profuse salivation , choreoathetosis, increased startle response, and terminal chronic seizures (71). The Material Safety Data Sheet (MSDS) for...irritation, headache, dizziness, nausea, epigastric pain, vomiting, anorexia, fatigue, twitching muscles, salivation , fluid in lungs, runny nose, and

Plyometric training improves voluntary activation and strength during isometric, concentric and eccentric contractions.

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Behrens, Martin; Mau-Moeller, Anett; Mueller, Karoline; Heise, Sandra; Gube, Martin; Beuster, Nico; Herlyn, Philipp K E; Fischer, Dagmar-C; Bruhn, Sven

2016-02-01

This study investigated effects of plyometric training (6 weeks, 3 sessions/week) on maximum voluntary contraction (MVC) strength and neural activation of the knee extensors during isometric, concentric and eccentric contractions. Twenty-seven participants were randomly assigned to the intervention or control group. Maximum voluntary torques (MVT) during the different types of contraction were measured at 110° knee flexion (180°=full extension). The interpolated twitch technique was applied at the same knee joint angle during isometric, concentric and eccentric contractions to measure voluntary activation. In addition, normalized root mean square of the EMG signal at MVT was calculated. The twitch torque signal induced by electrical nerve stimulation at rest was used to evaluate training-related changes at the muscle level. In addition, jump height in countermovement jump was measured. After training, MVT increased by 20Nm (95% CI: 5-36Nm, P=0.012), 24Nm (95% CI: 9-40Nm, P=0.004) and 27Nm (95% CI: 7-48Nm, P=0.013) for isometric, concentric and eccentric MVCs compared to controls, respectively. The strength enhancements were associated with increases in voluntary activation during isometric, concentric and eccentric MVCs by 7.8% (95% CI: 1.8-13.9%, P=0.013), 7.0% (95% CI: 0.4-13.5%, P=0.039) and 8.6% (95% CI: 3.0-14.2%, P=0.005), respectively. Changes in the twitch torque signal of the resting muscle, induced by supramaximal electrical stimulation of the femoral nerve, were not observed, indicating no alterations at the muscle level, whereas jump height was increased. Given the fact that the training exercises consisted of eccentric muscle actions followed by concentric contractions, it is in particular relevant that the plyometric training increased MVC strength and neural activation of the quadriceps muscle regardless of the contraction mode. Copyright © 2015 Sports Medicine Australia. Published by Elsevier Ltd. All rights reserved.

Effects of presynaptic muscarinic cholinoreceptor blockade on neuromuscular transmission as assessed by the train-of-four and the tetanic fade response to rocuronium.

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Kim, Yong Beom; Lee, Sangseok; Lee, Kyeong Chun; Kim, Ha Jung; Ro, Young Jin; Yang, Hong-Seuk

2017-07-01

This study investigated the effect of muscarinic M 1 and M 2 receptor antagonists on the rocuronium-induced train of four (TOF) fade and tetanic fade, respectively. Ex-vivo phrenic nerves and diaphragms were obtained from adult Sprague-Dawley rats and stabilized in Krebs buffer; the nerve-stimulated muscle TOF fade was observed at 20 s intervals. For the TOF study, phrenic nerves and diaphragms were incubated with pirenzepine (an M 1 blocker) at concentrations of 0 nmol L -1 (control), 10 nmol L -1 (PZP10), or 100 nmol L -1 (PZP100). Rocuronium was then administered incrementally until the first twitch tension had depressed by >95% during TOF stimulation. The mean TOF ratios were compared when the first twitch tensions were depressed by 40%-50%. For the tetanic fade study, 50 Hz/5 s tetani was applied initially, 30 min after the administration of a loading dose of rocuronium and methoctramine (an M 2 receptor blocker, loaded at 0 μmol L -1 [control], 1 μmol L -1 [MET1], or 10 μmol L -1 [MET10]). The EC 95 of rocuronium was significantly lower in the PZP10 group than in the control group. In the PZP10 group, the TOF ratios at 50% and first twitch tension depression were significantly lower than those in the control group (P=.02). During tetanic stimulation, the tetanic fade was significantly enhanced in the MET10 group compared to the other groups. This study shows that antagonists of muscarinic M 1 and M 2 receptors affect the rocuronium-induced neuromuscular block as demonstrated by the reduced EC 95 and TOF ratios (M 1 antagonist, pirenzepine) or the enhanced 50-Hz tetanic fade (M 2 antagonist, methoctramine). © 2017 The Authors. Clinical and Experimental Pharmacology and Physiology Published by John Wiley & Sons Australia, Ltd.

Altered Ca2+ kinetics associated with α-actinin-3 deficiency may explain positive selection for ACTN3 null allele in human evolution.

Directory of Open Access Journals (Sweden)

Stewart I Head

Full Text Available Over 1.5 billion people lack the skeletal muscle fast-twitch fibre protein α-actinin-3 due to homozygosity for a common null polymorphism (R577X in the ACTN3 gene. α-Actinin-3 deficiency is detrimental to sprint performance in elite athletes and beneficial to endurance activities. In the human genome, it is very difficult to find single-gene loss-of-function variants that bear signatures of positive selection, yet intriguingly, the ACTN3 null variant has undergone strong positive selection during recent evolution, appearing to provide a survival advantage where food resources are scarce and climate is cold. We have previously demonstrated that α-actinin-3 deficiency in the Actn3 KO mouse results in a shift in fast-twitch fibres towards oxidative metabolism, which would be more "energy efficient" in famine, and beneficial to endurance performance. Prolonged exposure to cold can also induce changes in skeletal muscle similar to those observed with endurance training, and changes in Ca2+ handling by the sarcoplasmic reticulum (SR are a key factor underlying these adaptations. On this basis, we explored the effects of α-actinin-3 deficiency on Ca2+ kinetics in single flexor digitorum brevis muscle fibres from Actn3 KO mice, using the Ca2+-sensitive dye fura-2. Compared to wild-type, fibres of Actn3 KO mice showed: (i an increased rate of decay of the twitch transient; (ii a fourfold increase in the rate of SR Ca2+ leak; (iii a threefold increase in the rate of SR Ca2+ pumping; and (iv enhanced maintenance of tetanic Ca2+ during fatigue. The SR Ca2+ pump, SERCA1, and the Ca2+-binding proteins, calsequestrin and sarcalumenin, showed markedly increased expression in muscles of KO mice. Together, these changes in Ca2+ handling in the absence of α-actinin-3 are consistent with cold acclimatisation and thermogenesis, and offer an additional explanation for the positive selection of the ACTN3 577X null allele in populations living in cold environments

Frequency-dependence of the slow force response.

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von Lewinski, Dirk; Zhu, Danan; Khafaga, Mounir; Kockskamper, Jens; Maier, Lars S; Hasenfuss, Gerd; Pieske, Burkert

2008-05-01

Stretch induces biphasic inotropic effects in mammalian myocardium. A delayed component (slow force response, SFR) has been demonstrated in various species, however, experimental conditions varied and the underlying mechanisms are controversial. The physiological relevance of the SFR is poorly understood. Experiments were performed in ventricular muscle strips from failing human hearts and non-failing rabbit hearts. Upon stretch, twitch force was assessed at basal conditions (1 Hz, 37 degrees C) and after changing stimulation frequency with and without blockade of the Na+/H+-exchanger-1 (NHE1) or reverse-mode Na+/Ca2+-exchange (NCX). Action potential duration (APD) was assessed using floating electrodes. Low stimulation rates (0.2 Hz) potentiated and higher stimulation rates (2 and 3 Hz) reduced the SFR. The extent of SFR inhibition by NHE1 or NCX inhibition was not affected by stimulation rate. APD decreased at 0.2 Hz but was not altered at higher stimulation rates. The data demonstrate frequency-dependence of the SFR with greater positive inotropic effects at lower stimulation rates. Subcellular mechanisms underlying the SFR are not fundamentally affected by stimulation rate. The SFR may have more pronounced physiological effects at lower heart rates.

Discovery and Synthesis of GS-5734, a Phosphoramidate Prodrug of a Pyrrolo[2,1 f][triazin-4-amino] Adenine C-Nucleoside for the Treatment of Ebola and Emerging Viruses

Science.gov (United States)

2017-01-26

10 mM KCl, 1 mM dithiothreitol (DTT), 0.1 mg/ml bovine serum albumin (BSA), 0.2 U/µl RNasin Plus RNase inhibitor (Promega),4 ng/µl sshRNA, 5 mM...for 90 min at 30 °C. After 90 min, 10 µl of the reaction mixture was spotted on DE81 anion exchange paper (Whatman, United Kingdom), which was then...washed with 3× Na2HPO4 (125 mM [pH 9]), 1× water, and 1× ethyl alcohol (EtOH). The filter paper was air-dried and exposed to the phosphorimager

Calcium currents in a fast-twitch skeletal muscle of the rat

OpenAIRE

1983-01-01

Slow ionic currents were measured in the rat omohyoid muscle with the three-microelectrode voltage-clamp technique. Sodium and delayed rectifier potassium currents were blocked pharmacologically. Under these conditions, depolarizing test pulses elicited an early outward current, followed by a transient slow inward current, followed in turn by a late outward current. The early outward current appeared to be a residual delayed rectifier current. The slow inward current was identified as a calci...

Oxidative potential of size-fractionated atmospheric aerosol in urban and rural sites across Europe.

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Shafer, Martin M; Hemming, Jocelyn D C; Antkiewicz, Dagmara S; Schauer, James J

2016-07-18

In this study we applied several assays, an in vitro rat alveolar macrophage model, a chemical ROS probe (DTT, dithiothreitol), and cytokine induction (TNFα) to examine relationships between PM-induced generation of reactive oxygen species (ROS) and PM composition, using a unique set of size-resolved PM samples obtained from urban and rural environments across Europe. From April-July 2012, we collected PM from roadside canyon, roadside motorway, and background urban sites in each of six European cities and from three rural sites spanning the continent. A Hi-Vol sampler was used to collect PM in three size classes (PM>7, PM7-3, PM3) and PM was characterized for total elements, and oxidative activity quantified in unfiltered and filtered PM extracts. We measured a remarkable uniformity in air concentrations of ROS and especially DTT activity across the continent. Only a 4-fold difference was documented for DTT across the urban sites and a similar variance was documented for ROS, implying that chemical drivers of oxidative activity are relatively similar between sites. The ROS and DTT specific activity was greater at urban background sites (and also rural sites) than at urban canyon locations. PM3 dominated the size distribution of both ROS activity (86% of total) and DTT activity (76% of total), reflecting both the large contribution of PM3 to total PM mass levels and importantly the higher specific oxidative activity of the PM3 in comparison with the larger particles. The soluble fraction of total activity was very high for DTT (94%) as well as for ROS (64%) in the PM3. However in the larger PM size fractions the contributions of the insoluble components became increasingly significant. The dominance of the insoluble PM drivers of activity was particularly evident in the TNFα data, where the insoluble contribution to cytokine production could be 100-fold greater than that from soluble components. ROS and DTT activity were strongly correlated in the PM3 (r = 0

The reducing agent Dithiothreitol (DTT) increases expression of c-myc and c- fos protooncogenes in human cells

DEFF Research Database (Denmark)

Skouv, J.; Sørensen, Ilona Kryspin; Frandsen, H.

1995-01-01

The objective of the present study was to assess the possible tumour promoting activity of the food mutagen 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-OH-PhIP), by studying its influence on the expression of three genes considered to be of relevance in the tumour promotion step...

Nested Potential Games

OpenAIRE

Hiroshi Uno

2007-01-01

This paper proposes a new class of potential games, the nested potential games, which generalize the potential games defined in Monderer and Shapley (1996), as well as the pseudo-potential games defined in Dubey et al. (2006). We show that each maximizer of a nested potential is a Nash equilibrium.

A wireless integrated circuit for 100-channel charge-balanced neural stimulation.

Science.gov (United States)

Thurgood, B K; Warren, D J; Ledbetter, N M; Clark, G A; Harrison, R R

2009-12-01

The authors present the design of an integrated circuit for wireless neural stimulation, along with benchtop and in - vivo experimental results. The chip has the ability to drive 100 individual stimulation electrodes with constant-current pulses of varying amplitude, duration, interphasic delay, and repetition rate. The stimulation is performed by using a biphasic (cathodic and anodic) current source, injecting and retracting charge from the nervous system. Wireless communication and power are delivered over a 2.765-MHz inductive link. Only three off-chip components are needed to operate the stimulator: a 10-nF capacitor to aid in power-supply regulation, a small capacitor (power and command reception. The chip was fabricated in a commercially available 0.6- mum 2P3M BiCMOS process. The chip was able to activate motor fibers to produce muscle twitches via a Utah Slanted Electrode Array implanted in cat sciatic nerve, and to activate sensory fibers to recruit evoked potentials in somatosensory cortex.

Structural Integration and Control of Peerless Human-like Prosthetic Hand

Science.gov (United States)

Dave, Ankit; Muthu, P.; Karthikraj, V.; Latha, S.

2018-04-01

Limb damage can create severe disturbance in movement and operative abilities wherein the prosthetic rehabilitation has the potential to replace function and enhance the quality of life. This paper presents a humanlike prosthetic hand using such unique design concept of hand model using artificial bones, ligaments, and tendons controlled using Arduino. Amongst various platforms available, Arduino is known for its adaptability, adoration and low cost. The design of prosthetic hand has a unique structure with all carpal, metacarpal, and phalanges which are bones of the hand. These bones are attached to each other following the pattern of human hand using the polymeric rubber as a functioning ligament. Furthermore, this structure of finger is driven by tendons attached to all fingers and passes through the ligaments working as pulley resulting in more degrees of freedom. The motor can twitch the tendons to achieve the action of fingers. Thus the servos, controlled by an Arduino, are used to regulate the movement mechanism of the prosthetic hand.

Lipoxygenase independent hexanal formation in isolated soy proteins induced by reducing agents.

Science.gov (United States)

Lei, Q; Boatright, W L

2008-08-01

Compared to corresponding controls, 6.5 mM dithiothreitol (DTT) elevated headspace hexanal level over aqueous slurries of both commercial isolated soy proteins (ISP) and laboratory ISP prepared with 80 degrees C treatment. Further analysis revealed that lipoxygenase (LOX) activity was not detected from these ISP, indicating that LOX is not involved in the observed hexanal increase. Levels of the induced headspace hexanal over the ISP aqueous slurries were proportional to the amount of DTT added in the range of 0 to 65 mM. Subsequent systematic investigations with model systems revealed that iron was required for the reducing agent-induced hexanal formation from linoleic acid. Erythorbate, another reducing agent, can also induce hexanal formation in both ISP and model systems. As a comparison, the LOX activity and hexanal synthesis in defatted soy flour were examined. The corresponding results showed that defatted soy flour maintained high LOX activities and that hexanal synthesis in such sample was significantly inhibited by high concentration DTT (above 130 mM). Data from the current investigation demonstrate the existence of LOX independent hexanal formation induced by reducing agents in ISP and the potential requirement of iron as a catalyst.

Assessment and Optimization of the GeneXpert Diagnostic Platform for Detection of Ebola Virus RNA in Seminal Fluid.

Science.gov (United States)

Pettitt, James; Higgs, Elizabeth; Fallah, Mosoka; Nason, Martha; Stavale, Eric; Marchand, Jonathan; Reilly, Cavan; Jensen, Kenneth; Dighero-Kemp, Bonnie; Tuznik, Kaylie; Logue, James; Bolay, Fatorma; Hensley, Lisa

2017-02-15

Recent studies have suggested that Ebola virus (EBOV) ribonucleic acid (RNA) potentially present in the semen of a large number of survivors of Ebola virus disease (EVD) in Western Africa may contribute to sexual transmission of EVD and generate new clusters of cases in regions previously declared EVD-free. These findings drive the immediate need for a reliable, rapid, user-friendly assay for detection of EBOV RNA in semen that is deployable to multiple sites across Western Africa. In this study, we optimized the Xpert EBOV assay for semen samples by adding dithiothreitol. Compared to the assays currently in use in Liberia (including Ebola Zaire Target 1, major groove binder real-time-polymerase chain reaction assays, and original Xpert EBOV assay), the modified Xpert EBOV assay demonstrated greater sensitivity than the comparator assays. Thus, the modified Xpert EBOV assay is optimal for large-scale monitoring of EBOV RNA persistence in male survivors. Published by Oxford University Press for the Infectious Diseases Society of America 2016. This work is written by (a) US Government employee(s) and is in the public domain in the US.

In Vitro Digestibility of Rapeseed and Bovine Whey Protein Mixtures.

Science.gov (United States)

Joehnke, Marcel Skejovic; Rehder, Alina; Sørensen, Susanne; Bjergegaard, Charlotte; Sørensen, Jens Christian; Markedal, Keld Ejdrup

2018-01-24

Partial replacement of animal protein sources with plant proteins is highly relevant for the food industry, but potential effects on protein digestibility need to be established. In this study, the in vitro protein digestibility (IVPD) of four protein sources and their mixtures (50:50 w/w ratio) was investigated using a transient pepsin hydrolysis (1 h) followed by pancreatin (1 h). The protein sources consisted of napin-rich rapeseed (Brassica napus L.) protein concentrates (RPCs; RP1, RP2) prepared in pilot scale and major bovine whey proteins (WPs; α-LA, alpha-lactalbumin; β-LG, beta-lactoglobulin). IVPD of individual protein sources was higher for WPs compared to RPCs. The RP2/β-LG mixture resulted in an unexpected high IVPD equivalent to β-LG protein alone. Protein mixtures containing RP1 showed a new IVPD response type due to the negative influence of a high trypsin inhibitor activity (TIA) level. Improved IVPD of RP1 alone and in protein mixtures was obtained by lowering the TIA level using dithiothreitol (DTT). These results showed that napin-rich protein products prepared by appropriate processing can be combined with specific WPs in mixtures to improve the IVPD.

Efficient system of artificial oil bodies for functional expression and purification of recombinant nattokinase in Escherichia coli.

Science.gov (United States)

Chiang, Chung-Jen; Chen, Hong-Chen; Chao, Yun-Peng; Tzen, Jason T C

2005-06-15

Nattokinase, a serine protease, and pronattokinase, when expressed in Escherichia coli, formed insoluble aggregates without enzymatic activity. For functional expression and purification, nattokinase or pronattokinase was first overexpressed in E. coli as an insoluble recombinant protein linked to the C terminus of oleosin, a structural protein of seed oil bodies, by an intein fragment. Artificial oil bodies were reconstituted with triacylglycerol, phospholipid, and the insoluble recombinant protein thus formed. Soluble nattokinase was subsequently released through self-splicing of intein induced by temperature alteration, with the remaining oleosin-intein residing in oil bodies and the leading propeptide of pronattokinase, when present, spontaneously cleaved in the process. Active nattokinase with fibrinolytic activity was harvested by concentrating the supernatant. Nattokinase released from oleosin-intein-pronattokinase exhibited 5 times higher activity than that released from oleosin-intein-nattokinase, although the production yields were similar in both cases. Furthermore, active nattokinase could be harvested in the same system by fusing pronattokinase to the N terminus of oleosin via a different intein linker, with self-splicing induced by 1,4-dithiothreitol. These results have shown a great potential of this system for bacterial expression and purification of functional recombinant proteins.

Potential Theory

CERN Document Server

Lukeš, Jaroslav; Netuka, Ivan; Veselý, Jiří

1988-01-01

Within the tradition of meetings devoted to potential theory, a conference on potential theory took place in Prague on 19-24, July 1987. The Conference was organized by the Faculty of Mathematics and Physics, Charles University, with the collaboration of the Institute of Mathematics, Czechoslovak Academy of Sciences, the Department of Mathematics, Czech University of Technology, the Union of Czechoslovak Mathematicians and Physicists, the Czechoslovak Scientific and Technical Society, and supported by IMU. During the Conference, 69 scientific communications from different branches of potential theory were presented; the majority of them are in­ cluded in the present volume. (Papers based on survey lectures delivered at the Conference, its program as well as a collection of problems from potential theory will appear in a special volume of the Lecture Notes Series published by Springer-Verlag). Topics of these communications truly reflect the vast scope of contemporary potential theory. Some contributions deal...

A Self-Paced Team Sport Match Simulation Results In Reductions In Voluntary Activation And Modifications To Biological, Perceptual And Performance Measures At Half-Time, And For Up To 96 Hours Post-Match.

Science.gov (United States)

Tofari, Paul; Kemp, Justin; Cormack, Stuart

2017-02-23

Assessing responses to soccer match-play is limited by match variability or unrealistic simulations. To address this, the biological, perceptual, and performance response were assessed using a self-paced, simulated soccer match protocol using a non-motorized treadmill. Twelve male team-sport athletes performed the 90-min simulation. Match activity; quadriceps twitch interpolation [voluntary activation (%VA) and potentiated twitch (POT)]; biochemical markers; strength and power performance; rating of perceived exertion (RPE) and self-report wellness were collected pre-, half-time, post-, and 2, 24, 48, 72 and 96-h post-match. Change compared to pre-match was calculated using effect size (ES) ±90% confidence limit, and relationships were assessed using regression analysis. Subjects covered 12445.8±768.7 m at 87.1±3.2% maximal HR (mean±SD). Reductions in %VA and POT was present at half-time (-0.38±0.46 and -0.79±0.30, respectively), and persisted post-match. Squat jump height decreased at half-time (-0.42±0.31) and was decreased until Post96. Perceptual fatigue, soreness (-0.92±0.88 and -1.49±0.76, respectively) and creatine kinase (CK, 1.11±0.43) peaked at Post24. Pre-test strength (N.kg) correlated with changes in CK (r=-0.58 to -0.81), peak oxygen consumption (V˙ O2peak) correlated with reduced perceived wellness at Post24 (r=0.44 to 0.58) and RPE post (r=-0.71±0.28). High-speed running correlated with soreness (r=0.42) and very high speed running with reduced POT (r=0.61). Previously unreported half-time reductions in %VA and POT plateaued by post-match, suggesting a role in regulating second-half performance. Perceptual and neuromuscular responses appear related to running intensity. Greater lower-body strength and V˙ O2peak were associated with less CK (i.e., muscle damage) and perceptual responses post-match, respectively, suggesting a training focus should be placed on these capacities.

Altered Elementary Calcium Release Events and Enhanced Calcium Release by Thymol in Rat Skeletal Muscle

OpenAIRE

Szentesi, Péter; Szappanos, Henrietta; Szegedi, Csaba; Gönczi, Monika; Jona, István; Cseri, Julianna; Kovács, László; Csernoch, László

2004-01-01

The effects of thymol on steps of excitation-contraction coupling were studied on fast-twitch muscles of rodents. Thymol was found to increase the depolarization-induced release of calcium from the sarcoplasmic reticulum, which could not be attributed to a decreased calcium-dependent inactivation of calcium release channels/ryanodine receptors or altered intramembrane charge movement, but rather to a more efficient coupling of depolarization to channel opening. Thymol increased ryanodine bind...

Caffeine, Adenosine Receptors and Estrogen in Toxin Models of Parkinson’s Disease

Science.gov (United States)

2010-10-30

creatine group, 77% in minocycline group); the most common adverse events included upper respiratory symptoms (26%), joint pain (19%), and nausea (17...fluttering movement of the fore paw); 3, orolingual AIMs (i.e., twitching of orofacial muscles, empty jaw movements, and tongue protrusion), as previously...General Hospital and NIH guidelines on the ethical use of animals, with adequate measures taken to minimize pain and discomfort. Mice were injected i.p

Il sistema calpaina/calpastatina in 3 muscoli dell’arto pelvico di struzzo African Black. Studio istochimico, immunoistochimico e morfometrico

OpenAIRE

Gomez De Ayala, Diego

2006-01-01

In the present research a description of the pelvic limb musculature of ostrich (African Black) was given. Muscle tissues were obtained from three muscles namely: Gastrocnemius pars externa, Tibialis .cranialis caput tibiale, Tbialis cranialis caput femorale. Haematoxilin-eosin technique was used to verify the morphologic aspect of the tissues (fibers integrity). The histochemical results revealed the presence of three fibre types only in the gastrocnemus pars extena uscle : slow-twitch o...

Experiments on Factors That Influence Muscular Function in Man.

Science.gov (United States)

1983-05-25

Publications resulting from A.F. Contract: The blood pressure response during isometric exercise in fast and slow twitch skeletal muscle in the cat... slow tiwtch muscle in the cat. J.S.Petrofsky and A.R. Lind. Pflugers Arch. 398: 149-154, 1981. The influence of fiber composition, recruitment order and...present indicating that p-adrenergic fibres mediated the vaso- constrictor response to the fatiguing muscles . Others have demonstrated changes in

Supersymmetric approach for Killingbeck radial potential plus noncentral potential in Schrodinger equation

International Nuclear Information System (INIS)

Cari, C.; Suparmi, A.; Yunianto, M.; Pratiwi, B. N.

2016-01-01

Killingbeck radial potential, which consists of harmonic oscillator, linier and Coulomb potentials, is combined with non-central potential. The solution of three dimensional Schrodinger equation for Killingbeck potential is combined with Poschl-Teller potential and Symmetrical Top non-central potentials are investigated using supersymmetry (SUSY) operator. The non-relativistic energy is obtained which is infuenced by potentials and the wave functions are produced by using SUSY operator. (paper)

Calculation of baryon chemical potential and strangeness chemical potential in resonance matter

International Nuclear Information System (INIS)

Fu Yuanyong; Hu Shouyang; Lu Zhongdao

2006-01-01

Based on the high energy heavy-ion collisions statistical model, the baryon chemical potential and strangeness chemical potential are calculated for resonance matter with net baryon density and net strangeness density under given temperature. Furthermore, the relationship between net baryon density, net strangeness density and baryon chemical potential, strangeness chemical potential are analyzed. The results show that baryon chemical potential and strangeness chemical potential increase with net baryon density and net strangeness density increasing, the change of net baryon density affects baryon chemical potential and strangeness chemical potential more strongly than the change of net strangeness density. (authors)

Pumping potential wells

International Nuclear Information System (INIS)

Hershkowitz, N.; Forest, C.; Wang, E.Y.; Intrator, T.

1987-01-01

Nonmonotonic plasma potential structures are a common feature of many double layers and sheaths. Steady state plasma potential wells separating regions having different plasma potentials are often found in laboratory experiments. In order to exist, such structures all must find a solution to a common problem. Ions created by charge exchange or ionization in the region of the potential well are electrostatically confined and tend to accumulate and fill up the potential well. The increase in positive charge should eliminate the well, but steady state structures are found in which the wells do not fill up. This means that it is important to take into account processes which 'pump' ions from the well. As examples of ion pumping of plasma wells, potential dips in front of a positively biased electron collecting anode in a relatively cold, low density multidipole plasma are considered. Pumping is provided by ion leaks from the edges of the potential dip or by oscillating the applied potential. In the former case the two dimensional character of the problem is shown to be important. (author)

Pumping potential wells

Science.gov (United States)

Hershkowitz, N.; Forest, C.; Wang, E. Y.; Intrator, T.

1987-01-01

Nonmonotonic plasma potential structures are a common feature of many double layers and sheaths. Steady state plasma potential wells separating regions having different plasma potentials are often found in laboratory experiments. In order to exist, such structures all must find a solution to a common problem. Ions created by charge exchange or ionization in the region of the potential well are electrostatically confined and tend to accumulate and fill up the potential well. The increase in positive charge should eliminate the well. Nevertheless, steady state structures are found in which the wells do not fill up. This means that it is important to take into account processes which 'pump' ions from the well. As examples of ion pumping of plasma wells, potential dips in front of a positively biased electro collecting anode in a relatively cold, low density multidipole plasma is considered. Pumping is provided by ion leaks from the edges of the potential dip or by oscillating the applied potential. In the former case the two dimensional character of the problem is shown to be important.

Pumping potential wells

International Nuclear Information System (INIS)

Hershkowitz, N.; Forest, C.; Wang, E.Y.; Intrator, T.

1987-01-01

Nonmonotonic plasma potential structures are a common feature of many double layers and sheaths. Steady state plasma potential wells separating regions having different plasma potentials are often found in laboratory experiments. In order to exist, all such structures must find a solution to a common problem. Ions created by charge exchange or ionization in the region of the potential well are electrostatically confined and tend to accumulate and fill up the potential well. The increase in positive charge should eliminate the well. Nevertheless, steady state structures are found in which the wells do not fill up. This means that it is important to take into account processes which pump ions from the well. As examples of ion pumping of plasma wells, potential dips in front of a positively biased electron collecting anode in a relatively cold, low density, multidipole plasma are considered. Pumping is provided by ion leaks from the edges of the potential dip or by oscillating the applied potential. In the former case the two-dimensional character of the problem is shown to be important

Dimethylarginine Dimethylaminohydrolase Overexpression enhances Insulin Sensitivity

Science.gov (United States)

Sydow, Karsten; Mondon, Carl E.; Schrader, Joerg; Konishi, Hakuoh; Cooke, John P.

2011-01-01

Objective Previous studies suggest that nitric oxide (NO) may modulate insulin-induced uptake of glucose in insulin-sensitive tissues. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of NO synthase (NOS). We hypothesized that a reduction in endogenous ADMA would increase NO synthesis and thereby enhance insulin sensitivity. Methods and Results To test this hypothesis we employed a transgenic mouse in which we overexpressed human dimethylarginine dimethylaminohydrolase (DDAH-I). The DDAH-I mice had lower plasma ADMA at all ages (22–70 weeks) by comparison to wild-type (WT) littermates. With a glucose challenge, WT mice showed a prompt increase in ADMA, whereas DDAH-I mice had a blunted response. Furthermore, DDAH-I mice had a blunted increase in plasma insulin and glucose levels after glucose challenge, with a 50% reduction in the insulin resistence index, consistent with enhanced sensitivity to insulin. In liver, we observed an increased Akt phosphorylation in the DDAH-I mice after i.p. glucose challenge. Incubation of skeletal muscle from WT mice ex vivo with ADMA (2μM) markedly suppressed insulin-induced glycogen synthesis in fast-twitch but not slow-twitch muscle. Conclusions These findings suggest that the endogenous NOS inhibitor ADMA reduces insulin sensitivity, consistent with previous observations that NO plays a role in insulin sensitivity. PMID:18239148

Relationship of self-esteem, manifest anxiety, and obsessive-compulsiveness to personal habits.

Science.gov (United States)

Joubert, C E

1993-10-01

75 women and 64 men responded to the Coopersmith Self-esteem Inventory, the Manifest Anxiety Scale, and the Maudsley Obsessional-compulsive Inventory in addition to responding to a questionnaire on personal habits. The results indicated that more frequent hair-pullers and nervous twitchers scored lower on self-esteem and higher on anxiety. People who giggled and those who bit their fingernails more often scored higher on obsessive-compulsiveness. Self-reported gigglers were higher on manifest anxiety. If the criterion of self-assessed seriousness of the behavior problem was used, people who bit their nails, picked their noses, pulled their hair, chewed on objects, giggled, ground their teeth, twitched nervously, and picked at scabs scored lower on self-esteem. Higher manifest-anxiety scores were found among the people who regarded their nail-biting, hair-pulling, object-chewing, nervous twitching, or giggling as serious problems. Finally, people who regarded their nail-biting as more serious tended to have higher obsessive-compulsive scores. The results in general suggest that the frequency of several of these behaviors is anxiety-related and that it is the person's assessments of these behaviors as problems rather than simply their frequency that is related to higher anxiety and lower self-esteem.

An Approach for Simulation of the Muscle Force Modeling It by Summation of Motor Unit Contraction Forces

Directory of Open Access Journals (Sweden)

Rositsa Raikova

2013-01-01

Full Text Available Muscle force is due to the cumulative effect of repetitively contracting motor units (MUs. To simulate the contribution of each MU to whole muscle force, an approach implemented in a novel computer program is proposed. The individual contraction of an MU (the twitch is modeled by a 6-parameter analytical function previously proposed; the force of one MU is a sum of its contractions due to an applied stimulation pattern, and the muscle force is the sum of the active MUs. The number of MUs, the number of slow, fast-fatigue-resistant, and fast-fatigable MUs, and their six parameters as well as a file with stimulation patterns for each MU are inputs for the developed software. Different muscles and different firing patterns can be simulated changing the input data. The functionality of the program is illustrated with a model consisting of 30 MUs of rat medial gastrocnemius muscle. The twitches of these MUs were experimentally measured and modeled. The forces of the MUs and of the whole muscle were simulated using different stimulation patterns that included different regular, irregular, synchronous, and asynchronous firing patterns of MUs. The size principle of MUs for recruitment and derecruitment was also demonstrated using different stimulation paradigms.

Influence of proprioceptive feedback on the firing rate and recruitment of motoneurons

Science.gov (United States)

De Luca, C. J.; Kline, J. C.

2012-02-01

We investigated the relationships of the firing rate and maximal recruitment threshold of motoneurons recorded during isometric contraction with the number of spindles in individual muscles. At force levels above 10% of maximal voluntary contraction, the firing rate was inversely related to the number of spindles in a muscle, with the slope of the relationship increasing with force. The maximal recruitment threshold of motor units increased linearly with the number of spindles in the muscle. Thus, muscles with a greater number of spindles had lower firing rates and a greater maximal recruitment threshold. These findings may be explained by a mechanical interaction between muscle fibres and adjacent spindles. During low-level (0% to 10%) voluntary contractions, muscle fibres of recruited motor units produce force twitches that activate nearby spindles to respond with an immediate excitatory feedback that reaches maximal level. As the force increases further, the twitches overlap and tend towards tetanization, the muscle fibres shorten, the spindles slacken, their excitatory firings decrease, and the net excitation to the homonymous motoneurons decreases. Motoneurons of muscles with greater number of spindles receive a greater decrease in excitation which reduces their firing rates, increases their maximal recruitment threshold, and changes the motoneuron recruitment distribution.

Effect of altered thyroid state on the in situ mechanical properties of adult cat soleus

Science.gov (United States)

Roy, R. R.; Zhong, H.; Hodgson, J. A.; Grossman, E. J.; Edgerton, V. R.

2003-01-01

To determine the responsiveness of cat hindlimb muscles to thyroid manipulation, adult female cats were made hypothyroid (thyroidectomy plus tapazole treatment), hyperthyroid (synthroid pellets), or maintained euthyroid. After 4 months, the hypothyroid soleus had slower time-to-peak (TPT, 80%) and half-relaxation (HRT) times, whereas the hyperthyroid soleus had faster TPT (20%) and HRT than euthyroid cats. The tension at low stimulation frequencies (5-15 Hz) was higher in hypothyroid and lower in hyperthyroid cats compared to euthyroid cats. Muscle weight, maximum twitch and tetanic (Po) tensions, and maximum rates of shortening (Vmax) were similar across groups. The soleus of hypothyroid cats was more fatigable than normal. The myosin heavy chain (MHC) composition, based on gel electrophoresis, was unaffected by thyroid hormone manipulation. Based on the reaction of monoclonal antibodies for specific MHCs, some fast fibers in the hypothyroid cats coexpressed developmental MHC. These data indicate that 4 months of an altered thyroid state result in changes in the isometric twitch speed properties of the cat soleus, but not the tension-related or isotonic properties. Further, a chronic decrease in thyroid hormone had a greater impact than a chronic increase in thyroid hormone on the mechanical properties of the adult cat soleus. Copyright 2003 S. Karger AG, Basel.

Systematic Evaluation of Protein Reduction and Alkylation Reveals Massive Unspecific Side Effects by Iodine-containing Reagents.

Science.gov (United States)

Müller, Torsten; Winter, Dominic

2017-07-01

Reduction and alkylation of cysteine residues is part of virtually any proteomics workflow. Despite its frequent use, up to date no systematic investigation of the impact of different conditions on the outcome of proteomics studies has been performed. In this study, we compared common reduction reagents (dithiothreitol, tris-(2-carboxyethyl)-phosphine, and β-mercaptoethanol) and alkylation reagents (iodoacetamide, iodoacetic acid, acrylamide, and chloroacetamide). Using in-gel digests as well as SAX fractionated in-solution digests of cytosolic fractions of HeLa cells, we evaluated 13 different reduction and alkylation conditions resulting in considerably varying identification rates. We observed strong differences in offsite alkylation reactions at 7 amino acids as well as at the peptide N terminus, identifying single and double adducts of all reagents. Using dimethyl labeling, mass tolerant searches, and synthetic peptide experiments, we identified alkylation of methionine residues by iodine-containing alkylation reagents as one of the major factors for the differences. We observed differences of more than 9-fold in numbers of identified methionine-containing peptide spectral matches for in-gel digested samples between iodine- and noniodine-containing alkylation reagents. This was because of formation of carbamidomethylated and carboxymethylated methionine side chains and a resulting prominent neutral loss during ESI ionization or in MS/MS fragmentation, strongly decreasing identification rates of methionine-containing peptides. We achieved best results with acrylamide as alkylation reagent, whereas the highest numbers of peptide spectral matches were obtained when reducing with dithiothreitol and β-mercaptoethanol for the in-solution and the in-gel digested samples, respectively. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Reactive oxygen species and fatigue-induced prolonged low-frequency force depression in skeletal muscle fibres of rats, mice and SOD2 overexpressing mice.

Science.gov (United States)

Bruton, Joseph D; Place, Nicolas; Yamada, Takashi; Silva, José P; Andrade, Francisco H; Dahlstedt, Anders J; Zhang, Shi-Jin; Katz, Abram; Larsson, Nils-Göran; Westerblad, Håkan

2008-01-01

Skeletal muscle often shows a delayed force recovery after fatiguing stimulation, especially at low stimulation frequencies. In this study we focus on the role of reactive oxygen species (ROS) in this fatigue-induced prolonged low-frequency force depression. Intact, single muscle fibres were dissected from flexor digitorum brevis (FDB) muscles of rats and wild-type and superoxide dismutase 2 (SOD2) overexpressing mice. Force and myoplasmic free [Ca(2+)] ([Ca(2+)](i)) were measured. Fibres were stimulated at different frequencies before and 30 min after fatigue induced by repeated tetani. The results show a marked force decrease at low stimulation frequencies 30 min after fatiguing stimulation in all fibres. This decrease was associated with reduced tetanic [Ca(2+)](i) in wild-type mouse fibres, whereas rat fibres and mouse SOD2 overexpressing fibres instead displayed a decreased myofibrillar Ca(2+) sensitivity. The SOD activity was approximately 50% lower in wild-type mouse than in rat FDB muscles. Myoplasmic ROS increased during repeated tetanic stimulation in rat fibres but not in wild-type mouse fibres. The decreased Ca(2+) sensitivity in rat fibres could be partially reversed by application of the reducing agent dithiothreitol, whereas the decrease in tetanic [Ca(2+)](i) in wild-type mouse fibres was not affected by dithiothreitol or the antioxidant N-acetylcysteine. In conclusion, we describe two different causes of fatigue-induced prolonged low-frequency force depression, which correlate to differences in SOD activity and ROS metabolism. These findings may have clinical implications since ROS-mediated impairments in myofibrillar function can be counteracted by reductants and antioxidants, whereas changes in SR Ca(2+) handling appear more resistant to interventions.

Generalized Rosenbluth potentials

International Nuclear Information System (INIS)

Hassan, M.H.A.

1977-05-01

It is shown that the coefficients of friction and diffusion of the Balescu-Lenard equation can be derived from two ''generalized Rosenbluth potentials'', which reduce to the standard Rosenbluth potentials if wave effects are neglected. The potentials are evaluated explicitly in the case of Maxwellian field particles. The dominant contribution of wave effects to the potentials is due to the interaction of electron field particles with ion sound waves

The Lambert-W step-potential – an exactly solvable confluent hypergeometric potential

Energy Technology Data Exchange (ETDEWEB)

Ishkhanyan, A.M., E-mail: aishkhanyan@gmail.com [Institute for Physical Research, NAS of Armenia, 0203 Ashtarak (Armenia); Armenian State Pedagogical University, 0010 Yerevan (Armenia); Institute of Physics and Technology, National Research Tomsk Polytechnic University, Tomsk 634050 (Russian Federation)

2016-02-15

We present an asymmetric step–barrier potential for which the one-dimensional stationary Schrödinger equation is exactly solved in terms of the confluent hypergeometric functions. The potential is given in terms of the Lambert W-function, which is an implicitly elementary function also known as the product logarithm. We present the general solution of the problem and consider the quantum reflection at transmission of a particle above this potential barrier. Compared with the abrupt-step and hyperbolic tangent potentials, which are reproduced by the Lambert potential in certain parameter and/or variable variation regions, the reflection coefficient is smaller because of the lesser steepness of the potential on the particle incidence side. Presenting the derivation of the Lambert potential we show that this is a four-parametric sub-potential of a more general five-parametric one also solvable in terms of the confluent hypergeometric functions. The latter potential, however, is a conditionally integrable one. Finally, we show that there exists one more potential the solution for which is written in terms of the derivative of a bi-confluent Heun function. - Highlights: • We introduce an asymmetric step-barrier potential for which the 1D Schrödinger equation is exactly solved in terms of confluent hypergeometric functions. • The potential is given in terms of the Lambert-function, which is an implicitly elementary function also known as the product logarithm. • This is a four-parametric specification of a more general five-parametric potential also solvable in terms of the confluent hypergeometric functions. • There exists one more potential the solution for which is written in terms of the derivative of a bi-confluent Heun function.

On the scalar potential models from the isospectral potential class

Energy Technology Data Exchange (ETDEWEB)

Lima, V. Gomes [Universidade Federal Rural do Rio de Janeiro, Seropedica, RJ (Brazil). Dept. de Fisica; Santos, V. Silva [Paraiba Univ., Campina Grande, PB (Brazil). Dept. de Engenharia Civil; Rodrigues, R. de Lima [Centro Brasileiro de Pesquisas Fisicas (CBPF), Rio de Janeiro, RJ (Brazil)]. E-mail: rafaelr@cbpf.br

2001-10-01

The static field classical configuration in (1+1)-dimensions for new non-linear potential models is investigated from an isospectral potential class and the concept of bosonic zero mode solution. One of the models considered here has a static nontopological configuration with a single vacuum state, whose potential in the stability equation corresponds to broken a supersymmetry. (author)

Super periodic potential

Science.gov (United States)

Hasan, Mohammd; Mandal, Bhabani Prasad

2018-04-01

In this paper we introduce the concept of super periodic potential (SPP) of arbitrary order n, n ∈I+, in one dimension. General theory of wave propagation through SPP of order n is presented and the reflection and transmission coefficients are derived in their closed analytical form by transfer matrix formulation. We present scattering features of super periodic rectangular potential and super periodic delta potential as special cases of SPP. It is found that the symmetric self-similarity is the special case of super periodicity. Thus by identifying a symmetric fractal potential as special cases of SPP, one can obtain the tunnelling amplitude for a particle from such fractal potential. By using the formalism of SPP we obtain the close form expression of tunnelling amplitude of a particle for general Cantor and Smith-Volterra-Cantor potentials.

A Novel Analysis of Performance Classification and Workload Prediction Using Electroencephalography (EEG) Frequency Data

Science.gov (United States)

2015-03-26

calming music to ease the individual before the start of the study [8]. EEG data contains noise ranging from muscle twitches, blinking and other functions...depict brain activity visually, Borghini et al was also able to note the trend of the supposed learning process using only the Theta EEG frequency...named Prediction of Operator Performance ( POP ). One of the assumptions of this model is that only a small number of cognitive activities can be

The Regulation of Skeletal Muscle Active Hyperemia: The Differential Role of Adenosine in Muscles of Varied Fiber Types

Science.gov (United States)

1986-04-21

cyclase mediates the coronary relaxation induced by adenosine. Adenosine-induced relaxation is accompanied by cyclic AMP accumulation in bovine ...and the reaction was started by adding 0.01 ml L-glutamic dehydrogenase ( bovine liver; 1200 U•ml-1 in SO% glycerol and vhosphate buffer; p~ 7.4...Physiol: London 68: 213-237, 1929. Dudley, G.A. and R.L. Terjung. Influence of acidosis on AMP deaTIIinase activity in contracting fast-twitch muscle

New Advances in Molecular Therapy for Muscle Repair after Diseases and Injuries

Science.gov (United States)

2008-04-01

PT, Zhang, CY, Wu, Z, Boss, O et al. (2002). Transcriptional co-activator PGC-1 alpha drives the formation of slow - twitch muscle fibres . Nature...Calcineurin and CaMK signaling pathways in fast -to- slow fiber type transformation of cultured mouse skeletal muscle fibers Xiaodong Mu, PhD The John...Surgery”). 3. Ectopic bone formation in fast and slow skeletal muscle (Meszaros L., “Influence of vascularity on muscle regeneration, fibrosis and

The effect of fast neutrons, as compared with X-rays upon mutation spectrum and mutation frequency in Arabidopsis thaliana (L.) Heynh. and Hordeum vulgare L. in relation to evaluation of the BARN-reactor

International Nuclear Information System (INIS)

Dellaert, L.M.W.

1980-01-01

Explanations were sought for the 'saturation' in mutant frequency, observed after relatively high irradiation doses (fast neutrons and X-rays) in Arabidopsis thaliana (L.) Heynh, when scoring for mutants is done in the siliques (Mueller's embryotest) of the 'main' inflorescence of M 1 -plants. Studies have been carried out on the effect of the presence of dithiothreitol (DTT) during irradiation, on fast neutron and X-ray induced M 1 -ovule sterility, M 2 -embryonic lethals, M 2 -chlorophyll mutants and M 2 -viable mutants in Arabidopsis thaliana. It was found that DTT provides considerable protection against both fast neutron and X-ray induced genetic damage. (Auth.)

Probing alpha-helical and beta-sheet structures of peptides at solid/liquid interfaces with SFG.

Science.gov (United States)

Chen, Xiaoyun; Wang, Jie; Sniadecki, Jason J; Even, Mark A; Chen, Zhan

2005-03-29

We demonstrated that sum frequency generation (SFG) vibrational spectroscopy can distinguish different secondary structures of proteins or peptides adsorbed at solid/liquid interfaces. The SFG spectrum for tachyplesin I at the polystyrene (PS)/solution interface has a fingerprint peak corresponding to the B1/B3 mode of the antiparallel beta-sheet. This peak disappeared upon the addition of dithiothreitol, which can disrupt the beta-sheet structure. The SFG spectrum indicative of the MSI594 alpha-helical structure was observed at the PS/MSI594 solution interface. This research validates SFG as a powerful technique for revealing detailed secondary structures of interfacial proteins and peptides.

ICRF enhanced potentials

International Nuclear Information System (INIS)

Nelson, B.A.

1987-01-01

Ion-confining potentials in the Phaedrus tandem mirror are shown to be enhanced over Boltzmann-relations predicted values by radio-frequency (rf) waves in the ion cyclotron range of frequencies (ICRF). The ICRF enhanced potential is larger in the end cell with a lower passing density. Peak potential values decrease with increasing ion endloss current (or central cell density) for a constant rf capacitor bank voltage, and increase with increasing rf-capacitor bank voltage, for a constant ion endloss value (or central cell density). In fully axisymmetric operation, a potential peak is produced in an end cell by the central-cell rf, (with-out end-cell rf) and is found only in the end cell nearer the central-cell antenna. ICRF enhanced potentials are explained as an equilibrium between the electron-collisional filling-in rate and the electron pumping out rate provided by axial time-varying electric fields. Thermal barrier-like potential structures were found in the transition regions between the central cell and end cells, in the fully axisymmetric Phaedrus. Central-cell ICRF trapping effects combined with end-cell μΔ B forces create and pump the barrier potential wells

Potential bags

International Nuclear Information System (INIS)

Ferreira, P.L.; Tomio, L.

1992-01-01

In this paper, relativistic confining potential models, endowed with bag constants associated to volume energy terms, are investigated. In contrast to the usual bag model, these potential bags are distinguished by having smeared bag surfaces. Based on the dynamical assumptions underlying the fuzzy bag model, these bag constants are derived from the corresponding energy-momentum tensor. Explicit expressions for the single-quark energies and for the nucleon bag constant are obtained by means of an improved analytical version of the saddle-point variational method for the Dirac equation with confining power-law potentials of the scalar plus vector (S + V) or pure scalar (S) type

Chronic β2 -adrenoceptor agonist treatment alters muscle proteome and functional adaptations induced by high intensity training in young men.

Science.gov (United States)

Hostrup, Morten; Onslev, Johan; Jacobson, Glenn A; Wilson, Richard; Bangsbo, Jens

2018-01-15

While several studies have investigated the effects of exercise training in human skeletal muscle and the chronic effect of β 2 -agonist treatment in rodent muscle, their effects on muscle proteome signature with related functional measures in humans are still incompletely understood. Herein we show that daily β 2 -agonist treatment attenuates training-induced enhancements in exercise performance and maximal oxygen consumption, and alters muscle proteome signature and phenotype in trained young men. Daily β 2 -agonist treatment abolished several of the training-induced enhancements in muscle oxidative capacity and caused a repression of muscle metabolic pathways; furthermore, β 2 -agonist treatment induced a slow-to-fast twitch muscle phenotype transition. The present study indicates that chronic β 2 -agonist treatment confounds the positive effect of high intensity training on exercise performance and oxidative capacity, which is of interest for the large proportion of persons using inhaled β 2 -agonists on a daily basis, including athletes. Although the effects of training have been studied for decades, data on muscle proteome signature remodelling induced by high intensity training in relation to functional changes in humans remains incomplete. Likewise, β 2 -agonists are frequently used to counteract exercise-induced bronchoconstriction, but the effects β 2 -agonist treatment on muscle remodelling and adaptations to training are unknown. In a placebo-controlled parallel study, we randomly assigned 21 trained men to 4 weeks of high intensity training with (HIT+β 2 A) or without (HIT) daily inhalation of β 2 -agonist (terbutaline, 4 mg dose -1 ). Of 486 proteins identified by mass-spectrometry proteomics of muscle biopsies sampled before and after the intervention, 32 and 85 were changing (false discovery rate (FDR) ≤5%) with the intervention in HIT and HIT+β 2 A, respectively. Proteome signature changes were different in HIT and HIT+β 2 A (P�

Functional and conformational properties of phaseolin (Phaseolus vulgris L.) and kidney bean protein isolate: a comparative study.

Science.gov (United States)

Yin, Shou-Wei; Tang, Chuan-He; Wen, Qi-Biao; Yang, Xiao-Quan

2010-03-15

Kidney bean (Phaseolus vulgris L.) seed is an underutilised plant protein source with good potential to be applied in the food industry. Phaseolin (also named G1 globulin) represents about 50 g kg(-1) of total storage protein in the seed. The aim of the present study was to characterise physicochemical, functional and conformational properties of phaseolin, and to compare these properties with those of kidney bean protein isolate (KPI). Compared with kidney bean protein isolate (KPI), the acid-extracted phaseolin-rich protein product (PRP) had much lower protein recovery of 320 g kg(-1) (dry weight basis) but higher phaseolin purity (over 950 g kg(-1)). PRP contained much lower sulfhydryl (SH) and disulfide bond contents than KPI. Differential scanning calorimetry analyses showed that the phaseolin in PRP was less denatured than in KPI. Thermal analyses in the presence or absence of dithiothreitol, in combination with SH and SS content analyses showed the contributions of SS to the thermal stability of KPI. The analyses of near-UV circular dichroism and intrinsic fluorescence spectra indicated more compacted tertiary conformation of the proteins in PRP than in KPI. PRP exhibited much better protein solubility, emulsifying activity index, and gel-forming ability than KPI. The relatively poor functional properties of KPI may be associated with protein denaturation/unfolding, with subsequent protein aggregation. The results presented here suggest the potential for acid-extracted PRP to be applied in food formulations, in view of its functional properties.

Application of potential

International Nuclear Information System (INIS)

Petris, L.

1979-01-01

This report summarizes the results obtained with the nucleon-nucleon potential presented previously for: 1) the deuteron properties and wave function, 2) a Hartree-Fock calculation on O 16 , and 3) the perturbation V-matrix and G-matrix results for the potential in relation to the results of other potentials and to saturation

Quantum potential theory

CERN Document Server

Schürmann, Michael

2008-01-01

This volume contains the revised and completed notes of lectures given at the school "Quantum Potential Theory: Structure and Applications to Physics," held at the Alfried-Krupp-Wissenschaftskolleg in Greifswald from February 26 to March 10, 2007. Quantum potential theory studies noncommutative (or quantum) analogs of classical potential theory. These lectures provide an introduction to this theory, concentrating on probabilistic potential theory and it quantum analogs, i.e. quantum Markov processes and semigroups, quantum random walks, Dirichlet forms on C* and von Neumann algebras, and boundary theory. Applications to quantum physics, in particular the filtering problem in quantum optics, are also presented.

Effective potentials from complex simulations: a potential-matching algorithm and remarks on coarse-grained potentials

International Nuclear Information System (INIS)

Toth, Gergely

2007-01-01

The projection of complex interactions onto simple distance-dependent or angle-dependent classical mechanical functions is a long-standing theoretical challenge in the field of computational sciences concerning biomolecules, colloids, aggregates and simple systems as well. The construction of an effective potential may be based on theoretical assumptions, on the application of fitting procedures on experimental data and on the simplification of complex molecular simulations. Recently, a force-matching method was elaborated to project the data of Car-Parrinello ab initio molecular dynamics simulations onto two-particle classical interactions (Izvekov et al 2004 J. Chem. Phys. 120 10896). We have developed a potential-matching algorithm as a practical analogue of this force-matching method. The algorithm requires a large number of configurations (particle positions) and a single value of the potential energy for each configuration. We show the details of the algorithm and the test calculations on simple systems. The test calculation on water showed an example in which a similar structure was obtained for qualitatively different pair interactions. The application of the algorithm on reverse Monte Carlo configurations was tried as well. We detected inconsistencies in a part of our calculations. We found that the coarse graining of potentials cannot be performed perfectly both for the structural and the thermodynamic data. For example, if one applies an inverse method with an input of the pair-correlation function, it provides energetics data for the configurations uniquely. These energetics data can be different from the desired ones obtained by all atom simulations, as occurred in the testing of our potential-matching method

Does metabosensitive afferent fibers activity differ from slow- and fast-twitch muscles?

Science.gov (United States)

Caron, Guillaume; Decherchi, Patrick; Marqueste, Tanguy

2015-09-01

This study was designed to investigate the metabosensitive afferent response evoked by electrically induced fatigue (EIF), lactic acid (LA) and potassium chloride (KCl) in three muscle types. We recorded the activity of groups III-IV afferents originating from soleus, gastrocnemius and tibialis anterior muscles. Our data showed a same pattern of response in the three muscles after chemical injections, i.e., a bell curve with maximal discharge rate at 1 mM for LA injections and a linear relationship between KCl concentrations and the afferent discharge rate. Furthermore, a stronger response was recorded after EIF in the gastrocnemius muscle compared to the two other muscles. The change in afferent discharge after 1 mM LA injection was higher for the gastrocnemius muscle compared to the response obtained with the corresponding concentration applied in the two other muscles, whereas changes to KCl injections did not dramatically differ between the three muscles. We conclude that anatomical (mass, phenotype, vascularization, receptor and afferent density…) and functional (flexor vs. extensor) differences between muscles could explain the amplitude of these responses.

Characterization of ryanodine receptor and Ca2+-ATPase isoforms in the thermogenic heater organ of blue marlin (Makaira nigricans).

Science.gov (United States)

Morrissette, Jeffery M; Franck, Jens P G; Block, Barbara A

2003-03-01

A thermogenic organ is found beneath the brain of billfishes (Istiophoridae), swordfish (Xiphiidae) and the butterfly mackerel (Scombridae). The heater organ has been shown to warm the brain and eyes up to 14 degrees C above ambient water temperature. Heater cells are derived from extraocular muscle fibers and express a modified muscle phenotype with an extensive transverse-tubule (T-tubule) network and sarcoplasmic reticulum (SR) enriched in Ca(2+)-ATPase (SERCA) pumps and ryanodine receptors (RyRs). Heater cells have a high mitochondria content but have lost most of the contractile myofilaments. Thermogenesis has been hypothesized to be associated with release and reuptake of Ca(2+). In this study, Ca(2+) fluxes in heater SR vesicles derived from blue marlin (Makaira nigricans) were measured using fura-2 fluorescence. Upon the addition of MgATP, heater SR vesicles rapidly sequestered Ca(2+). Uptake of Ca(2+) was thapsigargin sensitive, and maximum loading ranged between 0.8 micro mol Ca(2+) mg(-1) protein and 1.0 micro mol Ca(2+) mg(-1) protein. Upon the addition of 10 mmol l(-1) caffeine or 350 micro mol l(-1) ryanodine, heater SR vesicles released only a small fraction of the loaded Ca(2+). However, ryanodine could elicit a much larger Ca(2+) release event when the activity of the SERCA pumps was reduced. RNase protection assays revealed that heater tissue expresses an RyR isoform that is also expressed in fish slow-twitch skeletal muscle but is distinct from the RyR expressed in fish fast-twitch skeletal muscle. The heater and slow-twitch muscle RyR isoform has unique physiological properties. In the presence of adenine nucleotides, this RyR remains open even though cytoplasmic Ca(2+) is elevated, a condition that normally closes RyRs. The fast Ca(2+) sequestration by the heater SR, coupled with a physiologically unique RyR, is hypothesized to promote Ca(2+) cycling, ATP turnover and heat generation. A branch of the oculomotor nerve innervates heater organs

Compensatory axon sprouting for very slow axonal die-back in a transgenic model of spinal muscular atrophy type III.

Science.gov (United States)

Udina, Esther; Putman, Charles T; Harris, Luke R; Tyreman, Neil; Cook, Victoria E; Gordon, Tessa

2017-03-01

Smn +/- transgenic mouse is a model of the mildest form of spinal muscular atrophy. Although there is a loss of spinal motoneurons in 11-month-old animals, muscular force is maintained. This maintained muscular force is mediated by reinnervation of the denervated fibres by surviving motoneurons. The spinal motoneurons in these animals do not show an increased susceptibility to death after nerve injury and they retain their regenerative capacity. We conclude that the hypothesized immaturity of the neuromuscular system in this model cannot explain the loss of motoneurons by systematic die-back. Spinal muscular atrophy (SMA) is a common autosomal recessive disorder in humans and is the leading genetic cause of infantile death. Patients lack the SMN1 gene with the severity of the disease depending on the number of copies of the highly homologous SMN2 gene. Although motoneuron death in the Smn +/- transgenic mouse model of the mildest form of SMA, SMA type III, has been reported, we have used retrograde tracing of sciatic and femoral motoneurons in the hindlimb with recording of muscle and motor unit isometric forces to count the number of motoneurons with intact neuromuscular connections. Thereby, we investigated whether incomplete maturation of the neuromuscular system induced by survival motoneuron protein (SMN) defects is responsible for die-back of axons relative to survival of motoneurons. First, a reduction of ∼30% of backlabelled motoneurons began relatively late, at 11 months of age, with a significant loss of 19% at 7 months. Motor axon die-back was affirmed by motor unit number estimation. Loss of functional motor units was fully compensated by axonal sprouting to retain normal contractile force in four hindlimb muscles (three fast-twitch and one slow-twitch) innervated by branches of the sciatic nerve. Second, our evaluation of whether axotomy of motoneurons in the adult Smn +/- transgenic mouse increases their susceptibility to cell death demonstrated

Muscle and bone follow similar temporal patterns of recovery from muscle-induced disuse due to botulinum toxin injection.

Science.gov (United States)

Manske, Sarah L; Boyd, Steven K; Zernicke, Ronald F

2010-01-01

If muscle force is a primary source for triggering bone adaptation, with disuse and reloading, bone changes should follow muscle changes. We examined the timing and magnitude of changes in muscle cross-sectional area (MCSA) and bone architecture in response to muscle inactivity following botulinum toxin (BTX) injection. We hypothesized that MCSA would return to baseline levels sooner than bone properties following BTX injection. Female BALB mice (15 weeks old) were injected with 20 muL of BTX (1 U/100 g body mass, n=18) or saline (SAL, n=18) into the posterior calf musculature of one limb. The contralateral limb (CON) served as an internal control. MCSA and bone properties were assessed at baseline, 2, 4, 8, 12, and 16 weeks post-injection using in vivo micro-CT at the tibia proximal metaphysis (bone only) and diaphysis. Muscles were dissected and weighed after sacrifice. Significant GroupxLegxTime interactions indicated that the maximal decrease in MCSA (56%), proximal metaphyseal BV/TV (38%) and proximal diaphyseal Ct.Ar (7%) occurred 4 weeks after injection. There was no delay prior to bone recovery as both muscle and bone properties began to recover after this time, but MCSA and BV/TV remained 15% and 20% lower, respectively, in the BTX-injected leg than the BTX-CON leg 16 weeks post-injection. Gastrocnemius mass (primarily fast-twitch) was 14% lower in the BTX-injected leg than the SAL-injected leg, while soleus mass (primarily slow-twitch) was 15% greater in the BTX group than the SAL group. Our finding that muscle size and bone began to recover at similar times after BTX injection was unexpected. This suggested that partial weight-bearing and/or return of slow-twitch muscle activity in the BTX leg may have been sufficient to stimulate bone recovery. Alternatively, muscle function may have recovered sooner than MCSA. Our results indicated that muscle cross-sectional area, while important, may not be the primary factor associated with bone loss and recovery

The potential of renewable energy

International Nuclear Information System (INIS)

Piot, M.

2007-01-01

This article presents and comments on definitions of the potential of renewable forms of energy and, in a second part, takes a look at the potentials mentioned in the energy perspectives published by the Swiss Federal Office of Energy (SFOE). The following potentials are looked at: technical potential, ecological potential, economic potential, exploitable and expected potentials, technical, economic and ecological expansion potentials, potential of particular technologies in Switzerland, exploitable and expected expansion potential. Four scenarios for expansion potential are briefly described

Eulerian velocity reconstruction in ideal atmospheric dynamics using potential vorticity and potential temperature

Science.gov (United States)

Blender, R.

2009-04-01

An approach for the reconstruction of atmospheric flow is presented which uses space- and time-dependent fields of density ?, potential vorticity Q and potential temperature Î& cedil;[J. Phys. A, 38, 6419 (2005)]. The method is based on the fundamental equations without approximation. The basic idea is to consider the time-dependent continuity equation as a condition for zero divergence of momentum in four dimensions (time and space, with unit velocity in time). This continuity equation is solved by an ansatz for the four-dimensional momentum using three conserved stream functions, the potential vorticity, potential temperature and a third field, denoted as ?-potential. In zonal flows, the ?-potential identifies the initial longitude of particles, whereas potential vorticity and potential temperature identify mainly meridional and vertical positions. Since the Lagrangian tracers Q, Î&,cedil; and ? determine the Eulerian velocity field, the reconstruction combines the Eulerian and the Lagrangian view of hydrodynamics. In stationary flows, the ?-potential is related to the Bernoulli function. The approach requires that the gradients of the potential vorticity and potential temperature do not vanish when the velocity remains finite. This behavior indicates a possible interrelation with stability conditions. Examples with analytical solutions are presented for a Rossby wave and zonal and rotational shear flows.

On Artificial and Animal Electricity : Alessandro Volta vs. Luigi Galvani

OpenAIRE

Soeiro, Diana

2013-01-01

Two Italians, Alessandro Volta (1745–1827), a physicist, and Luigi Galvani (1737–1798), an obstetrician and physiologist, separately conducted experiments on dead frogs using metals that made their legs twitch. Volta concluded that electricity was an artificial and external phenomenon, dependent on the metals and unrelated with the frog’s body; Galvani concluded that the frog’s movement proved that there was such a thing as animal electricity that, even after life, remained stored in nerves a...

Anatomical and Physiological Characteristics of the Ferret Lateral Rectus Muscle and Abducena Nucleus

Science.gov (United States)

2007-01-25

from the ferret LR Slow Resistant group is larger than the typically powerful Fast Fatigable motor units in the cat. Whole Muscle Contractile...623-632, 1990. 21. HESS A and PILAR G. SLOW FIBRES IN THE EXTRAOCULAR MUSCLES OF THE CAT. J Physiol 169: 780-798, 1963. 22. Jacoby J, Chiarandini DJ...were split between the LR and retractor bulbi (RB) muscle slips. In addition to individual motor units, the whole LR muscle was evaluated for twitch

Reperfusion Strategies in the Management of Extremity Vascular Injury with Ischaemia

Science.gov (United States)

2012-01-01

28: 1026–1031. 45 Gürke L, Marx A, Sutter PM, Stierli P, Harder F, Heberer M. Function of fast - and slow - twitch rat skeletal muscle following ischemia...influence muscle necrosis, including temperature, muscle fibre type, muscle location and residual blood flow15,16. The earliest effects of limb...Crawford et al.11 reported that in a murine model of limb ischaemia ethyl pyruvate pretreatment resulted in a lower percentage of injured muscle fibres

Nucleus--nucleus potential

International Nuclear Information System (INIS)

Jaqaman, H.R.

1977-01-01

The nucleus--nucleus interaction is studied within the framework of the generator coordinate method that permits an easy incorporation of the full effects of antisymmetrization. It is found that the interaction, as far as the elastic scattering problem is concerned, can be described by a simple effective potential that is equivalent to the original many-body (and hence non-local) problem. The potential is obtained by dividing the wavefunction into a long-range part and a short-range part and requiring the former to satisfy a Schroedinger equation. This enables avoiding dealing with the troublesome short-range part of the wavefunction and provides a direct link with the optical model so that the potential obtained here is equivalent to the real part of the optical potential (the imaginary part is not investigated). The effective potential is found to consist of three parts: an interaction term between the nucleons belonging to different nuclei, a kinetic energy term due to the change in the intrinsic kinetic energy of the system as a result of the antisymmetrization, and finally an l-dependent part. The kinetic energy term is found to be very repulsive and effectively gives a hard core, and is calculated for the α--α and 16 O-- 16 O cases. The full potential is calculated for the α--α case for the S, D, and G partial waves and then used to calculate the corresponding phase shifts that are then compared with experimental results and other microscopic calculations. Finally, some recent results and analyses of fusion and deep inelastic reactions are reviewed that seem to indicate the presence of a hard core in the nucleus--nucleus potential. Such a hard core is present in the potential obtained in the sudden approximation

Potential of a polygonal prism and lamina; Takakuchu men no potential

Energy Technology Data Exchange (ETDEWEB)

Koyama, S

1996-05-01

With the use of rectangular coordinates O-XYZ, the potential of a calculation point P is expressed in the form of a triple repeated integral of a density {sigma} at point Q in the mass. The potential at the density {sigma} assumed to be 1 is named the potential of a polygonal prism. Further, a double repeated integral with an integral concerning Z removed from the triple integral is named the potential of polygonal lamina. This potential can be expressed in a quadratic form (linear form) with 2nd order partial derivative (1st order partial derivative) as a coefficient. On the contrary, in order to extract the 1st/2nd order partial derivatives from this potential by partial differential, it requires partial differentiation with these partial derivatives considered to be a constant. The reason that they can be realized is attributable to the zero result of the linear form which has as the coefficient a 3rd order partial derivative concerning the variable of integration in a primitive function. If this relation is used, the integral calculation and description may be simplified. An explanation was given with examples enumerated so that these conditions might be understood.

Soils - Potential Runoff

Data.gov (United States)

Kansas Data Access and Support Center — This digital spatial data set provides information on the spatial distribution of potential runoff-contributing areas in Kansas. Potential runoff-contributing areas...

Selective antibacterial activity of patchouli alcohol against Helicobacter pylori based on inhibition of urease.

Science.gov (United States)

Yu, Xiao-Dan; Xie, Jian-Hui; Wang, Yong-Hong; Li, Yu-Cui; Mo, Zhi-Zhun; Zheng, Yi-Feng; Su, Ji-Yan; Liang, Ye-er; Liang, Jin-Zhi; Su, Zi-Ren; Huang, Ping

2015-01-01

The aim of this study is to evaluate the antibacterial activity and urease inhibitory effects of patchouli alcohol (PA), the bioactive ingredient isolated from Pogostemonis Herba, which has been widely used for the treatment of gastrointestinal disorders. The activities of PA against selected bacteria and fungi were determined by agar dilution method. It was demonstrated that PA exhibited selective antibacterial activity against Helicobacter pylori, without influencing the major normal gastrointestinal bacteria. Noticeably, the antibacterial activity of PA was superior to that of amoxicillin, with minimal inhibition concentration value of 78 µg/mL. On the other hand, PA inhibited ureases from H.pylori and jack bean in concentration-dependent fashion with IC50 values of 2.67 ± 0.79 mM and 2.99 ± 0.41 mM, respectively. Lineweaver-Burk plots indicated that the type of inhibition was non-competitive against H.pylori urease whereas uncompetitive against jack bean urease. Reactivation of PA-inactivated urease assay showed DL-dithiothreitol, the thiol reagent, synergistically inactivated urease with PA instead of enzymatic activity recovery. In conclusion, the selective H.pylori antibacterial activity along with urease inhibitory potential of PA could make it a possible drug candidate for the treatment of H.pylori infection. Copyright © 2014 John Wiley & Sons, Ltd.

An improved yeast transformation method for the generation of very large human antibody libraries.

Science.gov (United States)

Benatuil, Lorenzo; Perez, Jennifer M; Belk, Jonathan; Hsieh, Chung-Ming

2010-04-01

Antibody library selection by yeast display technology is an efficient and highly sensitive method to identify binders to target antigens. This powerful selection tool, however, is often hampered by the typically modest size of yeast libraries (approximately 10(7)) due to the limited yeast transformation efficiency, and the full potential of the yeast display technology for antibody discovery and engineering can only be realized if it can be coupled with a mean to generate very large yeast libraries. We describe here a yeast transformation method by electroporation that allows for the efficient generation of large antibody libraries up to 10(10) in size. Multiple components and conditions including CaCl(2), MgCl(2), sucrose, sorbitol, lithium acetate, dithiothreitol, electroporation voltage, DNA input and cell volume have been tested to identify the best combination. By applying this developed protocol, we have constructed a 1.4 x 10(10) human spleen antibody library essentially in 1 day with a transformation efficiency of 1-1.5 x 10(8) transformants/microg vector DNA. Taken together, we have developed a highly efficient yeast transformation method that enables the generation of very large and productive human antibody libraries for antibody discovery, and we are now routinely making 10(9) libraries in a day for antibody engineering purposes.

State-dependent classical potentials

International Nuclear Information System (INIS)

D'Amico, M.

2001-01-01

As alternative treatment to the potential operators of standard quantum mechanics is presented. The method is derived from Bohm's mechanics. The operator scalar (V) and vector (A) potential functions are replaced by a quantum potential. It is argued that the classical potential is a special limiting case of a more general quantum potential. The theory is illustrated by deriving an equivalent single-particle equation for the i-th particle of an n-body Bohmian system. The resulting effective state-dependent potential holds the interaction between the single-particle self-wave ψ s and the environment wave ψ e of the n - 1 remaining particles. The effective state-dependent potential is offered as a resolution to the Aharonov-Bohm effect where the phase difference is shown to result from the presence of ψ e . Finally, the interaction between ψ s and ψ e is illustrated graphically

Reactive oxygen species induced by heat stress during grain filling of rice (Oryza sativa L.) are involved in occurrence of grain chalkiness.

Science.gov (United States)

Suriyasak, Chetphilin; Harano, Keisuke; Tanamachi, Koichiro; Matsuo, Kazuhiro; Tamada, Aina; Iwaya-Inoue, Mari; Ishibashi, Yushi

2017-09-01

Heat stress during grain filling increases rice grain chalkiness due to increased activity of α-amylase, which hydrolyzes starch. In rice and barley seeds, reactive oxygen species (ROS) produced after imbibition induce α-amylase activity via regulation of gibberellin (GA) and abscisic acid (ABA) levels during seed germination. Here, we examined whether ROS is involved in induction of grain chalkiness by α-amylase in developing rice grains under heat stress. To elucidate the role of ROS in grain chalkiness, we grew post-anthesis rice plants (Oryza sativa L. cv. Koshihikari) under control (25°C) or heat stress (30°C) conditions with or without antioxidant (dithiothreitol) treatment. The developing grains were analyzed for expression of NADPH oxidases, GA biosynthesis genes (OsGA3ox1, OsGA20ox1), ABA catabolism genes (OsABA8'OH1, OsABA8'OH2) and an α-amylase gene (OsAmy3E), endogenous H 2 O 2 content and the grain quality. In grains exposed to heat stress, the expression of NADPH oxidase genes (especially, OsRbohB, OsRbohD, OsRbohF and OsRbohI) and the ROS content increased. Heat stress also increased the expression of OsGA3ox1, OsGA20ox1, OsABA8'OH1, OsABA8'OH2 and OsAmy3E. On the other hand, dithiothreitol treatment reduced the effects of heat stress on the expression of these genes and significantly reduced grain chalkiness induced by heat stress. These results suggest that, similar to cereal seed germination mechanism, ROS produced under heat stress is involved in α-amylase induction in maturating rice grains through GA/ABA metabolism, and consequently caused grain chalkiness. Copyright © 2017 Elsevier GmbH. All rights reserved.

Oxidation of the diphosphoinositol polyphosphate phosphohydrolase-like Nudix hydrolase Aps from Drosophila melanogaster induces thermolability--A possible regulatory switch?

Science.gov (United States)

Winward, Lucinda; Whitfield, William G F; McLennan, Alexander G; Safrany, Stephen T

2010-07-01

Unlike mammalian cells, Drosophila melanogaster contains only a single member of the diphosphoinositol polyphosphate phosphohydrolase subfamily of the Nudix hydrolases, suggesting that functional specialisation has not occurred in this organism. In order to evaluate its function, Aps was cloned and characterized. It hydrolyses a range of (di)nucleoside polyphosphates, the most efficient being guanosine 5'-tetraphosphate (K(m)=11 microM, k(cat)=0.79 s(-1)). However, it shows a 5-fold preference for the hydrolysis of diphosphoinositol pentakisphosphate (PP-InsP(5), K(m)=0.07 microM, k(cat)=0.024 s(-1)). Assayed at 26 degrees C, Aps had an alkaline pH optimum and required a divalent ion: Mg(2+) (10-20 mM) or Mn(2+) (1 mM) were preferred for nucleotide hydrolysis and Mg(2+) (0.5-1 mM) or Co(2+) (1-100 microM) for PP-InsP(5) hydrolysis. GFP-fusions showed that Aps was predominantly cytoplasmic, with some nuclear localization. In the absence of dithiothreitol Aps was heat labile, rapidly losing activity even at 36 degrees C, while in the presence of dithiothreitol, Aps was heat stable, surviving for 5 min at 76 degrees C. Heat lability was restored by H(2)O(2) and mass spectrometric analysis suggested that this was due to reversible dimerisation involving two inter-molecular disulphides between Cys23 and Cys25. Aps expression was highest in embryos and declined throughout development. The ratio of PP-InsP(5) to inositol hexakisphosphate also decreased throughout development, with the highest level of PP-InsP(5) found in embryos. These data suggest that the redox state of Aps may play a role in controlling its activity by altering its stability, something that could be important for regulating PP-InsP(5) during development. Copyright 2010 Elsevier Ltd. All rights reserved.

Improvement of economic potential estimation methods for enterprise with potential branch clusters use

Directory of Open Access Journals (Sweden)

V.Ya. Nusinov

2017-08-01

Full Text Available The research determines that the current existing methods of enterprise’s economic potential estimation are based on the use of additive, multiplicative and rating models. It is determined that the existing methods have a row of defects. For example, not all the methods take into account the branch features of the analysis, and also the level of development of the enterprise comparatively with other enterprises. It is suggested to level such defects by an account at the estimation of potential integral level not only by branch features of enterprises activity but also by the intra-account economic clusterization of such enterprises. Scientific works which are connected with the using of clusters for the estimation of economic potential are generalized. According to the results of generalization it is determined that it is possible to distinguish 9 scientific approaches in this direction: the use of natural clusterization of enterprises with the purpose of estimation and increase of region potential; the use of natural clusterization of enterprises with the purpose of estimation and increase of industry potential; use of artificial clusterization of enterprises with the purpose of estimation and increase of region potential; use of artificial clusterization of enterprises with the purpose of estimation and increase of industry potential; the use of artificial clusterization of enterprises with the purpose of clustering potential estimation; the use of artificial clusterization of enterprises with the purpose of estimation of clustering competitiveness potential; the use of natural (artificial clusterization for the estimation of clustering efficiency; the use of natural (artificial clusterization for the increase of level at region (industries development; the use of methods of economic potential of region (industries estimation or its constituents for the construction of the clusters. It is determined that the use of clusterization method in

Myofilament calcium sensitivity: Role in regulation of in vivo cardiac contraction and relaxation

Directory of Open Access Journals (Sweden)

Jae-Hoon Chung

2016-12-01

Full Text Available Myofilament calcium sensitivity is an often-used indicator of cardiac muscle function, often assessed in disease states such as hypertrophic cardiomyopathy (HCM and dilated cardiomyopathy (DCM. While calcium sensitivity measurement provides important insights into the mechanical force-generating capability of a muscle at steady-state, the dynamic behavior of the muscle cannot be sufficiently assessed with a force-pCa curve alone. The dissociation constant (Kd of the force-pCa curve depends on the ratio of the apparent on-rate (kon and apparent off-rate (koff of calcium on TnC and as a stand-alone parameter cannot provide an accurate depiction of the dynamic contraction and relaxation behavior without the additional quantification of kon or koff, or actually measuring dynamic twitch kinetics in an intact muscle. In this review, we examine the effect of length, frequency, and beta-adrenergic stimulation on myofilament calcium sensitivity and dynamic contraction, the effect of membrane permeabilization on calcium sensitivity, and the dynamic consequences of various myofilament protein mutations with potential implications in contractile and relaxation behavior.

Relationships between motor unit size and recruitment threshold in older adults: implications for size principle.

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Fling, Brett W; Knight, Christopher A; Kamen, Gary

2009-08-01

As a part of the aging process, motor unit reorganization occurs in which small motoneurons reinnervate predominantly fast-twitch muscle fibers that have lost their innervation. We examined the relationship between motor unit size and the threshold force for recruitment in two muscles to determine whether older individuals might develop an alternative pattern of motor unit activation. Young and older adults performed isometric contractions ranging from 0 to 50% of maximal voluntary contraction in both the first dorsal interosseous (FDI) and tibialis anterior (TA) muscles. Muscle fiber action potentials were recorded with an intramuscular needle electrode and motor unit size was computed using spike-triggered averaging of the global EMG signal (macro EMG), which was also obtained from the intramuscular needle electrode. As expected, older individuals exhibited larger motor units than young subjects in both the FDI and the TA. However, moderately strong correlations were obtained for the macro EMG amplitude versus recruitment threshold relationship in both the young and older adults within both muscles, suggesting that the size principle of motor unit recruitment seems to be preserved in older adults.

Factors associated with night-time calf muscle cramps: a case-control study.

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Hawke, Fiona; Chuter, Vivienne; Burns, Joshua

2013-03-01

Although highly prevalent and painful, night-time calf muscle cramping is poorly understood, and no treatment has shown consistent efficacy or safety. One hundred sixty adults were recruited from New South Wales, Australia, including 80 who had night-time calf cramping at least once per week and 80 age- and gender-matched adults who did not. Participants were assessed using reliable tests of lower limb strength, flexibility, morphometrics, circulation, and sensation, and were questioned about health and lifestyle factors, diet, medications, exercise, symptomatology, sleeping habits, and footwear. Conditional logistic regression identified 3 factors independently associated with night-time calf muscle cramps: muscle twitching (OR 4.6, 95% CI 1.6-15.5, P = 0.01); lower limb tingling (OR 4.1, 95% CI 1.6-10.3, P = 0.003); and foot dorsiflexion weakness (OR 1.02, 95% CI 1.01-1.03, P = 0.002), which represented other measures of lower limb weakness in the model. Night-time calf muscle cramps were associated with markers of neurological dysfunction and potential musculoskeletal therapeutic targets. Copyright © 2012 Wiley Periodicals, Inc., a Wiley company.

Effective Kratzer and Coulomb potentials as limit cases of a multiparameter exponential-type potential

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García-Ravelo, J., E-mail: g.ravelo@hotmail.com [Departamento de Física, Escuela Superior de Física y Matemáticas, Instituto Politécnico Nacional, Edificio 9, Unidad Profesional Adolfo López Mateos, México D.F., 07738 (Mexico); Menéndez, A.; García-Martínez, J. [Departamento de Física, Escuela Superior de Física y Matemáticas, Instituto Politécnico Nacional, Edificio 9, Unidad Profesional Adolfo López Mateos, México D.F., 07738 (Mexico); Schulze-Halberg, A. [Department of Mathematics and Actuarial Science and Department of Physics, Indiana University Northwest, 3400 Broadway, Gary, IN 46408 (United States)

2014-06-13

We show that the effective Kratzer and Coulomb potentials can be obtained by taking particular limits of a multiparameter exponential potential that was studied recently. Moreover, we demonstrate that the bound state solutions of the exponential potential reduce correctly to their well-known counterparts associated with the Kratzer and Coulomb potentials. As a byproduct, we obtain a new limit relation for the hypergeometric function. - Highlights: • Kratzer and Coulomb potentials are limit cases of an exponential-type potential. • From exact s-waves, approximate solutions for l-waves are obtained. • l-waves of the potential tend to the solutions of the Kratzer and Coulomb potentials. • A non-evident identity between hypergeometric functions is demonstrated.

Effective Kratzer and Coulomb potentials as limit cases of a multiparameter exponential-type potential

International Nuclear Information System (INIS)

García-Ravelo, J.; Menéndez, A.; García-Martínez, J.; Schulze-Halberg, A.

2014-01-01

We show that the effective Kratzer and Coulomb potentials can be obtained by taking particular limits of a multiparameter exponential potential that was studied recently. Moreover, we demonstrate that the bound state solutions of the exponential potential reduce correctly to their well-known counterparts associated with the Kratzer and Coulomb potentials. As a byproduct, we obtain a new limit relation for the hypergeometric function. - Highlights: • Kratzer and Coulomb potentials are limit cases of an exponential-type potential. • From exact s-waves, approximate solutions for l-waves are obtained. • l-waves of the potential tend to the solutions of the Kratzer and Coulomb potentials. • A non-evident identity between hypergeometric functions is demonstrated

Crown Fire Potential

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Earth Data Analysis Center, University of New Mexico — Crown fire potential was modeled using FlamMap, an interagency fire behavior mapping and analysis program that computes potential fire behavior characteristics. The...

Retail Spending Potential

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City and County of Durham, North Carolina — This map shows the average household spending potential for retail goods in the United States in 2012. Spending potential data measures household consumer spending...

Virulence factors in Proteus bacteria from biofilm communities of catheter-associated urinary tract infections.

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Hola, Veronika; Peroutkova, Tereza; Ruzicka, Filip

2012-07-01

More than 40% of nosocomial infections are those of the urinary tract, most of these occurring in catheterized patients. Bacterial colonization of the urinary tract and catheters results not only in infection, but also various complications, such as blockage of catheters with crystalline deposits of bacterial origin, generation of gravels and pyelonephritis. The diversity of the biofilm microbial community increases with duration of catheter emplacement. One of the most important pathogens in this regard is Proteus mirabilis. The aims of this study were to identify and assess particular virulence factors present in catheter-associated urinary tract infection (CAUTI) isolates, their correlation and linkages: three types of motility (swarming, swimming and twitching), the ability to swarm over urinary catheters, biofilm production in two types of media, urease production and adherence of bacterial cells to various types of urinary tract catheters. We examined 102 CAUTI isolates and 50 isolates taken from stool samples of healthy people. Among the microorganisms isolated from urinary catheters, significant differences were found in biofilm-forming ability and the swarming motility. In comparison with the control group, the microorganisms isolated from urinary catheters showed a wider spectrum of virulence factors. The virulence factors (twitching motility, swimming motility, swarming over various types of catheters and biofilm formation) were also more intensively expressed. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Caffeine-induced increase in voluntary activation and strength of the quadriceps muscle during isometric, concentric and eccentric contractions.

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Behrens, Martin; Mau-Moeller, Anett; Weippert, Matthias; Fuhrmann, Josefin; Wegner, Katharina; Skripitz, Ralf; Bader, Rainer; Bruhn, Sven

2015-05-13

This study investigated effects of caffeine ingestion (8 mg/kg) on maximum voluntary torque (MVT) and voluntary activation of the quadriceps during isometric, concentric and eccentric contractions. Fourteen subjects ingested caffeine and placebo in a randomized, controlled, counterbalanced, double-blind crossover design. Neuromuscular tests were performed before and 1 h after oral caffeine and placebo intake. MVTs were measured and the interpolated twitch technique was applied during isometric, concentric and eccentric contractions to assess voluntary activation. Furthermore, normalized root mean square of the EMG signal was calculated and evoked spinal reflex responses (H-reflex evoked at rest and during weak isometric voluntary contraction) as well as twitch torques were analyzed. Caffeine increased MVT by 26.4 N m (95%CI: 9.3-43.5 N m, P = 0.004), 22.5 N m (95%CI: 3.1-42.0 N m, P = 0.025) and 22.5 N m (95%CI: 2.2-42.7 N m, P = 0.032) for isometric, concentric and eccentric contractions. Strength enhancements were associated with increases in voluntary activation. Explosive voluntary strength and voluntary activation at the onset of contraction were significantly increased following caffeine ingestion. Changes in spinal reflex responses and at the muscle level were not observed. Data suggest that caffeine ingestion induced an acute increase in voluntary activation that was responsible for the increased strength regardless of the contraction mode.

Somatic symptoms of anxiety and nonadherence to statin therapy.

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Korhonen, Maarit Jaana; Pentti, Jaana; Hartikainen, Juha; Kivimäki, Mika; Vahtera, Jussi

2016-07-01

The association between anxiety and nonadherence to preventive therapies remains unclear. We investigated whether somatic symptoms of anxiety predict statin nonadherence. This is a prospective cohort study of 1924 individuals who responded to a questionnaire survey on health status and initiated statin therapy after the survey during 2008-2010. We followed the cohort for nonadherence, defined as the proportion of days covered pain upon anger or emotion, sweating without exercise, flushing, tremor of hands or voice, muscle twitching) before the statin initiation, and 16% had experienced at least one symptom on average weekly to daily. 49% of respondents were nonadherent. Weekly to daily occurrence of these symptoms predicted a 33% increase in the risk of nonadherence (risk ratio [RR] 1.33, 95% confidence interval, CI, 1.13-1.57) compared to no symptoms when adjusted for sociodemographics, lifestyle risks, cardiovascular comorbidities, and depression. Particularly, chest pain upon anger or emotion (RR 1.21, 95% CI 1.01-1.46) and muscle twitching (RR 1.24, 95% CI 1.08-1.42) predicted an increased risk of nonadherence to statin therapy. Psychological symptoms of anxiety were not associated with nonadherence when adjusted for somatic symptoms. Somatic anxiety-related symptoms predicted nonadherence to statin therapy. Information on pre-existing somatic symptoms may help identifying patients at increased risk of statin nonadherence. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

EMG analysis tuned for determining the timing and level of activation in different motor units.

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Lee, Sabrina S M; Miara, Maria de Boef; Arnold, Allison S; Biewener, Andrew A; Wakeling, James M

2011-08-01

Recruitment patterns and activation dynamics of different motor units greatly influence the temporal pattern and magnitude of muscle force development, yet these features are not often considered in muscle models. The purpose of this study was to characterize the recruitment and activation dynamics of slow and fast motor units from electromyographic (EMG) recordings and twitch force profiles recorded directly from animal muscles. EMG and force data from the gastrocnemius muscles of seven goats were recorded during in vivo tendon-tap reflex and in situ nerve stimulation experiments. These experiments elicited EMG signals with significant differences in frequency content (p<0.001). The frequency content was characterized using wavelet and principal components analysis, and optimized wavelets with centre frequencies, 149.94 Hz and 323.13 Hz, were obtained. The optimized wavelets were used to calculate the EMG intensities and, with the reconstructed twitch force profiles, to derive transfer functions for slow and fast motor units that estimate the activation state of the muscle from the EMG signal. The resulting activation-deactivation time constants gave r values of 0.98-0.99 between the activation state and the force profiles. This work establishes a framework for developing improved muscle models that consider the intrinsic properties of slow and fast fibres within a mixed muscle, and that can more accurately predict muscle force output from EMG. Copyright © 2011 Elsevier Ltd. All rights reserved.

Administration of rocuronium based on real body weight versus fat-free mass in patients with lymphedema.

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Jing, Zhang; Muheremu, Aikeremujiang; Liu, Pengfei; Hu, Xiaoyun; Binjiang, Zhao

2017-12-01

Objective To compare the clinical pharmacokinetics of rocuronium when applied according to fat-free mass versus real body weight during anesthetic induction of patients with lymphedema. Methods Sixty patients with lymphedema (age, 18-60 years; American Society of Anesthesiologists physical status, I-II) undergoing elective surgery with general anesthesia were randomly divided into two groups. Rocuronium was administered based on the fat-free mass in 30 patients and real body weight in 30 patients. General anesthesia was induced with propofol and remifentanil by target-controlled infusion. Intubation was attempted when the onset time (T1) (time from end of bolus injection to 100% twitch depression) reached maximal inhibition, and respiratory support with mechanical ventilation was then applied. The T1, clinical duration (time from end of bolus injection to recovery of twitch tension to 25% of control), recovery index (time from 25% to 75% of recovery of T1), and dosage were recorded. Results Complete data were recorded for 59 patients, and there were no significant differences in the general condition, intubation condition, or median duration of action of rocuronium between the two groups. However, the median T1, recovery index, and dosage of rocuronium were significantly different. Conclusion Good intubation conditions and a shortened clinical duration can be obtained for patients with lymphedema when induction with rocuronium is based on the fat-free mass.

Nootropic activity of tuber extract of Pueraria tuberosa (Roxb).

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Rao, N Venkata; Pujar, Basavaraj; Nimbal, S K; Shantakumar, S M; Satyanarayana, S

2008-08-01

Nootropic effect of alcoholic (ALE; 50, 75, 100 mg/kg) and aqueous (AQE; 100, 200, 400 mg/kg) extracts of P. tuberosa was evaluated by using Elevated Plus Maze (EPM), scopolamine-induced amnesia (SIA), diazepam-induced amnesia (DIA), clonidine-induced (NA-mediated) hypothermia (CIH), lithium-induced (5-HT mediated) head twitches (LIH) and haloperidol-induced (DA- mediated) catalepsy (HIC) models. Piracetam was used as the standard drug. A significant increase in inflexion ratio (IR) was recorded in EPM, SIA and DIA models. A significant reversal effect was observed on rectal temperature in CIH model, reduction of head twitches in LIH models. However no significant reduction in catalepsy scores in HIC models were observed with test extracts and standard piracetam. The results indicate that nootropic activity observed with ALE and AQE of tuber extracts of P. tuberosa could be through improved learning and memory either by augmenting the noradrenaline (NA) transmission or by interfering with 5-hydroxytryptamine (5-HT) release. Further, the extracts neither facilitated nor blocked release of the dopamine (DA). Thus ALE and AQE elicited significant nootropic effect in mice and rats by interacting with cholinergic, GABAnergic, adrenergic and serotonergic systems. Phytoconstituents like flavonoids have been reported for their nootropic effect and these are present in both ALE and AQE extracts of tubers of P. tuberosa (Roxb) and these active principles may be responsible for nootropic activity.