Supramolecular Organization of the α121-α565 Collagen IV Network

Collagen IV is a family of 6 chains (α1-α6), that form triple-helical protomers that assemble into supramolecular networks. Two distinct networks with chain compositions of α121 and α345 have been established. These oligomerize into separate α121 and α345 networks by a homotypic interaction through their trimeric noncollagenous (NC1) domains, forming α121 and α345 NC1 hexamers, respectively. These are stabilized by novel sulfilimine (SN) cross-links, a covalent cross-link that forms between Met⁹³ and Hyl²¹¹ at the trimer-trimer interface. A third network with a composition of α1256 has been proposed, but its supramolecular organization has not been established. In this study we investigated the supramolecular organization of this network by determining the chain identity of sulfilimine-cross-linked NC1 domains derived from the α1256 NC1 hexamer. High resolution mass spectrometry analyses of peptides revealed that sulfilimine bonds specifically cross-link α1 to α5 and α2 to α6 NC1 domains, thus providing the spatial orientation between interacting α121 and α565 trimers. Using this information, we constructed a three-dimensional homology model in which the α565 trimer shows a good chemical and structural complementarity to the α121 trimer. Our studies provide the first chemical evidence for an α565 protomer and its heterotypic interaction with the α121 protomer. Moreover, our findings, in conjunction with our previous studies, establish that the six collagen IV chains are organized into three canonical protomers α121, α345, and α565 forming three distinct networks: α121, α345, and α121-α565, each of which is stabilized by sulfilimine bonds between their C-terminal NC1 domains.     

Enhancement of neuromuscular dynamics and strength behavior using extremely low magnitude mechanical signals in mice

Exercise in general, and mechanical signals in particular, help ameliorate the neuromuscular symptoms of aging and possibly other neurodegenerative disorders by enhancing muscle function. To better understand the salutary mechanisms of such physical stimuli, we evaluated the potential for low intensity mechanical signals to promote enhanced muscle dynamics. The effects of daily brief periods of low intensity vibration (LIV) on neuromuscular functions and behavioral correlates were assessed in mice. Physiological analysis revealed that LIV increased isometric force production in semitendinosus skeletal muscle. This effect was evident in both young and old mice. Isometric force recordings also showed that LIV reduced the fatiguing effects of intensive synaptic muscle stimulation. Furthermore, LIV increased evoked neurotransmitter release at neuromuscular synapses but had no effect on spontaneous end plate potential amplitude or frequency. In behavioral studies, LIV increased mouse grip strength and potentiated initial motor activity in a novel environment. These results provide evidence for the efficacy of LIV in producing changes in the neuromuscular system that translate into performance gains at a behavioral scale.     

Structural analysis of the starfish SALMFamide neuropeptides S1 and S2: The N-terminal region of S2 facilitates self-association

The neuropeptides S1 (GFNSALMFamide) and S2 (SGPYSFNSGLTFamide), which share sequence similarity, were discovered in the starfish Asterias rubens and are prototypical members of the SALMFamide family of neuropeptides in echinoderms. SALMFamide neuropeptides act as muscle relaxants and both S1 and S2 cause relaxation of cardiac stomach and tube foot preparations in vitro but S2 is an order of magnitude more potent than S1. Here we investigated a structural basis for this difference in potency using spectroscopic techniques. Circular dichroism spectroscopy showed that S1 does not have a defined structure in aqueous solution and this was supported by 2D nuclear magnetic resonance experiments. In contrast, we found that S2 has a well-defined conformation in aqueous solution. However, the conformation of S2 was concentration dependent, with increasing concentration inducing a transition from an unstructured to a structured conformation. Interestingly, this property of S2 was not observed in an N-terminally truncated analogue of S2 (short S2 or SS2; SFNSGLTFamide). Collectively, the data obtained indicate that the N-terminal region of S2 facilitates peptide self-association at high concentrations, which may have relevance to the biosynthesis and/or bioactivity of S2 in vivo.     

Ascaris lumbricoides: Coproporphyrinogen oxidase activity in eggs and muscle

The possible presence of coproporphyrinogen oxidase (EC 1.3.3.3), an oxygen-requiring enzyme in the porphyrin biosynthetic pathway, was investigated in supernatant fractions of homogenized Ascaris lumbricoides muscle and developing eggs, and in mitochondrial preparations of muscle. Compared with rat liver controls, low levels of enzyme activity were found in A. lumbricoides gut, 6-day eggs, and muscle mitochondria. Enzyme activity in muscle, 8-day, and 25-day eggs was not measurable under the conditions employed.     

Calcitonin gene-related peptide expression at endplates of different fibre types in muscles in rat hind limbs

Calcitonin gene-related peptide (CGRP) occurs only in some motoneurons. In this study, the presence of CGRP in motor endplates in relation to muscle fibre types was examined in slow (soleus muscle) and fast [tibialis anterior (TA) and extensor digitorum longus (EDL)] leg muscles of the rat. CGRP was detected by use of immunohistochemical methods, and staining for the mitochondrial-bound enzyme NADH-TR was used for demonstration of fibre types. The fibres showing low NADH-TR activity were interpreted as representing IIB fibres. All such fibres located in the superficial portion of TA were innervated by endplates displaying CGRP-like immunoreactivity (LI), whereas in the deep portion of TA some of these fibres lacked CGRP-LI at their endplates. Thirty per cent of the IIB fibres in EDL showed CGRP-LI at the endplates. All fibres in TA and EDL displaying high NADH-TR activity and interpreted as type-IIA fibres, lacked CGRP-LI in their motor innervation. One third of the fibres with intermediate NADH-TR activity in TA exhibited CGRP-LI at their endplates, whereas in EDL only few such fibres displayed CGRP-LI in the endplate formation. These fibres are likely to belong to type-IIX or type-I motor units. CGRP-LI was very rarely detected at the endplates in the soleus muscle. These observations show that distinct differences exist between the slow muscle, soleus, and the fast muscles, TA and EDL, but that there are also differences between the different types of fibres in TA and EDL with respect to presence of CGRP-LI at the endplates. As CGRP-LI was frequently detected at endplates of IIB fibres, it is likely that CGRP has a particular role related to the differentiation and maintenance of these fibres.     

血浆鸢尾素、生长分化因子-8在老年肌肉减少症患者中的表达及其诊断价值分析

摘要 目的：分析血浆鸢尾素、生长分化因子-8在老年肌肉减少症患者中的表达及其诊断价值。方法：选择2020年4月至2022年4月在我院接受体检的620例老年体检者作为研究对象,其中238例肌肉减少症者作为观察组,382例非肌肉减少症者作为对照组。检测两组血浆鸢尾素、生长分化因子-8表达水平,观察血浆鸢尾素、生长分化因子-8在不同严重程度的老年肌肉减少症患者中差异性,使用Pearson相关性分析血浆鸢尾素、生长分化因子-8与全身骨骼肌质量、四肢骨骼肌质量、相对骨骼肌指数、握力和步速的关系,通过ROC下面积（AUC）评价血浆鸢尾素、生长分化因子-8对老年肌肉减少症的诊断效能。结果：观察组血浆鸢尾素表达水平低于对照组,生长分化因子-8表达水平高于对照组（P＜0.05）;肌肉减少症前期、肌肉减少症期、重度肌肉减少症期的患者血浆鸢尾素表达水平依次降低,生长分化因子-8表达水平依次升高（P＜0.05）;经Pearson相关性分析,老年肌肉减少症患者全身骨骼肌质量、四肢骨骼肌质量、相对骨骼肌指数、握力和步速均与子鸢尾素呈正相关(P＜0.05),与生长分化因子-8呈负相关（P＜0.05）;经ROC曲线分析,血浆鸢尾素、生长分化因子-8诊断老年肌肉减少症的敏感度为78.49 %,特异度为90.67 %,AUC为0.914。结论：老年肌肉减少症患者血浆鸢尾素表达明显下调、生长分化因子-8表达明显上调,两者与病情严重程度密切相关,联合诊断此病的效能较好,值得临床予以重视应用。     

Nystatin-, mycoheptin- and levorin-induced conductance in the membrane of frog skeletal muscle fibres

The effects of the polyene antibiotics nystatin (2 × 10^–5–10^–4 mol/l), mycoheptin (1.3 × 10^–6–10^–5 mol/l) and levorin (10^–8–5 × 10^–5 mol/l)on isolated frog skeletal muscle fibres and whole sartorius muscles of the frog have been investigated. Cation conductance was measured under current clamp conditions using a double sucrosegap technique. Cation effluxes were studied by means of flame emission photometry. All three antibiotics increased the cation conductance and efflux rates; however, differences between the polyenes were found in the steady state values of induced cation transport at a given concentration. The values of both induced conductance g_A and efflux rate constants K_A formed the following sequence: levorin > mycoheptin > nystatin, demonstrating a correlation with the order of antifungal activities. The dose-response curves of lg polyene-induced cation transport against lg of antibiotic concentration in our experiments had slope values which were much lower than those in bilayers: 1.7 and 1.3 for nystatin and mycoheptin, respectively, whereas the aromatic heptaene levorin had an even smaller concentration dependence. The decline in the equilibrium conductance caused by nystatin- and mycoheptin removal was very fast (during the first minute = 0.74 and 2.39 min, respectively). In contrast, levorin-induced conductance was irreversible. It is proposed that the processes which limit the rate of channel formation are different in biological and model membranes. Correspondence to: N. E. Shvinka     

A dynamometer for evaluation of dynamic muscle work

The validation of a new dynamometer for evaluation of dynamic muscle work is presented. The device was based on a precise measurement of load displacements of any machine using gravitational loads as external resistance. It allowed, through a sensor consisting of an infrared photo interrupter, the calculation of velocity, force and power during concentric, eccentric and stretch-shortening cycle activity. To validate the dynamometer 33 male and female track and field athletes (12 throwers and 21 jumpers) participated in the study. The throwers (4 women and 8 men) were asked to perform half-squat exercises on a slide machine with a load of 100% of the subject's body mass. The day-to-day reproducibility of half-squat exercises gave a correlation coefficient ofr = 0.88, 0.97 and 0.95 for average push-off force (AF), average push-off velocity (AV), and average push-off power (AP) respectively. Comparison of half-squat measurements was performed against jumping and running test evaluation by the jumpers (7 women and 14 men). The interrelationships among the different variables studied demonstrated a strong correlation between AF, AV and AP and sprinting and jumping parameters (r = 0.53–0.97;P < 0.05–0.001). Using values of AF, AV and AP developed in half-squat exercises executed with different loads, ranging from 35% to 210% of the subject's body mass, it was also possible to establish the force-velocity and power-velocity relationships for both male and female jumpers. In any individual case, the maximal error due to the measurement system was calculated to be less than 0.3%, 0.9% and 1.2% for AF, AV, and AP respectively. Given the accuracy of the ergometer, the high reliability found between 2 days of measurements, and the specificity of the results it is suggested that the dynamic dynamometer would be suitable for evaluation of athletes performing specific skills. In addition, because single and multiple joint movements involving appropriate muscle groups can be easily performed, physiological characteristics could be evaluated for both athletic and rehabilitation purposes. Therefore, because of its simplicity of use and application, and its low cost the dynamometer would be suitable for both laboratory and field conditions.     

Alteration of regulatory enzyme activities in fast-twitch and slow-twitch muscles and muscle fibres in low-intensity endurance-trained rats

The effect of progressive, low-intensity endurance training on regulatory enzyme activities in slow-twitch (ST) and fast-twitch (FT) muscle fibres was studied in 32 rats. Of those rats 16 were trained on a treadmill at a running speed of 10m · min^–1 5 days a week over an 8-week period. Running time was progressively increased from 15 min to 2 h · day^–1. Of the rats 4 trained and 4 sedentary rats were also subjected to acute exhausting exercise. Enzyme activities of phosphofructokinase 1 (PFKI) from glycolysis, -ketoglutarate dehydrogenase (-KGDH) from the Krebs cycle and carnitine palmitoyltransferase (CPT I and II) from fatty acid metabolism in soleus, tibialis anterior and gastrocnemius muscles were measured in trained and sedentary rats. Enzyme activities of individual ST and FT fibres were measured from the freeze-dried gastrocnemius muscle of 8 trained and 8 sedentary rats. In the sedentary rats the activity of PFK1 in tibialis anterior and soleus muscles was 141% and 41% of the activity in gastrocnemius muscle, respectively. The activity of -KGDH in tibialis anterior and soleus muscles was 164% and 278% of the activity in gastrocnemius muscle, respectively. The activity of CPT I in tibialis anterior and gastrocnemius muscles were at the same level, but in soleus muscle the activity was 127% of that in mixed muscle. Endurance training increased enzyme activities of -KGDH and CPT I significantly (P < 0.05) in gastrocnemius muscle but not in soleus or tibialis anterior muscle. After training both -KGDH and CPT II activities were elevated significantly (P < 0.05) in the ST fibres of gastrocnemius muscle, whereas in FT fibres only -KGDH was increased. For PFK1 activity no significant change was observed in ST or FT fibres. After acute exercise, activities of mitochondrial enzymes -KGDH and CPT I tended to be elevated in all muscles. Thus, low-intensity endurance training induced significant peripheral changes in regulatory enzyme activities in oxidative and fatty acid metabolism in individual ST or FT muscle fibres.