Effects of different movement directions on electromyography recorded from the shoulder muscles while passing the target positions

PurposeWe compared electromyography (EMG) recorded from the shoulder joint muscles in the same position for different movement directions.MethodsFifteen healthy subjects participated. They performed shoulder elevation from 0° to 120°, shoulder depression from 120° to 0°, shoulder horizontal adduction from ?15° to 105°, and shoulder horizontal abduction from 105° to ?15°. The target positions were 90° shoulder elevation in the 0°, 30°, 60°, and 90° planes (0°, 30°, 60°, and 90° positions). EMG signals were recorded from the supraspinatus (SSP) muscle by fine-wire electrodes. EMG signals from the infraspinatus (ISP), anterior deltoid, middle deltoid, and posterior deltoid muscles were recorded using active surface electrodes.ResultsDuring elevation and horizontal abduction, the SSP showed significantly higher activity than that shown during depression and during horizontal adduction in the 0°, 30°, and 60° positions. During elevation, the ISP showed significantly higher activity than during depression and during horizontal adduction in the 90° position. During horizontal abduction, the ISP showed significantly higher activity than during depression in the 90° position.ConclusionsWhen the movement tasks were performed in different movement directions at the same speed, each muscle showed characteristic activity.     

Paternity analysis in a progeny test of Cryptomeria japonica revealed adverse effects of pollen contamination from outside seed orchards on morphological traits

To evaluate the effects of pollen contamination from outside of Cryptomeria japonica seed orchard on the growth performance (height and diameter at breast height, DBH) and morphological traits (stem straightness and basal stem straightness), paternity testing using seven microsatellite markers was performed in a progeny test. In the studied progeny test, high rates of inconsistency were found between the observed and expected genotypes. The average rates of pollen contamination from outside the orchard and self-fertilization were 58.47% and 0.65%, respectively. We divided the individuals of the studied progeny test into two groups based on their genotype data, for which: (1) both parents were elite trees and (2) only the mother trees were elite trees, and then compared them with respect to the growth performance and morphological traits of progenies using data at 20 and 30 years old. Significant adverse effects of contaminating pollen were detected in relation to straightness, but not tree height and DBH. The results suggest that the genetic gains for straightness generally show higher narrow-sense heritability than growth traits, which should be increased by reduction of pollen contamination. Breeding with paternal analysis (BWPA) is an effective approach for evaluating breeding materials based on maternal and paternal information revealed by DNA markers. The use of BWPA in progeny test allows effective forward- and backward selection without laborious and time-consuming tasks. In this study, we also suggest that the significant pollen contamination and paternal deviation found in the open-pollinated progeny test are serious impediments for BWPA.     

Nup358, a nucleoporin, functions as a key determinant of the nuclear pore complex structure remodeling during skeletal myogenesis

The nuclear pore complex (NPC) is the only gateway for molecular trafficking across the nuclear envelope. The NPC is not merely a static nuclear-cytoplasmic transport gate; the functional analysis of nucleoporins has revealed dynamic features of the NPC in various cellular functions, such as mitotic spindle formation and protein modification. However, it is not known whether the NPC undergoes dynamic changes during biological processes such as cell differentiation. In the present study, we evaluate changes in the expression levels of several nucleoporins and show that the amount of Nup358/RanBP2 within individual NPCs increases during muscle differentiation in C2C12 cells. Using atomic force microscopy, we demonstrate structural differences between the cytoplasmic surfaces of myoblast and myotube NPCs and a correlation between the copy number of Nup358 and the NPC structure. Furthermore, small interfering RNA-mediated depletion of Nup358 in myoblasts suppresses myotube formation without affecting cell viability, suggesting that NUP358 plays a role in myogenesis. These findings indicate that the NPC undergoes dynamic remodeling during muscle cell differentiation and that Nup358 is prominently involved in the remodeling process.     

Elucidations of the Catalytic Cycle of NADH-Cytochrome b5 Reductase by X-ray Crystallography: New Insights into Regulation of Efficient Electron Transfer

NADH-Cytochrome b₅ reductase (b5R), a flavoprotein consisting of NADH and flavin adenine dinucleotide (FAD) binding domains, catalyzes electron transfer from the two-electron carrier NADH to the one-electron carrier cytochrome b₅ (Cb5). The crystal structures of both the fully reduced form and the oxidized form of porcine liver b5R were determined. In the reduced b5R structure determined at 1.68 Å resolution, the relative configuration of the two domains was slightly shifted in comparison with that of the oxidized form. This shift resulted in an increase in the solvent-accessible surface area of FAD and created a new hydrogen-bonding interaction between the N5 atom of the isoalloxazine ring of FAD and the hydroxyl oxygen atom of Thr66, which is considered to be a key residue in the release of a proton from the N5 atom. The isoalloxazine ring of FAD in the reduced form is flat as in the oxidized form and stacked together with the nicotinamide ring of NAD⁺. Determination of the oxidized b5R structure, including the hydrogen atoms, determined at 0.78 Å resolution revealed the details of a hydrogen-bonding network from the N5 atom of FAD to His49 via Thr66. Both of the reduced and oxidized b5R structures explain how backflow in this catalytic cycle is prevented and the transfer of electrons to one-electron acceptors such as Cb5 is accelerated. Furthermore, crystallographic analysis by the cryo-trapping method suggests that re-oxidation follows a two-step mechanism. These results provide structural insights into the catalytic cycle of b5R.     

Regulated DNA rearrangement during sporulation in Bacillus weihenstephanensis KBAB4

Temperate phages can integrate their genomes into a specific region of a host chromosome to produce lysogens (prophage). During genome insertion, prophages may interrupt the gene coding sequence. In Bacillus subtilis, the sigma factor gene sigK is interrupted by a 48 kb prophage‐like element. sigK is a composite coding sequence from two partial genes during sporulation. For over two decades, however, no further examples of DNA element‐mediated gene reconstitution other than sigK have been identified in spore formers. Here we report that the gene for dipicolinic acid (DPA) synthetase β subunit spoVFB in B. weihenstephanensis KBAB4 is interrupted by a prophage‐like element named vfbin. DPA is synthesized in the mother cell and required for maintaining spore dormancy. We found that spoVFB was a composite coding sequence generated in the mother cell via chromosomal rearrangement that excised vfbin. Furthermore, vfbin caused excision after phage‐inducer treatment, but vfbin appeared to be defective as a prophage. We also found various spore‐forming bacteria in which sporulation‐related genes were disrupted by prophage‐like DNA elements. These results demonstrate the first example of a similar mechanism that affects a sporulation gene other than sigK and suggest that this prophage‐mediated DNA rearrangement is a common phenomenon in spore‐forming bacteria.     

Enzymatic Maceration Mechanism in Biochemical Pulping of Mitsumata (Edgeworthia papyrifera Sieb. et Zucc) Bast

In a comparison of the crude enzyme secreted by the bacterium Erwinia carotovora FERM P-7576 with the corresponding purified overall endo-pectate lyase (endo-PATE) pI (isoelectric point)-isozymes on a basis of equal activity and constitution, the former was found to have higher maceration activity for biochemical pulping of caustic soda-presoaked mitsumata (Edgeworthia papyrifera Sieb. et Zucc) bast than the latter. These results indicate that some additional enzyme (s) other than endo-PATE are required for the maceration. Focusing on the maceration activity, the intensive purification of the crude enzyme was conducted and another maceration factor was successfully isolated, which was identified as endo-pectin lyase (endo-PNTE). The isolated endo-PNTE was found to have a molecular weight of 39,000, pI of 8.2 and a specific activity of 7,911 units/mg. The enzymatic maceration of this bast fiber was concluded to be due only to a concerted action of these two enzymes.     

Egg viability of the rotifer Brachionus urceolaris after ingestion by the predatory cladoceran Leptodora kindtii

Egg resistance against the digestive process of a predator is an effective strategy for zooplankton to compensate population loss due to predation. Parthenogenetic eggs of the rotiferan Brachionus urceolaris, which were ingested by the predatory cladoceran Leptodora kindtii, were expelled from the feeding basket of the predator without digestion. We found a negative correlation between the unconsumed ratio of eggs after ingestion and body length of the predator. As high as 75% of the unconsumed eggs successfully hatched and the hatch ratio was independent of body length of L. kindtii. Our results indicate that the rotifer has an effective strategy to maintain its population in the environment with abundant invertebrate predators.     

Effects of reconstituted HDL on charge-based LDL subfractions as characterized by capillary isotachophoresis

Modified LDL in human plasma including small, dense LDL (sdLDL) and oxidized LDL carries a more negative charge than unmodified LDL and is atherogenic. We examined the effects of apolipoprotein A-I (apoA-I)/POPC discs on charge-based LDL subfractions as determined by capillary isotachophoresis (cITP). Three normal healthy subjects and seven patients with metabolic disorders were included in the study. LDL in human plasma was separated into two major subfractions, fast- and slow-migrating LDL (fLDL and sLDL), by cITP. Normal LDL was characterized by low fLDL, and mildly oxidized LDL in vitro and mildly modified LDL in human plasma were characterized by increased fLDL. Moderately oxidized LDL in vitro and moderately modified LDL in a patient with hypertriglyceridemia and HDL deficiency were characterized by both increased fLDL and a new LDL subfraction with a faster mobility than fLDL [very-fast-migrating LDL as determined by cITP (vfLDL)]. cITP LDL subfractions with faster electrophoretic mobility (fLDL vs. sLDL, vfLDL vs. fLDL) were associated with an increased content of sdLDL. Incubation of a plasma fraction with d>1.019 g/ml (depleted of triglyceride-rich lipoproteins) in the presence of apoA-I/POPC discs at 37 degrees C greatly decreased vfLDL and fLDL but increased sLDL. Incubation of whole plasma from patients with an altered distribution of cITP LDL subfractions in the presence of apoA-I/POPC discs also greatly decreased fLDL but increased sLDL. ApoA-I/POPC discs decreased the cITP fLDL level, the free cholesterol concentration, and platelet-activating factor acetylhydrolase activity in the sdLDL subclasses (d=1.040-1.063 g/ml) and increased the size of LDL. ApoA-I/POPC discs reduced charge-modified LDL in human plasma by remodeling cITP fLDL into sLDL subfractions.     

Bmi1 cooperates with Dnmt1-associated protein 1 in gene silencing

Polycomb group (PcG) proteins are involved in gene silencing through chromatin modifications. Among polycomb repressive complexes (PRCs), PRC1 exhibits H2A-K119 ubiquitin E3 ligase activity. However, the molecular mechanisms underlying PRC1-mediated gene silencing remain largely obscure. In this study, we found that Bmi1 directly interacts with Dnmt-associated protein 1 (Dmap1), which has been characterized to associate with the maintenance DNA methyltransferase, Dnmt1. Bmi1 was demonstrated to form a ternary complex with Dmap1 and Dnmt1 with Dmap1 in the central position. Chromatin immunoprecipitations confirmed the ternary complex formation within the context of the PRC1 at the Bmi1 target loci. Loss of Dmap1 binding to the Bmi1 target loci was tightly associated with derepressed gene expression in Bmi1-/- cells. Dmap1 knockdown exhibited the same impact as Bmi1 knockout did on the expression of Bmi1 targets, including Hox genes. Collectively, our findings suggest that Bmi1 incorporates Dmap1 in polycomb gene silencing.