Activation of Lactate Receptor Positively Regulates Skeletal Muscle Mass in Mice.

G protein-coupled receptor 81 (GPR81), a selective receptor for lactate, expresses in skeletal muscle cells, but the physiological role of GPR81 in skeletal muscle has not been fully elucidated. As it has been reported that the lactate administration induces muscle hypertrophy, the stimulation of GPR81 has been suggested to mediate muscle hypertrophy. To clarify the contribution of GPR81 activation in skeletal muscle hypertrophy, in the present study, we investigated the effect of GPR81 agonist administration on skeletal muscle mass in mice. Male C57BL/6J mice were randomly divided into control group and GPR81 agonist-administered group that received oral administration of the specific GPR81 agonist 3-Chloro-5-hydroxybenzoic acid (CHBA). In both fast-twitch plantaris and slow-twitch soleus muscles of mice, the protein expression of GPR81 was observed. Oral administration of CHBA to mice significantly increased absolute muscle weight and muscle weight relative to body weight in the two muscles. Moreover, both absolute and relative muscle protein content in the two muscles were significantly increased by CHBA administration. CHBA administration also significantly upregulated the phosphorylation level of p42/44 extracellular signal-regulated kinase-1/2 (ERK1/2) and p90 ribosomal S6 kinase (p90RSK). These observations suggest that activation of GRP81 stimulates increased the mass of two types of skeletal muscle in mice in vivo. Lactate receptor GPR81 may positively affect skeletal muscle mass through activation of ERK pathway.

Soymilk-fermented with Lactobacillus delbrueckii subsp. delbrueckii TUA4408L improves immune-health in pigs.

Lactobacillus delbrueckii subsp. delbrueckii TUA4408L has the ability to grow and ferment soymilk and is able to modulate the innate immune response of intestinal epithelial cells in vitro. These two properties prompt us to evaluate whether the soymilk fermented with the TUA4408L strain can induce beneficial immunomodulatory effects in vivo. For this purpose, pigs were selected as a preclinical model. The studies performed here demonstrated that the L. delbrueckii subsp. delbrueckii TUA4408L-fermented soymilk (TUA4408L FSM) reduced blood markers of inflammation and differentially regulated the expression of inflammatory and regulatory cytokines in the intestinal mucosa. These immunological changes induced by the TUA4408L FSM were associated to an enhanced resistance to pathogenic Escherichia coli and an improved grow performance and meat quality of pigs. The experiments and analysis in our study indicate that the immunobiotic TUA4408L FSM could be an interesting non-dairy functional food to beneficially modulate the intestinal immune system, improve protection against pathogens and reduce inflammatory damage. The preclinical study carried out here in pigs could have a better correlation in humans, compared to a rodent model. However, the clinical relevance of these findings still needs to be confirmed by further research, for example, in controlled human challenge studies.

Three-Dimensional Proton Beam Writing of Optically Active Coherent Vacancy Spins in Silicon Carbide.

Constructing quantum devices comprises various challenging tasks, especially when concerning their nanoscale geometry. For quantum color centers, the traditional approach is to fabricate the device structure after the nondeterministic placement of the centers. Reversing this approach, we present the controlled generation of quantum centers in silicon carbide (SiC) by focused proton beam in a noncomplex manner without need for pre- or postirradiation treatment. The generation depth and resolution can be predicted by matching the proton energy to the material's stopping power, and the amount of quantum centers at one specific sample volume is tunable from ensembles of millions to discernible single photon emitters. We identify the generated centers as silicon vacancies through their characteristic magnetic resonance signatures and demonstrate that they possess a long spin-echo coherence time of 42 ± 20 µs at room temperature. Our approach hence enables the fabrication of quantum hybrid nanodevices based on SiC platform, where spin centers are integrated into p-i-n diodes, photonic cavities, and mechanical resonators.

Development of an in vitro immunobiotic evaluation system against rotavirus infection in bovine intestinal epitheliocytes.

The bovine intestinal epithelial cell line (BIE cells) expresses the Toll-like receptor (TLR)3 and is able to mount an antiviral immune response after the stimulation with poly(I:C). In the present study, we aimed to further characterise the antiviral defence mechanisms in BIE cells by evaluating the innate immune response triggered by rotavirus (RV) infection. In addition, we attempted to determine whether immunobiotic bifidobacteria are able to confer protection of BIE cells against RV infection by beneficially modulating the antiviral immune response. RV OSU (porcine) and UK (bovine) effectively infected BIE cells, while a significant lower capacity to infect BIE cells was observed for human (Wa) and murine (EW) RV. We observed that viral infection in BIE cells triggered TLR3/RIG-I-mediated immune responses with activation of IRF3 and TRAF3, induction of interferon beta (IFN-ß) and up-regulation of inflammatory cytokines. Our results also demonstrated that preventive treatments with Bifidobacterium infantis MCC12 or Bifidobacterium breve MCC1274 significantly reduced RV titres in infected BIE cells and differentially modulated the innate immune response. Of note, both strains significantly improved the production of the antiviral factor IFN-ß in RV-infected BIE cells. In conclusion, this work provides comprehensive information on the antiviral immune response of BIE cells against RV, that can be further studied for the development of strategies aimed to improve antiviral defences in bovine intestinal epithelial cells. Our results also demonstrate that BIE cells could be used as a newly immunobiotic evaluation system against RV infection for application in the bovine host.

Modulation of porcine intestinal epitheliocytes immunetranscriptome response by Lactobacillus jensenii TL2937.

In order to evaluate probiotic strains applicable for the beneficial immunomodulation of the porcine gut (immunobiotics), we previously developed a porcine intestinal epitheliocyte cell line (PIE cells). Here, transcriptomic studies using PIE cells were performed considering that this information would be valuable for understanding the mechanisms involved in the protective activity of the immunobiotic strain Lactobacillus jensenii TL2937 against intestinal inflammatory damage in pigs. In addition, those studies would provide criteria for selecting biomarkers for the screening of new immunobiotic strains. We performed microarray analysis to investigate the transcriptomic response of PIE cells to the challenge with heat-stable enterotoxigenic Escherichia coli (ETEC) pathogen-associated molecular patterns (PAMPs) and, the changes induced by L. jensenii TL2937 in that response. The approach allowed us to obtain a global overview of the immune genes involved in the response of PIE cells to heat-stable ETEC PAMPs. We observed that L. jensenii TL2937 differently modulated gene expression in ETEC PAMPs-challenged PIE cells. Microarray and RT-PCR analysis indicated that the most remarkable changes in PIE cells transcriptomic profile after heat-stable ETEC PAMPs challenge were observed in chemokines, adhesion molecules, complement and coagulation cascades factors. In addition, an anti-inflammatory effect triggered by TL2937 strain in PIE cells was clearly demonstrated. The decrease in the expression of chemokines (CCL8, CXCL5, CXCL9, CXCL10, and CXCL11), complement (C1R, C1S, C3, and CFB), and coagulation factors (F3) by L. jensenii TL2937 supports our previous reports on the immunoregulatory effect of this strain. These results provided clues for the better understanding of the mechanism underlying host-immunobiotic interaction in the porcine host. The comprehensive transcriptomic profiles of PIE cells provided by our analyses successfully identified a group of genes, which could be used as prospective biomarkers for the screening and evaluation of new anti-inflammatory immunobiotics for the prevention of inflammatory intestinal disorders in pigs.

Hanging drop monoculture for selection of optimal antioxidants during in vitro maturation of porcine oocytes.

We analysed the effect of three antioxidants that have different functional mechanisms on the in vitro maturation (IVM) of porcine oocytes. Single oocyte monoculture using the hanging drop (HD) system has some advantages such as improving analysis efficiency brought by the smaller number of samples than the number of oocytes cultured in one drop. Direct effects of ligands on single oocytes could also be detected without considering the effects of paracrine factors from other oocytes. After 22 h of pre-culture, denuded oocytes were cultured for 22 h with 0.01 and 0.1 µg/ml of L-carnitine (LC), lactoferrin (LF) or sulforaphane (SF) in the presence/non-presence of oxidant stress induced by H2O2 supplementation to evaluate the reducing effects against oxidative stress on nuclear maturation. As a result, compared with LC and SF, LF showed effective reduction in oxidative stress at a lower concentration (0.01 µg/ml), suggesting that LF is a more effective antioxidant in porcine oocyte IVM. Additionally, LF also increased maturation rate even in culture without H2O2. Our results clearly suggest that the HD monoculture system is useful for screening the substances that affect porcine oocyte culture.

Is greater than 145{degrees} of deep knee flexion under weight-bearing conditions safe after total knee arthroplasty?: a fluoroscopic analysis of Japanese-style deep knee flexion.

We investigated the characteristics of patients who achieved Japanese-style deep flexion (seiza-sitting) after total knee replacement (TKR) and measured three-dimensional positioning and the contact positions of the femoral and tibial components. Seiza-sitting was achieved after surgery by 23 patients (29 knees) of a series of 463 TKRs in 341 patients. Pre-operatively most of these patients were capable of seiza-sitting, had a lower body mass index and a favourable attitude towards the Japanese lifestyle (27 of 29 knees). According to two-/three-dimensional image registration analysis in the seiza-sitting position, flexion, varus and internal rotation angles of the tibial component relative to the femoral component had means of 148° (SD 8.0), 1.9° (SD 3.2) and 13.4° (SD 5.9), respectively. Femoral surface contact positions tended to be close to the posterior edge of the tibial polyethylene insert, particularly in the lateral compartment, but only 8.3% (two of 24) of knees showed femoral subluxation over the posterior edge. The mean contact positions of the femoral cam on the tibial post were located 7.8 mm (sd 1.5) proximal to the lowest point of the polyethylene surface and 5.5 mm (SD 0.9) medial to the centre of the post, indicating that the post-cam contact position translated medially during seiza-sitting, but not proximally. Collectively, the seiza-sitting position seems safe against component dislocation, but the risks of posterior edge loading and breakage of the tibial polyethylene post remain.

Absorption, distribution and excretion of radioactivity by male rats after a single intravenous or subcutaneous dose of 14C-azacitidine.

The absorption, distribution and excretion of radioactivity were investigated in male Sprague-Dawley rats after a single intravenous or subcutaneous dose of 14C-azacitidine. After subcutaneous administration, 14C-azacitidine was rapidly absorbed and the radioactivity was distributed to the tissues. The absorption of radioactivity after subcutaneous administration was 76.6% of that observed after intravenous administration. There were no marked differences in the tissue distribution of the radioactivity between administration routes. The concentrations of radioactivity in most tissues, including the spleen and bone marrow, which are sites of action of azacitidine, were higher than those in the plasma. Particularly high concentrations of radioactivity were detected in the spleen, kidney and liver. The accumulation of radioactivity in blood cells increased from 0.5 to 48 h. The binding of azacitidine to serum protein was low at <9%, and the cumulative urinary and fecal excretion of radioactivity for 168 h after intravenous or subcutaneous administration was >95% of the administered dose, indicating that radioactivity did not accumulate in the tissues. The radioactivity was mainly excreted in the urine.

Contributions of FASN and SCD gene polymorphisms on fatty acid composition in muscle from Japanese Black cattle.

The fatty acid synthase (FASN) and stearoyl-CoA desaturase (delta-9-desaturase) (SCD) genes affect fatty acid composition. This study evaluated the contributions of polymorphisms of these genes on fatty acid composition in muscle in two different populations: 1189 and 1058 Japanese Black cattle from the Miyagi and the Yamagata populations respectively. We sampled intramuscular fat from the longissimus thoracis muscle in the Miyagi population and from the trapezius muscle in the Yamagata population. The collective contributions of FASN and SCD polymorphisms to total additive genetic variance for oleic acid were 13.46% in the Miyagi population and 16.29% in the Yamagata population and to phenotypic variance were 5.45% and 6.54% respectively. Although the individual effects of FASN and SCD polymorphisms on fatty acid composition were small, overall gene substitution may effectively improve fatty acid composition. In addition, we found that gene polymorphism contributions of fatty acids varied by population even in the same breed.

Evaluation of silica gel-immobilized phosphorylcholine columns for size exclusion chromatography and their application in the analysis of the subunit structures of fish-egg lectins.

Columns of phosphorylcholine (PC) immobilized on silica gel were shown to be useful for size exclusion chromatography (SEC) of proteins. The columns provided good separation of proteins in 50mM sodium phosphate buffer (pH 6.9) containing 0.25 M NaCl, and there was a linear relationship between the retention times and the logarithmic values of the molecular weights with a correlation coefficient (R(2)) of 0.978-0.992. The columns were used in analyzing the subunit structures of the rhamnose-binding lectins CSL1, CSL2, and CSL3, isolated from chum salmon (Oncorhynchus keta) eggs. Although the lectins, which are a group of carbohydrate-binding and hydrophobic proteins, behaved anomalously in SEC with conventional matrices, they could be eluted from the immobilized PC columns without non-size-related retention, thereby allowing their molecular weights to be reliably estimated.

Quantitative trait loci analysis on Sus scrofa chromosome 7 for meat production, meat quality, and carcass traits within a Duroc purebred population.

Many QTL analyses related to meat production and meat quality traits have been carried out using an F(2) resource population produced by crossing 2 genetically different breeds. This experiment was intended to investigate whether these QTL were segregating in a purebred Duroc population that had been selected for meat production and meat quality traits during 7 generations. Sus scrofa chromosome 7, for which significant QTL of intramuscular fat and many other traits have already been reported, was studied. The polymorphism of 10 microsatellite markers that were arranged at about 20-cM intervals was investigated on 1,004 pigs. In the selected population, 954 progeny were produced from mating of 99 sires and 286 dams. The QTL analysis for a full-sib family population was examined with the multigeneration pedigree structure of the population. Variance component analysis was used to detect QTL in this population and was examined for the multigeneration pedigree population. In this study, multigenerational pedigree estimated identical by descent coefficients among sibs were produced using Markov chain Monte Carlo methods. The maximum likelihood of odds score was found at the 70-cM position for the LM area, at the 0-cM position for the pork color standard, and at the 120-cM position for the number of thoracic vertebra, but no significant QTL for intramuscular fat were detected on SSC 7. These results indicate that QTL analysis via a variance component method within a purebred population was effective to determine that QTL were segregating in a population of purebred Durocs.

[How to treat infected pacemaker leads in very elderly patients].

Infection of pacemaker leads is one of the difficult problems after implantation, especially in elderly patients. Three octogenarians were referred to our hospital due to infection of pacemaker leads which had been resistant to antibiotic treatments. The patients were 81 approximately 88 years old including 1 male and 2 females. Primary implantation of pacemaker had been performed 9 months approximately 16 years before and infection had started 7 months approximately 7 years before. The infected leads were partially resected in an 88-year-old woman who had serious dementia and poor nutrition status. The infected leads were completely removed under cardiopulmonary bypass (on-beating) in an 81-year-old woman who had undergone 3 previous surgical treatments. The infected lead was partially removed in an 82-year-old man, but infection was not perfectly controlled. Therefore, the lead was completely removed under cardiopulmonary bypass with cardiac arrest. In conclusion, complete removal of the whole pacemaker system is desirable, while palliative surgery might be an option for very elderly patients with high risks.

Controlling the dissociative ionization of ethanol with 800 and 400 nm two-color femtosecond laser pulses.

Ethanol molecules were irradiated with a pair of temporally overlapping ultrashort intense laser pulses (10(13)-10(14) Wcm(2)) with different colors of 400 and 800 nm, and the dissociative ionization processes have been investigated. The yield ratio of the C-O bond breaking with respect to the C-C bond breaking was varied in the range of 0.17-0.53 sensitively depending on the delay time between the two laser pulses, and the absolute value of the yield of the C-O bond breaking was found to be increased largely when the Fourier-transform limited 800 nm laser pulse overlaps the stretched 400 nm laser pulse, demonstrating an advantage of the two-color intense laser fields in controlling chemical bond breaking processes.

The lateral wedged insole with subtalar strapping significantly reduces dynamic knee load in the medial compartment gait analysis on patients with medial knee osteoarthritis.

OBJECTIVE: Two lateral wedged insoles were compared: one with, and the other without, subtalar strapping. METHODS: Twenty-one patients (age 58-83, mean 72) with medial knee osteoarthritis (OA) were enrolled. Thirty-seven knees in the patients were divided into three groups based on the Kellgren and Lawrence OA grading system; grades 2 (cases=20), 3 (cases=11), and 4 (cases=6). The subjects were tested during walking barefoot and during walking with a silicon rubber lateral wedged insole with elevation of 10 mm attached to a barefoot. Gait analysis was performed on a 10 m walkway for each subject under three different walking conditions; barefoot, wearing a conventional insole, and a subtalar strapping insole. Peak knee varus moment during gait was measured under each condition, and compared between the three conditions and between the OA grades. RESULTS: On the whole (cases=37), the peak varus moment was significantly reduced by wearing either of the insoles, compared to walking barefoot. The reduction was more obvious with the strapping insole (-13%, P<0.01), compared with the conventional insole (-8%, P<0.05). In moderate OA patients (grades 2 and 3), the moments were significantly lower with the strapping insole, compared with the conventional insole (P=0.0048 and 0.005, respectively). However, no significant difference was detected in severe OA patients (grade 4) between the two types of insoles (P=0.4). CONCLUSIONS: Both lateral wedged insoles significantly reduced the peak medial compartment load during gait. The subtalar strapping insole had a greater effect than the conventional insole, particularly in patients with moderate medial knee OA.

Dissociative ionization of ethanol by 400 nm femtosecond laser pulses.

The dissociative ionization of ethanol in short-pulsed laser fields at approximately 400 nm is investigated. The yield ratio of the C-O bond breaking with respect to the C-C bond breaking increases sharply as the temporal width increases from 60 to 400 fs, and the yield ratio is two to three times as large as that at 800 nm in the entire pulse-width range of 60-580 fs. The enhancement of the C-O bond breaking of singly charged ethanol at 400 nm and the bond elongation prior to the Coulomb explosion of doubly charged ethanol occurring in the relatively weak light field intensity of 10(12)-10(13) W cm(2) is interpreted by the efficient light-induced coupling among the electronic states at the shorter wavelength of 400 nm. From the double pulse experiment, in which ethanol is irradiated with a pair of short pulses (<80 fs), the most efficient coupling occurs at Deltat=160 fs that is much earlier than Deltat=250 at 800 nm, where Deltat denotes the temporal separation of the two pulses, indicating that the nonadiabatic field-induced potential crossings of singly charged ethanol occurs much earlier at 400 nm than at 800 nm.

Infected Charcot spine.

STUDY DESIGN: Case report of an infected Charcot spine following spinal cord injury. OBJECTIVE: To describe this very rare pathological condition and the results of surgical treatment. SETTING: A department of orthopaedic surgery in Japan. METHODS: A 44-year-old man presented with a destructive lesion in the lumbo-sacral spine and a fistula in his back. Anterior bone graft, percutaneous external spinal fixation, and suction/irrigation of the wound were performed. After 4 months, posterior spinal instrumentation surgery was carried out. RESULTS: Primary closure of the fistula and complete bone fusion was achieved after the operation. CONCLUSION: Infection of a Charcot spine, although a rare clinical entity, should be considered as a diagnostic possibility in the spinal cord-injured patients. External spinal fixation is a useful method for the unstable spinal lesion with infection.

[Natural Y-grafting using internal thoracic artery branches for off-pump coronary artery bypass].

The excellent long-term patency rates achieved utilizing the internal thoracic arteries (ITAs) have stimulated a variety of approaches to expand the use of these conduits in coronary revascularization. The ITA divides at the level of the fifth or sixth intercostal space into the superior epigastric and musculophrenic branches. If these terminal branches of the ITA are large and long enough for grafting, they can be used to construct a "Y" anastomosis to the coronary arteries. We experienced 2 patients who underwent multivessel off-pump coronary artery bypass (OPCAB) using these ITA branches. In both patients the bilateral ITAs and the right gastroepiploic artery were taken down using the skeletonization technique as in situ grafts. One patient had 5 grafts including the right ITA terminal branches which were used for the left anterior descending branch (LAD) and diagonal branch. Another patient had 6 grafts including the left ITA terminal branches which were used for the posterolateral branch (PL)1 and PL2. Postoperative angiography revealed widely patent grafts in both patients. We believe that one of the advantages of applying the off-pump technique when the ITA branches are used is that it might be easier to arrange the angle of constructing a natural "Y" configuration compared with conventional coronary artery bypass grafting (CABG). In conclusion, the terminal branches of the ITA, if of suitable size, should be considered for "Y" grafts to diseased coronary arteries when performing complete arterial revascularization with only in situ grafts.

Conservative treatment of isolated posterior cruciate ligament injury in professional baseball players: a report of two cases.

Conservative treatment is currently recommended for most isolated posterior cruciate ligament (PCL) injuries in athletes. However, it is not known whether conservative treatment is applicable even in high performance athletes with isolated PCL injury. The results in two extremely high performance athletes, professional baseball players with isolated acute PCL injury treated conservatively are reported. A catcher and an out fielder, who were regular players, hurt their knees in baseball games. Magnetic resonance images of the knee detected complete PCL rupture. Following a carefully guided physical therapy program, a 3-week period of immobilization of the knee in full extension was achieved with a knee brace, while performing hard quadriceps muscle strengthening exercise, and then running exercise was started. Six to eight weeks after injury, they were able to return fully to their original sporting activity despite tibial posterior translation on posterior drawer test, and to sustain this activity over 2 years. Switching of weight-bearing to non-weight-bearing in a deep knee flexion is considered to contribute to subjective instability in athletes with PCL-deficiency. Probably because our cases, even though extremely high performance athletes were infrequently subjected to such a situation while playing baseball, they were able to return to their pre-injury level of athletic performance without severe subjective instability through conservative treatment.

[How much can off-pump coronary artery bypass grafting be performed for complete revascularization using only in situ arterial conduits?].

With the rapid advance of percutaneous catheter intervention, the direction taken by surgeons is not only to make coronary artery bypass grafting (CABG) less invasive but also to obtain better long-term results by using more arterial conduits. However, off-pump CABG (OPCAB) with only in situ arterial conduits such as the left and right internal thoracic arteries (ITAs) and the right gastroepiploic artery (RGEA) is technically demanding for the surgeon. The purpose of this study was to demonstrate the feasibility and safety of this technique. From January 2002 to December 2002, 57 patients underwent isolated CABG without cardiopulmonary bypass, excluding minimally invasive direct coronary artery bypass (MIDCAB) cases. There were 44 men and 13 women with a mean age of 64.1 years. Most of the patients (95%) underwent postoperative angiography within 1 month after surgery. The mean number of anastomoses was 3.8 +/- 1.4 (range 2-8). One patient suffered from mediastinitis and another from stroke. All patients were discharged from the hospital. Fifty patients (88%) received only in situ grafts for complete revascularization. Forty-one patients received bilateral ITAs and 27 received bilateral ITAs and RGEA. The mean number of anastomoses using bilateral ITAs and RGEA was 4.5 +/- 1.2 (range 3-8). The patency rate was 98%. Complete arterial revascularization with only in situ arterial conduits was associated with minimal operative risk and a high early graft patency rate, even in the off-pump situation. These results indicate that complete revascularization can be achieved in OPCAB with the exclusive use of in situ arterial conduits in patients with diffuse coronary arterial disease.

[Multivessel off-pump coronary aortic bypass grafting with an impaired and severely dilated left ventricle using the Starfish heart positioner].

Off-pump coronary artery bypass grafting (OPCAB) is gaining acceptance and is now more widely used for coronary revascularization worldwide. Accessing the vessels, particularly on the lateral and inferior walls of the heart, can sometimes be challenging while the heart is beating, especially in hearts with an impaired and severely dilated left ventricle. The Starfish heart positioner was designed to achieve optimal exposure of the lateral and inferior walls of the heart with minimal hemodynamic compromise. This device was used in a patient with a severely impaired and dilated left ventricle to successfully position the heart for quadruple bypass grafting using all in situ arterial grafts, such as bilateral international thoracic arteries (ITAs) and the right gastroepiploic artery. The degree of hematoma caused by the pressure of the suction cup of this device was minimal. This case report suggests that the use of Starfish heart positioner will facilitate and expand the ability of surgeons to easily and consistently perform OPCAB.