Quercetin attenuates adipogenesis and fibrosis in human skeletal muscle.

Age-associated increase in ectopic fat degeneration and fibrosis in the skeletal muscle contribute to muscle degradation and weakness. Quercetin is a bioactive flavonoid with anti-inflammatory and anti-obesity effects. Thus, we aimed to investigate the effects of quercetin on adipogenesis and fibrosis in the human skeletal muscle, which have not yet been elucidated. Human muscle-derived PDGFRα+/CD201+ cells (mesenchymal progenitors) were incubated with various concentrations of quercetin (0, 0.3, 1, and 3 µM) under adipogenic or fibrogenic conditions. Lipid accumulation was visualized via Oil Red O staining. The expression of genes implicated in adipocyte or fibroblast differentiation and activation of signaling pathways was analyzed. The quercetin-treated PDGFRα+/CD201+ cells showed attenuated lipid accumulation and adipogenic gene expression (CEBPA and ADIPOQ) via the inhibition of CREB phosphorylation under adipocyte differentiation conditions. Additionally, quercetin treatment significantly attenuated the expression of fibrogenic genes (TIMP1, ACTA2, COL1A1 and COL3A1) by inhibiting Smad2 phosphorylation. Quercetin suppressed the differentiation of muscle-derived PDGFRα+/CD201+ cells to adipocytes and fibroblasts at concentrations achievable by dietary and dietary supplement intake, which indicated its preventive or therapeutic effect against the loss of muscle quality.

Assessment of spinal alignment in standing position using Biplanar X-ray images and three-dimensional vertebral models.

We developed two methods for three-dimensional (3D) evaluation of spinal alignment in standing position by image matching between biplanar x-ray images and 3D vertebral models. One used a Slot-Scanning 3D x-ray Imager (sterEOS) to obtain biplanar x-ray images, and the other used a conventional x-ray system and a rotating table. The 3D vertebral model was constructed from the CT scan data. The spatial position of the vertebral model was determined by minimizing the contour difference between the projected image of the model and the biplanar x-ray images. Verification experiments were conducted using a torso phantom. The relative positions of the upper vertebrae to the lowest vertebrae of the cervical, thoracic, and lumbar vertebrae were evaluated. The mean, standard deviation, and mean square error of the relative position were less than 1° and 1 mm in all cases for sterEOS. The maximum mean squared errors of the conventional x-ray system and the rotating table were 0.7° and 0.4 mm for the cervical spine, 1.0° and 1.2 mm for the thoracic spine, and 1.1° and 1.2 mm for the lumbar spine. Therefore, both methods could be useful for evaluating the spinal alignment in standing position.

Experimental application of pulsed laser-induced water jet for endoscopic submucosal dissection: mechanical investigation and preliminary experiment in swine.

BACKGROUND AND AIM: A current drawback of endoscopic submucosal dissection (ESD) for early-stage gastrointestinal tumors is the lack of instruments that can safely assist with this procedure. We have developed a pulsed jet device that can be incorporated into a gastrointestinal endoscope. Here, we investigated the mechanical profile of the pulsed jet device and demonstrated the usefulness of this instrument in esophageal ESD in swine. METHODS: The device comprises a 5-Fr catheter, a 14-mm long stainless steel tube for generating the pulsed water jet, a nozzle and an optical quartz fiber. The pulsed water jet was generated at pulse rates of 3 Hz by irradiating the physiological saline (4°C) within the stainless steel tube with an holmium-doped yttrium-aluminum-garnet (Ho:YAG) laser at 1.1 J/pulse. Mechanical characteristics were evaluated using a force meter. The device was used only for the part of submucosal dissection in the swine ESD model. Tissues removed using the pulsed jet device and a conventional electrocautery device, and the esophagus, were histologically examined to assess thermal damage. RESULTS: The peak impact force was observed at a stand-off distance of 40 mm (1.1 J/pulse). ESD using the pulsed jet device was successful, as the tissue specimens showed precise dissection of the submucosal layer. The extent of thermal injury was significantly lower in the dissected bed using the pulsed jet device. CONCLUSION: The results showed that the present endoscopic pulsed jet system is a useful alternative for a safe ESD with minimum tissue injury.

l-Anserine Increases Muscle Differentiation and Muscle Contractility in Human Skeletal Muscle Cells.

l-Anserine, an imidazole peptide, has a variety of physiological activities, but its effects on skeletal muscle differentiation and muscle contractile force remain unknown. Thus, in this study, we investigated the effect of l-anserine on muscle differentiation and muscle contractile force in human skeletal muscle cells. In two-dimensional culture, 1 µM l-anserine significantly increased the myotube diameters (26.5 ± 1.71, 27.7 ± 1.08, and 28.8 ± 0.85 µm with 0, 0.1, and 1 µM l-anserine, respectively) and the expression levels of genes involved in muscle differentiation and the sarcomere structure. In three-dimensional culture, 1 µM l-anserine significantly increased the contractile force of engineered human skeletal muscle tissues cultured on a microdevice (1.99 ± 0.30, 2.17 ± 0.62, 2.66 ± 0.39, and 3.28 ± 0.85 µN with 0, 0.1, 0.5, and 1 µM l-anserine, respectively). l-Anserine also increased the myotube diameters and the proportion of myotubes with sarcomere structures in the cultured tissues. Furthermore, the histamine receptor 1 (H1R) antagonist attenuated the l-anserine-induced increase in the contractile force, suggesting the involvement of H1R in the mechanism of action of l-anserine. This study showed for the first time that l-anserine enhances muscle differentiation and muscle contractility via H1R.

Effects of Combining Docosahexaenoic Acid and Eicosapentaenoic Acid with Sesame Lignan on Vascular Endothelial Function.

Vascular endothelial cells produce vasoactive substances, such as nitric oxide (NO), to regulate vascular relaxation and contraction. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) enhance NO production in endothelial cells, and sesamin, a sesame lignan contained in sesame seeds, also promotes NO production. This study examined DHA, EPA, and sesamin's combined effects since it was expected that combining them would further enhance NO production in endothelial cells. Using a human umbilical vein endothelial cell (HUVEC), the NO amount secreted in the culture supernatant was analyzed. Sesamin metabolite (SC1) was used in the experiments because it is a major metabolite in human blood after sesamin absorption. When cells were treated with DHA or EPA alone, they increased NO production in a concentration-dependent manner, whereas no change in NO production was observed for SC1. NO production increased when DHA and EPA were treated in combination with SC1, although the low DHA and EPA concentrations showed no difference in NO production. In the concentrations in which the combined effect was observed, SC1 activated eNOS via calcium signaling, whereas DHA and EPA activated eNOS via alterations in the membrane lipid environment. The combined effect of the two pathways was considered to have enhanced the eNOS activity. These results suggested that combining DHA, EPA, and sesamin might improve vascular endothelial function.

Effects of Long-Chain Polyunsaturated Fatty Acids in Combination with Lutein and Zeaxanthin on Episodic Memory in Healthy Older Adults.

Arachidonic acid (ARA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA), which are long-chain polyunsaturated fatty acids (LCPUFAs), as well as lutein (L) and zeaxanthin (Z), can potentially improve brain function. However, the effect of a combination of these components (LCPUFAs + LZ) on memory function in healthy older individuals remains unclear. This study aimed to determine if LCPUFAs + LZ-supplemented food could improve memory function. Exploratory and confirmatory trials (Trials 1 and 2, respectively) were conducted in healthy older Japanese individuals with memory complaints. We conducted randomized, double-blind, placebo-controlled, parallel-group trials. Participants were randomly allocated to two groups: placebo or LCPUFAs + LZ. LCPUFAs + LZ participants were provided with supplements containing ARA, DHA, EPA, L, and Z for 24 weeks in Trial 1 and 12 weeks in Trial 2. Memory functions were evaluated using Cognitrax before and after each trial. Combined analyses were performed for subgroups of participants with cognitive decline in Trials 1 and 2. The results showed that supplementation with LCPUFAs + LZ did not significantly affect memory function in healthy, non-demented, older individuals with memory complaints whereas it improved memory function in healthy, non-demented, older individuals with cognitive decline.

Sesamin Metabolites Suppress the Induction of Cellular Senescence.

Cellular senescence induces inflammation and is now considered one of the causes of organismal aging. Accumulating evidence indicates that age-related deterioration of mitochondrial function leads to an increase in reactive oxygen species (ROS) and DNA damage, which in turn causes cellular senescence. Thus, it is important to maintain mitochondrial function and suppress oxidative stress in order to inhibit the accumulation of senescent cells. Sesamin and its isomer episesamin are types of lignans found in sesame oil, and after being metabolized in the liver, their metabolites have been reported to exhibit antioxidant properties. However, their effects on cellular senescence remain unknown. In this study, the effects of sesamin, episesamin, and their metabolites SC1 and EC1-2 on replicative senescence were evaluated using human diploid lung fibroblasts, and TIG-3 cells. The results showed that sesamin and episesamin treatment had no effect on proliferative capacity compared to the untreated late passage group, whereas SC1 and EC1-2 treatment improved proliferative capacity and mitigated DNA damage of TIG-3 cells. Furthermore, other cellular senescence markers, such as senescence-associated secretory phenotype (SASP), mitochondria-derived ROS, and mitochondrial function (ROS/ATP ratio) were also reduced by SC1 and EC1-2 treatment. These results suggest that SC1 and EC1-2 can maintain proper mitochondrial function and suppress the induction of cellular senescence.

Identification of the metabolites of episesamin in rat bile and human liver microsomes.

Episesamin is an isomer of sesamin, resulting from the refining process of non-roasted sesame seed oil. Episesamin has two methylendioxyphenyl groups on exo and endo faces of the bicyclic skeleton. The side methylendioxyphenyl group was metabolized by cytochrome-P450. Seven metabolites of episesamin were found in rat bile after treatment with glucuronidase/arylsulfatase and were identified using NMR and MS. The seven metabolites were (7α,7'ß,8α,8'α)-3,4-dihydroxy-3',4'-methylenedioxy-7,9':7',9-diepoxylignane (EC-1-1), (7α,7'ß,8α,8'α)-3,4-methylenedioxy-3',4'-dihydroxy-7,9':7',9-diepoxylignane (EC-1-2) and (7α,7'ß,8α,8'α)-3,4:3',4'-bis(dihydroxy)-7,9':7',9-diepoxylignane (EC-2), (7α,7'ß,8α,8'α)-3-methoxy-4-hydroxy-3',4'-methylenedioxy-7,9':7',9-diepoxylignane (EC-1m-1), (7α,7'ß,8α,8'α)-3,4-methylenedioxy-3'-methoxy-4'-hydroxy-7,9':7',9-diepoxylignane (EC-1m-2), (7α,7'ß,8α,8'α)-3-methoxy-4-hydroxy-3',4'-dihydroxy-7,9':7',9-diepoxylignane (EC-2m-1) and (7α,7'ß,8α,8'α)-3,4-dihydroxy-3'-methoxy-4'-hydroxy-7,9':7',9-diepoxylignane (EC-2m-2). EC-1-1, EC-1-2 and EC-2 were also identified as metabolites of episesamin in human liver microsomes. These results suggested that similar metabolic pathways of episesamin could be proposed in rats and humans.

Effects of Quercetin Glycoside Supplementation Combined With Low-Intensity Resistance Training on Muscle Quantity and Stiffness: A Randomized, Controlled Trial.

Objective: Aging of skeletal muscle is characterized not only by a decrease of muscle quantity but also by changes in muscle quality, such as an increase in muscle stiffness. The present study aimed to investigate the effects of supplementation with quercetin glycosides (QGs), well-known polyphenolic flavonoids, combined with resistance exercise on muscle quantity and stiffness. Materials and Methods: A randomized, controlled trial was conducted in community-dwelling, Japanese people aged 50-74 years who were randomly allocated to exercise with placebo or 200 or 500 mg of QG supplementation. All participants performed low-intensity resistance training mainly targeting thigh muscles with 40% of 1-repetition maximum, 3 days per week for 24 weeks. Muscle cross-sectional area (CSA), lean mass, and vastus lateralis (VL) muscle stiffness were measured before and after the 24-week intervention. Results: Forty-eight subjects completed the 24-week intervention. There were no significant group × time interactions in thigh CSA for primary outcome, as well as lean mass. VL muscle stiffness in the stretched position was significantly lower in both the 200 mg and 500 mg QG groups than in the placebo group after the 24-week intervention (p < 0.05). No significant correlation was observed between changes of VL muscle CSA and stiffness during the 24-week intervention. Conclusion: Quercetin glycoside supplementation combined with low-intensity resistance exercise improved passive muscle stiffness independently of muscle quantity. Clinical Trial Registration: [www.umin.ac.jp/ctr/], identifier [UMIN000037633].

The association between long-chain polyunsaturated fatty acid intake and changes in brain volumes among older community-dwelling Japanese people.

The relationship between age-related brain atrophy and long-chain polyunsaturated fatty acid (LCPUFA) intake is not fully understood. This study investigated the association of docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and arachidonic acid (ARA) intake and brain atrophy in non-demented older Japanese people (n = 810, aged 60-89 years) using data sets of a 2-year longitudinal study. Brain volumes were measured using 3D-MRI in the baseline and follow-up periods. The associations of multivariate-adjusted changes in brain volumes with baseline LCPUFA intake were assessed using a general linear model. Higher ARA intake was associated with a smaller decrease in frontal cortex volumes, which was accompanied by a lower risk of cognitive decline among the participants. In the subgroup analysis for low DHA and EPA intake, accounting for one-third of Japanese intake, DHA and EPA intake was positively correlated with preservation of the temporal cortex volume. These findings suggest that appropriate intake of LCPUFA may decelerate age-related brain atrophy and lead to the maintenance of brain health in older people.

The activation of P2X7 receptor impairs lysosomal functions and stimulates the release of autophagolysosomes in microglial cells.

Recently, autophagy has been associated with the TLR signaling pathway to eliminate intracellular pathogens in the innate immune system. However, it is unknown if other pathways regulate autophagy during the immunologic response. Given the critical role of the purinergic P2X7 receptor (P2X7R) pathway during various immunologic functions (i.e., caspase activation and IL-1beta secretion), the principal objective here was to determine whether the P2X7R pathway may regulate autophagy in immune cells. We observed in both MG6 mouse microglial cells and primary microglia that activation of P2X7R by ATP increases the expression of microtubule-associated protein 1 light chain 3 (LC3)-II, the autophagosomal membrane-associated form of LC3, in an extracellular Ca(2+)-dependent manner. Consistent with this, immunohistochemistry showed extensive formation of LC3-immunopositive dots, and electron microscopy demonstrated accumulation of autophagosomes and autophagolysosomes in ATP-treated cells. Importantly, the up-regulation of LC3-II by P2X7R activation was not affected by autophagy inhibitors, such as 3-methyladenine and PI3K inhibitors. Furthermore, while lysosomal functions were impaired by ATP treatment, autophagolysosomal components were released into the extracellular space. Similarly, a phagocytosis assay using Escherichia coli BioParticles showed that phagosome maturation was impaired in ATP-treated cells and a robust release of LC3-immunopositive phagolysosomes was induced along with a radial extension of microtubule bundles. Taken together, the data suggest a novel mechanism whereby the P2X7R signaling pathway may negatively regulate autophagic flux through the impairment of lysosomal functions, leading to stimulation of a release of autophagolysosomes/phagolysosomes into the extracellular space.

Differences in the effect of surface texturing on the wear loss of ß-type Ti-Nb-Ta-Zr and (α+ß)-type Ti-6Al-4V ELI alloys in contact with zirconia in physiological saline solution.

The effectiveness of dimple surface texturing via picosecond pulsed laser processing for reducing wear loss was investigated using two types of titanium alloys, ß-type Ti-29Nb-13Ta-4.6Zr (TNTZ) and α+ß-type Ti-6Al-4V ELI (Ti64). The two alloys showed different modes of wear against zirconia. As the sliding distance increased, the wear loss was observed to increase for Ti64, but not necessarily for TNTZ. The wear debris of Ti64 acted as abrasive particles, but that of TNTZ easily adhered to the surface, and the adhered wear debris turned into a hard wear-protective layer. Therefore, the dependence of wear loss on the sliding distance for these two titanium alloys could be attributed to the difference in the roles of wear debris between each titanium alloy and zirconia. Further, depending on this difference in wear mode, the effect of dimple surface texturing on the wear was found to be different in Ti64 and TNTZ. As the dimples can trap the wear debris, they are effective for reducing wear in Ti64 but are detrimental in TNTZ.

Protective role of endogenous gangliosides for lysosomal pathology in a cellular model of synucleinopathies.

Gangliosides may be involved in the pathogenesis of Parkinson's disease and related disorders, although the precise mechanisms governing this involvement remain unknown. In this study, we determined whether changes in endogenous ganglioside levels affect lysosomal pathology in a cellular model of synucleinopathy. For this purpose, dementia with Lewy body-linked P123H beta-synuclein (beta-syn) neuroblastoma cells transfected with alpha-synuclein were used as a model system because these cells were characterized as having extensive formation of lysosomal inclusions bodies. Treatment of these cells with D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), an inhibitor of glycosyl ceramide synthase, resulted in various features of lysosomal pathology, including compromised lysosomal activity, enhanced lysosomal membrane permeabilization, and increased cytotoxicity. Consistent with these findings, expression levels of lysosomal membrane proteins, ATP13A2 and LAMP-2, were significantly decreased, and electron microscopy demonstrated alterations in the lysosomal membrane structures. Furthermore, the accumulation of both P123H beta-syn and alpha-synuclein proteins was significant in PDMP-treated cells because of the suppressive effect of PDMP on the autophagy pathway. Finally, the detrimental effects of PDMP on lysosomal pathology were significantly ameliorated by the addition of gangliosides to the cultured cells. These data suggest that endogenous gangliosides may play protective roles against the lysosomal pathology of synucleinopathies.

Expression of basigin in reproductive tissues of estrogen receptor-{alpha} or -{beta} null mice.

Basigin plays important roles in both male and female reproduction because basigin (Bsg) null male and female mice are infertile. The aim of the present study was to determine whether basigin expression in reproductive organs requires estrogen receptor-alpha (ESR1, ERalpha) or -beta (ESR2, ERbeta). Expression of basigin protein in the testis, ovary, and male and female reproductive tracts was studied in adult wild-type (WT), Esr1-null (alphaERKO), and Esr2-null (betaERKO) mice by immunohistochemistry and immunoblotting. Basigin mRNA levels in ovary and uterus were examined by quantitative RT-PCR. In females, basigin protein expression was observed mainly in granulosa and interstitial cells of the ovary and epithelial cells of the proximal oviduct in all genotypes. Basigin protein was also expressed in the uterine epithelium at proestrus and estrus in WT and betaERKO mice but not in alphaERKO mice. However, a higher level of basigin mRNA was observed in uteri of alphaERKO mice compared with WT and betaERKO mice. In males, basigin was expressed in Leydig cells and all germ cells except spermatogonia in all genotypes. Basigin was present in epithelial cells lining the efferent ductules in WT and betaERKO mice, but expression was greatly reduced in alphaERKO mice. In epididymal ducts, basigin expression was observed in epithelial cells in the caput and cauda in all genotypes. These data suggest that expression of basigin protein requires ESR1, but not ESR2, in the uterus and efferent ductules, but is independent of estrogen receptor in the ovary, oviduct, testis, and epididymis.

The mode of biofilm formation on smooth surfaces by Campylobacter jejuni.

Many microorganisms produce extracellular polymers referred to collectively as "slime" or glycocalyx, and form biofilms on solid surfaces in natural ecosystems. Campylobacter jejuni, one of the most important foodborne pathogens, also has the ability to form biofilm on stainless steel, glass, or polyvinyl chloride in vitro. However, the issue of biofilm formation by Campylobacter species has not been extensively examined. The present study was performed to examine the mode of adhesion of C. jejuni to a smooth surface. When bacterial suspensions in Brucella broth were incubated in microplate wells with a glass coverslip, microcolonies 0.5~2 mm in diameter were formed on the coverslip within 2 hr from the start of incubation. These microcolonies gradually grew and formed a biofilm of net-like connections within 6 hr. Transmission electron microscopy indicated that massive amounts of extracellular material masked the cell surface, and this material bound ruthenium red, suggesting the presence of a polysaccharide moiety. Scanning electron microscopy indicated that the flagella acted as bridges, forming net-like connections between the organisms. To determine the genes associated with biofilm formation, aflagellate (flaA(-)) and flagellate but non-motile (motA(-)) mutants were constructed from strain 81-176 by natural transformation-mediated allelic exchange. The flaA(-) and motA(-) mutants did not form the biofilm exhibited by the wild-type strain. These findings suggest that flagella-mediated motility as well as flagella is required for biofilm formation in vitro.

Effects of the antlered form of Ganoderma lucidum on tumor growth and metastasis in cyclophosphamide-treated mice.

We examined the alleviation of cyclophosphamide-induced immunodepression by the antlered form of Ganoderma lucidum (G. lucidum AF) and also evaluated the anti-tumor and anti-metastatic effects of G. lucidum AF in cyclophosphamide-treated mice. G. lucidum AF alleviated cyclophosphamide-induced decrease in body weight, natural killer (NK) activity, interferon (IFN)-gamma production, and cytotoxic T lymphocyte (CTL) activity, and inhibited the abnormal increase and decrease in interleukine (IL)-4 level due to cyclophosphamide administration. Post-treatment with cyclophosphamide and G. lucidum AF significantly inhibited tumor growth in MM 46-bearing mice. When Lewis lung carcinoma cells were injected into mice after a cyclophosphamide administration, metastasis of these cells to the lung was increased, but G. lucidum AF suppressed it. The anti-tumor and anti-metastatic effects of the combination of G. lucidum AF and cyclophosphamide might influence the modulatory effects of G. lucidum AF on both cellular and humoral immunity. These findings suggest that G. lucidum AF would be beneficial in alleviating the reduction of immune response by chemotherapeutic anti-cancer drugs.

Plasma levels of DJ-1 as a possible marker for progression of sporadic Parkinson's disease.

DJ-1 is a multifunctional protein whose loss of function by gene mutations may play a causative role for familial Parkinson's disease (PD). A recent study has shown that the expression of this molecule is upregulated in both brains and cerebrospinal fluids (CSF) in various neurological disorders, including sporadic PD, Alzheimer's disease (AD) and stroke, raising a possibility that DJ-1 could be a potential biomarker for these diseases. In this context, the main objective of the present study was to determine if DJ-1 was increased in the plasma of PD patients. For this purpose, blood plasma samples collected from sporadic PD patients, dementia with Lewy bodies (DLB) and healthy age-matched controls were analyzed by immunoblotting and enzyme-linked immunosorbent assay. The results showed that the plasma DJ-1 levels in PD (n=104) were higher than those in control (n=80) (p<0.05). Moreover, the plasma DJ-1 levels in the advanced stage of PD (n=52, Yahr III-IV) were higher than those in the early stage of PD (n=52, Yahr I-II) (p<0.05), demonstrating that the plasma DJ-1 was correlated with the disease severity in PD. Plasma DJ-1 levels were also significantly higher in DLB (n=30) compared with both controls and early stage of PD (p<0.01). Taken together, these results suggest that the plasma DJ-1 could be a useful biomarker for the evaluation of the disease severity in PD and possibly in other Lewy body diseases.

Tissue distribution of basigin and monocarboxylate transporter 1 in the adult male mouse: a study using the wild-type and basigin gene knockout mice.

Basigin (Bsg) is a transmembrane protein that is responsible for targeting of monocarboxylate transporters (MCTs) to the cell membrane. The present study was conducted to determine whether or not Bsg was required for the proper localization of MCT isoform 1 (MCT1) in a wide range of tissues in adult male mice. The tissue distributions of Bsg and MCT1 in wild-type (WT) mice, the tissue distribution of MCT1 in Bsg gene knockout (Bsg-KO) mice, and the protein and mRNA levels of MCT1 in both genotypes were studied. Immunohistochemistry demonstrated that Bsg colocalized with MCT1 in the cerebrum, retina, skeletal and cardiac muscle, duodenal epithelium, hepatic sinusoid, proximal uriniferous tubules, Leydig cells, and efferent ductule epithelium in WT mice. Bsg was absent but MCT1 was present in Sertoli cells, cauda epididymis, myoepithelial cells and duct of the mandibular gland, surface epithelium of the stomach and bronchioles. In Bsg-KO mice, with the exception of Leydig cells, MCT1 immunostaining was greatly reduced in intensity and its distribution was altered in tissues that expressed both Bsg and MCT1 in WT mice. Levels of the protein and mRNA for MCT1 in these tissues did not change significantly in Bsg-KO mice. On the other hand, immunostaining patterns in cells in which Bsg was absent but MCT1 was present in WT mice remained unchanged in Bsg-KO mice. These observations suggest that Bsg is required for the proper localization of MCT1 in a wide range of cells but not in every cell type.

Phlorotannins as radical scavengers from the extract of Sargassum ringgoldianum.

To screen algal phlorotannins with antioxidative activities, 50% ethanol extracts of 25 Japanese marine algae were evaluated. Scavenging activity against superoxide anion radicals was frequently found with a high content of total phenolic compounds. Among these, the extract from the brown seaweed, Sargassum ringgoldianum, showed the strongest scavenging activity. The active fraction contained a mixture of high molecular weight polyphenols, phlorotannins that were found to be polymerized bifuhalol, as analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The scavenging activity of the fraction against superoxide anion radicals was estimated to be 1.0 microg/ml (IC(50)), which were approximately five times stronger than that of catechin.

Epididymal stone formation and decreased sperm production in roosters vaccinated with a killed strain of avian infectious bronchitis virus.

Our objective was to determine if vaccination with killed avian infectious bronchitis virus (AIBV) causes epididymal calcium stones in the rooster as is seen following vaccination with live attenuated AIBV. Specific-pathogen-free roosters were divided into three groups: nonvaccinated (NONVAC), live attenuated AIBV-vaccinated (LVAC), and killed AIBV-vaccinated (KVAC) groups. Roosters were vaccinated at 2, 6, 10, and 14 wk of age and the epididymal region was observed at 27 wk of age. Epididymal stones were present in 13% of NONVAC, 50% of KVAC, and 64% of LVAC roosters. Histologically, immune cells were seen in the interstitium of efferent ductules containing stones. We conclude that use of a killed vaccine does not reduce the incidence of epididymal stones.