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The extracellular signal-regulated kinase (ERK) is a serine/threonine kinase that is known to regulate cellular events such as cell proliferation and differentiation. The ERK signaling pathway is activated by fibroblast growth factors, and is considered to be indispensable for the differentiation of primitive endoderm cells, not only in mouse preimplantation embryos, but also in embryonic stem cell (ESC) culture. To monitor ERK activity in living undifferentiated and differentiating ESCs, we established EKAREV-NLS-EB5 ESC lines that stably express EKAREV-NLS, a biosensor based on the principle of fluorescence resonance energy transfer. Using EKAREV-NLS-EB5, we found that ERK activity exhibited pulsatile dynamics. ESCs were classified into two groups: active cells showing high-frequency ERK pulses, and inactive cells demonstrating no detectable ERK pulses during live imaging. Pharmacological inhibition of major components in the ERK signaling pathway revealed that Raf plays an important role in determining the pattern of ERK pulses. [ABSTRACT FROM AUTHOR]

Anhydrobiosis, one of the most extensively studied forms of cryptobiosis, is induced in certain organisms as a response to desiccation. Anhydrobiotic species has been hypothesized to produce substances that can protect their biological components and/or cell membranes without water. In extremotolerant tardigrades, highly hydrophilic and heat-soluble protein families, cytosolic abundant heat-soluble (CAHS) proteins, have been identified, which are postulated to be integral parts of the tardigrades' response to desiccation. In this study, to elucidate these protein functions, we performed in vitro and in vivo characterizations of the reversible self-assembling property of CAHS1 protein, a major isoform of CAHS proteins from Ramazzottius varieornatus, using a series of spectroscopic and microscopic techniques. We found that CAHS1 proteins homo-oligomerized via the C-terminal α-helical region and formed a hydrogel as their concentration increased. We also demonstrated that the overexpressed CAHS1 proteins formed condensates under desiccation-mimicking conditions. These data strongly suggested that, upon drying, the CAHS1 proteins form oligomers and eventually underwent sol–gel transition in tardigrade cytosols. Thus, it is proposed that the CAHS1 proteins form the cytosolic fibrous condensates, which presumably have variable mechanisms for the desiccation tolerance of tardigrades. These findings provide insights into molecular strategies of organisms to adapt to extreme environments. [ABSTRACT FROM AUTHOR]

Fabry disease results from a deficiency of the lysosomal enzyme ⍺‐Galactosidase‐A (⍺‐Gal A) and is estimated to occur in approximately 1:4100 live births. Characteristic of the disease is the accumulation of α‐Gal‐A substrates, primarily the glycosphingolipids (GSLs) globotriaosylceramide and globotriaosylsphingosine. Thrombotic events are a significant concern for Fabry patients, with strokes contributing to a significant decrease in overall lifespan. Currently, the mechanisms underlying the increased risk of thrombotic events experienced by Fabry patients are incompletely defined. Using a rat model of Fabry disease, we provide an improved understanding of the mechanisms linking GSL accumulation to thrombotic risk. We found that ⍺‐Gal A‐deficient rats accumulate myeloid‐derived leukocytes at sites of GSL accumulation, including in the bone marrow and circulation, and that myeloid‐derived leukocyte and megakaryocyte populations were prominent among cell types that accumulated GSLs. In the circulation, ⍺‐Gal A‐deficient rats had increases in cytokine‐producing cell types and a corresponding elevation of pro‐inflammatory cytokines. Lastly, circulating platelets from ⍺‐Gal A‐deficient rats accumulated a similar set of ⍺‐Galactosidase‐A substrates as was observed in megakaryocytes in the bone marrow, and exhibited increased platelet binding to fibrinogen in microfluidic and flow cytometric assays. [ABSTRACT FROM AUTHOR]

Bow-tie architecture is a layered network structure that has a narrow middle layer with multiple inputs and outputs. Such structures are widely seen in the molecular networks in cells, suggesting that a universal evolutionary mechanism underlies the emergence of bow-tie architecture. The previous theoretical studies have implemented evolutionary simulations of the feedforward network to satisfy a given input-output goal and proposed that the bow-tie architecture emerges when the ideal input-output relation is given as a rank-deficient matrix with mutations in network link intensities in a multiplicative manner. Here, we report that the bow-tie network inevitably appears when the link intensities representing molecular interactions are small at the initial condition of the evolutionary simulation, regardless of the rank of the goal matrix. Our dynamical system analysis clarifies the mechanisms underlying the emergence of the bow-tie structure. Further, we demonstrate that the increase in the input-output matrix reduces the width of the middle layer, resulting in the emergence of bow-tie architecture, even when evolution starts from large link intensities. Our data suggest that bow-tie architecture emerges as a side effect of evolution rather than as a result of evolutionary adaptation. [ABSTRACT FROM AUTHOR]

Abstract An exceptionally extensive bloom of the dinoflagellate Karenia mikimotoi caused mass mortality of aquaculture fishes leading the severe economic loss in 2017 in Imari Bay, Japan. Development of the 2017 bloom completely differed from other recent blooms (in 2012 and 2014). Bloom dynamics and its controlling factors were examined by field survey, numerical modeling and satellite data analysis. The bloom initially developed in the eastern area of Imari Bay as in the 2012 and 2014 blooms. Subsequently, the bloom broadly expanded to the western area and cell density increased to over 10,000 cells ml−1. Horizontal expansion of the bloom resulted in mass mortality of aquaculture fishes. Horizontal expansion and increase of cell density occurred accompanying a temperature decrease and nutrient increase in the mid and lower layers of the central area. Results of the numerical model indicated two factors related to the passage of a typhoon that promoted the bloom; horizontal bloom expansion induced by the wind-driven current and nutrient supply from the typhoon-induced coastal upwelling. This implies that the effect of the typhoon differentiated the 2017 bloom from previous blooms. The risk of harmful blooms as in the 2017 Karenia bloom in Imari Bay potentially will increase due to the effects of global warming. [ABSTRACT FROM AUTHOR]

The prototype for this database is available at https://microglycodb.test.glycosmos.org.The GlySpace Alliance is an alliance of glycan data portals GlyCosmos ([3]), GlyGen ([4]), and Glycomics@Expasy ([2]). The second workshop was devoted to GlyCosmos repositories, databases, and tools, and the third workshop was devoted to the GlySpace Alliance, including GlyGen, Glycomics@Expasy, and GlyCosmos. MicroGlycoDB is a database of microbial glycans and glycan pathways developed in collaboration with the University of Lille (France), the CCRC at the University of Georgia (USA), and others. [Extracted from the article]

To examine the environmental factors controlling the inshore recruitment dynamics of Anguilla japonica in the Oyodo River, Miyazaki Prefecture, Kyushu, glass eel samplings were carried out using fyke nets during winter (November-March) of 1994-2014. The peak CPUEs (catch per unit effort) were observed between November and February, but differed from year to year. The yearly CPUE was extremely high in 2002, when the sea surface temperature (SST) in the offshore area of the Oyodo River was the lowest in winter of all the sampling years. The negative SST anomaly of less than − 0.5 °C was sustained in the offshore area during the recruitment season in 2002, which was caused by two combined factors; low air temperature and the Kuroshio path. The oceanographic data showed that the dominant path of the Kuroshio was displaced eastward at 31°N in 2002, which was different from the average Kuroshio path. The eastward displacement of the Kuroshio induced a cyclonic mesoscale eddy in the offshore area of the Oyodo River, resulting in the entrainment of the cold seawater into coastal waters from deep water. The oceanographic condition in relation to the continuous low SST could be favorable for local recruitment of glass eels. [ABSTRACT FROM AUTHOR]

Living tissues undergo deformation during morphogenesis. In this process, cells generate mechanical forces that drive the coordinated cell motion and shape changes. Recent advances in experimental and theoretical techniques have enabled in situ measurement of the mechanical forces, but the characterization of mechanical properties that determine how these forces quantitatively affect tissue deformation remains challenging, and this represents a major obstacle for the complete understanding of morphogenesis. Here, we proposed a non-invasive reverse-engineering approach for the estimation of the mechanical properties, by combining tissue mechanics modeling and statistical machine learning. Our strategy is to model the tissue as a continuum mechanical system and to use passive observations of spontaneous tissue deformation and force fields to statistically estimate the model parameters. This method was applied to the analysis of the collective migration of Madin-Darby canine kidney cells, and the tissue flow and force were simultaneously observed by the phase contrast imaging and traction force microscopy. We found that our monolayer elastic model, whose elastic moduli were reverse-engineered, enabled a long-term forecast of the traction force fields when given the tissue flow fields, indicating that the elasticity contributes to the evolution of the tissue stress. Furthermore, we investigated the tissues in which myosin was inhibited by blebbistatin treatment, and observed a several-fold reduction in the elastic moduli. The obtained results validate our framework, which paves the way to the estimation of mechanical properties of living tissues during morphogenesis. [ABSTRACT FROM AUTHOR]

We developed two new FRET imaging measures for intramolecular FRET biosensors, called linearly proportional (LP) and highly contrasting (HC) measures, which can be easily calculated by the fluorescence intensities of donor and acceptor as a ratio between their weighted sums. As an alternative to the conventional ratiometric measure, which non-linearly depends on the fraction of active molecule, we first developed the LP measure, which is linearly proportional to the fraction of active molecules. The LP measure inherently unmixes bleed-through signals and is robust against fluorescence noise. By extending the LP measure, we furthermore designed the HC measure, which provides highly contrasting images of the molecular activity, more than the ratiometric measure. In addition to their advantages, these measures are insensitive to the biosensor expression level, which is a fundamental property of the ratiometric measure. Using artificial data and FRET imaging data, we showed that the LP measure effectively represents the fraction of active molecules and that the HC measure improves visual interpretability by providing high contrast images of molecular activity. Therefore, the LP and HC measures allow us to gain more quantitative and qualitative insights from FRET imaging than the ratiometric measure. [ABSTRACT FROM AUTHOR]

Spring neap variations in stratification and chlorophyll fluorescence were studied during the summers of 2011–2014 in a tidal sea (Yatsushiro Sea, Japan) using monitoring data and hydrodynamic models. Vertical profiles of salinity, temperature and chlorophyll fluorescence were collected nearly weekly from nine stations in this sea during the same period. Composite analysis using vertical profiles of density clearly indicated enhancement of the stratification during the neap tide and a vertically mixed water column during the spring tide in the tidal area. Interregional differences were revealed in the variation of chlorophyll fluorescence with the spring neap tidal cycle. More notable increases in chlorophyll fluorescence were observed during the neap tide in the tidal area around the narrow strait than in the inner area. Temporal stratification led to an increase in the chlorophyll fluorescence in the tidal strait during the neap tide. [ABSTRACT FROM AUTHOR]

The response to the mechanical loading of bone tissue has been extensively investigated; however, precisely how much strain intensity is necessary to promote bone formation remains unclear. Combination studies utilizing histomorphometric and numerical analyses were performed using the established murine maxilla loading model to clarify the threshold of mechanical strain needed to accelerate bone formation activity. For 7 days, 191 kPa loading stimulation for 30 min/day was applied to C57BL/6J mice. Two regions of interest, the AWAY region (away from the loading site) and the NEAR region (near the loading site), were determined. The inflammatory score increased in the NEAR region, but not in the AWAY region. A strain intensity map obtained from $$\upmu \hbox {CT}$$ images was superimposed onto the images of the bone formation inhibitor, sclerostin-positive cell localization. The number of sclerostin-positive cells significantly decreased after mechanical loading of more than $$150\,{\upmu }{\upvarepsilon }$$ in the AWAY region, but not in the NEAR region. The mineral apposition rate, which shows the bone formation ability of osteoblasts, was accelerated at the site of surface strain intensity, namely around $$170\,{\upmu }{\upvarepsilon }$$ , but not at the site of lower surface strain intensity, which was around $$80\,{\upmu }{\upvarepsilon }$$ in the AWAY region, thus suggesting the existence of a strain intensity threshold for promoting bone formation. Taken together, our data suggest that a threshold of mechanical strain intensity for the direct activation of osteoblast function and the reduction of sclerostin exists in a murine maxilla loading model in the non-inflammatory region. [ABSTRACT FROM AUTHOR]

Tooth extraction causes bone resorption of the alveolar bone volume. Although recombinant human bone morphogenetic protein 2 (rhBMP-2) markedly promotes de novo bone formation after tooth extraction, the application of high-dose rhBMP-2 may induce side effects, such as swelling, seroma, and an increased cancer risk. Therefore, reduction of the necessary dose of rhBMP-2 which can still obtain sufficient bone mass is necessary by developing a new osteogenic reagent. Recently, we showed that the systemic administration of OP3-4 peptide, which was originally designed as a bone resorption inhibitor, had osteogenic ability both in vitro and in vivo . This study evaluated the ability of the local application of OP3-4 peptide to promote bone formation in a murine tooth extraction model with a very low-dose of BMP. The mandibular incisor was extracted from 10-week-old C57BL6/J male mice and a gelatin hydrogel containing rhBMP-2 with or without OP3-4 peptide (BMP/OP3-4) was applied to the socket of the incisor. Bone formation inside the socket was examined radiologically and histologically at 21 days after the extraction. The BMP/OP3-4-group showed significant bone formation inside the mandibular extraction socket compared to the gelatin-hydrogel-carrier-control group or rhBMP-2-applied group. The BMP/OP3-4-applied mice showed a lower reduction of alveolar bone and fewer osteoclast numbers, suggesting that the newly formed bone inside the socket may prevent resorption of the cortical bone around the extraction socket. Our data revealed that OP3-4 peptide promotes BMP-mediated bone formation inside the extraction socket of mandibular bone, resulting in preservation from the loss of alveolar bone. [ABSTRACT FROM AUTHOR]

In previous studies, to address the problem caused by eutrophication and oligotrophication in coastal waters, nutrient cycles have been estimated using bio-geophysical models. These studies strictly evaluated land-derived nutrient loading but not the nutrient loading from offshore. This study demonstrated the quantitation of the offshore-derived nutrient-loading based on in-situ data not based on a bio-geochemical model in the semi-enclosed Tokyo Bay, Japan affected by oligotrophication and discusses about the contribution of a decadal change in the inward nutrient flux to oligotrophication. The inward water flux from offshore was calculated using Ferry-mounted current profiler data. The dissolved inorganic nutrient concentrations of the inward water were estimated using the in-situ sea temperature following the negative correlation relationship between the sea temperature and nutrient concentration. Estimated inward nutrient flux from the Pacific Ocean peaks in March (spring) and the amount of the inward nitrogen has the same order of magnitude as the land-derived input. Notable variability of the inward nutrient flux was confirmed during 2010–2019. The significant decadal change of the inward water mass flux was not confirmed because of the lack of significant decadal changes in the in-situ controlling factors identified by the hydrodynamic model. The decadal decreasing trend in the total nitrogen of inward water suggests a decrease in the inward nutrient flux which contributes to the oligotrophication in Tokyo Bay. • Inward nutrient flux from offshore was estimated by in-situ data. • Maximum inward nutrient flux is the same order as land-derived input in Tokyo Bay. • Notable variability of inward nutrient fluxes was confirmed. • Decadal decrease of offshore-derived nutrient contributes to oligotrophication. [ABSTRACT FROM AUTHOR]

Mosquitoes are important vectors of parasitic and viral diseases with Anopheles gambiae transmitting malaria and Aedes aegypti spreading yellow and Dengue fevers. Using two different approaches (solid-phase extraction and reversed-phase or hydrophilic interaction HPLC fractionation followed by MALDI-TOF MS or permethylation followed by NSI-MS), we examined the N-glycans of both A. gambiae and A. aegypti larvae and demonstrate the presence of a range of paucimannosidic glycans as well as bi- and tri-antennary glycans, some of which are modified with fucose or with sulphate or glucuronic acid residues; the latter anionic modifications were also found on N-glycans of larvae from another dipteran species ( Drosophila melanogaster ). The sulphate groups are attached primarily to core α-mannose residues (especially the α1,6-linked mannose), whereas the glucuronic acid residues are linked to non-reducing β1,3-galactose. Also, O-glycans were found to possess glucuronic acid and sulphate as well as phosphoethanolamine modifications. The presence of sulphated and glucuronylated N-glycans is a novel feature in dipteran glycomes; these structures have the potential to act as additional anionic glycan ligands involved in parasite interactions with the vector host. [ABSTRACT FROM AUTHOR]

Mass mortality of Seriola quinqueradiata was caused by a Chattonella bloom in summer 2010 in the southern area of the Yatsushiro Sea. We analyzed the diurnal records of the mortality occurrence and cell density of C. marina to examine the short-term mortality dynamics. In addition, a hydrodynamic model and Lagrangian particle tracking method were used to examine interregional differences in the mortalities. Mortalities were concentrated along the southern coast of Shishi Island and western coast of Nagashima Island in early July. In late July, mortalities occurred along the southern coast of Shishi Island and eastern coast of Nagashima Island. Severe mortalities occurred where the bloom (>100 cells ml of Chattonella marina) appeared in both early and late July. Model results show that interregional difference of the bloom was controlled by the advection system in the Yatsushiro Sea. The consistent spatio-temporal pattern between fish mortalities and bloom dynamics indicates the potential benefits of real-time monitoring of harmful algae and assessing factors controlling hydrographic conditions to mitigate mortalities. [ABSTRACT FROM AUTHOR]

The relationship between harmful blooms of raphidophycean flagellate Chattonella spp. and preceding meteorological conditions was investigated in the Yatsushiro Sea, Japan, using data over a 25-year period from 1988 to 2012. The occurrence date of Chattonella blooms was significantly correlated with the average air temperature from February through April and with the onset date of Baiu, the Japanese name for the rainy season in early summer. These two meteorological factors showed significant differences between occurrence and non-occurrence years of Chattonella blooms in the Yatsushiro Sea. The Chattonella blooms tended to occur in years that were relatively warm from winter to spring and had a relatively late onset of Baiu. The duration of Chattonella blooms was significantly correlated with the interval between the closing date of Baiu and the occurrence date of the Chattonella blooms, suggesting an important role of Baiu as nutrient input affecting the magnitude of the bloom event. The East Asian monsoon, which controls local meteorological conditions, is likely involved in the population dynamics of Chattonella through its life cycle and physiological characteristics. [ABSTRACT FROM AUTHOR]

We demonstrate that disulfide bond replacement is an efficient strategy for engineering therapeutic peptides. In previous work, short peptide fragments, known as WP9QY, with sequence homology with the predicted ligand contact surface of the receptor activator of NF-κB (RANK) were crosslinked through intramolecular disulfide bonds to inhibit RANK ligand (RANKL)-induced signaling, osteoclastogenesis, bone resorption in vitro, and bone loss in vivo. We report that replacement of the disulfide bond of WP9QY with an amine cross-linkage results in a significant improvement in enzymatic stability, with only a slight loss of bone resorption-blocking activity in vitro. Furthermore, the WP9QY derivative inhibits bone loss significantly in vivo, whereas the native form of WP9QY was not effective under the same conditions. [ABSTRACT FROM AUTHOR]

Summary: The extracellular signal-regulated kinase (ERK) plays a central role in the signaling cascades of cell growth. Here, we show that stochastic ERK activity pulses regulate cell proliferation rates in a cell density-dependent manner. A fluorescence resonance energy transfer (FRET) biosensor revealed that stochastic ERK activity pulses fired spontaneously or propagated from adjacent cells. Frequency, but not amplitude, of ERK activity pulses exhibited a bell-shaped response to the cell density and correlated with cell proliferation rates. Consistently, synthetic ERK activity pulses generated by a light-switchable CRaf protein accelerated cell proliferation. A mathematical model clarified that 80% and 20% of ERK activity pulses are generated by the noise and cell-to-cell propagation, respectively. Finally, RNA sequencing analysis of cells subjected to the synthetic ERK activity pulses suggested the involvement of serum responsive factor (SRF) transcription factors in the gene expression driven by the ERK activity pulses. [Copyright &y& Elsevier]

The progress in imaging technology with fluorescent proteins has uncovered a wide range of biological processes in developmental biology. In particular, genetically-encoded biosensors based on the principle of fluorescence resonance energy transfer ( FRET) have been used to visualize spatial and temporal dynamics of intracellular signaling in living cells. However, development of sensitive FRET biosensors and their application to developmental biology remain challenging tasks, which has prevented their widespread use in developmental biology. In this review, we first overview general procedures and tips of imaging with FRET biosensors. We then describe recent advances in FRET imaging - namely, the use of optimized backbones for intramolecular FRET biosensors and transposon-mediated gene transfer to generate stable cell lines and transgenic mice expressing FRET biosensors. Finally, we discuss future perspectives of FRET imaging in developmental biology. [ABSTRACT FROM AUTHOR]

Abstract: Infection by Opisthorchis viverrini (liver fluke) is a major public health problem in southeastern Asia, resulting in hepatobiliary disease and cholangiocarcinoma. Fluke surface glycoconjugates are prominently presented to the host, thereby constituting a crucial immunological interface that can determine the parasite’s success in establishing infection. Therefore, N- and O-linked glycoprotein glycan profiles of the infective metacercarial stage and of the mature adult were investigated by nanospray ionisation-linear ion trap mass spectrometry (NSI-MS n ). Glycan immunogenicity was investigated by immunoblotting with serum from infected humans. Metacercariae and adult parasites exhibit similar glycan diversity, although the prevalence of individual glycans and glycan classes varies by stage. The N-glycans of the metacercaria are mostly high mannose and monofucosylated, truncated-type oligosaccharides (62.7%), with the remainder processed to complex and hybrid type glycans (37.3%). The N-linked glycan profile of the adult is also dominated by high mannose and monofucosylated, truncated-type oligosaccharides (80.0%), with a smaller contribution from complex and hybrid type glycans (20.0%). At both stages, complex and hybrid type glycans are detected as mono-, bi-, tri-, or tetra-antennary structures. In metacercariae and adults, O-linked glycans are detected as mono- to pentasaccharides. The mucin type core 1 structure, Galβ1–3GalNAc, predominates in both stages but is less prevalent in the adult than in the metacercaria. Immunogenic recognition of liver fluke glycoproteins is reduced after deglycosylation but infected human serum was unable to recognise glycans released from peptides. Therefore, the most potent liver fluke antigenic epitopes are mixed determinants, comprised of glycan and polypeptide elements. [Copyright &y& Elsevier]

Abstract: A harmful bloom due to the raphidophycean flagellate, Chattonella antiqua, was found in the Yatsushiro Sea, western Kyushu, Japan, from the end of July to the beginning of August 2009. The bloom resulted in enormous economic damage to cultured finfish production in aquaculture farms concentrated in the southwestern area. To investigate the factors controlling the spatio-temporal distribution of the bloom, data analysis and numerical simulations were conducted using field monitoring data and a three-dimensional hydrodynamic model coupled to a Lagrangian particle-tracking model. Results of the monitoring data analysis showed that the initial development of the C. antiqua bloom occurred in Kusuura Bay and the northeastern area near the mouth of the Kuma River, and subsequently the bloom expanded rapidly to the whole area. The simulation results indicated that the source region of the widespread bloom was not Kusuura Bay but the northeastern area. The southwestward evolution of the bloom was primarily controlled by the passive transport due to the surface residual current driven by fresh water discharge from the Kuma River and northeasterly winds. On the favorable conditions of river discharge and wind, the massive bloom of C. antiqua that formed in the northeastern area was quickly transported southwestward within a few days. [Copyright &y& Elsevier]

The abundance and structural diversity of glycans on glycoproteins and glycolipids are highly regulated and play important roles during vertebrate development. Because of the challenges associated with studying glycan regulation in vertebrate embryos, we have chosen to study mouse embryonic stem (ES) cells as they differentiate into embryoid bodies (EBs) or into extraembryonic endodermal (ExE) cells as a model for cellular differentiation.WeprofiledN- andO-glycan structures isolated from these cell populations and examined transcripts encoding the corresponding enzymatic machinery for glycan biosynthesis in an effort to probe the mechanisms that drive the regulation of glycan diversity. During differentiation from mouse ES cells to either EBs or ExE cells, general trends were detected. The predominance of high mannose N-glycans in ES cells shifted to an equal abundance of complex and high mannose structures, increased sialylation, and increased α-Gal termination in the differentiated cell populations. Whereas core 1 O-glycan structures predominated in all three cell populations, increased sialylation and increased core diversity characterized the O-glycans of both differentiated cell types. Increased polysialylation was also found in both differentiated cell types. Differences between the two differentiated cell types included greater sialylation of N-glycans in EBs, whereas α-Gal-capped structures were more prevalent in ExE cells. Changes in glycan structures generally, but not uniformly, correlated with alterations in transcript abundance for the corresponding biosynthetic enzymes, suggesting that transcriptional regulation contributes significantly to the regulation of glycan expression. Knowledge of glycan structural diversity and transcript regulation should provide greater understanding of the roles of protein glycosylation in vertebrate development. [ABSTRACT FROM AUTHOR]

Gangliosides—sialylated glycosphingolipids—are the major glycoconjugates of nerve cells. The same four structures—GM1, GD1a, GD1b and GT1b—comprise the great majority of gangliosides in mammalian brains. They share a common tetrasaccharide core (Galβ1–3GalNAcβ1-4Galβ1-4Glcβ1-1′Cer) with one or two sialic acids on the internal galactose and zero (GM1 and GD1b) or one (GD1a and GT1b) α2–3-linked sialic acid on the terminal galactose. Whereas the genes responsible for the sialylation of the internal galactose are known, those responsible for terminal sialylation have not been established in vivo. We report that St3gal2 and St3gal3 are responsible for nearly all the terminal sialylation of brain gangliosides in the mouse. When brain ganglioside expression was analyzed in adult St3gal1-, St3gal2-, St3gal3- and St3gal4-null mice, only St3gal2-null mice differed significantly from wild type, expressing half the normal amount of GD1a and GT1b. St3gal1/2-double-null mice were no different than St3gal2-single-null mice; however, St3gal2/3-double-null mice were >95% depleted in gangliosides GD1a and GT1b. Total ganglioside expression (lipid-bound sialic acid) in the brains of St3gal2/3-double-null mice was equivalent to that in wild-type mice, whereas total protein sialylation was reduced by half. St3gal2/3-double-null mice were small, weak and short lived. They were half the weight of wild-type mice at weaning and displayed early hindlimb dysreflexia. We conclude that the St3gal2 and St3gal3 gene products (ST3Gal-II and ST3Gal-III sialyltransferases) are largely responsible for ganglioside terminal α2-3 sialylation in the brain, synthesizing the major brain gangliosides GD1a and GT1b. [ABSTRACT FROM PUBLISHER]

Abstract: Peptides are attractive as novel therapeutic reagents, since they are flexible in adopting and mimicking the local structural features of proteins. Versatile capabilities to perform organic synthetic manipulations are another unique feature of peptides compared to protein-based medicines, such as antibodies. On the other hand, a disadvantage of using a peptide for a therapeutic purpose is its low stability and/or high level of aggregation. During the past two decades, numerous peptides were developed for the treatment of bone diseases, and some peptides have already been used for local applications to repair bone defects in the clinic. However, very few peptides have the ability to form bone themselves. We herein summarize the effects of the therapeutic peptides on bone loss and/or local bone defects, including the results from basic studies. We also herein describe some possible methods for overcoming the obstacles associated with using therapeutic peptide candidates. [Copyright &y& Elsevier]

Abstract: Recently an increase in the serum levels of a bone formation marker after anti-tumor necrosis factor (TNF)-α therapy in rheumatoid arthritis patients has been reported. However, there remains no direct evidence that TNF-α antagonist could accelerate bone formation under inflammatory condition. Cavity-induced allosteric modification (CIAM) compound, F002, is a newly developed-TNF-α antagonist, which was designed by using the structure of TNF type 1 receptor, thus preventing TNF-α-induced signaling. In this study, we examined whether the CIAM compound can promote bone formation under inflammatory condition induced by lipopolysaccharide (LPS). Thirty-six 10-week-old male mice (C57BL/6J) were used. Half of the mice received 10mg/kg LPS, while the other half received PBS. Thereafter, incisor extraction was performed at 4days after either LPS or PBS injection. Intraperitoneal injections of 2mg/kg/day, 4mg/kg/day CIAM, or vehicle (10% DMSO) were performed once a day from day 0 to day 7 after incisor tooth extraction. The mice were sacrificed at 21days after the extraction. The regenerated bone mineral density (BMD) in the tooth socket, and the mineral apposition rate and the bone formation rate, direct evidences of bone formation, were significantly decreased in the LPS-injected group compared to the PBS-injected group. CIAM compound dose-dependently prevented the decrease of BMD under the LPS-injected condition, and promoted both the mineral apposition rate and the bone formation rate significantly compared to the LPS-injected group. We did not observe any significant differences among the PBS-injected groups. Taken together, TNF-α antagonist CIAM compound, was found to accelerate bone formation under LPS-induced inflammatory condition. [Copyright &y& Elsevier]

The mitogen-activated protein (MAP) kinase pathway is comprised of a three-tiered kinase cascade. The distributive kinetic mechanism of two-site MAP kinase phosphorylation inherently generates a nonlinear switch-like response. However, a linear graded response of MAP kinase has also been observed in mammalian cells, and its molecular mechanism remains unclear. To dissect these input-output behaviors, we quantitatively measured the kinetic parameters involved in the MEK (MAPK/ERK kinase)-ERK MAP kinase signaling module in HeLa cells. Using a numerical analysis based on experimentally determined parameters, we predicted in silico and validated in vivo that ERK is processively phosphorylated in HeLa cells. Finally, we identified molecular crowding as a critical factor that converts distributive phosphorylation into processive phosphorylation. We proposed the term quasi-processive phosphorylation to describe this mode of ERK phosphorylation that is operated under the physiological condition of molecular crowding. The generality of this phenomenon may provide a new paradigm for a diverse set of biochemical reactions including multiple posttranslational modifications. [ABSTRACT FROM AUTHOR]

Cryptococcal meningoencephalitis is an AIDS-defining illness caused by the opportunistic pathogen Cryptococcus neoformans. This organism possesses an elaborate polysaccharide capsule that is unique among pathogenic fungi, and the glycobiology of C. neoformans has been a focus of research in the field. The capsule and other cellular glycans and glycoconjugates have been described, but the machinery responsible for their synthesis remains largely unexplored. We recently discovered Xpt1p, an enzyme with the unexpected activity of generating a xylose-phosphate-mannose linkage. We now demonstrate that this novel activity is conserved throughout the C. neoformans species complex, localized to the Golgi apparatus, and functions in the O-glycosylation of proteins. We also present the first survey of O-glycans from C. neoformans. [ABSTRACT FROM AUTHOR]

Dystroglycanopathies are a subset of congenital muscular dystrophies wherein α-dystroglycan (α-DG) is hypoglycosylated. α-DG is an extensively O-glycosylated extracellular matrix-binding protein and a key component of the dystrophin-glycoprotein complex. Previous studies have shown α-DG to be post-translationally modified by both O-GalNAc- and O-mannose-initiated glycan structures. Mutations in defined or putative glycosyltransferase genes involved in O-mannosylation are associated with a loss of ligand-binding activity of α-DG and are causal for various forms of congenital muscular dystrophy. In this study, we sought to perform glycomic analysis on brain O-linked glycan structures released from proteins of three different knock-out mouse models associated with O-mannosylation (POMGnT1, LARGE (Myd), and DAG1-/-). Using mass spectrometry approaches, we were able to identify nine O-mannose-initiated and 25 O-GalNAc-initiated glycan structures in wild-type littermate control mouse brains. Through our analysis, we were able to confirm that POMGnT1 is essential for the extension of all observed O-mannose glycan structures with β1,2-linked GlcNAc. Loss of LARGE expression in the Myd mouse had no observable effect on the O-mannose-initiated glycan structures characterized here. Interestingly, we also determined that similar amounts of O-mannose-initiated glycan structures are present on brain proteins from α-DG-lacking mice (DAG1) compared with wild-type mice, indicating that there must be additional proteins that are O-mannosylated in the mammalian brain. Our findings illustrate that classical β1,2-elongation and β1,6-GlcNAc branching of O-mannose glycan structures are dependent upon the POMGnT1 enzyme and that O-mannosylation is not limited solely to α-DG in the brain. [ABSTRACT FROM AUTHOR]

The actin cytoskeleton undergoes rapid changes in its architecture during mitosis. Here, we demonstrate novel actin assembly dynamics in M phase. An amorphous cluster of actin filaments appears during prometaphase, revolves horizontally along the cell cortex at a constant angular speed, and fuses into the contractile ring after three to four revolutions. Cdk1 activity is required for the formation of this mitotic actin cluster and its revolving movement. Rapid turnover of actin in the filaments takes place everywhere in the cluster and is also required for its cluster rotation during mitosis. Knockdown of Arp3, a component of the actin filament-nucleating Arp2/3 complex, inhibits the formation of the mitotic actin cluster without affecting other actin structures. These results identify Arp2/3 complex as a key factor in the generation of the dynamic actin cluster during mitosis. [ABSTRACT FROM AUTHOR]

Abstract: Microbial activity has been investigated for the bentonite buffer and surrounding host rock (granodiorite) at the Kamaishi Mine in Iwate, Northeast Japan. For the host rock, the total number of bacteria and viable microorganisms was enumerated for deep groundwater in granodiorite which was sampled from six hydraulically separated zones. The six groundwater samples were taken from two horizontal boreholes drilled from drift KD-90. Using acridine orange direct staining, the total number of bacteria was 8.6×102 to 1.2×106 cells/ml. From the most probable number (MPN) technique, heterotrophs numbered in the range of 5.3×102 to 3.9×104 CFU/ml. Nitrate-reducing bacteria dominated, in the range of 4.9×102 to 5.4×104 MPN/ml, while the presence of sulphate-reducing bacteria and denitrifying bacteria was also confirmed. The coupled thermo-hydro-mechanical (T-H-M) experiments named “engineered barrier experiments” were carried out to examine the in situ performance of buffer material. At the end of the heating and cooling phases, bentonite samples were taken for microbial analysis to determine if the naturally present microbial population in the buffer material survived the conditions (i.e. compaction, heat and desiccation) in a simulated vault environment. Heterotrophic aerobic bacteria were enumerated by the pour plate method using R2A media. The results confirmed the existence of heterotrophs, which disappeared in bentonite samples with low water content (<12%). These results suggest that microbial activity is severely limited near waste container in the vault for some time after disposal, due to desiccation as a result of the heat output of the waste container. Such knowledge will be useful in assessing the potential effects of microbial activity on deep geological disposal of high level radioactive waste. [Copyright &y& Elsevier]

The article presents a study which determines whether zwitterionic glycosphingolipids (ZGLs) from the phytopathogenic filamentous fungus Trichoderma viride have functional roles in plant-fungus interactions. It notes that ZGLs were tested to identify if they would induce defense responses in cultured rice cells. It concludes that T. viride's ZGLs elicited expression of the PAL and PBZ1 genes, which are associated with pathogen resistance.

The reactivation mechanisms of coseismic surface ruptures associated with the 2011 M w 6.7 Fukushima-ken Hamadori earthquake in Japan are investigated using in-situ controlled hydraulic injections in subsurface boreholes. Two fault segments were selected for reactivation studies, one across a coseismic rupture, the Shionohira site, and one across a non-coseismically ruptured segment, the Minakami-kita site. A series of water injections in sealed sections of boreholes set across the fault progressively bring the fault to rupture by a step-by-step decrease of the effective normal stress clamping the fault. While the fault is rupturing during these hydraulic stimulations, borehole displacements, fluid pressure and injection flowrate are continuously monitored. Then, the tests were analyzed using fully coupled hydromechanical modeling. The model was calibrated on field data, and a parametric study was conducted to examine the modes of fault reactivation. Coseismic surface rupture of the Shionohira fault showed a pure dilatant slip response to hydraulic tests, while the tectonically un-activated Itozawa fault (South) indicated a complex hybrid response to tests related to both a higher frictional and cohesive strengths of the fault. The analysis of the induced Shionohira slip event showed that it is reasonably modeled as a Coulomb rupture with an eventual dependency of friction on slip velocity, in good accordance with laboratory-derived rate-and-state friction data on the Shinohira gouge samples. In contrast, the Itozawa fault reactivation mechanism appears dominated by tensile failure with limited Coulomb shear failure. Thus, the applied protocol proves to be able to isolate significant differences in fault physical properties and rupture mechanisms between two segments of the same fault system, opening perspectives to better assess near-surface rupture effects, and therefore the safety of structures such as dams or nuclear waste repositories subject to large earthquakes. • Coseismically and non-coseismically ruptured fault segments are reactivated in situ. • Fault displacements are measured during fluid injection in subsurface boreholes. • Reactivation of the coseismic rupture show a rate and state dilatant slip response. • The un-activated segment reactivation's mode is different from the coseismic's one. • The method helps quantifying fault displacement triggered by a designed earthquake. [ABSTRACT FROM AUTHOR]

Advances in time-lapse fluorescence microscopy have enabled us to directly observe dynamic cellular phenomena. Although the techniques themselves have promoted the understanding of dynamic cellular functions, the vast number of images acquired has generated a need for automated processing tools to extract statistical information. A problem underlying the analysis of time-lapse cell images is the lack of rigorous methods to extract morphodynamic properties. Here, we propose an algorithm called edge evolution tracking (EET) to quantify the relationship between local morphological changes and local fluorescence intensities around a cell edge using time-lapse microscopy images. This algorithm enables us to trace the local edge extension and contraction by defining subdivided edges and their corresponding positions in successive frames. Thus, this algorithm enables the investigation of cross-correlations between local morphological changes and local intensity of fluorescent signals by considering the time shifts. By applying EET to fluorescence resonance energy transfer images of the Rho-family GTPases Rac1, Cdc42, and RhoA, we examined the crosscorrelation between the local area difference and GTPase activity. The calculated correlations changed with time-shifts as expected, but surprisingly, the peak of the correlation coefficients appeared with a 6-8 min time shift of morphological changes and preceded the Rac1 or Cdc42 activities. Our method enables the quantification of the dynamics of local morphological change and local protein activity and statistical investigation of the relationship between them by considering time shifts in the relationship. Thus, this algorithm extends the value of time-lapse imaging data to better understand dynamics of cellular function. [ABSTRACT FROM AUTHOR]

Appropriate glycoprotein O-glycosylation is essential for normal development and tissue function in multicellular organisms. To comprehensively assess the developmental and functional impact of altered O-glycosylation, we have extensively analyzed the nonglycosaminoglycan, O-linked glycans expressed in Drosophila embryos. Through multidimensional mass spectrometric analysis of glycans released from glycoproteins by β-elimination, we detected novel as well as previously reportedO-glycans that exhibit developmentally modulated expression. The core 1 mucin-type disaccharide (Galβ1-3GalNAc) is the predominant glycan in the total profile. HexNAcitol, hexitol, xylosylated hexitol, and branching extension of core 1 with HexNAc (to generate core 2 glycans) were also evident following release and reduction. After Galβ1- 3GalNAc, the next most prevalent glycans were a mixture of novel, isobaric, linear, and branched forms of a glucuronyl core 1 disaccharide. Other less prevalent structures were also extended with HexA, including anO-fucose glycan. Although the expected disaccharide product of the Fringe glycosyl transferase, (GlcNAcβ1- 3)fucitol, was not detectable in whole embryos, mass spectrometry fragmentation and exoglycosidase sensitivity defined a novel glucuronyl trisaccharide as GlcNAcβ1-3(GlcAβ1-4)fucitol. Consistent with the spatial distribution of the Fringe function, the GlcAextended form of the Fringe product was enriched in the dorsal portion of the wing imaginal disc. Furthermore, loss of Fringe activity reduced the prevalence of the O-Fuc trisaccharide. Therefore, O-Fuc glycans necessary for the modulation of important signaling events in Drosophila are, as in vertebrates, substrates for extension beyond the addition of a single HexNAc. [ABSTRACT FROM AUTHOR]

The article presents a study which examined the relationship between baseball field tests and a testing protocol for baseball players. It involved 43 college baseball players, who performed the field tests of physical fitness, such as batting, fielding and running. It is found that players with high evaluation scores have better test results than those with low evaluation scores. Results have also shown a significant correlation between physical fitness found in the field tests and performance.

The practical realization of Chu's minimum-sized antenna is investigated using self-complementary antennas. Rigorous bandwidth formulas for broadband antennas are derived for broadband antennas based on Chu's fundamental mode equivalent circuit. By comparing input impedances and radiation patterns, the fabricated self-complementary antenna used for Japan's ultra-wideband higher band (7.25-10.25 GHz) is found approximated by the minimum-sized antenna. Using the rigorous bandwidth formulas above, the fabricated antenna is shown to be smaller than the minimum-sized antenna. This breakthrough is achieved by using a more complicated matching circuit for broadband antennas instead of the single-stage inductor used in bandwidth estimation with the Q factor. [ABSTRACT FROM AUTHOR]

The local accumulation of phosphatidylinositol (3,4,5) trisphosphate (PIP[sub 3]) and resulting activation of Rac1/ Cdc42 play a critical role in nerve growth factor (NGF)-induced neurite outgrowth. To further explore the mechanism, we visualized PIP[sub 3], phosphatidylinositol (3,4) bisphosphate, and Rac1/Cdc42 activities by fluorescence resonance energy transfer (FRET) imaging in NGF-stimulated PC12 cells. Based on the obtained FRET images, and with the help of in silico kinetic reaction model, we predicted that PI-5-phosphatase negatively regulates PIP[sub 3] upon NGF stimulation. In agreement with this model, depletion of Src homology 2 domain--containing inositol polyphosphate 5-phosphatase 2 (SHIP2) markedly potentiated NGF-induced Rac1/Cdc42 activation and PIP[sub 3] accumulation and considerably increased the number and the length of neurites in phosphate and tensin homologue--depleted PC12 cells. Further refinement of the computational model predicted Rac1 regulation of P13-kinase and SHIP2, which was also validated experimentally. We propose that the SHIP2-mediated negative feedback on PIP[sub 3] coordinately works with the PI3-kinase--mediated positive feedback to form an initial protrusive pattern and, later, to punctuate the PIP[sub 3] accumulation to maintain proper neurite outgrowth. [ABSTRACT FROM AUTHOR]

The structural diversity of glycoprotein N-linked oligosaccharides is determined by the expression and regulation of glycosyltransferase activities and by the availability of the appropriate acceptor/donor substrates. Cells in different tissues and in different developmental stages utilize these control points to manifest unique glycan expression patterns in response to their surroundings. The activity of a Toll-like receptor, called Tollo/Toll-8, induces a pattern of incompletely defined, but neural specific, glycan expression in the Drosophila embryo. Understanding the full extent of the changes in glycan expression that result from altered Tollo/Toll-8 signaling requires characterization of the complete N-linked glycan profile of both wild-type and mutant embryos. N-Linked glycans harvested from wild-type or mutant embryos were subjected to direct structural analysis by analytic and preparative high pressure liquid chromatography, by multidimensional mass spectrometry, and by exoglycosidase digestion, revealing a predominance of high mannose and paucimannose glycans. Di-, mono-, and nonfucosylated forms of hybrid, complex biantennary, and triantennary glycans account for 12% of the total wild-type glycan profile. Two sialylated glycans bearing N-acetylneuraminic acid were detected, the first direct demonstration of this modification in Drosophila. Glycan profiles change during normal development consistent with increasing α-mannosidase II and core fucosyltransferase enzyme activities, and with decreasing activity of the Fused lobes processing hexosaminidase. In tollo/toll-8 mutants, a dramatic, expected loss of difucosylated glycans is accompanied by unexpected decreases in monofucosylated and nonfucosylated hybrid glycans and increases in some nonfucosylated paucimannose and biantennary glycans. Therefore, tollo/toll-8 signaling influences flux through several processing steps that affect the maturation of N-linked glycans. [ABSTRACT FROM AUTHOR]

Activating receptor activator of NF-kappaB (RANK) and TNF receptor (TNFR) promote osteoclast differentiation. A critical ligand contact site on the TNFR is partly conserved in RANK. Surface plasmon resonance studies showed that a peptide (WP9QY) that mimics this TNFR contact site and inhibits TNF-alpha-induced activity bound to RANK ligand (RANKL). Changing a single residue predicted to play an important role in the interaction reduced the binding significantly. WP9QY, but not the altered control peptide, inhibited the RANKL-induced activation of RANK-dependent signaling in RAW 264.7 cells but had no effect on M-CSF-induced activation of some of the same signaling events. WP9QY but not the control peptide also prevented RANKL-induced bone resorption and osteoclastogenesis, even when TNFRs were absent or blocked. In vivo, where both RANKL and TNF-alpha promote osteoclastogenesis, osteoclast activity, and bone loss, WP9QY prevented the increased osteoclastogenesis and bone loss induced in mice by ovariectomy or low dietary calcium, in the latter case in both wild-type and TNFR double-knockout mice. These results suggest that a peptide that mimics a TNFR ligand contact site blocks bone resorption by interfering with recruitment and activation of osteoclasts by both RANKL and TNF. [ABSTRACT FROM AUTHOR]

Abstract: Rho-family GTPases play a central role in the regulation of neuronal morphogenesis. In growth cones, for example, Rho GTPases transduce extracellular stimuli into structural changes such as filopodia and lamellipodia. Although it is generally accepted that Rac1/Cdc42 and RhoA are positive and negative regulators of neurite outgrowth, respectively, the role of each Rho-family member in neuronal morphogenesis may change according to the cell context. At present, the mechanism underlying this complexity is largely unknown. In growth cones, this is partly due to a lack of information on the distribution of active Rho GTPases. Here, we visualized RhoA/Rac1/Cdc42 activities during laminin-induced growth cone advance of DRG neurons and N1E-115 neuroblastoma cells using probes based on fluorescence/Förster resonance energy transfer. The Rac1 and Cdc42 activities were high in the peripheral domain (P-domain) of growth cones. Active Rac1 was uniformly detected throughout the P-domain, whereas Cdc42 activity increased gradually toward the growth cone edge. Against a model involving RhoA down-regulation at the periphery of protruding growth cones, we found that the RhoA activity was higher in the P-domain than in the central domain and axon shaft, and that a high level of RhoA activity was maintained in the extending part of growth cones. In lysophosphatidic acid-treated N1E-115 cells, well-developed neurites with growth cones showed RhoA activation, but sustained their extended morphology until they were drawn toward the contracting somata. On the other hand, suppression of RhoA activity by C3 exoenzyme led to loss or deformation of actin bundles in the growth cones. Thus, RhoA activation in the shaft results in neurite retraction, whereas high RhoA activity in the P-domain is necessary to retain the spread morphology of nerve growth cone. [Copyright &y& Elsevier]

Abstract: GFP-based fluorescence resonance energy transfer (FRET) probes that visualize local activity-changes of Ras and Rho GTPases in living cells are now available for examining the spatio-temporal regulation of these proteins. This article describes principles and strategies to develop intramolecular FRET probes for Ras- and Rho-family GTPases. The procedure for characterizing candidate probes, and image acquisition and processing are also explained. An optimal FRET probe should have (i) a wide dynamic range (which means a high sensitivity), (ii) a high fluorescence intensity, (iii) target specificity, and (iv) a minimal perturbation to endogenous signaling cascades. Although an improvement of FRET probes should be executed in a trial-and-error manner, practical tips for optimization are provided here. In addition, we illustrate some applications of FRET probes for neuronal cells, which are composed of diverse subcellular compartments with different functions; thus, tools to decipher the dynamics of GTPase activity in each compartment have long been desired. [Copyright &y& Elsevier]

We found for the first time that Zygomycetes species showed resistance to Aureobasidin A, an antifungal agent. A novel family of neutral glycosphingolipids (GSLs) was found in these fungi and isolated from Mucor hiemalis, which is a typical Zygomycetes species. Their structures were completely determined by compositional sugar, fatty acid, and sphingoid analyses, methylation analysis, matrix-assisted laser desorption ionization time-of-flight/mass spectrometry, and ¹H NMR spectroscopy. They were as follows: Galβ1-6Galβ1-1Cer (CDS), Galα1-6Galβ1-6Galβ1-1Cer (CTS), Galα1-6Galα1-6Galβ1-6Galβ1-1Cer (CTeS), and Galα1-6Galα1-6Galα1-6Gal-6Galβ1-1Cer (CPS). The ceramide moieties of these GSLs consist of 24:0, 25:0, and 26:0 2-hydroxy acids as major fatty acids and 4-hydroxyoctadecasphinganine (phytosphingosine) as the sole sphingoid. However, the glycosylinositolphosphoceramide families that are the major GSLs components in fungi were not detected in Zygomycetes at all. This seems to be the reason that Aureobasidin A is not effective for Zygomycetes as an antifungal agent. Our results indicate that the biosynthetic pathway for GSLs in Zygomycetes is significantly different from those in other fungi and suggest that any inhibitor of this pathway may be effective for mucormycosis, which is a serious pathogenic disease for humans. [ABSTRACT FROM AUTHOR]

Bone destruction is a pathological hallmark of several chronic inflammatory diseases, including rheumatoid arthritis and periodontitis. Inflammation-induced bone loss of this sort results from elevated numbers of bone-resorbing osteoclasts. Gene targeting studies have shown that the transcription factor nuclear factor-?B (NF-?B) has a crucial role in osteoclast differentiation, and blocking NF-?B is a potential strategy for preventing inflammatory bone resorption. We tested this approach using a cell-permeable peptide inhibitor of the I?B-kinase complex, a crucial component of signal transduction pathways to NF-?B. The peptide inhibited RANKL-stimulated NF-?B activation and osteoclastogenesis both in vitro and in vivo. In addition, this peptide significantly reduced the severity of collagen-induced arthritis in mice by reducing levels of tumor necrosis factor-a and interleukin-1ß, abrogating joint swelling and reducing destruction of bone and cartilage. Therefore, selective inhibition of NF-?B activation offers an effective therapeutic approach for inhibiting chronic inflammatory diseases involving bone resorption. [ABSTRACT FROM AUTHOR]

Neurite outgrowth is an important process in the formation of neuronal networks. It is widely accepted that Rac1 and Cdc42, members of the Rho GTPase family, positively regulate neurite extension through reorganization of the actin cytoskeleton; however, it remains largely unknown when and where Rac1 and Cdc42 are activated during neuritogenesis. This study visualized the spatio-temporal regulation of Rac1 and Cdc42 activities during nerve growth factor (NGF)-induced neurite outgrowth in living PC12 cells by using probes based on the principle of fluorescence resonance energy transfer (FRET). Immediately after the addition of NGF, Rac1 and Cdc42 were transiently activated in broad areas of the cell periphery; a repetitive activation and inactivation cycle was then observed at the motile tips of protrusions. This localized activation, which was more evident in PC12 cells treated with NGF for more than 24 h, might facilitate neurite extension, because the expression of constitutively active mutants of Rac1 and Cdc42 abrogated NGF-induced neurite outgrowth. FRET imaging also delineated a difference between the localization of activated Rac1 and that of Cdc42 within the neurite tips. Experiments with dominant-negative mutants suggested that Rac1 and Cdc42 were activated by a common guanine nucleotide exchange factor(s) in an early stage of the activation phase. Therefore, the difference between Rac1- and Cdc42-activated areas possibly came from the differential localization between Rac1-specific GTPase-activation proteins (GAPs) and Cdc42-specific GAPs. It was concluded that the localized activation of Rac1 and Cdc42 was caused by both guanine nucleotide exchange factors and GAPs, and was important for neurite extension. [ABSTRACT FROM AUTHOR]

Characterizes alpha-galactosidase enzyme from the medicinal mushroom Ganoderma lucidum. Culture medium; Amino acid sequence; Molecular mass; Thermostability; Glycosidase activities in the extract of fruiting bodies; Enzyme homogeneity; Effects of various compounds and metal ions on enzyme activity.

Purpose: This study investigated the relationship between vertical and horizontal jumping ability and change of direction (COD) to measure athletic performance in 51 elite male handball players.Scope: Countermovement jump (CMJ), peak power, and standing long jump (SLJ) were measured. Participants performed a 20-m sprint test (time measured at 5, 10, and 20 m) and a zigzag test (COD: 135°, 90°, and 45°). The COD deficit, an index of the time required for COD, was calculated. The correlations between CMJ height and zigzag test times were relatively large (at 135°, r =  - 0.607; at 90°, r =  - 0.594; at 45°, r =  - 0.613; p < 0.01), whereas those between CMJ height and COD deficit were moderate (at 135°, r =  - 0.399, p < 0.01; at 90°, r =  - 0.350, p < 0.05; at 45°, r =  - 0.323, p < 0.05). SLJ showed a negative moderate correlation with COD deficit (at 135°, r =  - 0.439, p < 0.01; at 90°, r =  - 0.469, p < 0.01; at 45°, r =  - 0.380, p < 0.01).Conclusions: This study is the first to analyse SLJ ability and COD deficit parameters of handball players. We found that SLJ ability is moderately related to COD time and deficit; therefore, SLJ measurement may be a useful predictor of athletic performance. [ABSTRACT FROM AUTHOR]

If we assume that the state of suppression of pneumatic cells is the result of suppression of pneumatic cell growth by inflammatory stimulation in the middle ear pneumatic space, it is possible to improve the state of suppression by performing sufficient treatment during the growth period of the pneumatic cells. We indwelt a tympanic membrane ventilation tube (hereinafter referred to as tube) for treatment of otitis media with effusion (OME) in child patients aged 3-13 years and investigated the following points: i) relationship between the severity of inflammation of the lamina propria of middle ear mucosal specimens (hereinafter referred to as lamina propria) collected at the time of tube indwelling and the degree of growth of the pneumatic space; and ii) changes in the pneumatic space associated with treatment by tube indwelling, which was studied by comparing the above-described mucosal severity with the pneumatic space area of 2 years after tube indwelling, and with increase in the pneumatic space volume measured periodically after tube indwelling. The results indicated that mastoid cell growth suppression is higher in patients with a higher degree of inflammatory changes in the lamina propria. In association with treatment by tube indwelling, effusion accumulated in the pneumatic space and mucosal swelling disappeared early after the treatment, or 2 months of tube indwelling. After that, in patients with severe mucosal lesion, a long time, 1.5-2 years, was found to be required for repneumatization accompanying regrowth of the temporal bone. We confirmed that the severity of inflammation of the lamina propria is deeply involved in the growth and repneumatization of the pneumatic cells. [ABSTRACT FROM AUTHOR]

Glycans are one of the fundamental classes of macromolecules and are involved in a broad range of biological phenomena. A large variety of glycan structures can be synthesized depending on tissue or cell types and environmental changes. Here, we developed a comprehensive glycosylation mapping tool, termed GlycoMaple, to visualize and estimate glycan structures based on gene expression. We informatically selected 950 genes involved in glycosylation and its regulation. Expression profiles of these genes were mapped onto global glycan metabolic pathways to predict glycan structures, which were confirmed using glycomic analyses. Based on the predictions of N-glycan processing, we constructed 40 knockout HEK293 cell lines and analyzed the effects of gene knockout on glycan structures. Finally, the glycan structures of 64 cell lines, 37 tissues, and primary colon tumor tissues were estimated and compared using publicly available databases. Our systematic approach can accelerate glycan analyses and engineering in mammalian cells. [Display omitted] • A comprehensive glycosylation mapping tool, termed GlycoMaple, was developed • GlycoMaple could visualize and estimate glycan structures based on gene expression • A cell library of knocked out genes related to N-linked glycosylation was constructed • Glycan changes between normal and diseased tissues were estimated using GlycoMaple Diverse glycan structures are synthesized depending on cell types. Huang et al. have developed a comprehensive glycosylation mapping tool based on gene expression, named GlycoMaple, for visualizing glycosylation pathways and estimating glycan structures synthesized in cells. The tool could contribute to supporting glycan analysis, biomarker development, and glycoengineering. [ABSTRACT FROM AUTHOR]

To examine the environmental factors controlling the inshore recruitment dynamics of Anguilla japonica in the Oyodo River, Miyazaki Prefecture, Kyushu, glass eel samplings were carried out using fyke nets during winter (November-March) of 1994-2014. [ABSTRACT FROM AUTHOR]