HSP70 and HSP90 Differentially Regulate Translocation of Extracellular Antigen to the Cytosol for Cross-Presentation

Antigens (Ag) from cancer or virus-infected cells must be internalized by dendritic cells (DCs) to be presented to CD8+ T cells, which eventually differentiate into Ag-specific cytotoxic T lymphocytes (CTLs) that destroy cancer cells and infected cells. This pathway is termed cross-presentation and is also implicated as an essential step in triggering autoimmune diseases such as Type I diabetes. Internalized Ag locates within endosomes, followed by translocation through a putative pore structure spanning endosomal membranes into the cytosol, where it is degraded by the proteasome to generate antigen peptides. During translocation, Ag is believed to be unfolded since the pore size is too narrow to accept native Ag structure. Here, we show that paraformaldehyde-fixed, structurally inflexible Ag is less efficient in cross-presentation because of diminished translocation into the cytosol, supporting the “unfolded Ag” theory. We also show that HSP70 inhibitors block both endogenous and cross-presentation. ImageStream analysis revealed that the inhibition in cross-presentation is not due to blocking of Ag translocation because a HSP70 inhibitor rather facilitates the translocation, which is in marked contrast to the effect of an HSP90 inhibitor that blocks Ag translocation. Our results indicate that Ag translocation to the cytosol in cross-presentation is differentially regulated by HSP70 and HSP90

社会人ラグビーフットボール選手におけるポジション別栄養アセスメント

A rugby football team consists of 15 players: 8 forwards and 7 backs. Rugby football players must improve their performance according to role of their position. The purpose of this study was to assess nutritional status based on body composition and clinical parameters to compare the forward and back positions in rugby football players. Forty-one male amateur rugby football players （23 forwards and 18 backs, age 24±4 years） playing in the Top West A1 league participated in this study. Body composition and circumference were measured in the preseason. The following were significantly higher in forwards than in backs: height; body weight; body fat percentage; lean body mass; visceral fat area（VFA）; and neck, chest, flexed arm, forearm, hip, thigh, and calf circumference. Six forwards and one back had a VFA of over 100 cm^2. Hemoglobin, hematocrit, alanine aminotransferase （ALT）, and gamma-glutamyl transpeptidase were significantly higher in forwards than in backs. In some forwards, clinical parameters such as triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, or ALT were outside the normal range. The result of nutritional assessment based on body composition and these clinical parameters suggests that physical characteristics such as body weight, lean body mass, body fat percentage, and circumference are higher in rugby football forwards compared with backs. However, some players have higher abdominal visceral fat accumulation, indicating higher risk of lifestyle-related diseases, regardless of positions

EEG Correlates of the Flow State: A Combination of Increased Frontal Theta and Moderate Frontocentral Alpha Rhythm in the Mental Arithmetic Task

Flow experience is a subjective state experienced during holistic involvement in a certain activity, which has been reported to function as a factor promoting motivation, skill development, and better performance in the activity. To verify the positive effects of flow and develop a method to utilize it, the establishment of a reliable measurement of the flow state is essential. The present study utilized an electroencephalogram (EEG) during an experimentally evoked flow state and examined the possibility of objective measurement of immediate flow. A total of 16 participants (10 males, 6 females) participated in the experiment that employed a mental arithmetic task developed in a previous study. Post-trial self-report of the flow state and EEG during task execution were measured and compared among three conditions (Boredom, Flow, and Overload) that had different levels of task difficulty. Furthermore, the correlations between subjective flow items and EEG activity were examined. As expected, the ratings on the subjective evaluation items representing the flow state were the highest in the Flow condition. Regarding the EEG data, theta activities in the frontal areas were higher in the Flow and the Overload conditions than in the Boredom condition, and alpha activity in the frontal areas and the right central area gradually increased depending on the task difficulty. These EEG activities correlated with self-reported flow experience, especially items related to the concentration on the task and task difficulty. From the results, the flow state was characterized by increased theta activities in the frontal areas and moderate alpha activities in the frontal and central areas. The former may be related to a high level of cognitive control and immersion in task, and the latter suggests that the load on the working memory was not excessive. The findings of this study suggest the possibility of distinguishing the flow state from other states using multiple EEG activities and indicate the need for other physiological indicators corresponding to the other aspects of flow experience

Structure-Correlation NMR Spectroscopy for Macromolecules Using Repeated Bidirectional Photoisomerization of Azobenzene

Control over macromolecular structure offers bright potentials for manipulation of macromolecular functions. We here present structure-correlation NMR spectroscopy to analyze the correlation between polymorphic macromolecular structures driven by photoisomerization of azobenzene. The structural conversion of azobenzene was induced within the mixing time of a NOESY experiment using a colored light source, and the reverse structural conversion was induced during the relaxation delay using a light source of another color. The correlation spectrum between <i>trans</i>- and <i>cis</i>-azobenzene was then obtained. To maximize the efficiency of the bidirectional photoisomerization of azobenzene-containing macromolecules, we developed a novel light-irradiation NMR sample tube and method for irradiating target molecules in an NMR radio frequency (rf) coil. When this sample tube was used for photoisomerization of an azobenzene derivative at a concentration of 0.2 mM, data collection with reasonable sensitivity applicable to macromolecules was achieved. We performed isomerization of an azobenzene-cross-linked peptide within the mixing time of a NOESY experiment that produced cross-peaks between helix and random-coil forms of the peptide. Thus, these results indicate that macromolecular structure manipulation can be incorporated into an NMR pulse sequence using an azobenzene derivative and irradiation with light of two types of wavelengths, providing a new method for structural analysis of metastable states of macromolecules

Development of an in vivo-mimic silkworm infection model with Mycobacterium avium complex

In vivo-mimic silkworm infection models with Mycobacterium avium and Mycobacterium intracellulare were newly established to evaluate the therapeutic effects of anti-M. avium complex (MAC) antibiotics. Silkworms raised at 37°C died within 72 hours of an injection of M. avium or M.intracellulare (2.5 × 107 colony-forming unit (CFU)/larva·g) into the hemolymph. Clinical anti-mycobacterial (tuberculosis) antibiotics were evaluated under these conditions. Clarithromycin, kanamycin, streptomycin, amikacin, and ciprofloxacin exerted therapeutic effects in a dose-dependent manner, which was consistent with those in the mouse model. Furthermore, three effective actinomycete culture broths were selected in the screening program of our microbial broth library using the silkworm model, and four active metabolites, ohmyungsamycins A and B (1 and 2), chartreusin (3), and griseoviridin (4), were identified. Among these compounds, 1 showed the lowest 50% effective dose (ED50) value (8.5 µg/larva·g), while 3 had the best ED50/minimum inhibitory concentration (MIC) ratio (7.4). These results indicate that silkworm models are a useful tool for identifying anti-MAC antibiotics candidates with veritable therapeutic effects

High-cell density culture of poly(lactate-co-3-hydroxybutyrate)-producing Escherichia coli by using glucose/xylose-switching fed-batch jar fermentation

Poly(lactate-co-3-hydroxybutyrate) [P(LA-co-3HB)] is produced in engineered Escherichia coli harboring the genes encoding an LA-polymerizing enzyme (LPE) and monomer-supplying enzymes. In this study, high cell-density fed-batch jar fermentation was developed using xylose and/or glucose as the carbon source. Fed-batch fermentation was initially performed with 20 g/L sugar during the batch phase for 24 h, and subsequent sugar feeding from 24 to 86 h. The feeding rate was increased in a stepwise manner. When xylose alone was used for cultivation, the cells produced the polymer at 11.6 g/L, which was higher than the 4.3 g/L obtained using glucose as the sole carbon source. However, in the first 24 h the growth in the glucose culture was greater than in the xylose culture. Based on these results, glucose was used for cell growth (at the initial stage) and xylose was used for polymer production (at the feeding stage). As expected, in the glucose/xylose switching fermentation method, polymer production was significantly enhanced, eventually reaching 26.7 g/L. The enhanced polymer production obtained by using xylose was presumably due to overflow metabolism. In fact, during xylose feeding, acetic acid excretion was greater than that in case of the glucose grown culture, suggesting the channeling of the metabolic flux from acetyl-CoA towards polymer production over into the tricarboxylic acid cycle in the xylose-fed cultures. Therefore, this sequential glucose/xylose feed strategy is potentially useful for production of acetyl-CoA derived compounds in E. coli