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We analyzed the relationship between the nanometer-scale structure of amylopectin and physicochemical properties such as thermal properties of starch granules and higher-order internal structure of starch granules evaluated by using synchrotron radiation X-ray scattering and scanning electron microscopy. In this study, a line of allelic mutants of rice lacking in starch branching enzyme IIb (BEIIb) activity in their endosperm were used as starch sources. The crystal lattice changed from A-type to C- and B-type. The nanoscale cluster size, and higher-order structures of the starch granules varied among mutant lines. The thermal gelatinization properties of the rice-derived starch samples, as well as the dependence of this process on their crystal structure, were evaluated using differential scanning calorimetry and synchrotron X-ray scattering. After gelatinization, several nm-scaled density fluctuations owing to the formation of gel balls were observed. These fluctuations of all starch samples correlated with the cluster size decreased with the temperature.

IntroductionThe most effective method of assessing sarcopenia has yet to be determined, whether by single muscle or by whole muscle segmentation. The purpose of this study was to compare the prognostic value of these two methods using computed tomography (CT) images in patients with oral squamous cell carcinoma (OSCC).Materials and methodsSex- and age-adjusted Cox proportional hazards models were employed for each parameter of sarcopenia related to overall survival, disease-free survival, and disease-specific survival. Harrell’s concordance index was calculated for each model to assess discriminatory power.ResultsIn this study including 165 patients, a significant correlation was found between the CT-based assessment of individual muscles and their cross-sectional area. Single muscle assessments showed slightly higher discriminatory power in survival outcomes compared to whole muscle assessments, but the difference was not statistically significant, as indicated by overlapping confidence intervals for the C-index between assessments. To further validate our measurements, we classified patients into two groups based on intramuscular adipose tissue content (P-IMAC) of the spinous process muscle. Analysis showed that the higher the P-IMAC value, the poorer the survival outcome.ConclusionOur findings indicate a slight advantage of single-muscle over whole-muscle assessment in prognostic evaluation, but the difference between the two methods is not conclusive. Both assessment methods provide valuable prognostic information for patients with OSCC, and further studies involving larger, independent cohorts are needed to clarify the potential advantage of one method over the other in the prognostic assessment of sarcopenia in OSCC.

Major advances have been made in cancer treatment, but the prognosis for elderly cancer patients with sarcopenia and frailty remains poor. Myokines, which are thought to exert preventive effects against sarcopenia, have been reported to be associated with the prognosis of various cancers, but their effect on head and neck squamous cell carcinoma (HNSCC) is unknown. The aim of this study was to clarify the influence of exercise on the control of HNSCC and to examine the underlying mechanism involved. Mice were injected with HSC-3-M3 cells, a human cell line of highly metastatic and poorly differentiated tongue cancer, at the beginning of the study. Just prior to transplantation, blood was collected from the mice, and the levels of myokines were measured by ELISA. Oncostatin M (OSM), a selected myokine, was added to HSC-3-M3 cells, after which the cell proliferation ability, cell cycle, and protein expression were analyzed in vitro. Tumor cell viability was lower (control: 100%, exercise: 75%), tumors were smaller (control: 26.2 mm3, exercise: 6.4 mm3), and survival was longer in the exercise group than in the control group in vivo. OSM inhibited HSC-3-M3 cell proliferation in a concentration-dependent manner in vitro. The addition of OSM increased the proportion of cells in the G0/G1 phase, decreased the proportion of cells in the G2/M phase, and increased the expression of the CDK inhibitors p21 and p27. These results indicate that exercise may directly inhibit the proliferation of HNSCC cell lines via OSM.

Background: Better prognostic biomarkers for oral squamous cell carcinoma (OSCC) must be developed, particularly within the realm of clinically and frequently administered tests, to advise appropriate clinical therapy and follow-up. In this study, we retrospectively investigated which of the several inflammation-nutrition indicators might predict the prognosis of patients with OSCC.Methods: The preoperative neutrophil-lymphocyte ratio (NLR), lymphocyte–monocyte ratio (LMR), platelet–lymphocyte ratio (PLR), CRP-albumin ratio (CAR), Glasgow prognostic score (GPS), modified GPS (mGPS), prognostic nutritional index (PNI), controlling nutrition status (CONUT), and modified CONUT (mCONUT) were retrospectively evaluated using blood samples collected 1–5 days before surgery. To estimate the effect on the prognosis of tumor progression, the mean values of the markers between stages I/II and III/IV were used for subgroup analysis. The multivariate Cox proportional hazards model included all independent variables significantly associated with survival in the univariate analysis to determine the independent variables.Results: A total of 112 patients (69 males and 43 females) with primary OSCC who underwent surgical treatment at our hospital were included. There were statistically significant differences in the mean values of monocytes, platelets, and albumin between stages I/II and III/IV. According to the multivariate Cox proportional hazards regression, a low PNI was associated with shorter overall survival (OS) and disease-free survival (DFS); women were associated with shorter DFS.Conclusion: The pretreatment PNI had excellent predictive value for the 5-year OS and DFS of patients with OSCC. Future large-scale prospective studies with a high sample size are needed to verify our findings in OSCC patients.

Most rhodophytes synthesize semi-amylopectin as a storage polysaccharide, whereas some species in the most primitive class (Cyanidiophyceae) make glycogen. To know the roles of isoamylases in semi-amylopectin synthesis, we investigated the effects of isoamylase gene (CMI294C and CMS197C)-deficiencies on semi-amylopectin molecular structure and starch granule morphology in Cyanidioschyzon merolae (Cyanidiophyceae). Semi-amylopectin content in a CMS197C-disruption mutant (ΔCMS197C) was not significantly different from that in the control strain, while that in a CMI294C-disruption mutant (ΔCMI294C) was much lower than those in the control strain, suggesting that CMI294C is essential for semi-amylopectin synthesis. Scanning electron microscopy showed that the ΔCMI294C strain contained smaller starch granules, while the ΔCMS197C strain had normal size, but donut-shaped granules, unlike those of the control strain. Although the chain length distribution of starch from the control strain displayed a semi-amylopectin pattern with a peak around degree of polymerization (DP) 11–13, differences in chain length profiles revealed that the ΔCMS197C strain has more short chains (DP of 3 and 4) than the control strain, while the ΔCMI294C strain has more long chains (DP ≥12). These findings suggest that CMI294C-type isoamylase, which can debranch a wide range of chains, probably plays an important role in semi-amylopectin synthesis unique in the Rhodophyta.

Abstract Yeast mannan (YM) is an indigestible water-soluble polysaccharide of the yeast cell wall, with a notable prebiotic effect on the intestinal microbiota. We previously reported that YM increased Bacteroides thetaiotaomicron abundance in in vitro rat faeces fermentation, concluding that its effects on human colonic microbiota should be investigated. In this study, we show the effects of YM on human colonic microbiota and its metabolites using an in vitro human faeces fermentation system. Bacterial 16S rRNA gene sequence analysis showed that YM administration did not change the microbial diversity or composition. Quantitative real-time PCR analysis revealed that YM administration significantly increased the relative abundance of Bacteroides ovatus and B. thetaiotaomicron. Moreover, a positive correlation was observed between the relative ratio (with or without YM administration) of B. thetaiotaomicron and B. ovatus (r = 0.92), suggesting that these bacteria utilise YM in a coordinated manner. In addition, YM administration increased the production of acetate, propionate, and total short-chain fatty acids. These results demonstrate the potential of YM as a novel prebiotic that selectively increases B. thetaiotaomicron and B. ovatus and improves the intestinal environment. The findings also provide insights that might be useful for the development of novel functional foods.

Abstract Background Although fatigue is a common and distressing symptom in cancer survivors, the mechanism of fatigue is not fully understood. Therefore, this study aims to investigate the relation between the fatigue and mindfulness of breast cancer survivors using anxiety, depression, pain, loneliness, and sleep disturbance as mediators. Methods Path analysis was performed to examine direct and indirect associations between mindfulness and fatigue. Participants were breast cancer survivors who visited a breast surgery department at a university hospital in Japan for hormonal therapy or regular check-ups after treatment. The questionnaire measured cancer-related-fatigue, mindfulness, anxiety, depression, pain, loneliness, and sleep disturbance. Demographic and clinical characteristics were collected from medical records. Results Two-hundred and seventy-nine breast cancer survivors were registered, of which 259 answered the questionnaire. Ten respondents with incomplete questionnaire data were excluded, resulting in 249 participants for the analyses. Our final model fit the data well (goodness of fit index = .993; adjusted goodness of fit index = .966; comparative fit index = .999; root mean square error of approximation = .016). Mindfulness, anxiety, depression, pain, loneliness, and sleep disturbance were related to fatigue, and mindfulness had the most influence on fatigue (β = − .52). Mindfulness affected fatigue not only directly but also indirectly through anxiety, depression, pain, loneliness, and sleep disturbance. Conclusions The study model helps to explain the process by which mindfulness affects fatigue. Our results suggest that mindfulness has both direct and indirect effects on the fatigue of breast cancer survivors and that mindfulness can be used to more effectively reduce their fatigue. It also suggests that health care professionals should be aware of factors such as anxiety, depression, pain, loneliness, and sleep disturbance in their care for fatigue of breast cancer survivors. Trial registration This study was registered in the University Hospital Medical Information Network Clinical Trials Registry (UMIN number. 000027720 ) on June 12, 2017.

d-amino acids produced by Lactobacillus are thought to contribute to the taste quality and health functions; however, no studies have comprehensively evaluated the concentrations of the D- and L-forms of amino acids separately in individual Lactobacillus strains. To gain insight into amino acid concentrations in Lactobacillus, we evaluated amino acid concentrations in culture broth of Lactobacillus separately for the D- and L-forms. Lactobacillus strains were cultured in culture broth, and the amino acid concentrations in supernatant were assessed. The amino acid concentrations obtained by liquid chromatography-tandem mass spectrometry (LC-MS/MS) were subjected to cluster analysis based on Bray-Curtis distance with Ward's minimum variance method. In the analysis of amino acid concentrations under culture with different monosaccharides, the distances among strains cultured with the same monosaccharide were significantly greater than those among cultures of the same strain under different monosaccharides (p

Lactobacillus gasseri CP2305 (CP2305) is a paraprobiotic that exhibits beneficial effects on the intestinal function and microbiota, and increases resistance to psychological stress. The stress response mechanism mainly involves the hypothalamic–pituitary–adrenal axis, which is influenced by the gut–brain axis. Furthermore, the gut–brain axis also communicates bidirectionally with the intestinal microbiota. Additionally, the hypothalamic–pituitary–adrenal and hypothalamic–pituitary–gonadal axes share a common route that affects both mental and health aspects in women. This double-blind, placebo-controlled, parallel-group clinical trial aimed to analyze the influence of the intake of CP2305 on mild symptoms associated with menopause. Eighty women aged 40–60 years ingested CP2305 or placebo tablets for six consecutive menstrual cycles. Assessment was based on the observation of climacteric symptoms with two validated questionnaires—the Simplified Menopausal Index (SMI) and the Greene Climacteric Scale (GCS). The results showed that CP2305 provided significant relief in the SMI total score, SMI vasomotor score, SMI psychological score, GCS total score, GCS somatic score, and GCS vasomotor score compared to the placebo. The percentage of women with symptom relief for the SMI total score was 75.0%, with 30 of 40 women in the CP2305 group, and 55.0%, with 22 of 40 women in the placebo group (p = 0.0594). These findings provide new insights into the function of paraprobiotic CP2305 in relieving mild climacteric symptoms in women.

It is known that one of starch branching enzyme (BE) isoforms, BEIIb, plays a specific role not only in the synthesis of distinct amylopectin cluster structure, but also in the formation of the internal structure of starch granules in rice endosperm because in its absence the starch crystalline polymorph changes to the B-type from the typical A-type found in the wild-type (WT) cereal endosperm starch granules. In the present study, to examine the contribution of BEIIb to the amylopectin cluster structure, the chain-length distributions of amylopectin and its phosphorylase-limit dextrins (Φ-LD) from endosperm and culm of a null be2b mutant called amylose-extender (ae) mutant line, EM10, were compared with those of its WT cultivar, Kinmaze, of japonica rice. The results strongly suggest that BEIIb specifically formed new short chains whose branch points were localized in the basal part of the crystalline lamellae and presumably in the intermediate between the crystalline and amorphous lamellae of amylopectin clusters in the WT endosperm, whereas in its absence branch points which were mainly formed by BEI were only located in the amorphous lamellae of amylopectin. These differences in the cluster structure of amylopectin between Kinmaze and EM10 endosperm were considered to be responsible for the differences in the A-type and B-type crystalline structures of starch granules between Kinmaze and EM10, respectively. The changes in internal structure of starch granules caused by BEIIb were analyzed by wide angle X-ray diffraction, small-angle X-ray scattering, solid state 13C NMR, and optical sum frequency generation spectroscopy. It was noted that the size the amylopectin cluster in ae endosperm (approximately 8.24 nm) was significantly smaller than that in WT endosperm (approximately 8.81 nm). Based on the present results, we proposed a model for the cluster structure of amylopectin in WT and ae mutant of rice endosperm. We also hypothesized the role of BEIIa in amylopectin biosynthesis in culm where BEIIb was not expressed and instead BEIIa was the major BE component in WT of rice.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Accumulating evidence has shown that sarcopenia in patients with oral squamous cell carcinoma (OSCC) is at a risk of poor prognosis. There is no universal consensus on how to assess sarcopenia in patients with OSCC in daily practice. It is important to validate the usefulness of sarcopenia assessment from cervical muscles, which are frequently used in routine clinical practice in patients with OSCC. In this study, we investigated whether preoperative lumbar (L3) skeletal muscle mass and adiposity in OSCC patients were associated with cervical (C3) skeletal muscle mass and adiposity from CT measurements. We also investigated whether skeletal muscle mass and adiposity in the C3 muscles were associated with survival rates in patients with OSCC. We demonstrated that both the quality and quantity of muscle between the C3 and L3 levels were positively correlated with each other. We also demonstrated that the survival rates in patients with low sternocleidomastoid muscle mass index, high processus spinosus muscle-intramuscular adipose tissue content, and the combination of both were significantly lower than those in the controls. These results suggest that the assessment of sarcopenia from multiple neck muscles by preoperative CT measurements may be useful in predicting the prognosis of patients with OSCC.

Three starch branching enzyme (BE) isozymes, BEI, BEIIa, and BEIIb, are involved in starch biosynthesis in rice endosperm. Past in vivo and in vitro studies have suggested that each BE isozyme plays a distinct role in forming the fine structure of amylopectin. To elucidate more details of their roles, we prepared DNA constructs in which all the possible combinations of the expressions of these three isozymes were suppressed in developing rice endosperm. Analysis of the chain-length distributions of amylopectin produced under these various conditions confirmed the contributions of the individual BE isozymes to the fine structure of amylopectin in rice endosperm. Among these isozymes, the impact of loss of BEIIb activity on amylopectin fine structure was most remarkable and indicated that it plays a specific role in the synthesis of short chains with a 6–13 degree of polymerization (DP). The contribution of BEI to the amylopectin synthesis was unclear when only BEI activity was reduced. It was clear, however, when both BEI and BEIIb activities were substantially inhibited. The DP11-22 intermediate chains were markedly reduced in the ΔBEI/BEIIb line compared with the ΔBEIIb line, indicating that BEI plays a distinct role in the synthesis of these intermediate chains. Although no substantial change in amylopectin chain profile was detected in the ΔBEIIa line, the role of BEIIa could be deciphered by analyzing amylopectin fine structure from the ΔBEI/BEIIa/BEIIb line in comparison to that from ΔBEI/BEIIb line. This strongly suggests that BEIIa compensates for the role of BEI, rather than that of BEIIb, by forming intermediate chains of DP11-22. In addition, the new possibility that BEIIa is involved in the formation of starch granules in rice endosperm was suggested because the onset temperature for gelatinization of starch granules in the ΔBEIIa/BEIIb line was significantly higher than that in the ΔBEIIb line. In summary, the present study highlights the distinct roles of BEI, BEIIa, and BEIIb in the synthesis of amylopectin in developing rice endosperm.

Yeast mannan (YM) is an indigestible water-soluble polysaccharide of the yeast cell wall. In vitro fecal fermentation studies showed that YM could exhibit a notable prebiotic effect. The aim of this randomized, double-blind, placebo-controlled study was to assess the efficacy of YM intake on the intestinal environment and skin condition. One hundred and ten healthy female subjects aged 30–49 years were supplemented with YM or placebo for eight weeks. Skin dryness was set as the primary endpoint. No side effects were observed during the study. Microbiota analyses revealed that YM intake selectively increased the relative abundance of Bacteroides thetaiotaomicron and Bacteroides ovatus compared to that by placebo. Feces and urine analyses showed that YM intake lowered the concentration of fecal p-cresol, indole, and skatole, and elevated urinal equol levels compared to those in placebo. Furthermore, YM supplementation ameliorated subjective skin dryness. This study suggests that YM intake could promote beneficial Bacteroides and improve the intestinal environment and skin condition.

The impact of preoperative malnutrition and sarcopenia on survival in oral squamous cell carcinoma (OSCC) patients remains controversial. We investigated the effects of the preoperative nutritional status and abnormalities in body composition on the mortality of OSCC patients. A retrospective study involving 103 patients with OSCC was conducted. Disease-specific survival (DSS) according to the preoperative psoas muscle mass index (PMI) and intramuscular adipose tissue content (IMAC) was evaluated. Univariate and multivariate analyses were performed to determine the predictive performance of the covariates with respect to DSS. The DSS rate in patients with high IMAC and low PMI was significantly lower than that in controls. Multivariate analysis revealed that a low preoperative Prognostic Nutritional Index (PNI) and high IMAC were independent risk factors. We demonstrated that preoperative malnutrition and abnormal body composition, such as preoperative skeletal muscle quality, are associated with DSS in OSCC patients. Our study suggests that the evaluation of preoperative malnutrition and skeletal muscle quality would be useful for predicting mortality in patients with OSCC.

Despite the fact that gut microbiota is closely associated with obesity, few studies have focused on the influences of paraprobiotics as food ingredients on both obesity prevention and the gut microbial community. In this study, we evaluated the effects of fragmented Lactobacillus amylovorus CP1563 (CP1563) as a paraprobiotic for obesity prevention and investigated its effects on the gut microbial community in pre-obese subjects. One hundred sixty-nine healthy subjects with a body mass index from 25.0 to 29.9 kg/m2 ingested beverages with or without the fragmented CP1563 containing 10-hydroxyoctadecanoic acid (10-HOA) for 12 weeks. The changes in abdominal, total, visceral, and subcutaneous fatty areas were significantly lower in the CP1563-10-HOA group than in the placebo group at 12 weeks. Furthermore, 16S rRNA gene sequencing of fecal DNA revealed that the changes in the abundances of the genera Roseburia and Lachnospiraceae;g were significantly greater in the CP1563-10-HOA group than in the placebo group, and the changes in the abundances of the genus Collinsella was significantly smaller in the CP1563-10HOA group than in the placebo group. Our results showed that continuous ingestion of the fragmented CP1563 containing 10-HOA reduced abdominal body fat and affected the gut microbial community in pre-obese healthy subjects. Our findings may contribute to the understanding of the relationship between the anti-obesity effect of paraprobiotics and gut microbiota.

The authors previously confirmed the serum uric acid-lowering effects of the combination of glycine and tryptophan in subjects with mild hyperuricemia. This study examined whether combined supplementation with glycine and tryptophan suppressed the elevation in serum uric acid levels caused by purine ingestion and accelerated urinary uric acid excretion in subjects with lower urate excretion using a randomized, single-blind, placebo-controlled, crossover clinical trial design. Healthy Japanese adult males with lower urate excretion ingested water containing purines in addition to dextrin (placebo), tryptophan, glycine, or a glycine and tryptophan mixture. The combined supplementation with glycine and tryptophan significantly reduced the elevated serum uric acid levels after purine ingestion. Glycine alone and in combination with tryptophan significantly increased urinary uric acid excretion and urate clearance compared with the effects of the placebo. Urinary pH increased by the ingestion of the mixture. These results suggested that the improved water solubility of uric acid due to increased urinary pH contributed to the increase of urinary uric acid excretion.

We determined the serum uric acid-lowering effects of combined daily supplementation of glycine and tryptophan in patients with mild hyperuricemia using a randomized, double-blind, placebo-controlled, crossover clinical trial design. Japanese healthy adult males and females with mild hyperuricemia (fasting serum uric acid of 6.6–7.9 mg/dL) ingested a powder mixture containing 3.0 g of glycine and 0.2 g of tryptophan or a placebo powder once daily at bedtime for 6 weeks. Combined supplementation with glycine and tryptophan significantly decreased serum uric acid levels (from 7.1 mg/dL to 6.7 mg/dL, p = 0.004) before and after the trial. Serum uric acid concentrations significantly decreased in the subjects supplemented with the amino acid mixture compared with those in placebo-treated subjects (p = 0.028). In addition, the combination treatment with glycine and tryptophan decreased serum triglyceride levels (from 119 mg/dL to 86 mg/dL, p = 0.002). Increased solubility of uric acid caused by urinary pH were likely contributors to the serum uric acid-lowering effects of the amino acid mixture.

It has been believed that isoamylase (ISA)-type α-glucan debranching enzymes (DBEs) play crucial roles not only in α-glucan degradation but also in the biosynthesis by affecting the structure of glucans, although molecular basis on distinct roles of the individual DBEs has not fully understood. In an attempt to relate the roles of DBEs to their chain-length specificities, we analyzed the chain-length distribution of DBE enzymatic reaction products by using purified DBEs from various sources including rice, cyanobacteria, and bacteria. When DBEs were incubated with phytoglycogen, their chain-length specificities were divided into three groups. First, rice endosperm ISA3 (OsISA3) and Eschericia coli GlgX (EcoGlgX) almost exclusively debranched chains having degree of polymerization (DP) of 3 and 4. Second, OsISA1, Pseudomonas amyloderamosa ISA (PsaISA), and rice pullulanase (OsPUL) could debranch a wide range of chains of DP≧3. Third, both cyanobacteria ISAs, Cyanothece ATCC 51142 ISA (CytISA) and Synechococcus elongatus PCC7942 ISA (ScoISA), showed the intermediate chain-length preference, because they removed chains of mainly DP3-4 and DP3-6, respectively, while they could also react to chains of DP5-10 and 7-13 to some extent, respectively. In contrast, all these ISAs were reactive to various chains when incubated with amylopectin. In addition to a great variation in chain-length preferences among various ISAs, their activities greatly differed depending on a variety of glucans. Most strikingly, cyannobacteria ISAs could attack branch points of pullulan to a lesser extent although no such activity was found in OsISA1, OsISA3, EcoGlgX, and PsaISA. Thus, the present study shows the high possibility that varied chain-length specificities of ISA-type DBEs among sources and isozymes are responsible for their distinct functions in glucan metabolism.

The structural changes in the plastid-amyloplast system in the parenchyma cells of sweet potato tuberous roots during thickening were examined by electron microscopy. In the tuberous roots, proplastids and plastids that contain starch granules propagated in young parenchyma cells adjacent to the meristem, but amyloplasts did not in parenchyma cells. It was suggested that the number of amyloplasts in a parenchyma cell is determined by the propagation of the proplastids and plastids. The form of amyloplasts and the number, size and form of starch granules in them were various. Tubular membranes containing the electron-dense substance were formed in plastids and extended from the envelope membranes of plastids to the starch granules. The electron-dense substance also existed around the starch granules. Tubular membranes are converted into membrane-bound inclusion bodies as a result of loading of the electron-dense substance into these tubes in the plastids. The inclusion bodies were also at the periphery of the amyloplasts. In this study, the functions of both tubular membrane and the inclusion were discussed. The localization of starch branching enzyme in tuberous roots was examined by immunogold electron microscopy. The label for branching enzyme was localized predominantly throughout the surface of each starch granule, suggesting that this is the branching for amylopectin synthesis, but not throughout the stroma in the plastid-amyloplast system. Small and round starch granules were often formed at parts of the periphery in the amyloplast. Dense labeling for the enzyme was detected around the granules. The increase of the number of starch granules in an amyloplast is certainly made by means of the formation of new starch granules at the periphery of the amyloplast. It is likely that the new granules are intensively formed there.

Staphylococcus aureus uses two-component systems (TCSs) to adapt to stressful environmental conditions. To colonize a host, S. aureus must resist bacteriocins produced by commensal bacteria. In a comprehensive analysis using individual TCS inactivation mutants, the inactivation of two TCSs, graRS and braRS, significantly increased the susceptibility to the class I bacteriocins, nukacin ISK-1 and nisin A, and inactivation of vraSR slightly increased the susceptibility to nukacin ISK-1. In addition, two ABC transporters (BraAB and VraDE) regulated by BraRS and one transporter (VraFG) regulated by GraRS were associated with resistance to nukacin ISK-1 and nisin A. We investigated the role of these three TCSs of S. aureus in co-culture with S. warneri, which produces nukacin ISK-1, and Lactococcus lactis, which produces nisin A. When co-cultured with S. warneri or L. lactis, the braRS mutant showed a significant decrease in its population compared with the wild-type, whereas the graRS and vraSR mutants showed slight decreases. Expression of vraDE was elevated significantly in S. aureus co-cultured with nisin A/nukacin ISK-1-producing strains. These results suggest that three distinct TCSs are involved in the resistance to nisin A and nukacin ISK-1. Additionally, braRS and its related transporters played a central role in S. aureus survival in co-culture with the strains producing nisin A and nukacin ISK-1.

Introduction: The most effective method of assessing sarcopenia has yet to be determined, whether by single muscle or by whole muscle segmentation. The purpose of this study was to compare the prognostic value of these two methods using computed tomography (CT) images in patients with oral squamous cell carcinoma (OSCC). Materials and methods: Sex- and age-adjusted Cox proportional hazards models were employed for each parameter of sarcopenia related to overall survival, disease-free survival, and disease-specific survival. Harrell’s concordance index was calculated for each model to assess discriminatory power. Results: In this study including 165 patients, a significant correlation was found between the CT-based assessment of individual muscles and their crosssectional area. Single muscle assessments showed slightly higher discriminatory power in survival outcomes compared to whole muscle assessments, but the difference was not statistically significant, as indicated by overlapping confidence intervals for the C-index between assessments. To further validate our measurements, we classified patients into two groups based on intramuscular adipose tissue content (P-IMAC) of the spinous process muscle. Analysis showed that the higher the P-IMAC value, the poorer the survival outcome. Conclusion: Our findings indicate a slight advantage of single-muscle over whole-muscle assessment in prognostic evaluation, but the difference between the two methods is not conclusive. Both assessment methods provide valuable prognostic information for patients with OSCC, and further studies involving larger, independent cohorts are needed to clarify the potential advantage of one method over the other in the prognostic assessment of sarcopenia in OSCC. [ABSTRACT FROM AUTHOR]

This study examined the achievements and issues in online case development and online practice in elderly care facilities, which were implemented as alternative measures during COVID-19, in addition to regular hospital care. When said achievements were assessed using the gerontological nursing core curriculum scale, hospital practice, which included patient interaction, scored higher than online practice in “risk assessment of health disorders” and “comprehensive assessment of elderly people.” However, the online case development practice, where nurses could work at their own pace, scored as high as hospital practice in the “exploration of nursing to allow an individual to live true to themselves.” Furthermore, when the advantages and disadvantages of each form of practice were examined using quantitative text analysis, online practice caused anxiety due to the inability to grasp the progress of others, as well as difficulties in discussions. These factors hinder active learning, such as group work, which is necessary for team medicine and multidisciplinary cooperation in integrated community care. [ABSTRACT FROM AUTHOR]

Regarding a PTFE (polytetrafluoroethylene) nozzle ablation due to arc radiation exposure in SF6 circuit-breaker, the ablated mass of the nozzle was formulated as a function of an arc current and an internal radius of the nozzle surface considering the radiation power absorbed on the surface. Furthermore, this formulation contributed to subsequent development of calculation method for evaluating gas pressure rise process during current interruption. And, verification experiments of the calculationmethodwere performedwith a model SF6 circuitbreaker equipped a puffer chamber. Calculation results derived from the developed calculation method showed good agreement with measured gas pressure rise process. On the basis of the verified results, a discussion wasmade to explain pressure rise mechanism. [ABSTRACT FROM AUTHOR]

Background: The mixture of glycine and tryptophan exhibited serum uric acid-lowering effects in our previous clinical trial. Objective: Using a randomized, double-blind, placebocontrolled, and parallel study design, this current study aimed to examine whether this mixture enhanced the estimated glomerular filtration rate (eGFR) as an indicator of renal function in healthy individuals. Methods: Healthy Japanese adult males and females ingested a powder mixture containing 3.0 g of glycine and 0.2 g of tryptophan or a placebo powder once daily at bedtime for 8 weeks. Results: After 8 weeks of continual ingestion, the combined glycine and tryptophan supplementation significantly enhanced eGFR. It also decreased serum uric acid levels, consistent with our previous reports. Meanwhile, the continual ingestion of the mixture had no influence on serum total or essential amino acids. Conclusions: The current study demonstrated that the combined oral administration of glycine and tryptophan. significantly elevated the eGFR of healthy participants. However, further investigation is required to elucidate the detailed mechanisms underlying the potential therapeutic or preventive effect of combined glycine and tryptophan supplementation. Nevertheless, the uric acid-lowering effect of glycine and tryptophan mixture has the potential to directly influence renal function. [ABSTRACT FROM AUTHOR]

Alzheimer's disease is the most common form of dementia and is characterized by neuritic plaques in the brain that contain amyloid β (Aβ) peptide. Deposition of Aβ plaques affects synaptic function and is associated with learning and memory impairment. The aim of this study was to evaluate the effect of lactononadecapeptide (LNDP; NIPPLTQTPVWPPFLQPE), derived from β-casein, on an Aβ25-35-induced Alzheimer's disease model and to investigate the mechanism of action of LNDP. We investigated the effect of LNDP on memory impairment induced by an intracerebroventricular injection of Aβ25-35 and neurotrophic gene expression in ddY mice. LNDP (0.01, 0.1 and 1 mg/kg b.w.) was orally administered once daily from 7 days before to 7 days after the Aβ25-35 injection, and the Y-maze test was performed 7 days after the Aβ25-35 injection (n =: 15 per group). We subsequently examined the effect of LNDP on the gene expression of brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF) and neurotrophin-3 (NT-3) in the hippocampus using real-time polymerase chain reaction. Intracerebroventricular injection of Aβ25-35 induced memory impairment in the mice, whereas oral administration of LNDP improved memory impairment without significantly affecting the locomotor response. We also demonstrated that oral administration of LNDP increased BDNF, NGF and NT-3 mRNA expression in the hippocampus. These results suggest that LNDP may be useful in the prevention or treatment of neurodegenerative disorders such as Alzheimer's disease and may be linked to BDNF, NGF and NT-3 expression in the hippocampus. [ABSTRACT FROM AUTHOR]

During the last couple of decades, rapid progress has been made in increasing our understanding of the specific contributions of the most important enzymes to starch biosynthesis in cereal endosperms, particularly in rice endosperm. Now, we can basically predict how and to what extent the starch-related phenotypes can be modified in the endosperm in accordance with alterations of activities of single or simultaneously multiple starch biosynthetic isozymes. Thus, starch bioengineering in planta is now a most particularly promising field of plant biotechnology. Here, an overview of the present status of biochemical, molecular, and genetic research is presented, regarding starch biosynthesis and its engineering for the generation of new cultivars with improved cooking or novel functional properties of starch in rice endosperm. A huge number and variety of cultivars have been developed and cultivated mainly in paddy fields of Asian countries because rice is the most important staple food grain there. The availability of such invaluable genetic resources, as well as databases for various types of mutants, complete genome sequences, and omics tools should ensure further progress in the rice starch biotechnology research field. [ABSTRACT FROM AUTHOR]

Starch granule morphology differs markedly among plant species. However, the mechanisms controlling starch granule morphology have not been elucidated. Rice (Oryza sativa) endosperm produces characteristic compound-type granules containing dozens of polyhedral starch granules within an amyloplast. Some other cereal species produce simple-type granules, in which only one starch granule is present per amyloplast. A double mutant rice deficient in the starch synthase (SS) genes SSIIIa and SSIVb (ss3a ss4b) produced spherical starch granules, whereas the parental single mutants produced polyhedral starch granules similar to the wild type. The ss3a ss4b amyloplasts contained compound-type starch granules during early developmental stages, and spherical granules were separated from each other during subsequent amyloplast development and seed dehydration. Analysis of glucan chain length distribution identified overlapping roles for SSIIIa and SSIVb in amylopectin chain synthesis, with a degree of polymerization of 42 or greater. Confocal fluorescence microscopy and immunoelectron microscopy of wild-type developing rice seeds revealed that the majority of SSIVb was localized between starch granules. Therefore, we propose that SSIIIa and SSIVb have crucial roles in determining starch granule morphology and in maintaining the amyloplast envelope structure. We present a model of spherical starch granule production. [ABSTRACT FROM AUTHOR]

Amylopectin is a highly branched, organized cluster of glucose polymers, and the major component of rice starch. Synthesis of amylopectin requires fine co-ordination between elongation of glucose polymers by soluble starch synthases (SSs), generation of branches by branching enzymes (BEs), and removal of misplaced branches by debranching enzymes (DBEs). Among the various isozymes having a role in amylopectin biosynthesis, limited numbers of SS and BE isozymes have been demonstrated to interact via protein-protein interactions in maize and wheat amyloplasts. This study investigated whether protein-protein interactions are also found in rice endosperm, as well as exploring differences between species. Gel permeation chromatography of developing rice endosperm extracts revealed that all 10 starch biosynthetic enzymes analysed were present at larger molecular weights than their respective monomeric sizes. SSIIa, SSIIIa, SSIVb, BEI, BEIIb, and PUL co-eluted at mass sizes >700 kDa, and SSI, SSIIa, BEIIb, ISA1, PUL, and Pho1 co-eluted at 200-400 kDa. Zymogram analyses showed that SSI, SSIIIa, BEI, BEIIa, BEIIb, ISA1, PUL, and Pho1 eluted in high molecular weight fractions were active. Comprehensive co-immunoprecipitation analyses revealed associations of SSs-BEs, and, among BE isozymes, BEIIa-Pho1, and pullulanase-type DBE-BEI interactions. Blue-native-PAGE zymogram analyses confirmed the glucan-synthesizing activity of protein complexes. These results suggest that some rice starch biosynthetic isozymes are physically associated with each other and form active protein complexes. Detailed analyses of these complexes will shed light on the mechanisms controlling the unique branch and cluster structure of amylopectin, and the physicochemical properties of starch. [ABSTRACT FROM AUTHOR]

In a previous study, we reported that the common reed accumulates water-soluble Cd complexed with an a-glucan-like molecule, and that the synthesis of this molecule is induced in the stem of the common reed under Cd stress. We studied the metabolic background to ensure a-glucan accumulation under the Cd stress conditions that generally inhibit photosynthesis. We found that the common reed maintained an adequate CO2 assimilation rate, tended to allocate more assimilated 11C to the stem, and accumulated starch granules in its stem under Cd stress conditions. AGPase activity, which is the rate-limiting enzyme for starch synthesis, increased in the stem of common reed grown in the presence of Cd. Starch accumulation in the stem of common reed was not obvious under other excess metal conditions. Common reed may preferentially allocate assimilated carbon as the carbon source for the formation of Cd and a-glucan complexes in its stem followed by prevention of Cd transfer to leaves acting as the photosynthetic organ. These responses may allow the common reed to grow even under severe Cd stress conditions. [ABSTRACT FROM AUTHOR]

Background: Starch is the most important carbohydrate in plant storage tissues. Multiple isozymes in at least four enzyme classes are involved in starch biosynthesis. Some of these isozymes are thought to interact and form complexes for efficient starch biosynthesis. Of these enzyme classes, starch synthases (SSs) and branching enzymes (BEs) play particularly central roles. Results: We generated double mutant lines (ss1/be1 and ss1L/be2b) between SSI (the largest component of total soluble SS activity) and BEI or BEIIb (major BEs in developing rice endosperm) to explore the relationships among these isozymes. The seed weight of ss1/be1 was comparable to that of wild type, although most ss1/be2b seeds were sterile and no double recessive plants were obtained. The seed weight of the double recessive mutant line ss1L/be2b, derived from the leaky ss1 mutant (ss1L) and be2b, was higher than that of the single be2b mutant. Analyses of the chain-length distribution of amylopectin in ss1/be1 endosperm revealed additive effects of SSI and BEI on amylopectin structure. Chain-length analysis indicated that the BEIIb deficiency significantly reduced the ratio of short chains in amylopectin of ss1L/be2b. The amylose content of endosperm starch of ss1/be1 and ss1L/be2b was almost the same as that of wild type, whereas the endosperm starch of be2b contained more amylose than did that of wild type. SSI, BEI, and BEIIb deficiency also affected the extent of binding of other isozymes to starch granules. Conclusions: Analysis of the chain-length distribution in amylopectin of the double mutant lines showed that SSI and BEI or BEIIb primarily function independently, and branching by BEIIb is followed by SSI chain elongation. The increased amylose content in be2b was because of reduced amylopectin biosynthesis; however, the lower SSI activity in this background may have enhanced amylopectin biosynthesis as a result of a correction of imbalance between the branching and elongation found in the single mutant. The fact that a deficiency of SSI, BEI, or BEIIb affected the affinity of other starch biosynthetic isozymes for the starch granule implies that there is a close interaction among SSI, BEI and BEIIb during amylopectin biosynthesis in rice endosperm. [ABSTRACT FROM AUTHOR]

[Purpose] We investigated the effect on feelings of burden of care staff collaboratively working with physical therapists (PTs). [Subjects] The subjects were 28 care staff. [Method] We conducted a 13-item questionnaire survey, using the Japanese version of the Zarit Burden Interview with the items related to family removed, and compared the results between the group of care staff collaborating with PTs, and the group of caregivers who were not. [Results] The Zarit score of the group of care staff not collaborating with PTs was higher than that of the group of care staff collaborating with PTs. Furthermore, from within caregivers' burden, we extracted factors of service responses, service trends, and care staff feelings of burden. [Conclusion] To reduce the caregivers' burden of care staff, it is important to prevent the decline of the discretionary power of care staff through the establishment of team-style work performance. Furthermore, when PTs collaborate with care staff, promotion of the standardization of assistance methods, based on evaluation of vital functions and motor learning theory, is a useful approach. [ABSTRACT FROM AUTHOR]

The stems (leaf sheaths and culms) of rice plants (Oryza sativa L.) accumulate high levels of starch before heading, which is subsequently remobilized after heading to provide a carbon source for grain filling. To elucidate the molecular mechanism of starch synthesis in rice leaf sheath, a comprehensive expression analysis was conducted on the gene families encoding starch synthesis–related enzymes, ADP-glucose pyrophosphorylase (EC 2.7.7.27), starch synthase (EC 2.4.1.21) and branching enzyme (EC 2.4.1.18). The changes in the activities of these enzymes in leaf sheaths during the heading period were shown to be accompanied by a coordinated change in the transcript levels of particular members of the corresponding gene families. A similar change before and after heading was also found in the messenger RNA level of GPT2, one of the two genes encoding for a plastidial glucose-6-phosphate/phosphate translocator, suggesting that the capacity of both starch synthesis and its substrate import is under coordinated transcriptional regulation. The members of the gene families predominantly expressed in leaf sheaths are mostly different from those actively expressed in the developing endosperm. The time course for the transcript level of some of these genes appears to correlate to hexose level in the leaf sheaths, suggesting that hexose levels may be a controlling factor in their expression. [ABSTRACT FROM AUTHOR]

In maturing endosperms of a variety of sugary mutants of rice, phytoglycogen-like polysaccharides with highly branched α-glucans were accumulated instead of amylopectin, while the amylose content greatly decreased. Measurement of activities per endosperm of the 10 major enzymes involved in starch and sucrose metabolism revealed that the activity of starch debranching enzyme (R-enzyme) was specifically reduced in the sugary mutants. The activity of starch branching enzyme I (Q-enzyme 1) was also significantly decreased, but less so than the R-enzyme, in the mutants, suggesting some coordination of the expression of the genes coding for R-enzyme and Q-enzyme I. Western blot analysis showed that the sugary mutations of rice resulted in a decrease in the amount of R-enzyme protein, but not in major modification of the enzyme. The findings strongly suggest that R-enzyme plays a critical role in determining the amylopectin fine structure, since at the extremely low level of R-enzyme activity as compared with Q-enzyme activity, as found in sugary mutants, the rice endosperm produced phytoglycogen. We hypothesize that balance of activities or interaction between Q-enzyme and R-enzyme may be responsible for the fine structure of α-polyglucans in plant tissues. [ABSTRACT FROM AUTHOR]

The pattern of isoforms of starch branching enzyme II or Q‐enzyme II in the tuberous root of sweet potato was distinct from those of other organs; altogether 7 isoforms of QEII were contained in the sweet potato plant. The QEIIf isoform, one of the two major QEII isoforms in the tuberous root, was purified to homogeneity by using a variety of HPLC columns. The purified QEIIf was a monomeric protein with a molecular mass of about 85 kDa. Western blot analysis showed that the polyclonal antibodies raised against the purified QEIIf was significantly reactive to the rice endosperm QEI, but not to the rice endosperm QEIIa. Furthermore, the sweet potato QEIIf reacted with the antiserum raised against the rice endosperm QEI, but not with that against the rice endosperm QEIIa. The results suggest that the sweet potato QEIIf is more similar to the rice endosperm QEI than to the rice endosperm QEIIa. [ABSTRACT FROM AUTHOR]

ADP glucose pyrophosphorylase from developing rice (Oryza saliva) endosperm was purified. The final preparation yielded 6 major protein spots as separated by two-dimensional polyacrylamide electrophoresis. All 6 polypeptides had similar molecular weights of ca 50 kDa and cross-reacted with polyclonal antibodies raised against two main protein bands among them. The results suggest that the rice endosperm ADPglucose pyrophorsphorylase is tetrameric and composed of multiple subunits with similar amino acid structure. [ABSTRACT FROM AUTHOR]

Two isoforms of starch branching enzyme (Q-enzyme), QEI and QEII, have been purified to homogeneity from developing rice endosperm. QEI and QEII, with molecular weights of about 80 and 85 kDa, respectively, could be fully separated by anion-exchange or hydrophobic chromatography. The peptide maps obtained after V8 proteinase digestion were quite different between the two enzymes. Antibodies prepared against QEI showed no immunological cross-reaction with the QEII protein in Western blot experiments, and anti-QEII serum did not react with the QEI protein. The data indicate that QEI and QEII are distinct proteins encoded by different genes in rice plants. [ABSTRACT FROM AUTHOR]

In monocots, starch branching enzyme II (BEII) was functionally differentiated into BEIIa and BEIIb after separation from the dicots, and in cereals BEIIb plays a distinct role in amylopectin biosynthesis in the endosperm. The present study was conducted to examine to what extent a green algal BEII has an overlapping function with BEIIb in starch biosynthesis by introducing the Chlorella BEII gene into an amylose-extender (ae) mutant of rice. Chlorella BEII was found to complement the contribution of the rice endosperm BEIIb to the structures of amylopectin and starch granules because these mutated phenotypes were recovered almost completely to those of the wild type by the expression of Chlorella BEII. When the recombinant BE enzymes were incubated with the rice ae amylopectin, the branching pattern of Chlorella BEII was much more similar to that of rice BEIIb rather than rice BEIIa. Detailed analyses of BE reaction products suggests that BEIIb and Chlorella BEII only transfer chains with a degree of polymerization (DP) of 6 and 7, whereas BEIIa preferably transfers short chains with a DP of about 6–11. These results show that the Chlorella BEII is functionally similar to rice BEIIb rather than BEIIa. [ABSTRACT FROM AUTHOR]

Storage glucans were analyzed in the Porphyridiales which include the most primitive and phylogenetically diverged species in the Rhodophyta, to understand early evolution of the glucan structure in the Rhodophyta. The storage glucans of both Galdieria sulphuraria and Cyanidium caldarium consisted of glycogen, while those of Rhodosorus marinus, Porphyridium purpureum, P. sordidum and Rhodella violacea could be defined as semi-amylopectin. X-ray diffraction analysis of the glucans demonstrated variation in the crystalline structure: the patterns in P. purpureum and R. violacea were of A- and B-types, respectively, while α-glucans of R. marinus and P. sordidum displayed structures with lower crystallinity. Electron microscopic observations indicated that the α-glucans of P. sordidum consisted of two kinds of granules; a minor component of more dense granules with crystalline leaflets and a major component of softer ones without crystalline structure. Gel permeation chromatography showed that all the species containing the semi-amylopectin-type glucans also contained amylose, although the relative amounts of this fraction were different depending on the species. Our results are consistent with two distinct evolution scenarios defined either by the independent acquisition of semi-crystalline starch-like structures in the different plant lineages or more probably by the loss of starch and reversion to glycogen synthesis in cyanidian algae growing in hot and acid environments. [ABSTRACT FROM AUTHOR]