Two-dimensional water-molecule-cluster layers at nanobubble interfaces.

HYPOTHESIS: Bulk nanobubbles (NBs) have high surface charge densities and long lifetimes. Despite several attempts to understand the lifetime of NBs, their interfacial layer structure remains unknown. It is hypothesized that a specific interfacial layer exists with a hydrogen bond network that stabilizes NBs. EXPERIMENTS: In situ infrared reflectance-absorption spectroscopy and density functional theory were used to determine the interfacial layer structure of NBs. Furthermore, nuclear magnetic resonance spectroscopy was used to examine the interfacial layer hardness of bubbles filled with N2, O2, and CO2, which was expected to depend on the encapsulated gas species. FINDINGS: The interfacial layer was composed of three-, four-, and five-membered ring clusters of water molecules. An interface model was proposed in which a two-dimensional layer of clusters with large electric dipole moments is oriented toward the endohedral gas, and the hydrophobic surface is adjacent to the free water. The interfacial layer hardness was dependent on the interaction with the gas (N2 > O2 > CO2), which supports the proposed interface model. These findings can be generalized to the structure of water at gas-water interfaces.

Characterization of Bulk Nanobubbles Formed by Using a Porous Alumina Film with Ordered Nanopores.

Nanobubbles (NBs), with their unique physicochemical properties and promising applications, have become an important research topic. Generation of monodispersed bulk NBs with specified gas content remains a challenge. We developed a simple method for generating bulk NBs, using porous alumina films with ordered straight nanoscaled holes. Different techniques, such as nanoparticle tracking analysis (NTA), atomic force microscopy (AFM), and infrared absorption spectroscopy (IRAS), are used to confirm NB formation. The NTA data demonstrate that the minimum size of the NBs formed is less than 100 nm, which is comparable to the diameter of nanoholes in the porous alumina film. By generating NBs with different gases, including CO2, O2, N2, Ar, and He, we discovered that the minimum size of NBs negatively correlated with the solubility of encapsulated gases in water. Due to the monodispersed size of NBs generated from the highly ordered porous alumina, we determined that NB size is distributed discretely with a uniform increment factor of [Formula: see text]. To explain the observed characteristic size distribution of NBs, we propose a simple model in which two NBs of the same size are assumed to preferentially coalesce. This characteristic bubble size distribution is useful for elucidating the basic characteristics of nanobubbles, such as the long-term stability of NBs. This distribution can also be used to develop new applications of NBs, for example, nanoscaled reaction fields through bubble coalescence.

Host Cell Prediction of Exosomes Using Morphological Features on Solid Surfaces Analyzed by Machine Learning.

Exosomes are extracellular nanovesicles released from any cells and found in any body fluid. Because exosomes exhibit information of their host cells (secreting cells), their analysis is expected to be a powerful tool for early diagnosis of cancers. To predict the host cells, we extracted multidimensional feature data about size, shape, and deformation of exosomes immobilized on solid surfaces by atomic force microscopy (AFM). The key idea is combination of support vector machine (SVM) learning for individual exosome particles and their interpretation by principal component analysis (PCA). We observed exosomes derived from three different cancer cells on SiO2/Si, 3-aminopropyltriethoxysilane-modified-SiO2/Si, and TiO2 substrates by AFM. Then, 14-dimensional feature vectors were extracted from AFM particle data, and classifiers were trained in 14-dimensional space. The prediction accuracy for host cells of test AFM particles was examined by the cross-validation test. As a result, we obtained prediction of exosome host cells with the best accuracy of 85.2% for two-class SVM learning and 82.6% for three-class one. By PCA of the particle classifiers, we concluded that the main factors for prediction accuracy and its strong dependence on substrates are incremental decrease in the PCA-defined aspect ratio of the particles with their volume.

Daily step count and all-cause mortality in a sample of Japanese elderly people: a cohort study.

BACKGROUND: This study aimed to examine the relationship between pedometer-assessed daily step count and all-cause mortality in a sample of elderly Japanese people. METHODS: Participants included 419 (228 males and 191 females) physically independent, community-dwelling 71-year-old Japanese people. The number of steps per day was measured by a waist-mounted pedometer for seven consecutive days at baseline. Participants were divided into quartiles based on their average number of steps/day (first quartile, < 4503 steps/day; second quartile, 4503-6110 steps/day; third quartile, 6111-7971 steps/day; fourth quartile, > 7972 steps/day) and were followed up over a mean period of 9.8 years (1999-2010) for mortality. RESULTS: Seventy-six participants (18.1%) died during the follow-up period. The hazard ratios (adjusted for sex, body mass index, cigarette smoking, alcohol intake, and medication use) for mortality across the quartiles of daily step count (lowest to highest) were 1.00 (reference), 0.81 (95%CI, 0.43-1.54), 1.26 (95%CI, 0.70-2.26), and 0.46 (95%CI, 0.22-0.96) (P for trend = 0.149). Participants in the highest quartile had a significantly lower risk of death compared with participants in the lowest quartile. CONCLUSION: This study suggested that a high daily step count is associated with a lower risk of all-cause mortality in physically independent Japanese elderly people.

Fabrication of Au-Nanoparticle-Embedded Lipid Bilayer Membranes Supported on Solid Substrates.

We fabricated gold nanoparticle (Au-NP)-embedded supported lipid bilayers (SLBs) by two methods. In the vesicle-vesicle fusion method, vesicles with hydrophobized Au-NPs are ruptured and fused on SiO2/Si substrates. In the vesicle-membrane fusion method, SLBs without Au-NPs were preformed on the substrate and then vesicles with Au-NPs were fused into the preformed membranes. In the former method, Au-NP incorporation into the SLBs was observed as an increase in the membrane thickness in atomic force microscopy (AFM) images and directly observed by transmission electron microscopy. In the latter method, fusion of vesicles into the preformed membranes was confirmed by the fluorescent color change in the preformed membranes, and Au-NP incorporation was also confirmed by an increase in the membrane thickness in the AFM images. Key techniques for the successful vesicle-membrane fusion are hydrophobization of Au-NPs, approach control of vesicles by mixing the charged lipids, and destabilization of the lipid bilayers by adding lipids with a small polar headgroup.

Reconstitution of Human Ion Channels into Solvent-free Lipid Bilayers Enhanced by Centrifugal Forces.

Artificially formed bilayer lipid membranes (BLMs) provide well-defined systems for functional analyses of various membrane proteins, including ion channels. However, difficulties associated with the integration of membrane proteins into BLMs limit the experimental efficiency and usefulness of such BLM reconstitution systems. Here, we report on the use of centrifugation to more efficiently reconstitute human ion channels in solvent-free BLMs. The method improves the probability of membrane fusion. Membrane vesicles containing the human ether-a-go-go-related gene (hERG) channel, the human cardiac sodium channel (Nav1.5), and the human GABAA receptor (GABAAR) channel were formed, and the functional reconstitution of the channels into BLMs via vesicle fusion was investigated. Ion channel currents were recorded in 67% of the BLMs that were centrifuged with membrane vesicles under appropriate centrifugal conditions (14-55 × g). The characteristic channel properties were retained for hERG, Nav1.5, and GABAAR channels after centrifugal incorporation into the BLMs. A comparison of the centrifugal force with reported values for the fusion force revealed that a centrifugal enhancement in vesicle fusion was attained, not by accelerating the fusion process but by accelerating the delivery of membrane vesicles to the surface of the BLMs, which led to an increase in the number of membrane vesicles that were available for fusion. Our method for enhancing the probability of vesicle fusion promises to dramatically increase the experimental efficiency of BLM reconstitution systems, leading to the realization of a BLM-based, high-throughput platform for functional assays of various membrane proteins.

CD56(dim)CD16(high) and CD56(bright)CD16(-) cell percentages associated with maximum knee extensor strength and incidence of death in elderly.

Physical fitness is an indicator of systemic well-being in humans. Little is known about the role of physical fitness for maintaining systemic health in the elderly. Here, we study elderly subjects to determine the relationships between physical fitness and CD56 and CD16 surface NK cell markers on peripheral blood lymphocytes, as well as to analyze the relationship between the surface markers and incidence of death. We selected 253 independent elderly subjects (122 female; 131 male) who were 79-80 years old. Subjects having a higher proportion of CD56(dim)CD16(high) within CD56(+)CD16(+) cells, or ration of CD56(dim)CD16(high) and CD56(dim)CD16(-) cells had a significant positive correlation with maximum bilateral knee extensor strength/weight (kg) (r = 0.425; P < 0.0001 or r = 0.323; P < 0.0001). In contrast, an increased proportion of CD56(bright)CD16(-) cells within lymphocyte significantly negatively correlated with the maximum bilateral knee extensor strength/weight (kg) (r = -0.290; P = 0.0004); and these subjects had a significantly lower mortality during the 5 years following measurement of death. Therefore, we found that a synergistic effect of the right and left leg muscle strength was associated with proportion of matured NK and NKT cells and induced a low proportion of CD56(bright)CD16(-) cells within lymphocyte. Moreover, the low proportion of CD56(bright)CD16(-) cells was associated with incidence of death. In conclusion, measurements of physical fitness, the proportion of CD56(dim)CD16(high) within CD56(+)CD16(+) cells, the ratio of CD56(dim)CD56(high) and CD56(dim)CD16(-) cells, and the proportion of CD56(bright)C16(-) cells in lymphocytes are important indicators to check elderly health.

Tracking of Pedometer-Determined Physical Activity in Healthy Elderly Japanese People.

BACKGROUND: Tracking refers to the tendency for an individual to maintain their rank within a group over time. This study longitudinally investigated the 8-year tracking of pedometer-determined physical activity among physically independent elderly Japanese people aged 72 to 80 years. METHODS: Steps/day were measured when participants were aged 72 and assessed again after 2, 5, and 8 years. The number of participants with a 2-year follow-up (72 to 74 years) was 177 (101 males, 76 females), with a 5-year follow-up (72 to 77 years) was 183 people (110 males, 73 females), and with an 8-year follow-up (72 to 80 years) was 145 people (91 males, 54 females). Step counts were continuously measured for 1 week in January, April, July, and October during each year of assessment. RESULTS: A high rank correlation coefficient for steps/day exceeding 0.60 was obtained at the 2-, 5- and 8-year follow-up examinations for both males and females. CONCLUSION: This study suggests that the rank within a group of pedometer-determined steps/day remains stable over up to 8 years in healthy Japanese people aged between 72 to 80 years old.

Characterization of citrates on gold and silver nanoparticles.

In this paper, we report different coordinations of citrates on gold (AuNP) and silver (AgNP) nanoparticles, as determined using Fourier transform infrared spectroscopy (FTIR) and molecular orbital (MO) calculations. AuNPs and AgNPs are found to have completely different interactions with the carboxylate anchoring groups, as indicated by their unique asymmetric stretching vibrations in the FTIR spectra. The ν(as) (COO(-)) of citrate exhibits a high-frequency shift resulting from the formation of a unidentate coordination on AuNPs, whereas this vibration exhibits a low-frequency shift as a result of ionic bond formation on AgNPs, as predicted from the MO calculations of the corresponding metal complex salts. The enhancement in the IR signals when their vibration direction was perpendicular to the nanoparticle surface revealed the influence of localized surface plasmons excited on the metal nanoparticles.

Reconstitution of human ether-a-go-go-related gene channels in microfabricated silicon chips.

This paper reports on the reconstitution of human ether-a-go-go-related gene (hERG) channels in artificial bilayer lipid membranes (BLMs) formed in micropores fabricated in silicon chips. The hERG channels were isolated from Chinese hamster ovary cell lines expressing the channels and incorporated into the BLMs formed by a process in which the two lipid monolayers were folded into the micropores. The characteristic features of hERG channels reported by the patch-clamp method, including single-channel conductance, voltage dependence, sensitivity to typical drugs and dependence on the potassium concentration, were investigated in the BLM reconstitution system. The BLM with hERG channels incorporated exhibited a lifetime of ~65 h and a tolerance to repetitive solution exchanges. Such stable BLMs containing biological channels have the potential for use in a variety of applications, including high-throughput drug screening for various ion-channel proteins.

Interactions between serum vitamin D levels and vitamin D receptor gene FokI polymorphisms for renal function in patients with type 2 diabetes.

BACKGROUND: We aimed to examine associations among serum 25-hydroxyvitamin D (25OHD) levels, 1,25-dihyroxyvitamin D (1,25OHD) levels, vitamin D receptor (VDR) polymorphisms, and renal function based on estimated glomerular filtration rate (eGFR) in patients with type 2 diabetes. METHODS: In a cross-sectional study of 410 patients, chronic kidney disease (CKD) stage assessed by eGFR was compared with 25OHD, 1,25OHD, and VDR FokI (rs10735810) polymorphisms by an ordered logistic regression model adjusted for the following confounders: disease duration, calendar month, use of angiotensin converting enzyme inhibitors/angiotensin receptor blockers or statins, and serum calcium, phosphate, and intact parathyroid hormone levels. RESULTS: 1,25OHD levels, rather than 25OHD levels, showed seasonal oscillations; peak levels were seen from May to October and the lowest levels were seen from December to February. These findings were evident in patients with CKD stage 3 ~ 5 but not stage 1 ~ 2. eGFR was in direct proportion to both 25OHD and 1,25OHD levels (P<0.0001), but it had stronger linearity with 1,25OHD (r = 0.73) than 25OHD (r = 0.22) levels. Using multivariate analysis, 1,25OHD levels (P<0.001), but not 25OHD levels, were negatively associated with CKD stage. Although FokI polymorphisms by themselves showed no significant associations with CKD stage, a significant interaction between 1,25OHD and FokITT was observed (P = 0.008). The positive association between 1,25OHD and eGFR was steeper in FokICT and CC polymorphisms (r = 0.74) than FokITT polymorphisms (r = 0.65). CONCLUSIONS: These results suggest that higher 1,25OHD levels may be associated with better CKD stages in patients with type 2 diabetes and that this association was modified by FokI polymorphisms.

Stable lipid bilayers based on micro- and nano-fabrication as a platform for recording ion-channel activities.

In this review, we will discuss our recent approaches for the formation of mechanically stable bilayer lipid membranes (BLMs) by combining with BLM formation and micro- and nano-fabrication techniques. BLMs were prepared across a microaperture fabricated in silicon (Si) chips or nanoporous alumina films using a minimized amount of organic solvent. Although BLMs spanned over the porous alumina film showed better electrical properties, such as background current noise and current transient, BLMs suspended in a thin Si(3)N(4) septum showed a much superior BLM stability. The BLMs showed tolerance to a high voltage of ±1 V, a membrane lifetime of >40 h, and tolerance to repetitive solution exchanges. Application to a drug screening system has been examined by using the human ether-a-go-go-related gene (hERG) potassium channel as an illustrative example. The potentiality of the present system as a platform of the high-throughput analysis for ion-channel protein is also discussed.

Comparison between ultrasonography and MR imaging for discriminating squamous cell carcinoma nodes with extranodal spread in the neck.

OBJECTIVE: To compare the diagnostic ability of ultrasonography (US) and MR imaging for discriminating squamous cell carcinoma (SCC) nodes with extranodal spread (ENS) in the neck. METHODS: US and MR imaging was retrospectively evaluated for differentiating ENS-positive (n=28) from ENS-negative (n=26) SCC nodes (>10mm short-axis diameter) in 50 patients with head and neck SCCs. We assessed nodal size on US and MR images; irregular nodal margin on US; and vanishing nodal border, flare, and shaggy nodal margin signs on T1-, fat-suppressed T2-, and contrast-enhanced T1-weighted MR images, respectively. US and MR images were analyzed by 3 radiologists in consensus and the results were compared between ENS-positive and ENS-negative SCC nodes. RESULTS: The nodal sizes of ENS-positive nodes (21±9 mm) were significantly larger than those of ENS-negative SCC nodes (14±4 mm) (p<0.001). Irregular nodal margins were more frequently observed in ENS-positive SCC nodes (75%) than in ENS-negative SCC nodes (12%). The vanishing nodal margin, flare, and shaggy nodal margin signs were more frequently observed in ENS-positive SCC nodes (93%, 89%, and 82%, respectively) than in ENS-negative nodes (46%, 19%, and 19%, respectively). A combination of size (≥22 mm) and imaging criteria (irregular margin or flare sign) best discriminated ENS-positive SCC nodes with 82% sensitivity, 89% specificity, and 85% accuracy for US and 89% sensitivity, 81% specificity, and 85% accuracy for MR imaging. CONCLUSION: US discriminated ENS-positive from ENS-negative SCC nodes with comparable accuracy and higher specificity than MR imaging.

Cinacalcet for hemodialyzed patients with or without a high PTH level to control serum calcium and phosphorus: ECO (evaluation of cinacalcet HCl outcome) study.

BACKGROUND: We investigated the influence of cinacalcet on serum Ca and P in hemodialyzed patients with or without a high PTH level, according to K/DOQI guideline, to control serum Ca and P levels. METHODS: We recruited 130 patients in this prospective cohort study and classified them into Group A (iPTH > 300 pg/ml), Group B (iPTH = 181 - 300 pg/ml), and Group C (iPTH ≤ 180 pg/ml). After 24 weeks on cinacalcet, serum Ca, P and iPTH were measured. RESULTS: The achievement rate of the target iPTH level in JSDT guideline was significantly higher in Group B compared with Group A. The achievement rate of serum Ca and P target levels in the Japanese Society for Dialysis Therapy (JSDT) guideline was higher in Group C. In Group A and Group C, the simultaneous achievement rates (Ca, P, and iPTH) in KDOQI guideline increased after treatment with cinacalcet (p < 0.01). There was no difference in the reduction of Ca and P among the groups, while the iPTH reduction was significantly lower in groups B and C compared with that in A. CONCLUSION: Administration of cinacalcet to patients with or without high PTH levels, according to the K/DOQI guideline, facilitates the control of Ca and P levels.

Physical fitness and 6.5-year mortality in an 85-year-old community-dwelling population.

Although poor physical fitness is known to be associated with increased mortality in adult and elderly populations, this association is not conclusive in very elderly. The purpose of the present study was to evaluate the association for a very old community-dwelling population. The participants (90 males, 117 females) were 85-year-old individuals residing in Fukuoka, Japan. Baseline examinations including muscle strength of the handgrip and leg extension, one-leg standing, leg stepping rate, and walking were performed in 2003 and these subjects were followed for 6.5 years. During the follow-up period, 81 individuals (49 males and 32 females) died. Handgrip strength and leg extension strength at age 85 were stronger in surviving men than in non-survivors. Total mortality adjusted for both gender and serum level of total cholesterol fell 5-6% with a 1-kg increase in the handgrip strength of a single hand or both hands. Total mortality also decreased 2% with a 1 kg increase in the leg extension strength of both legs. With adjustment for gender and total cholesterol, mortality fell by 57% in participants of the walking test and fell by 45% in participants of the stepping-rate test compared to mortality in nonparticipants. No association was found between mortality and participation in the handgrip strength test, leg extension strength test, or one-leg standing time test. In conclusion, not only poor muscle strength in handgrip or leg extension, but also nonparticipation in walking test or leg-stepping test were independent predictors of total mortality in a very elderly population.

Plant growth activities of aspyran, asperentin, and its analogues produced by the fungus Aspergillus sp.

Aspyran (1), a novel compound, and the known isocoumarin asperentin (2), also known as cladosporin, together with its analogues 3-6 were isolated from Aspergillus sp. and their structures established by spectroscopic methods including 2D NMR spectroscopy. The effects of 1-6 on plant growth were examined by bioassays using lettuce and rice seedlings. Compounds 1 and 3 promoted the root growth of the seedlings, while 2 and 5 were inhibitory. Compounds 4 and 6 did not show any effect on the growth of lettuce and rice seedlings, respectively.

Home systolic blood pressure on the morning of dialysis days has prognostic impact for hypertensive hemodialysis patients.

BACKGROUND: Hypertension is a leading cause of cardiovascular (CV) disease in the general population. Although hypertension is very common in maintenance hemodialysis (HD) patients, adequate blood pressure (BP) values and measurement timing have not been defined. METHODS: A total of 49 hypertensive HD patients were recruited. Average age was 63 ± 11 years, and duration of dialysis therapy was 6.2 ± 4.2 years. Dialysis unit BPs and various types of home BPs were separately measured, and which BPs were the most critical markers in evaluating the effect of hypertension on left ventricular hypertrophy and CV events was investigated. RESULTS: Predialysis systolic BPs were not correlated with any home BPs. Left ventricular mass index (LVMI) had a significant positive correlation with home BPs, especially morning systolic BPs on HD days (P < 0.01) and non-HD days (P < 0.05), on univariate and multivariate analysis. In contrast, predialysis BPs did not correlate with LVMI. During the follow-up period (47 ± 18 months), it was demonstrated that diabetes and home BPs, especially systolic BPs on the morning of HD days, were significant predictors of CV events on multivariate Cox regression analysis. A 10 mmHg increase in BP had a significantly elevated relative risk for CV events. CONCLUSIONS: Home BP, especially systolic BPs in the morning on HD days, can provide pivotal information for management of HD patients.

Nematicidal activities of 4-hydroxyphenylacetic acid and oidiolactone D produced by the fungus Oidiodendron sp.

Two nematicides, 4-hydroxyphenylacetic acid (4-HPA) (1) and oidiolactone D (2), were isolated from cultures of the fungus Oidiodendron sp., and their structures were identified by spectroscopic analyses. Compound 2 showed nematicidal activities against the root-lesion nematode, Pratylenchus penetrans, and the pine wood nematode, Bursaphelenchus xylophilus. Compound 1 was also active against these two nematodes but to a lesser extent.

Influence of dental occlusion on physical fitness decline in a healthy Japanese elderly population.

The aim of this study was to clarify the relationship between dental occlusion and physical fitness by a longitudinal survey. A sample of 348 subjects (171 men and 177 women) aged 71 was investigated by Eichner index (EI) as an occlusal condition (Class A: no loss; Class B: partial loss; Class C: complete loss) and five types of physical fitness tests and were re-examined 8 years later. The upper 50% were sampled to analyze the correlation between each physical fitness decline and the EI at the baseline by logistic regression models. Logistic regression analyses revealed that leg extensor power (Class B vs. Class A; odds ratio=OR=4.61, p=0.010) and one-leg standing time with eyes open (Class C vs. Class A; OR=4.27, p=0.031) showed significant correlations with the EI at the baseline. In this study, partial or complete loss of occlusion was associated with a decline in leg extensor power or a decrease in one-leg standing time with eyes open. These findings suggest that maintenance of dental occlusion may prevent a decrease in activities of daily living in the elderly.

Ag nanoparticle sheet as a marker of lateral remote photocatalytic reactions.

The lateral remote photocatalytic activity of TiO(2) nanotubes fabricated by anodic oxidation, was characterized by use of a 'silver nanosheet' as a marker. The silver nanosheet is a two-dimensional crystalline film composed of myristate-capped silver nanoparticles (d = 5 nm), which has a sharp localized plasmon absorption band at lambda(max) = 470 nm. A quartz substrate was coated with TiO(2) nanotubes to a coverage of 10% and then covered with a silver nanosheet. Upon UV irradiation a rapid decrease in the intensity of the 470 nm plasmon band and an increase in the long-wavelength absorption (550-1000 nm) band were clearly confirmed, indicating fusion of the nanoparticles due to decomposition of the myristate capping agent. Surface morphology measured by high-resolution scanning electron microscopy (SEM) and atomic force microscopy (AFM) revealed the decomposition and fusion of silver nanoparticles even in regions of the surface a micron away from the TiO(2) nanotubes.