Generation of Ultralong Liposome Tubes by Membrane Fusion beneath a Laser-Induced Microbubble on Gold Surfaces.

Membrane fusion (MF) is one of the most important and ubiquitous processes in living organisms. In this study, we developed a novel method for MF of liposomes. Our method is based on laser-induced bubble generation on gold surfaces (a plasmonic nanostructure or a flat film). It is a simple and quick process that takes about 1 min. Upon bubble generation, liposomes not only collect and become trapped but also fuse to form long tubes beneath the bubble. Moreover, during laser irradiation, these long tubes remain stable and move with a waving motion while continuing to grow, resulting in the creation of ultralong tubes with lengths of about 50 µm. It should be noted that the morphology of these ultralong tubes is analogous to that of a sea anemone. The behavior of the tubes was also monitored by fluorescence microscopy. The generation of these ultralong tubes is discussed on the basis of Marangoni convection and thermophoresis.

In Situ Hybridization to Identify Stem Cells in the Freshwater Sponge Ephydatia fluviatilis.

Sponges (Porifera) are a large phylum that includes an enormous number of species. They are classified into four classes. Among these four classes, class Demospongiae is the largest and contains more than 90% of sponge species. In the last decade, methodologies for molecular studies and sequencing resources in sponge biology have dramatically advanced and made it possible to clearly define particular types of cells based on the genes they are expressing. Here we describe in detail the method of high-resolution WISH (whole mount in situ hybridization) and dual color fluorescent detection of in situ hybridization (dual color FISH) that we have established to detect particular types of cells, especially their stem cells known as archeocytes, in juveniles of freshwater demosponge, E. fluviatilis.

Imidazoline- and Benzamidine-Based Trypanosome Alternative Oxidase Inhibitors: Synthesis and Structure-Activity Relationship Studies.

The trypanosome alternative oxidase (TAO), a mitochondrial enzyme involved in the respiration of the bloodstream form trypomastigotes of Trypanosoma brucei, is a validated drug target against African trypanosomes. Earlier series of TAO inhibitors having a 2,4-dihydroxy-6-methylbenzoic acid scaffold ("head") and a triphenylphosphonium or quinolin-1-ium cation as a mitochondrion-targeting group ("tail") were shown to be nanomolar inhibitors in enzymatic and cellular assays. We investigated here the effect of different mitochondrion-targeting cations and other scaffold modifications on the in vitro activity of this class of inhibitors. Low micromolar range activities were obtained, and the structure-activity relationship studies showed that modulation of the tail region with polar substituents is generally detrimental to the enzymatic and cellular activity of TAO inhibitors.

Distribution of Antimicrobial Resistance in Campylobacter Strains Isolated from Poultry at a Slaughterhouse and Supermarkets in Japan.

Fifty strains of Campylobacter jejuni/coli were detected in 108 specimens of chicken meat and organs sampled at six supermarkets and one poultry slaughterhouse (large scale) between April and October 2013 (isolation rates: 84.8% from the slaughterhouse, 29.3% from the supermarkets). 46/50 strains were successfully recovered and subjected to the E-test to examine their susceptibility to three fluoroquinolone antibacterial agents authorized for use in poultry in Japan: enrofloxacin (ERFX), ofloxacin (OFLX), and norfloxacin (NLFX). 29 isolates (63%) were resistant to all three agents and 2 isolates (4.3%) were resistant to two agents (ERFX and OFLX). The resistance rates of strains isolated fom the supermarkets and slaughterhouse were 61.9% and 72.0%, respectively. Because the chickens processed at the slaughterhouse were raised without the use of fluoroquinolone, the results did not suggest a positive relationship between the use of these agents and the distribution of antimicrobial-resistant bacteria. Susceptibility to macrolide antibiotics (erythromycin [EM]) was also tested in 42 strains, and one strain (2.4%), C. coli from a retailer sample, showed resistance. Previous studies have detected high rates of fluoroquinolone-resistant strains, suggesting an expanding distribution of resistant bacteria. The detection of EM-resistant bacteria downstream in the food distribution chain (i.e., closer to consumers) is a concern for human health.

Dynamic Transport and Cementation of Skeletal Elements Build Up the Pole-and-Beam Structured Skeleton of Sponges.

Animal bodies are shaped by skeletons, which are built inside the body by biomineralization of condensed mesenchymal cells in vertebrates [1, 2] and echinoderms [3, 4], or outside the body by apical secretion of extracellular matrices by epidermal cell layers in arthropods [5]. In each case, the skeletons' shapes are a direct reflection of the pattern of skeleton-producing cells [6]. Here we report a newly discovered mode of skeleton formation: assembly of sponges' mineralized skeletal elements (spicules) in locations distant from where they were produced. Although it was known that internal skeletons of sponges consist of spicules assembled into large pole-and-beam structures with a variety of morphologies [7-10], the spicule assembly process (i.e., how spicules become held up and connected basically in staggered tandem) and what types of cells act in this process remained unexplored. Here we found that mature spicules are dynamically transported from where they were produced and then pierce through outer epithelia, and their basal ends become fixed to substrate or connected with such fixed spicules. Newly discovered "transport cells" mediate spicule movement and the "pierce" step, and collagen-secreting basal-epithelial cells fix spicules to the substratum, suggesting that the processes of spiculous skeleton construction are mediated separately by specialized cells. Division of labor by manufacturer, transporter, and cementer cells, and iteration of the sequential mechanical reactions of "transport," "pierce," "raise up," and "cementation," allows construction of the spiculous skeleton spicule by spicule as a self-organized biological structure, with the great plasticity in size and shape required for indeterminate growth, and generating the great morphological diversity of individual sponges.

Longitudinal nutritional assessment in acute lymphoblastic leukemia during treatment.

BACKGROUND: A nutritional assessment of pediatric patients with cancer is important to improve their outcome. The number of longitudinal nutritional studies during treatment, however, is limited. The purpose of this study was to investigate the longitudinal changes in anthropometric measures and serum albumin level during chemotherapy in patients with acute lymphoblastic leukemia (ALL). METHODS: We retrospectively reviewed the charts of 23 patients (19 boys, four girls) with ALL from April 2007 to March 2010. The median age at diagnosis was 4.5 years. Bodyweight, height, and serum albumin levels were measured at the start and the end point of each chemotherapy phase. RESULTS: At diagnosis, two patients (8.7%) were underweight and five patients (21.7%) were overweight according to body mass index z-score, while five patients were underweight and three (13.0%) were overweight according to Waterlow score. The prevalence of malnourished patients did not change significantly throughout chemotherapy by either assessment. The absolute scores in either assessment were significantly reduced at the sanctuary treatment phase. Low serum albumin (<3.2 g/dL) was found in two patients at diagnosis. Mean albumin decreased significantly at the induction and the re-induction phases. CONCLUSIONS: Given that nutritional status under a similar chemotherapeutic regimen as assessed by anthropometric measures and albumin level differed among patients, careful observation of the nutritional status and intervention may be necessary at different phases of chemotherapy.

Adipocytokines in childhood cancer survivors and correlation with metabolic syndrome components.

BACKGROUND: Although there are several studies on the prevalence of metabolic syndrome (MetS) in childhood cancer survivors (CCS), the association between MetS components and serum adipocytokine level has not been elucidated. METHODS: The charts of 49 patients (27 male, 22 female) who had attended the CCS clinic of the Department of Pediatrics, Kyoto University Hospital, between April 2009 and March 2010 were retrospectively reviewed. Median age was 10.7 years, and the median interval since the completion of chemotherapy was 5.1 years. The diagnosis of MetS was made based on the Japanese criteria for either children or adults. RESULTS: Three (6.1%) of 49 patients fulfilled the criteria for MetS, and 28 (57.1%) had at least one component of MetS. High leptin level was seen in 18 patients (36.7%), and low total adiponectin level was seen in 20 (40.8%). The number of patients with high leptin was correlated with body mass index z-score (>2.0), abdominal circumference/height (≥0.5), diastolic blood pressure and fasting blood sugar. The number of patients with low total adiponectin was correlated with systolic blood pressure and triglyceride. When the patients were divided into three groups based on the number of positive MetS components (0, 1 and 2-4), leptin and adiponectin tended to be higher and lower in the third group, respectively. CONCLUSIONS: Adipocytokines may play a role in the pathogenesis of MetS occurring in CCS. It is recommended that adipocytokines be evaluated together with MetS components at the CCS follow-up clinics.

Evaluation of taste acuity by the filter-paper disc in Japanese young women: the relationship with micronutrients status.

The aim of the present study is to investigate the taste acuity in Japanese young women in relation to their micronutrient status. Thirty-eight healthy young women (mean age; 21.3, range; 19-27 years) were enrolled. Gustatory thresholds were estimated for four basic tastes: sweet (sucrose), salty (sodium chloride), sour (tartaric acid), and bitter (quinine hydrochloride) by a filter-paper disk method. Various concentrations at each taste were serially scored from disc number 1 (lowest) to number 5 (highest). The lowest concentration at which the quality of the taste was correctly identified was defined as the recognition threshold. The mean of three measurements for each test on three different days was calculated. We divided our participants into normal taste and hypogeusia groups based on the mean threshold disc numbers, <=3.5 and >3.5, respectively, according to previous literature using the same method. We also measured serum concentrations and dietary intakes of micronutrients including zinc, iron, copper, and selenium. The numbers of participants belonging to the hypogeusia group were 24 (63.2%) for sweet, 19 (50.0%) for sour, 17 (44.7%) for bitter, and 16 (42.1%) for salty taste. Although the hypogeusia group exhibited significantly lower serum iron concentrations, except for the salty taste, the other three micronutrients concentrations did not show any association with the four taste acuities. Dietary micronutrient intake did not show any association with the four taste acuities. This study indicates that in addition to zinc status, iron status should be considered in the study of taste acuity.

Fatigue in survivors of childhood acute lymphoblastic and myeloid leukemia in Japan.

BACKGROUND: Fatigue in cancer survivors is a serious problem in pediatric oncology, but reports on this issue are limited, especially in Asian countries. METHODS: Sixty-three patients with acute lymphoblastic leukemia and 18 patients with acute myeloid leukemia who attended a follow-up outpatient clinic were enrolled. Participants were required to be >8 years of age, in remission, and without any cancer treatment for at least the previous 1 year. A control group consisted of 243 subjects whose age and gender were matched with the patient group. A questionnaire consisting of 12 items was devised for fatigue measurement. RESULTS: Principal factor analysis identified three dimensions, defined as physical fatigue, decreased function, and altered mood. The mean total and the three fatigue dimension scores tended to be higher in the control group, but significant differences between the scores were seen only in the total and physical fatigue scores. Multiple regression analysis indicated an association of present older age or shorter duration after completion of treatment with total and physical fatigue, and an association of presence of total body irradiation with decreased function. CONCLUSION: Pediatric leukemia survivors in Japan experience equal or less fatigue compared with that of controls in different fatigue dimensions. Elucidation of underlying mechanisms of cancer-related fatigue including the differences of cultural background among different countries is necessary for future study of this issue.

Retrospective individual tracking of body mass index in obese and thin adolescents back to childhood.

The objective of this study is to track body mass index (BMI) in obese or thin adolescents from adolescence to childhood on an individual basis. This was performed at a single school with a 12-year combination education system in an urban city in Japan. A total of 617 students in the 3rd grade of senior high school (17 years old) during 2005-2009 were enrolled. Based on the Japanese BMI reference in childhood adjusted for age and gender, obesity and thinness were defined as >=90th percentile and <=5th percentile, respectively. Sixty-three (10.2%) and 84 (13.6%) students were found to be obese and thin, respectively. Complete annual tracking of BMI back to the 1st grade of elementary school (6 years old) (1994-1998) was possible in 47 obese (74.6%) and 67 thin students (80.0%). The most common ages when obesity was first detected were 6-8 years for males, and 12-14 years for females, and the most common ages when thinness was first detected were 12-14 years for males, and 15-17 years for females. Once obesity or thinness started, these conditions remained until 17 years old in most students. Obese students whose obesity started earlier tended to have higher BMIs at 17 years old in both genders. This will be the first tracking study of BMI in obese and thin adolescents on an individual basis. A longitudinal study of BMI during childhood is useful for establishing intervention programs to prevent obesity or thinness in adolescence.

Fatigue status in relation to lifestyle in healthy Japanese adolescents.

In order to investigate the prevalence of physical, mental, and chronic fatigue syndrome-(CFS-) related fatigue and its relation to lifestyle, 1,225 adolescents (591 males, 634 females) aged 11 to 16 years were asked to complete a self-reported questionnaire on fatigue status and lifestyle in the past one month. There was no gender difference in physical and mental fatigue scores, but CFS-related scores were significantly higher in females than in males. These scores were found to increase with the increase of age. After adjusting for age and gender, multiple regression analysis showed that physical and mental fatigue scores were associated with sleeping hours, extracurricular sports activity, food balance, the frequencies of snacks between regular meals, intake of sugar-sweetened beverages, and visits to the nurse's room. This paper is the first large cross-sectional study on fatigue in healthy adolescents in Japan, albeit there were numerous such studies in Western countries.