Acceleration-triggered animal-borne videos show a dominance of fish in the diet of female northern elephant seals.

Knowledge of the diet of marine mammals is fundamental to understanding their role in marine ecosystems and response to environmental change. Recently, animal-borne video cameras have revealed the diet of marine mammals that make short foraging trips. However, novel approaches that allocate video time to target prey capture events is required to obtain diet information for species that make long foraging trips over great distances. We combined satellite telemetry and depth recorders with newly developed date-/time-, depth- and acceleration-triggered animal-borne video cameras to examine the diet of female northern elephant seals during their foraging migrations across the eastern North Pacific. We obtained 48.2 h of underwater video, from cameras mounted on the head (n=12) and jaw (n=3) of seals. Fish dominated the diet (78% of 697 prey items recorded) across all foraging locations (range: 37-55°N, 122-152°W), diving depths (range: 238-1167 m) and water temperatures (range: 3.2-7.4°C), while squid comprised only 7% of the diet. Identified prey included fish such as myctophids, Merluccius sp. and Icosteus aenigmaticus, and squid such as Histioteuthis sp., Octopoteuthis sp. and Taningia danae Our results corroborate fatty acid analysis, which also found that fish are more important in the diet, and are in contrast to stomach content analyses that found cephalopods to be the most important component of the diet. Our work shows that in situ video observation is a useful method for studying the at-sea diet of long-ranging marine predators.

[Anesthetic Management of Emergency Thrombectomy for a Patient with Undiagnosed Polycythemia Vera].

Here we report successful anesthetic management of emergency thrombectomy for a patient with undiag- nosed polycythemia vera. A 67-year-old man com- plained of numbness of the right lower limb and was diagnosed with acute artery obstruction. Emergency thrombectomy was scheduled. Preoperative blood exam revealed hemoglobin 21.0 g · dl⁻¹ (hematocrit, 63.4%). During central venous catheter placement, we con- firmed high backflow blood viscosity; blood was diluted with plasma substitute. Hemoglobin was main- tained at 14-15 g · dl⁻¹ with continuous administration of plasma substitute. On re-perfusion of the right lower limb, potassium increased to 7.6 mEq · ml⁻¹, which responded to calcium carbonate, GI therapy, and furo- semide. Surgery was uneventful and the patient was diagnosed with polycythemia vera postoperatively. As perioperative management of polycythemia vera is challenging, particularly in undiagnosed and untreated cases, efforts should be made to avoid further throm- bosis and cardiac events.

Validity of brief food frequency questionnaire for estimation of dietary intakes of folate, vitamins B6 and B12, and their associations with plasma homocysteine concentrations.

BACKGROUND: Low nutrient intakes of folate, vitamins B(6) and B(12) have been associated with the risk of chronic diseases and neural tube defects. The aim of this study was to develop a brief food frequency questionnaire (FFQ) for the assessment of these nutrient intakes and to assess its validity. METHODS: We developed a brief FFQ designed specifically for estimating intakes of folate and vitamins B(6) and B(12) and covering 21 food items, and used it to carry out dietary surveys of 579 Japanese men and women. The associations of these vitamin intakes with their serum levels and homocysteine concentrations were examined using multiple linear regression analysis after adjusting for sex, age and cardiovascular risk factors. RESULTS: The Spearman correlation coefficients between vitamin intakes and their serum concentrations were 0.10 for folate, 0.08 for vitamin B(6) and 0.22 for B(12). Changes in plasma homocysteine concentrations associated with a one-standard-deviation increment were -0.03 for dietary folate, -0.03 for dietary vitamin B(6) and -0.03 for dietary vitamin B(12), and were -0.06 for serum folate, -0.03 for serum vitamin B(6) and -0.06 for serum vitamin B(12). CONCLUSION: Positive correlations between dietary intakes estimated from our brief FFQ and serum concentrations for folate and vitamin B(12) as well as negative correlations between dietary intakes and plasma homocysteine concentrations suggest the usefulness of this questionnaire.

Enantioselectivity-Evaluation of Chiral Copper(II) Complexes Coordinated by Novel Chiral Tetradentate Ligands for Free Amino Acids by Mass Spectrometry Coupled With the Isotopically Labeled Enantiomer Method.

A series of copper(II) complexes with chiral tetradentate ligands, N,N'-ethylene- bis(S-amino acid methyl amide or methyl ester) prepared from S-alanine, S-phenylalanine, S-valine or S-proline, was generated in methanol. The copper complexes provided three component complexes in the presence of a free chiral amino acid. The enantioselectivity for the amino acid was evaluated by electrospray ionization-mass spectrometry coupled with the deuterium-labeled enantiomer method and these copper complexes were found to exhibit high enantioselectivity for free amino acids having bulky side chains. This result suggests that steric interaction between the tetradentate ligand and free amino acid was a major factor in chiral recognition. The copper complex with a chiral tetradentate ligand prepared from S-proline showed opposite enantioselectivity to copper complexes consisting of tetradentate ligands prepared from other S-amino acids. The conformational difference of the tetradentate ligand in the copper complex was found to be significant for enantioselectivity.

Essential role for gene profiling analysis in the authentication of human cell lines.

Cross-contamination between cultured cell lines can result in the generation of erroneous scientific data. Hence, it is very important to eliminate cell lines that are of an origin different from that being claimed. Inter-species contamination can be detected by various established methods, such as karyotype and isozyme analyses. However, it has been impossible to detect intraspecies cross-contamination prior to the development of technology to detect differences between cell lines at the molecular level. Recently, profiling of short tandem repeat (STR) polymorphisms has been established as a method for the analyses of gene polymorphism. Gene profiling by STR polymorphism (STR profiling) is a simple and reliable method to identify individual cell lines. Each human cell line currently provided by the Cell Engineering Division of the RIKEN BioResource Center was analyzed by STR profiling to authenticate its identity. We found that more than 10 human cell lines out of approximately 400 were in fact identical to a different cell line deposited in the collection, and therefore had been misidentified. We conclude that STR profiling is a useful and powerful method for eliminating cell lines that have been misidentified by cross-contamination or by other causes. Hence, STR profiling of human cell lines used in published research will likely be a prerequisite for publication in the future, so that the problem of misidentification of cell lines can be eliminated.

Establishment of induced pluripotent stem cells from human neonatal tissues.

Following the success in establishing human induced pluripotent stem (iPS) cells, research into various applications of the cells derived from human iPS cells has begun in earnest. The use of iPS cell-derived cells in clinical therapies is one of the most exciting of the possible applications. However, the risk of tumorigenicity is the biggest potential obstacle to use iPS cell derivatives in the clinic. It should be noted that the human cells used to generate iPS cell lines may have acquired genetic mutations and these might influence the tumorigenicity of the cells. In particular, the cells of older people have a higher risk of genetic mutations than those of younger people. Here, we show that iPS cells could be derived from short-term cultures of neonatal tissues. The established human iPS cells expressed various markers of undifferentiated cells and formed teratoma in immunodeficient mice. The human iPS cells derived from neonatal tissues may represent a clinical material possessing less tumorigenicity.

Activation of focal adhesion kinase in detached human epidermal cancer cells and their long-term survival might be associated with cell surface expression of laminin-5.

While normal epithelial cells are anchorage-dependent, cancer cells are anchorage-independent. To elucidate the mechanism underlying the anchorage-independence of cancer cells, we cultured detached cells in medium containing elastase. Detached human epidermal cancer cells (DJM-1) survived for at least 3 weeks and focal adhesion kinase was still phosphorylated. In contrast, most detached keratinocytes underwent rapid apoptosis and focal adhesion kinase was not phosphorylated while the cells were alive. Thus, discontinuation of the phosphorylation of focal adhesion kinase preceded cell death. Immunostaining showed laminin-5 expression on the surface of detached DJM-1 cells, but not on detached keratinocytes. Receptors for laminin-5 (i.e. integrins) were detected on both detached DJM-1 cells and keratinocytes. Laminins are secreted proteins, so we speculated that laminin-5 adhered to the surface of secreting DJM-1 cells via integrins and evoked activation of focal adhesion kinase, with the resultant signalling cascade promoting cellular survival. If this hypothesis is correct, cell surface expression of laminin-5 may be used to explain the characteristics of cancer, immortality, tumour formation, metastasis and angiogenesis.

Trends in dietary intake of folate, vitamins B6, and B12 among Japanese adults in two rural communities from 1974 through 2001.

BACKGROUND: The 5th edition of the Japanese food composition table enables us to evaluate intakes of folate and vitamins B6 and B12, which are associated with risk of chronic diseases. METHODS: We investigated long-term trends in dietary intake of those nutrients in two rural communities; Ikawa from1974 through 2000, and Kyowa from 1982 through 2001. The 24-hour recall method was adopted. Intake of green tea interviewed from 1994 was used to examine food sources for these nutrients in the latest period, but not to evaluate long-term trends. Reduced intakes of nutrients due to cooking were not taken into account. RESULTS: Age-adjusted mean folate intake increased by 30% in Ikawa between the 1970's and 1980's, and then leveled off to the latest survey, while that in Kyowa did not change throughout the survey periods. The increased folate intake was primarily due to green/yellow vegetables. Mean vitamin B6 intake did not change except that it increased for Ikawa women in the 1980's and decreased for Kyowa men in the latest period. No secular trend was found for mean vitamin B12 intake. The largest source for folate intake was total vegetables (38-58% of total intake) and the second largest source was alcohol/beverages including green tea (11-24%). Fish/shellfish was the largest source for vitamins B6 (16-23%) and B12 (77-84%). CONCLUSIONS: Dietary intakes of folate, vitamins B6 and B12 showed no notable long-term trend, except for an increased folate intake between the 1970's and 1980's due to an increased intake of green/yellow vegetables.

GGA proteins associate with Golgi membranes through interaction between their GGAH domains and ADP-ribosylation factors.

ADP-ribosylation factors (ARFs) are a family of small GTPases that are involved in various aspects of membrane trafficking events. These include ARF1-ARF6, which are divided into three classes on the basis of similarity in the primary structure: Class I, ARF1-ARF3; Class II, ARF4 and ARF5; and Class III, ARF6. Previous studies identified a novel family of potential ARF effectors, termed GGA1-GGA3, which interact specifically with GTP-bound ARF1 and ARF3 and are localized to the trans-Golgi network (TGN) or its related compartment(s) (GGA is an abbreviation for Golgi-localizing, gamma-adaptin ear homology domain, ARF-binding protein). In the present study we have shown that ARF proteins belonging to the three classes, ARF1, ARF5 and ARF6, can interact with all GGA proteins in a yeast two-hybrid assay, in vitro and in vivo. Segmentation of GGA proteins and isolation of GGA mutants defective in ARF binding have revealed that a limited region within the GGA homology domain, which is conserved in the GGA family, is essential for ARF binding. Expression in cells of GTPase-restricted mutants of ARF1 and ARF5 blocks dissociation of GGA proteins from membranes induced by brefeldin A. However, neither of the ARF mutants recruits GGA mutants defective in ARF binding. On the basis of these observations, we conclude that at least ARF1 (Class I) and ARF5 (Class II) in their GTP-bound state cause recruitment of GGA proteins on to TGN membranes. In contrast, on the basis of similar experiments, ARF6 (Class III) may be involved in recruitment of GGA proteins to other compartments, possibly early endosomes.

Validity of the self-administered food frequency questionnaire used in the 5-year follow-up survey for the JPHC Study to assess folate, vitamin B6 and B12 intake: comparison with dietary records and blood level.

To validate a self-administered food frequency questionnaire (FFQ) for the estimation of dietary intake of folate, vitamins B6 and B12, we compared measures of these vitamins by the FFQ and plasma concentrations among 87 male volunteers aged 40-89 years in the Japan Public Health Center-based prospective study (JPHC Study). No men used folate, vitamins B6 or B12 as supplements. There was a moderate association between energy-adjusted dietary intake and plasma concentration for folate and vitamin B6. The Spearman correlation coefficient was 0.26 and 0.23, respectively, in the total samples. These correlation coefficients were slightly higher when men in the Ishikawa PHC area were excluded from the analysis; the correlation coefficient was 0.29 for folate and 0.26 for vitamin B6. No significant correlation was found between dietary vitamin B12 intake and serum B12 concentration; the correlation coefficient was 0.06 in the total sample and 0.15 when the Ishikawa men were excluded from the analysis. In conclusion, we found that the FFQ used for the 5-year follow-up survey of JPHC Study could reasonably rank individuals by dietary intake of folate and vitamin B6, but not vitamin B12, among Japanese community-dwelling population samples.

GAT (GGA and Tom1) domain responsible for ubiquitin binding and ubiquitination.

GGAs (Golgi-localizing, gamma-adaptin ear domain homology, ADP-ribosylation factor (ARF)-binding proteins) are a family of monomeric adaptor proteins involved in membrane trafficking from the trans-Golgi network to endosomes. The GAT (GGA and Tom1) domains of GGAs have previously been shown to interact with GTP-bound ARF and to be crucial for membrane recruitment of GGAs. Here we show that the C-terminal subdomain of the GAT domain, which is distinct from the N-terminal GAT subdomain responsible for ARF binding, can bind ubiquitin. The binding is mediated by interactions between residues on one side of the alpha3 helix of the GAT domain and those on the so-called Ile-44 surface patch of ubiquitin. The binding of the GAT domain to ubiquitin can be enhanced by the presence of a GTP-bound form of ARF. Furthermore, GGA itself is ubiquitinated in a manner dependent on the GAT-ubiquitin interaction. These results delineate the molecular basis for the interaction between ubiquitin and GAT and suggest that GGA-mediated trafficking is regulated by the ubiquitin system as endosomal trafficking mediated by other ubiquitin-binding proteins.