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Pollen developmental pathway in plants involving synchronized transferal of cellular divisions from meiosis (microsporogenesis) to mitosis (pollen mitosis I/II) eventually offers a unique “meiosis-mitosis shift” at pollen mitosis I. Since the cell type (haploid microspore) and fate of pollen mitosis I differ from typical mitosis (in meristem cells), it is immensely important to analyze the chromosomal distribution of phosphorylated H3S10 histone during atypical pollen mitosis I to comprehend the role of histone phosphorylation in pollen development. We investigated the chromosomal phosphorylation of H3S10 histone during pollen mitosis I in orchids using immunostaining technique. The chromosomal distribution of H3S10ph during pollen mitosis I revealed differential pattern than that of typical mitosis in plants, however, eventually following the similar trends of mitosis in animals where H3S10 phosphorylation begins in the pericentromeric regions first, later extending to the whole chromosomes, and finally declining at anaphase/early cytokinesis (differentiation of vegetative and generative cells). The study suggests that the chromosomal distribution of H3S10ph during cell division is not universal and can be altered between different cell types encoded for diverse cellular processes. During pollen development, phosphorylation of histone might play a critical role in chromosome condensation events throughout pollen mitosis I in plants.

Both coccolith-bearing cells (C-cells) and naked cells (N-cells) of the coccolithophorid Pleurochrysis haptonemofera can grow in salinities of more than 7 per thousand (about 20% of a "normal" sea water salinity [35 per thousand]), with the highest growth rates in salinities of more than 14 per thousand. Microscopic observations of cells suspended in 100 mM NaCl (7 per thousand) showed that, while N-cells were swelling uniformly all over the cell surface, C-cells were bulging the plasma membrane from the hole of the coccosphere at the apical (flagellar) pole of the cell. Effects of several cations and anions on the morphological change of C-cells under hypoosmotic pressure were investigated. When 100 mM K(+) was used, protoplasts were released from the coccosphere completely in almost all the cells. This phenomenon was shown with K(+) most effectively. The protoplasts could grow in the fresh medium and form the first coccolith within 9 h.

Studies of maize starch branching enzyme mutants suggest that the amylose extender high amylose starch phenotype is a consequence of the lack of expression of the predominant starch branching enzyme II isoform expressed in the endosperm, SBEIIb. However, in wheat, the ratio of SBEIIb and SBEIIa expression are inversely related to the expression levels observed in maize and rice. Analysis of RNA at 15 days post anthesis suggests that there are about 4-fold more RNA for SBE IIa than for SBE IIb. The genes for SBE IIa and SBE IIb from wheat are distinguished in the size of the first three exons, allowing isoform-specific antibodies to be produced. These antibodies were used to demonstrate that in the soluble fraction, the amount of SBE IIa protein is two to three fold higher than SBIIb, whereas in the starch granule, there is two to three fold more SBE IIb protein amount than SBE IIa. In a further difference to maize and rice, the genes for SBE IIa and SBE IIb are both located on the long arm of chromosome 2 in wheat, in a position not expected from rice-maize-wheat synteny.

Autosporulation is a common mode of propagation for unicellular algae. Autospore-forming species of Chlorellaceae, Chlorella vulgaris Beijerinck, C. sorokiniana Shihira et Krauss, C. lobophora Andreyeva, and Parachlorella kessleri (Fott et Nováková) Krienitz et al. have glucosamine as the main constituent of their rigid cell wall. Recent phylogenetic analyses have showed that the Chlorellaceae divided into two sister groups: the Chlorella-clade and the Parachlorella-clade. We compared the cell wall structure and synthesis of the daughter cell wall in the four species by electron microscopy using rapid freezing and freeze substitution methods. The cell wall of C. vulgaris, C. sorokiniana, and C. lobophora consisted of an electron-dense thin layer with an average thickness of 17-20, 22, and 19 nm, respectively. In these three species, daughter cell wall synthesis occurred on the outer surface of the plasma membrane in the early cell-growth phase. The cell wall of P. kessleri, however, was electron-transparent and 54-59 nm in thickness. Ruthenium red staining of P. kessleri indicated that ruthenium-red-specific polysaccharides accumulated over the outer surface of the plasma membrane. Immunoelectron microscopic observation with an anti-beta-1, 3-glucan antibody and staining with wheat germ agglutinin (WGA) indicated that the cell wall contained beta-1, 3-glucan and WGA specific N-acetyl-beta-D-glucosamine. In P. kessleri, daughter cell wall synthesis began after successive protoplast division. The daughter cell wall synthesis during autosporulation in the four species of Chlorellaceae can be classified into two types-the early and the late types.

Introduction of large-DNA fragments into cereals by Agrobacterium-mediated transformation is a useful technique for map-based cloning and molecular breeding. However, little is known about the organization and stability of large fragments of foreign DNA introduced into plant genomes. In this study, we produced transgenic rice plants by Agrobacterium-mediated transformation with a large-insert T-DNA containing a 92-kb region of the wheat genome. The structures of the T-DNA in four independent transgenic lines were visualized by fluorescence in situ hybridization on extended DNA fibers (fiber FISH). By using this cytogenetic technique, we showed that rearrangements of the large-insert T-DNA, involving duplication, deletion and insertion, had occurred in all four lines. Deletion of long stretches of the large-insert DNA was also observed in Agrobacterium.

SUMMARY The green macroalgal genus Ulva (incl. Entemmorpha) contains economically valuable species, is of relevance for coastal management (green tides), and certain taxa serve as experimental organisms for fundamental research in green algae. The nuclear genome size of Ulva (Entemmorpha) compressa Linnaeus was measured in propidium iodide stained nuclei using laser scanning cytometry. Nuclei of fixed gametes yielded reproducible values, whereas nuclei extracted from multicellular gametophytes were unsuitable. With nuclei of Arabidopsis thaliana (L.) Heynh and Saccharomyces cerevisiae Hansen as references, the haploid nuclear genome size of U. compressa was calculated as 135 ± 7 Mbp. This is the smallest genome so far known from any species of Ulva.

SUMMARY The small subunit of the chloroplast enzyme ribulose-1,5-bisphosphate carboxyase/oxygenase (Rubisco) is encoded by rbcS. We isolated and characterized three rbcS genes (NbrbcS1-1, NbrbcS1-2, and NbrbcS2) from the unicellular green alga Nannochloris bacillaris (Chlorophyta, Trebouxiophyceae). In the haploid N. bacillaris genome, each is a single-copy gene located on different chromosomes. Each mature peptide contains 140 amino acid residues. NbrbcS1-1 and NbrbcS1-2 are identical and share 80% identity with NbrbcS2, while the respective transit sequences share only approximately 48% identity with that of NbrbcS2. NbrbcS1-1 transcription was suppressed in the dark and recovered drastically after transfer to light. In contrast, NbrbcS1-2 and NbrbcS2 expression were not reduced after transfer from light to dark. In chlorophyllic tobacco cells containing green fluorescent protein fusion proteins of the transit sequences of each peptide, green fluorescent protein signals were localized on particles matching chloroplasts. The first introns of NbrbcS1-1 and NbrbcS1-2 are identical to the corresponding introns of 37 rbcS genes in eight embryophyte species. While the second intron is conserved in the green algae, the NbrbcS2 intron appears to involve sliding by one base pair. The NbrbcS1-1 and NbrbcS1-2 intron conserved in green algae and embryophytes might be an ancestral intron from nuclear-encoded rbcS.

Disodium cromoglycate (DSCG) is a widely used anti-asthmatic drug with few side effects. One of the mechanisms by which DSCG suppresses asthma is prevention of anaphylactic release of chemical mediators from mast cells. However, it has also been reported that DSCG attenuates late asthmatic response and airway hyperresponsiveness, the induction mechanisms of which can not be explained simply by release of chemical mediators from mast cells. Mechanisms of anti-inflammatory effects of DSCG other than its mast-cell stabilising effect have been unclear. In this study, we evaluated effects of DSCG on early and late asthmatic response in guinea pig. Guinea pigs were alternately sensitized and challenged by exposure to mists of ovalbumin+Al(OH)3 and ovalbumin, respectively. DSCG administered intratracheally before the fifth challenge dose-dependently inhibited late asthmatic response, but early asthmatic response was not affected. Interestingly, DSCG at 2 mg/kg potently suppressed increases in cysteinyl leukotrienes and thromboxane A2 in the lung during late asthmatic response. Eosinophilia was slightly reduced by DSCG. The inhibitory effect of DSCG on late asthmatic response is apparently due to inhibition of release of arachidonic acid metabolites.

Disodium cromoglycate is an anti-asthmatic drug that has mast cell-stabilizing effects and other anti-inflammatory effects. However, the mechanisms of its anti-inflammatory effects are unclear. In this study, we evaluated effects of disodium cromoglycate on eosinophilia, early and late asthmatic responses, and production of arachidonic acid metabolites in guinea pig lungs. Guinea pigs were alternately sensitized and challenged by exposure to mists of ovalbumin+Al(OH) 3 and ovalbumin, respectively. Disodium cromoglycate (0.5–2 mg/0.1 ml/animal) administered intratracheally before the fifth challenge dose-dependently inhibited asthmatic response, but early asthmatic response was not affected. Disodium cromoglycate at 2 mg/animal potently suppressed increases in cysteinyl leukotrienes (CysLTs) and thromboxane A 2 in the lung during late asthmatic response. Eosinophilia was slightly reduced by disodium cromoglycate. The inhibitory effect of disodium cromoglycate on late asthmatic response is apparently due to inhibition of the release of arachidonic acid metabolites, some of which may be derived from eosinophils that infiltrate the lung.

We identified and characterized elongationless3 (eln3-1), a restriction enzyme-mediated integration (REMI) mutation affecting fruiting body morphogenesis in Coprinus cinereus. The mutant produces an aberrant fruiting body in which the stipe hardly elongates during fruiting body maturation. In the wild type, cylindrical stipe cells, elongation growth of which is responsible for stipe elongation, make side-by-side contact with one another and run parallel to the stipe axis, whereas in the mutant, the organization of the stipe tissue is disturbed and much space is produced between stipe cells. This disorganization of the stipe tissue, together with reduced elongation of the stipe cells, causes the mutant stipe short and bulgy. After a plasmid rescue, the eln3 gene was identified as a DNA fragment that complements the eln3-1 mutation. The eln3 ORF is predicted to encode a protein of 927 amino acids with a general glycosyltransferase domain and to be located in the plasma membrane. Transcription of the eln3 gene is specifically activated in rapidly elongating stipes. Possible involvement of the putative Eln3 enzyme in cell-to-cell connection is discussed. Index Descriptors: Homobasidiomycete; Coprinus cinereus; Fruiting body morphogenesis; REMI mutant; Stipe elongation; Cell-to-cell connection; Membrane protein; Glycosyltransferase

Two FtsZ paralogues (NbFtsZ1 and NbFtsZ2) were isolated from the unicellular green alga Nannochloris bacillaris Naumann. These sequences encoded proteins of 435 and 439 amino acids with tubulin signature motifs (GGGTG[T/S]G), which are important for GTP binding activity. NbFtsZ1 and NbFtsZ2 had four and three introns, respectively, and two different putative core promoters; a TATA box (TATAAAA) and an initiator element (CCCAGG) were located 40 bp and 80 bp upstream of the coding regions of NbFtsZ1 and NbFtsZ2, respectively. Southern blot hybridization and contour-clamped homogeneous electric field electrophoresis showed that N. bacillaris contained at least one copy of each gene and that NbFtsZ1 was located on chromosome 5 and NbFtsZ2 on chromosome 3 or 4. Phylogenetically, NbFtsZ1 and NbFtsZ2 belong to the vascular plant protein families FtsZ1 and FtsZ2, respectively. The FtsZ1 proteins do not contain carboxy-terminal consensus sequences, whereas all FtsZ2 proteins possess the consensus sequence (I/V)PxFL(R/K)(K/R)(K/R). Our study has shown that NbFtsZ2 possesses a similar consensus sequence (VPDFLRRK), whereas NbFtsZ1 does not, further supporting their classification as FtsZ2 and FtsZ1. Escherichia coli ftsZ mutants transformed with cloned NbFtsZ1, and NbFtsZ2 cDNAs were restored for the capacity to divide by binary fission, suggesting that the proteins retained the ability to function in the bacterium. An anti-NbFtsZ2 antibody specifically recognized a single protein band of approximately 51 kDa on an immunoblot of N. bacillaris cellular proteins. Immunostaining of the algal cells with this antibody produced an intense fluorescent signal as a ring near the middle of the cell, which corresponded to the chloroplast division site.

The cell division mechanisms of seven strains from six species of Nannochloris Naumann were analyzed and compared with those of three species of Chlorella Beijerinck and Trebouxia erici Ahmadjian using differential interference microscopy and fluorescence microscopy. Nannochloris bacillaris Naumann divides by binary fission and N. coccoides Naumann divides by budding. Distinct triangular spaces or mother cell walls were found in the dividing autosporangia of the other five strains from four species of Nannochloris, three species of Chlorella, and T. erici. In an attempt to infer an evolutionary relationship between nonautosporic and autosporic species of Nannochloris, we constructed a phylogenetic tree of the actin genes using seven strains from six species of Nannochloris, three species of Chlorella, and T. erici. Nannochloris species were polyphyletic in the Trebouxiophyceae group. Two nonautosporic species of N. bacillaris and N. coccoides were monophyletic and positioned distally. Moreover, to determine their phylogenetic position within the Trebouxiophyceae, we constructed phylogenetic tree of 18S rRNA genes adding other species of Trebouxiophyceae. Nannochloris species were polyphyletic in the Trebouxiophyceae and appeared in two different lineages, a Chlorella-Nannochloris group and a Trebouxia- Choricystis group. The nonautosporic species, N. bacillaris and N. coccoides, and three autosporic species of Nannochloris belonged to the Chlorella-Nannochloris group. Nannochloris bacillaris and N. coccoides were also monophyletic and positioned distally in the phylogenetic tree of 18S rRNA genes. These results suggest that autosporulation is the ancestral mode of cell division in Nannochloris and that nonautosporulative mechanisms, such as binary fission and budding, evolved secondarily.

Conserved domains of two types of LTR retrotransposons, Ty1–copia- and Ty3–gypsy-like retrotransposons, were isolated from the dioecious plant Silene latifolia, whose sex is determined by X and Y chromosomes. Southern hybridization analyses using these retrotransposons as probes resulted in identical patterns from male and female genomes. Fluorescence in situ hybridization indicated that these retrotransposons do not accumulate specifically in the sex chromosomes. These results suggest that recombination between the sex chromosomes of S. latifolia has not been severely reduced. Conserved reverse transcriptase regions of Ty1–copia-like retrotransposons were isolated from 13 different Silene species and classified into two major families. Their categorization suggests that parallel divergence of the Ty1–copia-like retrotransposons occurred during the differentiation of Silene species. Most functional retrotransposons from three dioecious species, S. latifolia, S. dioica, and S. diclinis, fell into two clusters. The evolutionary dynamics of retrotransposons implies that, in the genus Silene, dioecious species evolved recently from gynodioecious species.Key words: retrotransposon, dioecious plant, sex chromosome.

Histone proteins and the nucleosomes along with DNA are the essential components of eukaryotic chromatin. Post-translational histone–DNA interactions and modifications eventually offer significant alteration in the chromatin environment and potentially influence diverse fundamental biological processes, some of which are known to be epigenetically inherited and constitute the “epigenetic code”. Such chromatin modifications evidently uncover remarkable diversity and biological specificity associated with distinct patterns of covalent histone marks. The past few years have witnessed major breakthroughs in plant biology research by utilizing chromatin modification-specific antibodies through molecular cytogenetic tools to ascertain hallmark signatures of chromatin domains on the chromosomes. Here, we survey current information on chromosomal distribution patterns of chromatin modifications with special emphasis on histone methylation, acetylation, phosphorylation, and centromere-specific histone 3 (CENH3) marks in plants using immuno-FISH as a basic tool. Major available information has been classified under typical and comparative cytogenetic detection of chromatin modifications in plants. Further, spatial distribution of chromatin environment that exists between different cell types such as angiosperm/gymnosperm, monocot/dicot, diploid/polyploids, vegetative/generative cells, as well as different stages, i.e., mitosis versus meiosis has also been discussed in detail. Several challenges and future perspectives of molecular cytogenetics in the grooming field of plant chromatin dynamics have also been addressed.

Gastric cancer is a unique malignancy, with definite geographic differences in incidence, pathology, treatment and outcome. While the incidence has been declining in the Western hemisphere, steady rates have been reported in Eastern countries, particularly South Korea and Japan. One of the most profound differences between the East and West centers around treatment strategies, with Western clinicians routinely adopting a neoadjuvant approach, prior to surgical resection. Eastern clinicians, however, favor primary surgical therapy and have pioneered many of the techniques currently used worldwide. From endoscopic therapies to minimally-invasive surgery, including laparoscopic and robotic techniques, the Eastern surgeons have studied their techniques with high-volumes of patients. Western surgeons, practicing in systems where gastric cancer care is not centralized, typically have performed less aggressive surgical resections, although generally see more advanced diseases. In the era where global care is becoming more standardized, however, the differences in surgical practice have lessened. This review compares the surgical techniques and outcomes for gastric cancer practiced in the East with those standard in the West.

Many animals migrate between breeding and wintering areas; however, whether each animal behaves consistently in space and time between consecutive years is less well understood. Furthermore, previous breeding state (successful or failed) is often not considered when attempting to understand consistent individual differences in behaviour that are likely to impact upon the subsequent behaviour. Between 2006 and 2010, we used geolocators to track the migratory movements of a pelagic seabird, the streaked shearwater Calonectris leucomelas, with individuals () being followed for two years or more, including 23 birds that had chicks in two seasons and 23 birds in just one season. All individuals, except for one bird, migrated to the same broad wintering areas, and their migratory route as well as the centre of wintering distribution did not change in relation to the previous breeding outcomes. Migration schedules (dates of departure from the breeding colony, southward and northward migrations, and first return to the colony) did not differ significantly between years for individuals that had chicks during both years, while failed individuals left the breeding colony and appeared to start the southward migration at an earlier date than the previous successful year. Nonetheless, the timing of the southward migration was consistent within individuals, including both males and females, over successive years regardless of the previous breeding outcome, and also the timing of first return back to the colony for females that had chicks in the both previous years and eggs in the both following season. This may imply the existence of individual-specific broad time schedules, possibly a circannual rhythm, though ecological conditions might affect the exact timing of the actual departure event. Our results present evidence for high levels of individually consistent behaviour for this pelagic seabird outside the breeding season.

Genes and cDNAs for starch-branching enzyme II (SBEII) have been isolated from libraries constructed from Aegilops tauschii and wheat (Triticum aestivum) endosperm, respectively. One class of genes has been termedwSBEII-DA1 and encodes the N terminus reported for an SBEII from wheat endosperm. On the basis of phylogenetic comparisons with other branching enzyme sequences, wSBEII-DA1 is considered to be a member of the SBEIIa class. ThewSBEII-DA1 gene consists of 22 exons with exons 4 to 21 being identical in length to the maize (Zea mays) SBEIIb gene, and the gene is located in the proximal region of the long arm of chromosome 2 at a locus designated sbe2a. RNA encoding SBEIIa can be detected in the endosperm from 6 d after flowering and is at its maximum level from 15 to 18 d after anthesis. Use of antibodies specific for SBEIIa demonstrated that this protein was present in both the soluble and granule bound fractions in developing wheat endosperm. We also report a cDNA sequence for SBEIIa that could arise by variant transcription/splicing. A second gene, termedwSBEII-DB1, was isolated and encodes an SBEII, which shows greater sequence identity with SBEIIb-type sequences than with SBEIIa-type sequences. Comparisons of SBEII gene structures among wheat, maize, and Arabidopsis indicate the lineage of the SBEII genes.

A single cell divides to multiply, but not all cells follow the same pattern of division. We studied cell division in seven strains from six species belonging to the genus Nannochloris Naumann and classified their modes of cell division into three types: binary fission (N. bacillaris Naumann), budding (N. coccoides Naumann), and autosporulation resulting in the formation of two to four daughter cells (N. maculata Butcher, N. sp. SAG 251-2, N. atomus Butcher CCAP 251/7 and SAG 14.87, and N. eucaryotum[Wilhelm et al.] Menzel and Wild). To determine the evolutionary relationships among these multiple modes of cell division, we investigated the strains' genome sizes, copy number of actin genes, and phylogeny. The genome sizes were determined by counter-clamped homogeneous electric fields electrophoresis and fluorimetry. The genomes are very small and range from 12.6 Mbp (N. maculata) to 47.4 Mbp (N. atomus SAG 14.87). The genomes of Nannochloris species seem to be among the smallest for free-living eukaryotes. Nannochloris bacillaris (binary fission), N. coccoides (budding), Nannochloris sp. (two-cell type of autosporulation), and N. eucaryotum (multicell type of autosporulation) contain a single actin gene, whereas N. maculata (two-cell type of autosporulation) and two strains of N. atomus (two-cell type of autosporulation) contain two actin genes. This suggests that the actin gene was duplicated in this eukaryote, which has a very small genome. Phylogenetic analyses of partial actin gene sequences suggest that autosporulation is the ancestral mode of cell division.

A 314-bp tandemly repeated DNA sequence, named pAc074, was characterized in Allium cepa by fluorescence in situ hybridization (FISH) analyses using random amplified fragment as probe. The nucleotide sequences of the clone pAc074 is partially homologous to the satellite DNA sequences, ACSAT1, ACSAT2, and ACSAT3, of A. cepa with 81%, 81% and 78% similarity, respectively. Our sequential C-banding and FISH with pAc074 probe also clearly showed a close relation between C-heterochromatin at telomeric region and pAc074 sequences on all the chromosomes except on chromosome 6. On the long arm of chromosome 7, pAc074 sequences appeared as interstitial band which did not correspond to C-heterochromatin bands. Instead, the C-heterochromatin bands corresponded with the 5S rDNA signals. This is the first evidence of simultaneous banding of the 5S rDNA and C-band in A. cepa.

Monolayers of amino acid derivatized diacetylene lipids on water were studied by Langmuir trough and epi-fluorescence microscopy (EFM). Examination of the amino acid derivatized diacetylene lipids showed a wide variety of surface characteristics with many novel properties. Many of these new properties are advantageous for film-based biosensor development. To characterize this family of molecules, pressure–area (π–A) isotherms were measured that demonstrate hydrophilic headgroup compounds allow for close-packed monomolecular layer formation in the solid state upon continuous compression. The effect of subphase pH and temperature on monolayer compressibility and phase transition was examined. Decrease of pH results in attenuation of LE–LC phase transition in Gln-PDA monolayers due to charge screening, while temperature affects the monolayers through regular thermal agitation, and the effect is opposite to the thermal anomaly observed for the non-derivatized diacetylene lipids. EFM studies reveal that only the lipids derivatized with hydrophilic headgroups exhibit phase transition and domain formation, while those with hydrophobic headgroups do not easily form polymerizable matrices. The shape and growth of the domains were found to be strongly headgroup dependent. Photo-polymerization of diacetylene lipid domains on water using EFM is discussed.

Marine Biology and Ecology Research Program, Extremobiosphere Research Center, Japan Agency for Marine-Earth Science and Technology, Yokosuka, Kanagawa 236-0061, Graduate School of Frontier Sciences, University of Tokyo, Kashiwa, Chiba 277-8562, Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, School of Marine Sciences and Technology, Tokai University, Shimizu-Orido, Shizuoka 424-8610, and Kazusa DNA Research Institute, Kisarazu, Chiba 292-0818, Japan

OBJECTIVE Although persistent hyperglycemia contributes greatly to the progression of diabetic micro- and macroangiopathy, microangiopathy progresses more rapidly than macroangiopathy in some type 2 diabetic patients, with the opposite being true in others. This study was conducted to identify factors responsible for such dissociation. RESEARCH DESIGN AND METHODS Patients with proliferative diabetic retinopathy and a carotid intima-media thickness (IMT) level ≤1.0 mm were classified as the microangiopathy group (MIG); those with an IMT level >1.1 mm and without retinopathy or with background retinopathy were assigned to the macroangiopathy group (MAG). Only middle-aged patients, 50–69 years old, were included in this study. There were 54 patients in the MIG and 68 patients in the MAG. RESULTS Patients in the MIG were significantly younger at the onset of diabetes, and those in the MAG had a significantly higher mean ratio of apoprotein (apo) B to apoAI. The percentage of patients with a family history of diabetes was significantly higher in the MIG. Maternal inheritance was common among these patients. Those with obesity, a family history of diabetes, and younger onset of hypertension were more common in the MAG. In the multiple logistic regression analyses, maternal inheritance and early onset of diabetes were independent risk factors for the acceleration of microangiopathy. A personal history of obesity and a family history of hypertension were independently related to the development of macroangiopathy. CONCLUSIONS Our results suggest that patients with early onset and maternal inheritance of diabetes may have a high risk for the progression of diabetic microangiopathy, while patients with hyperlipidemia, a history of obesity, and a family history of hypertension seem prone to the development of atherosclerosis.

The aim of the study was to examine the presence of psychological and mental health problems in patients with thalidomide embryopathy in Japan in order to develop and build future support systems.The present study examined the presence/absence of electroencephalographic abnormalities, intellectual/cognitive functions, and mental health problems in 22 participants (nine men, 13 women) with thalidomide embryopathy. Participants completed the electroencephalograph instrument. Participants were also assessed using the Wechsler Adult Intelligence Scale-III; the Autism-Spectrum Quotient; the General Health Questionnaire-28, and the Mini-International Neuropsychiatric Interview.The results suggest the following: (i) electroencephalographic abnormality observed in several thalidomide embryopathy participants is unlikely to be the direct result of thalidomide; (ii) the cognitive functions of working memory and processing speed are lower in thalidomide embryopathy patients than in healthy individuals; and (iii) 40.9% of the thalidomide embryopathy participants have possible mental disorders, with more mental problems observed than in healthy individuals.Deterioration of mental health in patients with thalidomide embryopathy is indicated. Anxiety, insomnia, and physical symptoms were especially remarkable and may have resulted in restriction of social activities. Therefore, careful examination and active support of patients' psychological and mental problems is essential.

Unresectable hepatic metastases from melanoma present a challenge to the physician. Although there have been advances in systemic therapies for patients with metastatic cutaneous melanoma to the liver, the overall prognosis is poor and is worse in patients with metastatic ocular melanoma. Isolated hepatic perfusion, open or percutaneous, allows high doses of cytotoxic chemotherapy to be delivered directly to the liver while minimizing systemic exposure, and has emerged as a viable treatment options for these complex patients.

Ocean-scale monitoring of pollution is challenging. Seabirds are useful indicators because they travel over a broad foraging range. Nevertheless, this coarse spatial resolution is not fine enough to discriminate pollution in a finer scale. Previous studies have demonstrated that pollution levels are higher in the Sea of Japan and South and East China Seas than the Northen Pacific Ocean. To test these findings in a wide-ranging animal, we tracked streaked shearwaters (Calonectris leucomelas) from four islands in Japan using global positioning system (GPS) and measured persistent organic pollutants (POPs) in the oil of their preen glands. The POPs did not change during 6 to 21 days when birds from Awashima were foraging only in the Sea of Japan, while it increased when they crossed to the Pacific through the Tsugaru Strait and foraged along the eastern coast of Hokkaido where industrial cities occur. These results indicate that POPs in the oil reflect relatively short-term exposure. Concentrations of POPs displayed greater variation among regions. Total polychlorinated biphenyls were highest in birds foraging in a small area of the semiclosed Seto Inland Sea surrounded by urbanized coast, p,p'-dichlorodiphenyltrichloroethane (DDT) was highest in birds foraging in the East China Sea, and total hexachlorocyclohexanes were highest in birds foraging in the Sea of Japan. All were lowest in birds foraging in the Pacific. This distribution of POPs concentration partly agrees with previous findings based on mussels, fish, and seawater and possibly reflects the mobility and emission sources of each type of POP. These results highlight the importance of information on the foraging area of highly mobile top predators to make them more effective monitors of regional marine pollution.

Summary Albatrosses are known to expend only a small amount of energy during flight. The low energy cost of albatross flight has been attributed to energy-efficient gliding (soaring) with sporadic flapping, although little is known about how much time and energy albatrosses expend in flapping versus gliding during cruising flight. Here, we examined the heart rates (used as an instantaneous index of energy expenditure) and flapping activities of free-ranging black-browed albatrosses (Thalassarche melanophrys) to estimate the energy cost of flapping as well as time spent in flapping activities. The heart rate of albatrosses during flight (144 beats min-1) was similar to that while sitting on the water (150 beats min-1). In contrast, heart rate was much higher during takeoff and landing (ca. 200 beats min-1). Heart rate during cruising flight was linearly correlated with the number of wing flaps per minute, suggesting an extra energy burden of flapping. Albatrosses expend only 4.6% ± 1.4% of their time in flapping during cruising flight, which was significantly lower than those during and shortly after takeoff (9.8% ± 3.5%). Flapping activity, which amounted to just 4.6% of the time in flight, accounted for 13.3% of the total energy expenditure during cruising flight. These results support the idea that albatrosses achieve energy-efficient flight by reducing the time spent in flapping activity, which is associated with high energy expenditure.

Background Esophageal cancer represents a major public health problem in the world. Several minimally invasive esophagectomy (MIE) techniques have been described and represent a safe alternative for the surgical management of esophageal cancer in selected centers with high volume and surgeons experienced in minimally invasive procedures. Methods The authors reviewed the most recent and largest studies published in the medical literature that reported the outcomes for MIE techniques. Results In larger series, MIE has proven to be equivalent in postoperative morbidity and mortality to the open esophagectomy. However, MIE has been associated with less blood loss, reduced postoperative pain, decreased time in the intensive care unit, and shortened length of hospital stay compared with the conventional open approaches. Despite limited data, no significant difference in survival stage for stage has been observed between open esophagectomy and MIE. Conclusions The myriad of MIE techniques complicates the debate for defining the optimal surgical approach for the treatment of esophageal cancer. Randomized controlled trials comparing MIE with conventional open esophagectomy are needed to clarify the ideal procedure with the lowest postoperative morbidity, best quality of life after surgery, and long-term survival.

Hydrothermal synthesis of ZnO nanorods from KOH and Zn(NO3)2 aqueous solutions was carried out at 673 K and 30 MPa for ca 0.03 s of reaction time with a flow apparatus without organic feed reagents. We examined effects of heat-up order of the starting solutions, concentrations, and flow rate on aspect ratio of ZnO crystals. Synthesized nanorods had an average length of 230 nm and width of 38 nm. The flow-through supercritical water (FT-SCW) method has a large potential for varying crystal morphology through control of nucleation and crystal growth in the heat-up, reaction, and cool-down cycles.

A certain chromosome of Aegilops triuncialis is known to cause chromosomal aberrations in common wheat. We studied how frequently the Ae. triuncialis chromosome induced chromosome mutations in chromosome 1R of rye which was substituted for chromosome 1B in a common wheat cultivar. Wheat-rye translocations and deletions in chromosome 1R were found in more than 10% of the plants examined, and most of them were stably transmitted in the subsequent generations. The possible use of this system for inducing wheat-alien translocations is discussed in relation to wheat breeding.

It becomes so important to observe a wild life for obtaining not only knowledge of its biological behaviors but also interactions with human beings in terms of geoenvironmental investigation and assessment. A sensor network is considered to be a suitable and powerful tool to monitor and observe a wild life in fields. In order to monitor/observe seabirds, a sensor network is deployed in Awashima island, Japan. A sensor platform is useful for early and quick deployment in fields. Atlas, a server — client type sensor platform, is used with several sensors, i.e., infrared sensors, thermometers within a nest, a sound sensor and network camera. Although the project has just started, useful data such as a leaving/returning pattern of seabirds are figured out by the sensor network.

Common wheat, Triticum aestivum, is an allohexaploid species consisting of three different genomes (A, B, and D). The three genomes were simultaneously discriminated with different colors. Biotinylated total genomic DNA of the diploid A genome progenitor Triticum urartu, digoxigenin-labeled total genomic DNA of the diploid D genome progenitor Aegilops squarrosa, and nonlabeled total genomic DNA of one of the possible B genome progenitors Ae. speltoides were hybridized in situ to metaphase chromosome spreads of Triticum aestivum cv. Chinese Spring. For detection, only two fluorochromes, fluorescein and rhodamine, were used. The A, B, and D genomes were simultaneously detected by their yellow, brown, and orange fluorescence, respectively. The genomic fluorescence in situ hybridization pattern of chromosome 4A of cv. Chinese Spring wheat showed that the distal 32% of the long arm was derived from a B genome chromosome. Furthermore, by using two highly repeated sequence probes, pSc 119.2 and pAsl, and two fluorochromes simultaneously, we were able to identify all B and D genome chromosomes and chromosomes 1A, 4A, and 5A of wheat.Key words: common wheat, in situ hybridization, multicolor fluorescence.

Objective. Primary cytoreductive surgery is well accepted in the initial management of ovarian cancer with a goal of maximal tumor reduction. The role of cytoreductive surgery at disease recurrence is controversial and guidelines are not standardized. We aimed to review cases of women with recurrent ovarian cancer who were collaboratively managed by two teams of oncologic surgeons with different areas of surgical expertise. Methods. A list of 616 patients with recurrent ovarian cancer from 1995 to 2009 was generated at a single institution. 20 cases of recurrent ovarian cancer were identified that were managed collaboratively. Data collected included date of diagnosis, initial treatment, recurrence date, location and number of sites of recurrence, secondary cytoreductive procedure performed, residual disease after surgery, pre-operative status, post-operative course, and pathologic findings. Results. Of the 20 cases that fit eligibility criteria, 11 were completely resected, 5 were incompletely resected, and 4 were biopsied only. Median disease-free interval following primary surgery was 18months (6–147). Median interval from diagnosis to collaborative cytoreduction was 63months (13–170). Our patients had metastatic disease to the liver (11), lymph nodes (8), the diaphragm (7), other locations including colon, pancreas, lung, adrenal, kidney (9). Two patients had additional miliary disease. All patients underwent joint surgical management by gynecologic and surgical oncologists. There were no deaths in the immediate post-operative period. The 5year survival rate was 45% following the joint surgical effort, with a median post-collaborative surgery survival duration of 42months. Conclusions. Previous studies document survival benefit of surgery for women with recurrent ovarian cancer when there has been a long disease-free interval, localized pelvic or intra-abdominal recurrences and an optimal performance status. Most gynecologic oncologists do not perform extensive liver or diaphragm resections or lymph node excision above the renal vessels; thus, collaboration with a surgical oncologist is a viable option. In this small descriptive study, the feasibility of this reasonably well-tolerated approach, with possible survival benefit, is documented.

International audience; Koalas operate on a tight energy budget and, thus, may not always display behavioral avoidance reaction when placed in a stressful condition. We investigated the physiological response of captive koalas Phascolarctos cinereus in a conservation centre to the presence of tourists walking through their habitat. We compared, using animal-attached data-recorders, the electrocardiogram activity of female koalas in contact with tourists and in a human-free area. One of the koalas in the tourist zone presented elevated heart rate values and variability throughout the recording period. The remaining female in the exhibit area showed a higher field resting heart rates during the daytime than that in the isolated area. In the evening, heart rate profiles changed drastically and both the koalas in the exhibit and in the tourist-free zones displayed similar field resting heart rates, which were lower than those during the day. In parallel, the autonomic nervous systems of these two individuals evolved from sympathetic-dominant during the day to parasympathetic-dominant in the evening. Our results report ECG of free-living koalas for the first time. Although they are preliminary due to the difficulty of having sufficient samples of animals of the same sex and age, our results stress out the importance of studies investigating the physiological reaction of animals to tourists.

Benzene and toluene are common organic solvents currently in worldwide industrial usage, which are metabolized mainly by hepatic cytochrome P450 2E1 (CYP2E1) in humans. Genetic polymorphism of CYP2E1 in 5'-flanking and coding regions has been found previously in Caucasian and Chinese populations. In this study, the effects of CYP2E1 alleles causing amino acid substitutions (CYP2E1*2, CYP2E1*3 and CYP2E1*4; wild-type, CYP2E1.1A) on benzene hydroxylation and toluene methylhydroxylation were studied using recombinant CYP2E1 enzymes of wild-type (CYP2E1.1) and variants (CYP2E1.2 having Arg76His, CYP2E1.3 having Val389Ile and CYP2E1.4 having Val179Ile) expressed in yeast cells. The K (m), V (max) and CL (int) values of CYP2E1.1 were 10.1 mM, 9.38 pmol/min/pmol CYP and 0.99 nL/min/pmol CYP for benzene hydroxylation, and 3.97 mM, 19.9 pmol/min/pmol CYP and 5.26 nL/min/pmol CYP for toluene methylhydroxylation, respectively. The K (m), V (max) and CL (int) values for benzene and toluene metabolism of CYP2E1.2, CYP2E1.3 and CYP2E1.4 were comparable to those of wild-type CYP2E1. These findings may mean that the polymorphic alleles of CYP2E1 causing amino acid substitutions are not directly associated with the metabolic activation of benzene and toluene. The information gained in this study should help to identify the variations in the toxicity of environmental pollutants.

Before the availability of hepatitis B immunoglobulin (HBIG) in hepatitis B-positive transplant recipients, the acute mortality was very high, in many centers up to 50% within 60 days post-transplant. The overall reinfection rate was approximately 60% within the initial 6 months, increasing to 80-90% within the initial 12 months and, in many cases, leading to allograft loss and death or retransplantation. These recurrent infections were often more severe and more rapidly progressing than the initial infection, probably due to high-dose immunosuppressive regimens. The poor prognosis before introduction of HBIG made hepatitis B liver disease an absolute contraindication for liver transplantation, leaving these patients with very limited treatment options. This changed in the late 1980s with the introduction of HBIG, which reduced the incidence of hepatitis B in the transplanted liver to approximately 15-50%, with concomitant improvement in graft and overall survival. The prognosis was further improved by a combination of long-term HBIG and antiviral therapy, in particular lamivudine, which reduced the reinfection rate, in most cases to between 0 and 5%. Owing to the cost and relative inconvenience of HBIG, some transplant centers have experimented with early discontinuation of HBIG and replacement with antiviral monotherapy. A number of studies, however, have found significantly higher recurrence rates associated with lamivudine monotherapy (40-50%) compared with combination therapy and, hence, lamivudine monotherapy is not recommended.

A terminal deletion in the short arm of chromosome 4B was obtained in the progeny of an alien monosomic addition line of common wheat (Triticum aestivum (L.) emend. Thell) with a chromosome from Aegilops cylindrica Host. Common wheat plants homozygous for the deletion were completely male sterile. The breakpoint was at about 84% of the length of the short arm from the centromere. Therefore, the male-fertility gene was located in the distal 16% region of the chromosome 4B short arm. This terminal deletion practically suppressed meiotic pairing between the short arms of the normal 4B and the deletion 4B chromosomes.

International audience; The characteristics of autonomic nervous activity were examined on captive great cormorants Phalacrocorax carbo hanedae, using a power spectral analysis of heart rate variability. Heart rates were calculated from recordings of the electrocardiograms of the birds via embarked data loggers. We investigated the effects of blockades of the sympathetic or parasympathetic nervous systems using the indices of autonomic nervous activity such as high frequency (0.061-1.5 Hz) component, low frequency (0.02-0.060 Hz) component and the low frequency power component to high frequency power component ratio. Resting heart rate (85.5 ± 6.1 bpm) was lower than the intrinsic heart rate (259.2 ± 15.3 bpm). The heart rate drastically increased after the injection of the parasympathetic nervous blocker, on the other hand it slightly decreased after the injection of the sympathetic nervous blocker. The sympathetic, parasympathetic and net autonomic nervous tones calculated from heart rate with and without blockades were 40.9 ± 27.6, -44.5 ± 7.4 and -29.5 ± 9.0%, respectively. The effect of the parasympathetic nervous blockade on low frequency and high frequency power was greater than that of the sympathetic nervous blockade. Those data suggested that the parasympathetic nervous activity was dominant for great cormorants.

Although the role of protein kinase C (PKC) has been suggested in agonist-induced bronchial smooth muscle contraction, the PKC isoform(s) involved in this phenomenon is not clear now. In the present study, the effects of three PKC inhibitors, GF1092603X, Gö6976 and rottlerin on acetylcholine (ACh)-induced bronchial smooth muscle contraction were examined to identify the PKC isoform(s) involved in the contraction. Bronchial smooth muscles were pretreated with each PKC inhibitor (10(-6) and 10(-5) M) 30 min before cumulative administration of ACh. In another series of experiments, the effects of PKC inhibitors on the maximal contraction induced by 10(-3) M ACh were determined: the inhibitors were cumulatively administered (10(-8)- 10(-5) M) after the ACh-induced contraction reached plateau. The ACh-induced bronchial smooth muscle contraction was significantly inhibited by GF109203X (inhibitor of PKC alpha, beta, gamma, delta and epsilon) but not by Gö6976 (PKC delta, beta and gamma inhibitor) and rottlerin (PKC delta and theta inhibitor). Moreover, mRNA and protein of PKC epsilon were detected in rat bronchial smooth muscle. Taken together, PKC epsilon might be involved in the ACh-induced bronchial smooth muscle contraction in rats.

Mucinous cystic neoplasms of the pancreas are rare tumors that tend to occur in young women. They are thought to be responsive to sex hormones. We report a case of a 32-year-old pregnant woman with a 7-month history of pain and a left upper quadrant abdominal mass. She arrived at the emergency department with exacerbation of her symptoms during week 15 of gestation. Findings from ultrasonography and magnetic resonance imaging suggested a pancreatic cyst. Percutaneous aspiration of the cyst fluid yielded mucinous fluid with an elevated carcinoembryonic antigen level. The patient underwent a spleen-preserving distal open pancreatectomy. We present herein a brief review of the current literature on mucinous cystic neoplasms during pregnancy. On the basis of our experience and the existing knowledge of this condition, we advocate resection during the second trimester with splenic preservation.

With the increasing usage of neoadjuvant chemotherapy (NAC) in locally advanced breast cancer (LABC), there is the need to investigate the routine axillary node dissections performed in this group of patients. Controversy exists about the utility of sentinel node biopsy (SNB), either before or after NAC. With the addition of trastuzumab in the treatment of Her2/neu-positive LABC patients, the validity of SNB in this subset population needs to be investigated. A retrospective study of 20 patients who underwent NAC for LABC was undertaken. The pathology of the axillary nodes, sentinel nodes, and primary tumor after neoadjuvant chemotherapy were examined. Twenty patients underwent NAC with doxorubicin and cyclophosphamide, followed sequentially by paclitaxel and carboplatin, with or without trastuzumab based on Her2/neu status. Post chemotherapy, 20 patients underwent mastectomy or lumpectomy with SNB with axillary node dissections. The overall accuracy of SNB was 95 per cent with a false-negative rate of 14 per cent (1/7). In Her2/neu-positive patients, overall accuracy was 100 per cent (8/8) and a false-negative rate of zero per cent. Sentinel node biopsy is a viable option in patients who have undergone NAC. Her2/neu-positive patients who had undergone NAC with trastuzumab had comparable accuracy for sentinel node biopsy in predicting axillary node status.

Hypothesis Dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) is a reliable and accurate method for monitoring primary tumor response in the breast and can be used as a surrogate to predict final axillary nodal status. Design Retrospective study (October 1, 2004, through February 28, 2006) of 46 patients with clinically staged locally advanced breast cancer. Setting Comprehensive cancer center. Patients Forty-six patients with locally advanced breast cancer. Interventions Neoadjuvant chemotherapy (NAC), DCE-MRI, mastectomy and lumpectomy, and axillary lymph node dissection. Main Outcome Measures The DCE-MRI results and pathologic response of the breast and axillary lymph nodes. Results Forty-six patients underwent NAC with doxorubicin hydrochloride and cyclophosphamide, followed by paclitaxel and carboplatin, with or without trastuzumab based on human epidermal growth factor receptor 2 (HER2/neu) status. Twenty-one patients (46%) had a complete pathologic response. For the HER2/neu-positive patients, the complete pathologic response rate was 70% (14/20). The accuracy, sensitivity, and specificity of the primary tumor response in predicting the axillary nodal status were 78%, 88%, and 72%, respectively. The accuracy, sensitivity, and specificity of the DCE-MRI–measured response in the primary tumor in predicting axillary nodal status were 74%, 62%, and 82%, respectively. For the HER2/neu-positive patients, the accuracy, sensitivity, and specificity improved to 80%, 75%, and 82%, respectively. Conclusions The results of DCE-MRI of the primary tumor can be predictive of axillary nodal status, especially in patients receiving trastuzumab who are HER2/neu positive. The HER2/neu-positive patients with a complete clinical response on DCE-MRI are highly unlikely to benefit from an axillary lymph node dissection. For HER2/neu-negative patients, sentinel lymph node sampling is warranted.

Cytochrome P450 2E1 (CYP2E1) is an enzyme of major toxicological interest because it metabolizes various drugs, precarcinogens and solvents to reactive metabolites. In this study, human and cynomolgus monkey CYP2E1 cDNAs (humCYP2E1 and monCYP2E1, respectively) were cloned, and the corresponding proteins were heterologously expressed in yeast cells to identify the functions of primate CYP2E1s. The enzymatic properties of CYP2E1 proteins were characterized by kinetic analysis of chlorzoxazone 6-hydroxylation and 4-nitrophenol 2-hydroxylation. humCYP2E1 and monCYP2E1 enzymes showed 94.3% identity in their amino acid sequences. The functional CYP content in yeast cell microsomes expressing humCYP2E1 was 38.4 pmol/mg protein. The level of monCYP2E1 was 42.7% of that of humCYP2E1, although no significant differences were statistically observed. The K(m) values of microsomes from human livers and yeast cells expressing humCYP2E1 for CYP2E1-dependent oxidation were 822 and 627 microM for chlorzoxazone 6-hydroxylation, and 422 and 514 microM for 4-nitrophenol 2-hydroxylation, respectively. The K(m) values of microsomes from cynomolgus monkey livers and yeast cells expressing monCYP2E1 were not significantly different from those of humans in any enzyme source. V(max) and V(max)/K(m) values of human liver microsomes for CYP2E1-dependent oxidation were 909 pmol/min/mg protein and 1250 nl/min/mg protein for chlorzoxazone 6-hydroxylation, and 1250 pmol/min/mg protein and 2990 nl/min/mg protein for 4-nitrophenol 2-hydroxylation, respectively. The kinetic parameter values of cynomolgus monkey livers were comparable to or lower than those of human liver microsomes (49.5-102%). In yeast cell microsomes expressing humCYP2E1, V(max) and V(max)/K(m) values for CYP2E1-dependent oxidation on the basis of CYP holoprotein level were 170 pmol/min/pmol CYP and 272 nl/min/pmol CYP for chlorzoxazone 6-hydroxylation, and 139 pmol/min/pmol CYP and 277 nl/min/pmol CYP for 4-nitrophenol 2-hydroxylation, respectively, and the kinetic parameters of monCYP2E1 exhibited similar values. These findings suggest that human and cynomolgus monkey CYP2E1 enzymes have high homology in their amino acid sequences, and that their enzymatic properties are considerably similar. The information gained in this study should help with in vivo extrapolation and to assess the toxicity of xenobiotics.