First evidence of asexual recruitment of Pocillopora acuta in Okinawa Island using genotypic identification.

Okinawa Island is located near the center of the Nansei Islands (∼24-31°N), at a relatively high latitude for coral reefs. Nevertheless, more than 80 coral genera (over 400 species) are abundant in the Nansei Islands. Since March, 2017, scleractinian corals have been held in an outdoor tank at the OIST Marine Science Station at Seragaki, Onna with natural sea water flow-through in order to be used in molecular biological and physiological studies. In January, 2018, we found small pocilloporid-like colonies suspected to have originated asexually. We collected 25 small colonies and measured their sizes and weights. Also, we validated the classification and clonality of the colonies using a mitochondrial locus and nine microsatellite loci. Almost all of the small colonies collected in the outdoor tank were ≤1 cm in both width and height. The weight of dried skeletons ranged from 0.0287 to 0.1807 g. Genetic analysis determined that they were, in fact, Pocillopora acuta. Only one mitochondrial haplotype was shared and two microsatellite multilocus genotypes were detected (20 colonies of one and four colonies of the other). The mitochondrial haplotype and one microsatellite multilocus genotype for 20 colonies corresponded to those of one P. acuta colony being kept in the tank. One small colony matched both multilocus genotypes. This may have been a chimeric colony resulting from allogenic fusion. These small colonies were not produced sexually, because the only potential parent in the tank was the aforementioned P. acuta colony. Instead, they were more likely derived from asexual planula release or polyp bail-out. Corals as Pocillopora acuta have the capacity to produce clonal offspring rapidly and to adapt readily to local environments. This is the first report of asexual reproduction by planulae or expelled polyps in P. acuta at Okinawa Island.

Ciliomotor circuitry underlying whole-body coordination of ciliary activity in the Platynereis larva.

Ciliated surfaces harbouring synchronously beating cilia can generate fluid flow or drive locomotion. In ciliary swimmers, ciliary beating, arrests, and changes in beat frequency are often coordinated across extended or discontinuous surfaces. To understand how such coordination is achieved, we studied the ciliated larvae of Platynereis dumerilii, a marine annelid. Platynereis larvae have segmental multiciliated cells that regularly display spontaneous coordinated ciliary arrests. We used whole-body connectomics, activity imaging, transgenesis, and neuron ablation to characterize the ciliomotor circuitry. We identified cholinergic, serotonergic, and catecholaminergic ciliomotor neurons. The synchronous rhythmic activation of cholinergic cells drives the coordinated arrests of all cilia. The serotonergic cells are active when cilia are beating. Serotonin inhibits the cholinergic rhythm, and increases ciliary beat frequency. Based on their connectivity and alternating activity, the catecholaminergic cells may generate the rhythm. The ciliomotor circuitry thus constitutes a stop-and-go pacemaker system for the whole-body coordination of ciliary locomotion.

An ancient role for nitric oxide in regulating the animal pelagobenthic life cycle: evidence from a marine sponge.

In many marine invertebrates, larval metamorphosis is induced by environmental cues that activate sensory receptors and signalling pathways. Nitric oxide (NO) is a gaseous signalling molecule that regulates metamorphosis in diverse bilaterians. In most cases NO inhibits or represses this process, although it functions as an activator in some species. Here we demonstrate that NO positively regulates metamorphosis in the poriferan Amphimedon queenslandica. High rates of A. queenslandica metamorphosis normally induced by a coralline alga are inhibited by an inhibitor of nitric oxide synthase (NOS) and by a NO scavenger. Consistent with this, an artificial donor of NO induces metamorphosis even in the absence of the alga. Inhibition of the ERK signalling pathway prevents metamorphosis in concert with, or downstream of, NO signalling; a NO donor cannot override the ERK inhibitor. NOS gene expression is activated late in embryogenesis and in larvae, and is enriched in specific epithelial and subepithelial cell types, including a putative sensory cell, the globular cell; DAF-FM staining supports these cells being primary sources of NO. Together, these results are consistent with NO playing an activating role in induction of A. queenslandica metamorphosis, evidence of its highly conserved regulatory role in metamorphosis throughout the Metazoa.

Psychosocial Determinants of Fruit and Vegetable Consumption in a Japanese Population.

There is limited evidence in Japan regarding the psychosocial determinants of fruit/vegetable intake. We performed a cross-sectional study of people aged 18 years or older in four regions of Japan; 2308 (men: 1012, women: 1296) individuals who completed the questionnaires were included. We found that 24.8% of people were aware of the current recommendations for vegetables and 13.2% for fruit and that "ability to design meals" and "availability when eating outside of the home" were the most important factors related to self-efficacy and barriers to fruit and vegetable intake, respectively. People with high self-efficacy (OR: 3.16; 95% CI: 2.17, 4.60 for fruit; OR: 4.52; 95% CI: 3.08, 6.64 for vegetables) were more likely to consume more fruit and vegetables. People with high scores on attitude (OR: 1.54; 95% CI: 1.06, 2.24) and social support (OR: 1.59; 95% CI: 1.11, 2.27) were more likely to consume more fruit. People with high perceived barriers (OR: 0.69; 95% CI: 0.48, 0.98) were less likely to consume fruit. This study suggests a need to increase the general population's awareness of the fruit and vegetable intake recommendations; facilitating positive attitudes, self-efficacy, and social support for individuals and strengthening the ability of individuals to design meals with more vegetables and fruit might be useful intervention programs.

Nitric oxide acts as a positive regulator to induce metamorphosis of the ascidian Herdmania momus.

Marine invertebrates commonly have a biphasic life cycle in which the metamorphic transition from a pelagic larva to a benthic post-larva is mediated by the nitric oxide signalling pathway. Nitric oxide (NO) is synthesised by nitric oxide synthase (NOS), which is a client protein of the molecular chaperon heat shock protein 90 (HSP90). It is notable, then, that both NO and HSP90 have been implicated in regulating metamorphosis in marine invertebrates as diverse as urochordates, echinoderms, molluscs, annelids, and crustaceans. Specifically, the suppression of NOS activity by the application of either NOS- or HSP90-inhibiting pharmacological agents has been shown consistently to induce the initiation of metamorphosis, leading to the hypothesis that a negative regulatory role of NO is widely conserved in biphasic life cycles. Further, the induction of metamorphosis by heat-shock has been demonstrated for multiple species. Here, we investigate the regulatory role of NO in induction of metamorphosis of the solitary tropical ascidian, Herdmania momus. By coupling pharmacological treatments with analysis of HmNOS and HmHSP90 gene expression, we present compelling evidence of a positive regulatory role for NO in metamorphosis of this species, in contrast to all existing ascidian data that supports the hypothesis of NO as a conserved negative regulator of metamorphosis. The exposure of competent H. momus larvae to a NOS inhibitor or an NO donor results in an up-regulation of NOS and HSP90 genes. Heat shock of competent larvae induces metamorphosis in a temperature dependent manner, up to a thermal tolerance that approaches 35°C. Both larval/post-larval survival and the appearance of abnormal morphologies in H. momus post-larvae reflect the magnitude of up-regulation of the HSP90 gene in response to heat-shock. The demonstrated role of NO as a positive metamorphic regulator in H. momus suggests the existence of inter-specific adaptations of NO regulation in ascidian metamorphosis.

Differences in heat shock protein 70 expression during larval and early spat development in the Eastern oyster, Crassostrea virginica (Gmelin, 1791).

For a variety of species, changes in the expression of heat shock proteins (HSP) have been linked to key developmental changes, i.e., gametogenesis, embryogenesis, and metamorphosis. Many marine invertebrates are known to have a biphasic life cycle where pelagic larvae go through settlement and metamorphosis as they transition to the benthic life stage. A series of experiments were run to examine the expression of heat shock protein 70 (HSP 70) during larval and early spat (initial benthic phase) development in the Eastern oyster, Crassostrea virginica. In addition, the impact of thermal stress on HSP 70 expression during these early stages was studied. C. virginica larvae and spat expressed three HSP 70 isoforms, two constitutive, HSC 77 and HSC 72, and one inducible, HSP 69. We found differences in the expression of both the constitutive and inducible forms of HSP 70 among larval and early juvenile stages and in response to thermal stress. Low expression of HSP 69 during early larval and spat development may be associated with the susceptibility of these stages to environmental stress. Although developmental regulation of HSP 70 expression has been widely recognized, changes in its expression during settlement and metamorphosis of marine invertebrates are still unknown. The results of the current study demonstrated a reduction of HSP 70 expression during settlement and metamorphosis in the Eastern oyster, C. virginica.

Immunohistochemical distribution of SP-D, compared with that of SP-A and KL-6, in interstitial pneumonias.

The immunohistochemical distribution of SP-D was compared with that of SP-A and KL-6 using a monoclonal antibody in lung tissues of 15 cases of collagen vascular disease-associated interstitial pneumonia (CVD-IP), 4 cases of hypersensitivity pneumonitis (CHP), and 6 cases of other diseases to determine their differences in distribution. In this study, the main targets were alveolar epithelial cells, especially those in the regenerating stage, as well as lymph vessels and stroma. The cytoplasm of type II alveolar epithelial cells and Clara cells was positive for SP-D, with sharp margins; interestingly, however, during the process of regeneration large positive cells were intermingled with relatively small negative cells, even in the same row of cells. In sharp contrast, staining for SP-A and KL-6 was positive in the cytoplasm of all the regenerating alveolar epithelial cells, as well as Clara cells. Staining for KL-6 was usually positive in the surface of air spaces in linear fashion. Staining for SP-A was also positive in elastic fibers in vascular walls. In areas of destruction of pulmonary structures, loose stroma and the endothelial cells of lymph vessels as well as their contents were distinctly positive for SP-A and/or KL-6 but not SP-D. Judging from these results in pulmonary tissues of CVD-IP and HP, SP-D might be a marker for maturity of regenerating epithelial cells. Both SP-A and KL-6 were detected in intimate relationship to the stage of regeneration of alveolar epithelial cells and were expressed before SP-D. In addition, the lymph vessels play a very important role in transfer of KL-6 into the bloodstream.

Detection method for genetically modified papaya using duplex PCR.

A simple and rapid method for the identification of genetically modified (GM) papaya, derived from Line 55-1, was developed by modifying the Japanese official PCR method. Genomic DNA was directly extracted from the fresh fruit without the lyophilization step, using a commercial silica-based kit. To develop a duplex PCR method which simultaneously detects the GM papaya-specific gene and the intrinsic papain gene, the papain 2-5'/3' (amplicon size; 184 bp) primer pair for the detection of the papain gene was newly designed within the region of the products (211 bp) amplified using the papain 1-5'/-3' primer pair adopted in the Japanese official PCR method. To detect the GM papaya-specific gene, the primer pair Nos C-5'/CaM N-3' described in the Japanese official method was used. The DNA sequences of the GM papaya gene and the intrinsic papain gene were co-amplified using the PCR method in a single tube. The developed duplex PCR method allows the simultaneous detection of the products by means of agarose gel electrophoresis or microchip electrophoresis. The proposed method for GM papaya identification is simple and rapid.

[Detection of short tandem repeat (STR) polymorphisms by microchip electrophoresis for individual identification of cattle].

A simple and rapid detection of short tandem repeat (STR) markers was studied as a screening test for individual identification of cattle. DNAs were extracted from eight commercial beef samples by a proteinase K-boil method followed by purification with 2-propanol precipitation. Five STR markers, known to be highly polymorphic, were amplified by PCR and analyzed both by a conventional sequencing analysis (SEQ) and by a proposed microchip electrophoresis (MEP). Every marker revealed high polymorphism, such as 5-9 alleles in SEQ analysis, and 4-6 alleles in MEP analysis. This simple and rapid MEP analysis is expected to be an effective screening tool with use of confirmatory SEQ analysis.

[Measurement of peak inspiratory flow with asthma: effect of inhaled beta2-stimulant].

This study was designed to evaluate peak inspiratory flow (PIF) and peak expiratory flow (PEF) in 24 patients with mild asthma. After inhalation of a beta2-stimulant (beta2), PIF significantly increased from 173.0 +/- 67.0 (range 70-300 L/min) to 194.0 +/- 66.7 after 1 minute, and to 199.3 +/- 63.0 after 15 minutes (p < 0.0025, and p < 0.008, respectively). PEF also significantly increased. The previous inhalation of beta2 improved the efficacy of inhalation of dry powder. This evidence should be considered in performing patient education for effective methods of inhalation.

[Effect of oral beta2-stimulant on pulmonary function in asthma patients treated with multiple medications].

This study was designed to evaluate the effect of discontinuing oral beta2-stimulant in patients with asthma who were being treated with multiple medications. Thirty-two asthmatics controlled under multiple medications who had a stable PEF were entered. Patient symptoms, PEF, FEV10, V75,V50, and V25 were evaluated before and after discontinuing beta2-stimulant. Results showed that after discontinuing beta2-stimulant, there was little change in symptoms, PEF, or FEV10, and only two patients had to be re-medicated with oral beta2-stimulant. However, deterioration of V50 and V25 were clearly observed, suggesting that oral beta2-stimulant had an affect on dilatation of the small airway. Based on our data, we should regard that when discontinuing oral beta2-stimulant from combined use, the lung function reflecting the small airway decreases even if no change of symptoms is observed.

[Acute effect of inhaled beta(2)-agonists with different type of intrinsic activity on airway resistance in healthy volunteers].

Inhaled beta(2)-agonists (long-acting as well as short acting) are used world-wide for the relief of asthma symptoms. However, there are few reports which have evaluated the additive effect of short-acting beta(2)-agonists to long-acting beta(2)-agonists on airway resistance measured by a plethysmography. This study was designed to evaluate the additive effect of inhaled short-acting beta(2)-agonists (protecarol) to long-acting beta(2)-agonists (salmeterol) on airway resistance in normal healthy volunteers (S+P group). In addition, to compare the effects of beta(2)-agonists which have different types of intrinsic activities, acute effect of inhaled procaterol adding to procaterol was also evaluated (P+P group). Seven healthy volunteers (all male and all non-smokers) were entered in this study. Pulmonary function was measured by a body plethysmography. Forced expiratory volume per 1 second (FEV1), the maximum flow rate at 25% (V(.) 25), the maximum flow rate at 50% of forced vital capacity (V(.) 50), and airway resistance were measured before and after inhalation of salmeterol (1 dry powder, 50 microg) or procaterol (2 puffs, 20 microg). Sixty minutes after inhalation of salmeterol, or 15 minutes after inhalation of procaterol, inhalation of procaterol (2 puffs, 20 microg) was added, and then pulmonary function was monitored. FEV1, V(.) 25, and V(.) 50 were significantly increased after inhalation of salmeterol as well as procaterol. In addition, airway resistance decreased significantly after inhalation of salmeterol as well as procaterol. In the S+P group, additional decrease of airway resistance after inhalation of procaterol was relatively small compared with the P+P group. In conclusion, although additional bronchodilatoric effects were observed in the S+P and P+P group, the effects seemed to be different based on the intrinsic activity of each beta(2)-agonist.

[Beta 2-adrenoceptor polymorphism and effects of inhaled beta 2-stimulant (procaterol) and an anti-cholinergic drug (oxitropium) on the airway resistance].

It has been suggested that the Arg 16/Gly 16 allele at codon 16 of beta 2-adrenoceptor polymorphism plays a role in down-regulating the stimulus of bronchodilatation caused by beta 2-agonists. This study was designed to evaluate the difference of bronchodilator responsiveness to beta 2-agonist (procaterol) and anti-cholinergic drug (oxitropium) between those who have Arg 16/Gly 16 allele (hetero type) and those who have Gly 16/Gly 16 allele (variant type) at codon 16 of in healthy women. Airway resistance and other pulmonary function tests were measured by a body plethysmography before and 5, 10, 15, 20, and 30 minutes after inhalation of procaterol or inhalation of procaterol and oxitropium. In healthy women inhaled procaterol, percent changes of respiratory airway resistance compared with values before inhalation were -2.8 after 5 minutes, -7.5 after 10 minutes, -11.2 after 15 minutes, -15.4 after 20 minutes, and -12.6 after 30 minutes. In healthy women inhaled porcaterol and oxitropium, percent changes of respiratory airway resistance compared with values before inhalation were -14.5 after 5 minutes, -18.9 after 10 minutes, -17.0 after 15 minutes, -20.8 after 20 minutes, and -20.4 after 30 minutes. Patterns of decrease of respiratory airway resistance differed between women who have Arg 16/Gly 16 allele (hetero type) and those who have Gly 16/Gly 16 allele (variant type). In women who have Gly 16/Gly 16 allele (variant type), although acute decrease of respiratory airway resistance was observed, the duration of bronchodilator effect by inhaled procaterol and oxitropium was shorter than those observed in Arg 16/Gly 16 allele (hetero type). The present study showed inhalation of procaterol and oxitropium had a differential bronchodilator effect in healthy women, depending on their genotype of beta 2-adrenoceptor polymorphism at codon 16.

Measurement of inferior vena cava diameter for evaluation of venous return in subjects on day 10 of a bed-rest experiment.

We evaluated the usefulness of measurements of the inferior vena cava (IVC) diameters on abdominal echograms as an indicator of changes of venous return in subjects with orthostatic intolerance (OI) induced by simulated microgravity. We performed a standing test and recorded the IVC diameters on abdominal echograms in 12 subjects placed on a 20-day 6 degrees head-down-tilt bed-rest experiment. We found that different patterns of changes in IVC diameter occurred in the standing test on day 10 of the experiment; in five subjects with a marginal decrease in pulse pressure, IVC diameters in the upright position were markedly decreased compared with those in the supine position. In five subjects with feelings of discomfort, the IVC diameters in the upright position distended or did not decrease from those in the supine position. These results suggested that the changes in IVC diameter on the standing test indicated the presence of various types of hemodynamic responses of OI caused by simulated microgravity. In this study, we also evaluated changes in body-water compartments by conducting multifrequency bioelectrical impedance analysis. Longitudinal data analysis showed that the total body-water-to-fat-free mass and extracellular fluid-to-fat-free mass ratios decreased during the experimental period and recovered thereafter, and that the ratio of intracellular fluid to fat-free mass decreased during the experiment. No significant difference in changes in body-water compartments was seen among subjects with different patterns of changes in IVC diameters. Measurement of IVC diameter was useful to estimate hemodynamic changes in subjects with OI.

[A case of carcinomatous lymphangitis of the lungs due to stomach cancer responding to TS-1].

We report a case of carcinomatous lymphangitis of the lungs due to and stomach cancer showing remarkable response to TS-1. The patient was a 51-year-old man whose chest x-ray and computed tomography (CT) revealed lymphangitis, and endoscopic examination showed stomach cancer on posterior wall of stomach body. Bone marrow metastasis was suspected because platelet count was 50/microliter, and myelocytes and metamyelocytes emerged in peripheral blood. TS-1 80 mg/day was administered orally for 28 days as 1 course. After 4 courses of TS-1, chest x-ray showed remarkable improvement, and platelet count was normalized. The patient survived for 10 months after the first visit. We suggest that TS-1 is an effective therapy for carcinomatous lymphangitis of the lungs due to stomach cancer.

[Case of pulmonary tuberculosis in late stage of pregnancy].

A 28-year-old woman who was a nurse was admitted to our hospital because her sputum was positive for M. tuberculosis. She was pregnancy of 35 weeks. First, she was administered INH, RFP, PZA and was treated with cesarean section on the 21st day after starting tuberculosis chemotherapy. The operation was done in operating room of negative pressure ventilation. The patient returned to the tuberculosis ward, and the newborn infant entered to a newborn nursery room after confirming negative tubercle bacilli in amnionic fluid by PCR examination. EB was added to the regimen of chemotherapy after childbirth. In general hospitals, infection control is an important issue as seen in this case.