Intraspecific variation in photosynthetic thermal acclimation in Fagus crenata seedlings from two populations growing at different elevations in northern Japan.

Plants can acclimate their photosynthesis to growth temperature, but the contribution of local adaptation to intraspecific variation in thermal acclimation of photosynthesis is not fully understood. Here, we experimentally investigated the photosynthetic thermal acclimation in Fagus crenata Blume seedlings from two populations growing at different elevations and temperature regimes (low- and high-elevation sites) in northern Japan. We acclimated seedlings for 14 to 23 days at daytime temperatures of either 22 °C (control) or 27 °C (warm treatment) and obtained photosynthetic temperature-response curves in the range of 19 to 32 °C. The optimum temperature of photosynthesis (Topt) was ~0.6 °C higher in seedlings acclimated at 27 °C than in those acclimated at 22 °C, and it was significantly lower in seedlings with higher stomatal sensitivity to leaf-to-air vapor pressure deficit than in those with lower sensitivity. The effects of warm treatment, population and treatment-population interaction on Topt were not significant in the two-way analysis of variance, but the effect of treatment became significant when stomatal sensitivity to leaf-to-air vapor pressure deficit was included as a covariate in the model. Structural equation modeling indicated that seedlings with lower root biomass had lower Topt because of the high stomatal sensitivity to leaf-to-air vapor pressure deficit. Structural equation modelling also indicated that the way of shifting the Topt differed between the two populations: seedlings from a high-elevation site depended on decreasing photosynthetic rates at low temperatures for the increase in Topt but seedlings from a low-elevation site did not. We suggest that the difference in thermal acclimation of photosynthesis between the two populations may reflect adaptation to different climate regimes and that belowground traits should be considered when investigating thermal acclimation capacity, especially in seedlings.

Rapid analysis of GBSS1 and Vinv genes expressed in potato tubers using microtubers produced in liquid culture medium.

KEY MESSAGE: This study established a rapid method for the gene expression analysis in potato tubers. The use of microtubers would be useful for primary evaluation of tuber-expressed genes. In the development of transgenic potato or of potato with other genome modifications (e.g., genome editing or RNA-directed DNA methylation (RdDM) and so on) to improve tuber traits, analysis of the target gene is often difficult because of the long cultivation cycle (3-4 months), large areas required, numerous materials for plant cultivation, and considerable efforts needed to obtain transgenic tubers. We demonstrate here rapid and convenient analysis of gene expression in potato microtubers. Enough microtubers for expression analysis can be induced over about 4 weeks in a simple liquid medium in an Erlenmeyer flask. High-quality RNA and protein can be easily prepared from microtubers and used for northern blot, qRT-PCR, and western blot analyses without further purification. We investigated the expression of two tuber-expressed genes (GBSS1 and Vinv) in microtubers derived from the wild-type and from lines derived from RdDM-mediated transcriptional gene silencing. As expected, the expression of both genes was similar between microtubers and normal tubers. Furthermore, we demonstrated that microtubers can be used in western blot and confocal immunofluorescent microscopy analyses. These results suggest that expression analysis using microtubers is a convenient tool for the analysis of tuber-expressed genes such as GBSS1 and Vinv in potato.

Assessments of fine-scale spatial patterns of SNPs in an old-growth beech forest.

The spatial patterns of non-neutral genetic variations at fine spatial scales and their possible associations with microenvironments have not been well-documented for tree populations. Based on 25-32 SNP markers, we examine whether non-neutral SNPs and their associations with microenvironments can be detected in FcMYB1603, a gene homologous to that encoding a protein induced by drought stress in Arabidopsis thaliana for the 166 adult trees in a 1-ha plot in a mature population of Fagus crenata. In the 83 individuals of a younger cohort of below canopy trees, the nonsynonymous SNP at locus FcMYB1603_684 exhibited a spatial signature representing a departure from the expected spatial patterns of neutral genetic variation. Evaluations of non-neutrality for this locus were robust against the potential risks of false positives due to the low number of SNP loci, a low criterion set for minor allele frequency, and any edge effect on the trees' spatial structure. An older cohort exhibited no signal of the existence of non-neutral genetic variation, suggesting that temporal fluctuation in the microenvironmental conditions on the forest floor may have exposed different cohorts to different magnitudes of selection pressure. Although genotypes of the locus showed a spatial association with a microenvironmental variable potentially related to soil moisture, the present study was subject to a limitation due to the generally low polymorphism of nonsynonymous loci within the single plot, which suggests that it will be important to replicate the study design in order to carry out research on fine-scale non-neutral genetic variations.

Methanobacterium kanagiense sp. nov., a hydrogenotrophic methanogen, isolated from rice-field soil.

A pure culture of an obligately anaerobic, hydrogenotrophic, methanogenic archaeon, designated strain 169(T), which grows with hydrogen and carbon dioxide as the sole energy and carbon sources, was isolated from an anaerobic propionate-oxidizing enrichment culture originally obtained as an inoculant from rice-field soil in Japan. Cells of strain 169(T) were non-motile, Gram-reaction-variable and rod-shaped or slightly curved rods with rounded ends (1.6-5.0 × 0.35-0.5 µm). Strain 169(T) had fimbriae at both ends of the cell (up to ~10 per cell) but did not possess flagella. Ultrathin sections showed a single-layered, electron-dense cell wall about 6 nm thick, which is typical of Gram-positive bacteria. Growth was observed at 15 °C-45 °C (optimum 40 °C), at pH  6.5-9.6 (optimum pH 7.5-8.5) and in 0-70 g NaCl l(-1) (0-1.2 M) (optimum 5 g NaCl l(-1); 0.086 M). Strain 169(T) utilized only hydrogen and carbon dioxide as energy and carbon sources. The DNA G+C content was 39.3 mol%. The results of 16S rRNA gene sequence analysis indicated that strain 169(T) was most closely related to Methanobacterium subterraneum DSM 11074(T) (96.8 % sequence similarity) and Methanobacterium formicicum DSM 1535(T) (96.4 %). On the basis of its morphological, physiological and phylogenetic characteristics, strain 169(T) ( = DSM 22026(T) = JCM 15797(T)) represents a novel species of the genus Methanobacterium, for which the name Methanobacterium kanagiense sp. nov. is proposed.

Noninvasive positive pressure ventilation in patients with perioperative negative pressure pulmonary edema.

Negative pressure pulmonary edema (NPPE) is a noncardiogenic pathological process that is treated with invasive ventilation via a tracheal tube. To investigate the feasibility and safety of noninvasive positive pressure ventilation (NPPV) as an alternative treatment for NPPE, we retrospectively reviewed charts of 15 perioperative NPPE patients. Eight patients were treated by NPPV and 7 were treated by invasive ventilation. Patient characteristics, duration of NPPV, duration of intensive care unit (ICU) stay, and maximum airway pressure were investigated for the NPPV-treated patients. All patients treated by NPPV had a patent airway after complete relief of the airway obstruction and recovered from NPPE symptoms within one postoperative day. Arterial blood gas analysis showed a significant improvement in the PaO(2)/FiO(2) ratio from 132 + or - 30 mmHg in the operating room to 282 + or - 77 mmHg at discontinuation of NPPV. Serious complications, such as ventilator-associated pneumonia or aspiration pneumonia, did not occur, and intubation was not required for any patient. Favorable outcomes in these cases suggest that NPPV could be a feasible and safe alternative for treating NPPE if the patency of the airway is restored.

Postoperative intensive care unit drug fever caused by dexmedetomidine.

Dexmedetomidine hydrochloride is a potent, highly selective alpha-2 adrenergic receptor agonist, broadly used as a sedative drug in intensive care units. We describe the case of a 59-yr-old patient who experienced drug fever caused by dexmedetomidine hydrochloride. The patient was transferred to the intensive care unit with an abdominal aortic aneurysm rupture. After initiation of sedation with dexmedetomidine hydrochloride, he developed pyrexia of more than 39 degrees C. This symptom improved rapidly 7 h after stopping dexmedetomidine hydrochloride. Other possible causes (such as infection) were sequentially eliminated.

Factors predicting successful noninvasive ventilation in acute lung injury.

PURPOSE: Noninvasive ventilation (NIV) has been successfully used to treat various forms of acute respiratory failure. It remains unclear whether NIV has potential as an effective therapeutic method in patients with acute lung injury (ALI). The aims of this study were to determine factors predicting the need for endotracheal intubation in ALI patients treated with NIV, and to promote the selection of patients suitable for NIV. METHODS: We conducted a retrospective study of all patients admitted to the intensive care unit (ICU) of the Nippon Medical School Hospital from 2000 to 2006 with a diagnosis of ALI, in whom NIV was initiated. RESULTS: A total of 47 patients with ALI received NIV, and 33 patients (70%) successfully avoided endotracheal intubation. Patients who required endotracheal intubation had a significantly higher Acute Physiology and Chronic Health Evaluation (APACHE) II score and a significantly higher Simplified Acute Physiology Score (SAPS) II, and a significantly lower arterial pH. The respiratory rate decreased significantly within 1 h of starting NIV only in patients successfully treated with NIV. An APACHE II score of more than 17 (P = 0.022) and a respiratory rate of more than 25 breaths x min(-1) after 1 h of NIV (P = 0.024) were independent factors associated with the need for endotracheal intubation. Patients who avoided endotracheal intubation had a significantly lower ICU mortality rate and in-hospital mortality rate than patients who required endotracheal intubation. CONCLUSION: We determined an APACHE II score of more than 17 and a respiratory rate of more than 25 breaths x min(-1) after 1 h of NIV as factors predicting the need for endotracheal intubation in ALI patients treated with NIV.

The efficacy of dexmedetomidine in patients with noninvasive ventilation: a preliminary study.

BACKGROUND: Agitation is associated with failure of noninvasive ventilation (NIV). We investigated the effect of dexmedetomidine in patients with NIV. METHODS: This was a prospective clinical investigation in an intensive care unit. Dexmedetomidine was infused in 10 patients in whom NIV was difficult because of agitation. RESULTS: Ramsay and Richmond Agitation-Sedation Scale scores were maintained at 2.94 +/- 0.94 and -1.23 +/- 1.30, respectively. All patients were successfully weaned from NIV, and the respiratory state was not worsened. CONCLUSION: This study shows that dexmedetomidine is an effective sedative drug for patients with NIV.

Clinical role and efficacy of landiolol in the intensive care unit.

Beta-adrenergic receptor blockers have proved to be effective for the management of various cardiovascular diseases and the prevention of perioperative cardiac events and cerebrovascular accidents. Landiolol is a short-acting beta-blocker, with high beta 1-selectivity and a short duration of action. We thought landiolol was valuable and suitable for intensive care unit (ICU) patients, and conducted a retrospective study. The records of 80 patients (58 post-surgical patients; group S and 22 internal medicine patients; group IM) were reviewed. Thirty-seven (64%) of the group S patients were post-coronary artery bypass graft surgery, and the IM group consisted mostly of patients with acute myocardial infarction. The most common indication for landiolol in group S was the prevention of myocardial ischemia (50%), and in group IM, it was atrial fibrillation (45%). The median infusion rate of landiolol was 5 microg.kg(-1).min(-1) and the median infusion time was 2 days. Twenty-six patients were continued on oral beta-adrenergic receptor blockers. Landiolol reduced heart rate significantly without reducing blood pressure, and stabilized hemodynamics. We confirmed that landiolol is valuable as a bridge to starting oral beta-adrenergic receptor blockers and as an anti-arrhythmic agent, and that it is suitable for ICU patients due to its high beta 1-selectivity and rapid onset and offset of action.

Isolation and characteristics of methanosaeta in paddy field soils.

The population of filamentous acetate-utilizing methanogens in paddy field soils was 2.0 x 10(4) MPN/g dry soil in the submerged condition. They were able to form colonies in a deep agar medium, but not in a roll tube. Filamentous acetate-utilizing methanogens isolated from Kanagi, Japan (strain K-5) and Tsukuba, Japan (strain T-3) were divided into two types based on length of filaments. One type, strain K-5, formed a short chain which was dispersed easily by weak shaking. The other type, strain T-3, formed a long chain, which formed cotton-like flocs and was not dispersed by weak shaking. They had sheaths composed of a pair of adjacent membranes on the outside of the cell membranes. The 16S rRNA gene similarities of strain T-3 and K-5 to Methanosaeta concilii strain Opfikon were 100% and 99.5% respectively. Filamentous acetate-utilizing methanogens were also isolated from paddy field soils in various other regions of Japan. Our results suggest that Methanosaeta is universal in paddy soils and that it plays an important role in methane production from acetate.

A case of cardiac tamponade following esophageal resection.

We report a rare case of cardiac tamponade after esophageal resection for esophageal cancer. A 69-year-old man underwent subtotal esophagectomy and reconstruction of the gastric tube with cervical anastomosis via the poststernal route and three-field lymphadectomy via a median sternotomy. On postoperative day 4, the patient developed dyspnea, chest oppression, and hemodynamic instability due to cardiac tamponade. Emergency percutaneous catheter drainage was performed to manage the cardiac tamponade. Acute pericarditis due to the original surgical procedure was suspected to be the cause of the tamponade. Although rare, cardiac tamponade should be considered as a cause of hemodynamic instability after esophageal resection.

Cannabinoid suppressed bicuculline-induced convulsion without respiratory depression in the brainstem-spinal cord preparation from newborn rats.

Previous studies have suggested that cannabinoid compounds are anticonvulsants and that these compounds depress respiratory activity. However, the anticonvulsant potential of cannabinoids and their depressive effect on respiration have not been evaluated simultaneously. In the present study, we used a brainstem-spinal cord preparation model to investigate changes in inspiratory activity and the anticonvulsant effects of a cannabinoid receptor agonist, WIN 55, 212-2, in bicuculline-induced convulsion. Application of 10 microM WIN 55, 212-2 caused no change in inspiratory activity (6.9+/- 0.89 bursts/min vs. 8.0+/- 1.3 bursts/min, not significant) and decreased bicuculline-induced seizure-like nerve activity (number of seizure-like activities in 10 min, 11+/- 7.4 bursts vs. 1.5+/- 1.6 bursts, P< 0.01; average duration of seizure-like activity, 8.9+/- 4.0 sec vs. 4.7+/- 2.1 sec, P> 0.01). Our results suggest that administration of an appropriate dose of cannabinoid receptor agonist WIN 55,212-2 has an anticonvulsant effect but does not cause respiratory depression.

Characteristics of an anaerobic, syntrophic, butyrate-degrading bacterium in paddy field soil.

The number of syntrophic butyrate-degrading bacteria in a flooded paddy field soil was 1.7 x 10(3) MPN/g dry soil. Butyrate was degraded to acetate and methane when paddy soils were incubated anaerobically with the addition of butyrate. However, butyrate degradation was completely suppressed by the addition of the specific inhibitor of methanogenesis, 2-bromoethanesulfonate (BES) to the soil. A hydrogen-using methanogen, strain TM-8, was isolated from flooded paddy field soil. Strain TM-8 was identified as Methanobacterium formicicum based on its physiology and phylogeny. Syntrophic butyrate-degrading bacteria were enumerated and isolated using strain TM-8. A syntrophic butyrate-degrading bacterium, strain TB-6, was isolated in coculture with strain TM-8 from paddy soil. The strain was Gram-negative, had curved rods, and grew on crotonate. Sulfate was not used as an electron acceptor. Strain TB-6 was closely related to S. wolfei subsp. wolfei. The relation between strain TB-6 and the members of Syntrophomonas are discussed.

Salicylate action on medullary inspiratory neuron activity in a brainstem-spinal cord preparation from newborn rats.

Salicylate affects central respiratory control. The inspiratory neurons are the most important component of the medullary respiratory control center because they modulate the final motor output via the phrenic nerve. We investigated changes in burst rate, intraburst firing frequency, and membrane properties of inspiratory neurons in the isolated brainstem after the administration of salicylate. Newborn rat brainstem-spinal cord preparations were superfused with salicylate. Whole-cell recordings were performed from inspiratory neurons. Application of 1 mM salicylate caused an increase in the inspiratory neuronal burst rate from 6.9 +/- 1.6 bursts/min to 8.2 +/- 1.9 bursts/min (P < 0.05). The inspiratory neuron burst rate decreased from 8.3 +/- 0.7 bursts/min to 4.5 +/- 1.1 bursts/min after the application of 10 mM salicylate (P < 0.01). The depressant effect of 10 mM salicylate was antagonized by the gamma-aminobutyric acid (GABA) receptor antagonist bicuculline (1 microM). Resting membrane potential and intraburst firing frequency did not change with the application of salicylate and bicuculline even when the burst rate did change. We conclude that the effects of salicylate on the medullary inspiratory neurons are mainly due to a presynaptic action. GABAergic mechanisms are probably involved in the salicylate-induced central respiratory depression.

Isolation of a subfamily of genes for R2R3-MYB transcription factors showing up-regulated expression under nitrogen nutrient-limited conditions.

Plant R2R3-MYB transcription factors are encoded by more than 100 copies of genes. In this study, we attempted to isolate some members of the R2R3-MYB superfamily involved in regulation of nitrogen fixation in legumes. A library of 300 recombinant plasmid clones containing the R2R3-MYB fragments of the superfamily was screened by differential hybridization to isolate R2R3-MYB genes whose expression was up-regulated under nitrogen nutrient-limited conditions. Two groups of clones were identified, each of which seemed to represent a gene responsive to nitrogen starvation. The entire coding regions for the genes were further isolated by PCR and were designated LjMYB101 and LjMYB102. By screening a genomic library of Lotus japonicus with a probe derived from LjMYB101, the third gene, LjMYB103, was isolated. In addition, a candidate for the soybean orthologue of LjMYB101 was isolated and designated GmMYB101. Sequence alignment of the genes with members of the plant R2R3-MYB superfamily showed that they all belonged to the subgroup 10 of the superfamily. The expression analysis of the genes showed that the organ-specific and nitrate-regulated expression profile of MYB101 was very similar to that of CHS in Lotus as well as in soybean, suggesting a possible role for MYB101 in regulation of flavonoid biosynthesis in response to nitrate starvation. On the other hand, an interesting relationship, in structure and function, was found between LjMYB101 and LjGln1, suggesting an alternative role for MYB101 in regulation of nitrogen metabolism.

Sequence divergence at chalcone synthase gene in pigmented seed coat soybean mutants of the Inhibitor locus.

Seed coat color in soybeans is determined by the I (Inhibitor) locus. The dominant I allele inhibits seed coat pigmentation, and it has been suggested that there is a correlation between the inhibition of pigmentation by the I allele and chalcone synthase (CHS) gene silencing in the seed coat. Analysis of spontaneous mutations from I to i has shown that these mutations are closely related to the deletion of one of the CHS genes (designated ICHS1). In soybeans with the I/I genotype (cv. Miyagi shirome), a truncated form of the CHS gene (CHS3) is located in an inverse orientation 680 bp upstream of ICHS1, and it was previously suggested that the truncated CHS3- ICHS1 cluster might be involved in CHS gene silencing in the seed coat. In the current study, the truncated CHS3- ICHS1 cluster was compared with the corresponding region of pigmented seed coat mutants in which I had changed to i in Miyagi shirome and in the strain Karikei 584. In the Karikei 584 mutant, the truncated CHS3-ICHS1 cluster was retained and the sequence diverged at a point immediately upstream (32 bp) of this cluster. The sequences upstream of the points of divergence in both mutants almost perfectly matched a part of the registered sequence in a soybean BAC clone containing the soybean cyst nematode resistance-associated gene, and inspection of the sequences suggested that the sequence divergence of the CHS gene in the Karikei 584 and Miyagi shirome mutants was due to an unequal crossing-over via 4-bp or 5-bp short repeats, respectively.