Frontiers of torenia research: innovative ornamental traits and study of ecological interaction networks through genetic engineering

Advances in research in the past few years on the ornamental plant torenia (Torenia spps.) have made it notable as a model plant on the frontier of genetic engineering aimed at studying ornamental characteristics and pest control in horticultural ecosystems. The remarkable advantage of torenia over other ornamental plant species is the availability of an easy and high-efficiency transformation system for it. Unfortunately, most of the current torenia research is still not very widespread, because this species has not become prominent as an alternative to other successful model plants such as Arabidopsis, snapdragon and petunia. However, nowadays, a more global view using not only a few selected models but also several additional species are required for creating innovative ornamental traits and studying horticultural ecosystems. We therefore introduce and discuss recent research on torenia, the family Scrophulariaceae, for secondary metabolite bioengineering, in which global insights into horticulture, agriculture and ecology have been advanced. Floral traits, in torenia particularly floral color, have been extensively studied by manipulating the flavonoid biosynthetic pathways in flower organs. Plant aroma, including volatile terpenoids, has also been genetically modulated in order to understand the complicated nature of multi-trophic interactions that affect the behavior of predators and pollinators in the ecosystem. Torenia would accordingly be of great use for investigating both the variation in ornamental plants and the infochemical-mediated interactions with arthropods

Development of temporal response properties and contrast sensitivity of V1 and V2 neurons in macaque monkeys

The temporal contrast sensitivity of human infants is reduced compared to that of adults. It is not known which neural structures of our visual brain sets limits on the early maturation of temporal vision. In this study we investigated how individual neurons in the primary visual cortex (V1) and visual area 2 (V2) of infant monkeys respond to temporal modulation of spatially optimized grating stimuli and a range of stimulus contrasts. As early as 2 wk of age, V1 and V2 neurons exhibited band-pass temporal frequency tuning. However, the optimal temporal frequency and temporal resolution of V1 neurons were much lower in 2- and 4-wk-old infants than in 8-wk-old infants or adults. V2 neurons of 8-wk-old monkeys had significantly lower optimal temporal frequencies and resolutions than those of adults. Onset latency was longer in V1 at 2 and 4 wk of age and was slower in V2 even at 8 wk of age than in adults. Contrast threshold of V1 and V2 neurons was substantially higher in 2- and 4-wk-old infants but became adultlike by 8 wk of age. For the first 4 wk of life, responses to high-contrast stimuli saturated more readily in V2. The present results suggest that although the early development of temporal vision and contrast sensitivity may largely depend on the functional maturation of precortical structures, it is also likely to be limited by immaturities that are unique to V1 and V

Heme oxygenase-1 induction in the brain during lipopolysaccharide-induced acute inflammation

Delirium occurs in 23% of sepsis patients, in which pro-inflammatory cytokines and nitric oxide are suggested to be involved. However, in animal experiments, even a subseptic dose of lipopolysaccharide (LPS) injection induces both pro-inflammatory cytokines and inducible nitric oxide synthase in the brain, suggesting that the brain oxidative reaction can be induced in the subseptic condition. Then, we evaluated the changes of heme oxygenase-1 (HO-1), a sensitive oxidative marker, as well as interleukin (IL)-1β, IL-6, and inductible nitric oxide synthase (iNOS) mRNA in the hypothalamus and hippocampus of rats using real-time PCR after peripheral injection of LPS (2.0 mg/kg). As a result, these four kinds of mRNAs were induced significantly in both areas after LPS injection. These results suggest that peripheral inflammation induces an oxidative reaction in the brain, even if the inflammation is not lethal. It is also considered that several pathways are involved in brain HO-1 induction

Strict De Novo Methylation of the 35S Enhancer Sequence in Gentian

A novel transgene silencing phenomenon was found in the ornamental plant, gentian (Gentiana triflora × G. scabra), in which the introduced Cauliflower mosaic virus (CaMV) 35S promoter region was strictly methylated, irrespective of the transgene copy number and integrated loci. Transgenic tobacco having the same vector did not show the silencing behavior. Not only unmodified, but also modified 35S promoters containing a 35S enhancer sequence were found to be highly methylated in the single copy transgenic gentian lines. The 35S core promoter (−90)-introduced transgenic lines showed a small degree of methylation, implying that the 35S enhancer sequence was involved in the methylation machinery. The rigorous silencing phenomenon enabled us to analyze methylation in a number of the transgenic lines in parallel, which led to the discovery of a consensus target region for de novo methylation, which comprised an asymmetric cytosine (CpHpH; H is A, C or T) sequence. Consequently, distinct footprints of de novo methylation were detected in each (modified) 35S promoter sequence, and the enhancer region (−148 to −85) was identified as a crucial target for de novo methylation. Electrophoretic mobility shift assay (EMSA) showed that complexes formed in gentian nuclear extract with the −149 to −124 and −107 to −83 region probes were distinct from those of tobacco nuclear extracts, suggesting that the complexes might contribute to de novo methylation. Our results provide insights into the phenomenon of sequence- and species- specific gene silencing in higher plants

Effects of perceptual learning on local stereopsis and neuronal responses of V1 and V2 in prism-reared monkeys.

Visual performance improves with practice (perceptual learning). In this study, we sought to determine whether or not adult monkeys reared with early abnormal visual experience improve their stereoacuity by extensive psychophysical training and testing, and if so, whether alterations of neuronal responses in the primary visual cortex (V1) and/or visual area 2 (V2) are involved in such improvement. Strabismus was optically simulated in five macaque monkeys using a prism-rearing procedure between 4 and 14 wk of age. Around 2 yr of age, three of the prism-reared monkeys ( trained monkeys) were tested for their spatial contrast sensitivity and stereoacuity. Two other prism-reared monkeys received no training or testing ( untrained monkeys). Microelectrode experiments were conducted around 4 yr of age. All three prism-reared trained monkeys showed improvement in stereoacuity by a factor of 7 or better. However, final stereothresholds were still approximately 10-20 times worse than those in normal monkeys. In V1, disparity sensitivity was drastically reduced in both the trained and untrained prism-reared monkeys and behavioral training had no obvious effect. In V2, the disparity sensitivity in the trained monkeys was better by a factor of approximately 2.0 compared with that in the untrained monkeys. These data suggest that the observed improvement in stereoacuity of the trained prism-reared monkeys may have resulted from better retention of disparity sensitivity in V2 and/or from learning by upstream neurons to more efficiently attend to residual local disparity information in V1 and V

Dysfunction of CD8+PD-1+T cells in type 2 diabetes caused by the impairment of metabolism-immune axis

The metabolic changes and dysfunction in CD8+T cells may be involved in tumor progression and susceptibility to virus infection in type 2 diabetes (T2D). In C57BL/6JJcl mice fed with high fat-high sucrose chow (HFS), multifunctionality of CD8+splenic and tumor-infiltrating lymphocytes (TILs) was impaired and associated with enhanced tumor growth, which were inhibited by metformin. In CD8+splenic T cells from the HFS mice, glycolysis/basal respiration ratio was significantly reduced and reversed by metformin. In the patients with T2D (DM), multifunctionality of circulating CD8+PD-1+T cells stimulated with PMA/ionomycin as well as with HLA-A*24:02 CMV peptide was dampened, while metformin recovered multifunctionality. Both glycolysis and basal respiration were reduced in DM, and glycolysis was increased by metformin. The disturbance of the link between metabolism and immune function in CD8+PD-1+T cells in T2D was proved by recovery of antigen-specific and non-specific cytokine production via metformin-mediated increase in glycolytic activity

Plasma angiotensin-converting enzyme 2 (ACE2) is a marker for renal outcome of diabetic kidney disease (DKD) (U-CARE study 3)

Introduction ACE cleaves angiotensin I (Ang I) to angiotensin II (Ang II) inducing vasoconstriction via Ang II type 1 (AT1) receptor, while ACE2 cleaves Ang II to Ang (1-7) causing vasodilatation by acting on the Mas receptor. In diabetic kidney disease (DKD), it is still unclear whether plasma or urine ACE2 levels predict renal outcomes or not. Research design and methods Among 777 participants with diabetes enrolled in the Urinary biomarker for Continuous And Rapid progression of diabetic nEphropathy study, the 296 patients followed up for 9 years were investigated. Plasma and urinary ACE2 levels were measured by the ELISA. The primary end point was a composite of a decrease of estimated glomerular filtration rate (eGFR) by at least 30% from baseline or initiation of hemodialysis or peritoneal dialysis. The secondary end points were a 30% increase or a 30% decrease in albumin-to-creatinine ratio from baseline to 1 year. Results The cumulative incidence of the renal composite outcome was significantly higher in group 1 with lowest tertile of plasma ACE2 (p=0.040). Group 2 with middle and highest tertile was associated with better renal outcomes in the crude Cox regression model adjusted by age and sex (HR 0.56, 95% CI 0.31 to 0.99, p=0.047). Plasma ACE2 levels demonstrated a significant association with 30% decrease in ACR (OR 1.46, 95% CI 1.044 to 2.035, p=0.027) after adjusting for age, sex, systolic blood pressure, hemoglobin A1c, and eGFR. Conclusions Higher baseline plasma ACE2 levels in DKD were protective for development and progression of albuminuria and associated with fewer renal end points, suggesting plasma ACE2 may be used as a prognosis marker of DKD.Trial registration number UMIN000011525

陰膳方式食物収集による日本人の栄養調査 : 血清脂質と栄養摂取量

Nutrient intake was surveyed by total food duplicate method in combination with serum biochemistry among adult Japanese women. The survey was conducted twice in the same 19 sites all over Japan, once in the years around 1980 (the first survey on 199 women) and then in 1991-6 (the second survey on 379 women). The participants 19 sites were classified into four groups of those in Hokkaido, urban areas, rural areas and Okinawa by the location of residence, and relation of findings serum lipid analyses with nutrient intakes was examined. In the Hokkaido group, cholesterol and triglyceride in serum, and intake of animal fat and saturated fatty acids (SFAs) were lower in the second survey than in the first survey. In the urban group, serum cholesterol, and intakes of animal fats and SFAs were the highest among the four groups both in the first and second surveys. In contrast, serum cholesterol, and intakes of animal fats and SFAs in the rural group were lower than the levels in other groups in the first survey, but the three groups in the second survey. In Okinawa, serum cholesterol was lower than others in the second survey. Intake of fat-based energy was high, but intakes of monounsaturated fatty acids and n-6 polyunsaturated fatty acids were also high. When all subjects were combined, the serum cholesterol level was 193mg/100ml (as an arithmetic mean) in the first survey and it was 202 mg/100 ml in the second survey, and this increase was paralleled by the increase in the number of aged survey subjects. Serum triglyceride level was 111mg/100ml in the first survey, and 99 mg/dl in the second survey. The reduction may be attributable to the reduced nutrient intake among young people. Serum cholesterol correlated significantly with intake of animal fats and SFAs, but the correlation with total energy intake was significant only in the second survey. BMI correlated significantly with serum triglyceride levels in both surveys, but the correlation coefficient was smaller in the second survey than in the first survey. There was no correlation between BMI and serum cholesterol in both surveys

日本人女性の食事からのカルシウム摂取量

This study was initiated 1. to examine if there was any secular changes in dietary calcium (Ca) intake among Japanese women in past 15 years, and 2. to identify the food groups insufficient intake of which may induce Ca insufficiency. 24-Hour food duplicate samples were collected from 433 and 760 adult women in the first (1977-1981) and the second survey (1991-1998), respectively, from various parts of Japan. Daily Ca intakes were estimated from weights of food items in the duplicate samples, taking advantages of food composition tables (FCT-based estimates). Possible difference in means was examined by Student's t-test and multiple comparison test (Scheffe), and that in distribution by chi-square test. The average dietary intake of Ca was 607 and 605 mg/day in the first and the second survey, respectively, with no significant difference. The subjects, separately for the two surveys, were classified by the Ca intake into 5 groups, i.e., those with intake of 419 or less, 420 to 539,540 to 659,660 to 999 and 1,000 or more mg Ca/day, and also by decade of ages. Reduced Ca intake from fish (including shellfish) and vegetables coupled with increased intake from milk (including milk products) were noted among the three low Ca intake groups when the intakes were compared between the two surveys, whereas no food group-specific changes were noted among the two high intake groups. Ca intake from milk was high in the latter two groups. Further analysis showed that low intakes of pulse, fish, milk, vegetables and algae were associated with insufficiency in Ca intakes. Classification by ages showed that Ca insufficiency among those at the ages of 20's was attributable to low intakes of pulse, fish and algae. When compared between the two surveys, reduced intakes of fish and vegetables and increased intake of milk were commonly observed in most age groups, and milk intake was substantially increased at advanced ages. FCT-based estimates and ICP-MS measurements gave 581 and 542 mg/day as averages for the former and the latter, respectively. The observation that the latter is 93% of the former suggests that Ca insufficiency should be taken more seriously than the estimates would suggest