Growth of diabetes drug expenditure decomposed-A nationwide analysis.

OBJECTIVES: The aim of this study was to quantify different factors underlying the growth of diabetes drug expenditure in Finland. METHODS: Data representing purchases of antidiabetic agents between 2003 and 2015 were extracted from a nationwide prescription register. By using Fisher's Ideal Indexes, the per capita expenditure growth for both insulins and non-insulin antidiabetic agents was decomposed into six different determinants: purchase volume, purchase size, switches between therapeutic classes, switches within therapeutic classes, unit costs and switches to generic alternatives. RESULTS: Between 2003 and 2015, the per capita expenditure on insulins increased by €8.64 and on non-insulins by €13.73. For insulins, holding other factors constant, change in the number of purchases represented a €4.67 increase in expenditure, change in the size of purchases a €4.33 increase and switches between therapeutic classes a €4.07 increase. For non-insulins, change in the number of purchases represented a €10.22 increase in expenditure and switches between therapeutic classes, a €10.17 increase. Changes in purchase size increased the non-insulin per capita expenditure by €1.48. For both insulins and non-insulins, changes in prices and product level switches had decreasing effects on expenditures. CONCLUSIONS: The main drivers of the growth in diabetes drug expenditure were volume growth and switches to newer and more expensive drugs. Price changes, however, had a decreasing effect on the overall diabetes drug expenditure.

Genome sequencing and population genomic analyses provide insights into the adaptive landscape of silver birch.

Silver birch (Betula pendula) is a pioneer boreal tree that can be induced to flower within 1 year. Its rapid life cycle, small (440-Mb) genome, and advanced germplasm resources make birch an attractive model for forest biotechnology. We assembled and chromosomally anchored the nuclear genome of an inbred B. pendula individual. Gene duplicates from the paleohexaploid event were enriched for transcriptional regulation, whereas tandem duplicates were overrepresented by environmental responses. Population resequencing of 80 individuals showed effective population size crashes at major points of climatic upheaval. Selective sweeps were enriched among polyploid duplicates encoding key developmental and physiological triggering functions, suggesting that local adaptation has tuned the timing of and cross-talk between fundamental plant processes. Variation around the tightly-linked light response genes PHYC and FRS10 correlated with latitude and longitude and temperature, and with precipitation for PHYC. Similar associations characterized the growth-promoting cytokinin response regulator ARR1, and the wood development genes KAK and MED5A.

Peroxidase-Generated Apoplastic ROS Impair Cuticle Integrity and Contribute to DAMP-Elicited Defenses.

Cuticular defects trigger a battery of reactions including enhanced reactive oxygen species (ROS) production and resistance to necrotrophic pathogens. However, the source of ROS generated by such impaired cuticles has remained elusive. Here, we report the characterization of Arabidopsis thaliana ohy1 mutant, a Peroxidase 57 (PER57) - overexpressing line that demonstrates enhanced defense responses that result both from increased accumulation of ROS and permeability of the leaf cuticle. The ohy1 mutant was identified in a screen of A. thaliana seedlings for oligogalacturonides (OGs) insensitive/hypersensitive mutants that exhibit altered growth retardation in response to exogenous OGs. Mutants impaired in OG sensitivity were analyzed for disease resistance/susceptibility to the necrotrophic phytopathogens Botrytis cinerea and Pectobacterium carotovorum. In the ohy1 line, the hypersensitivity to OGs was associated with resistance to the tested pathogens. This PER57 overexpressing line exhibited a significantly more permeable leaf cuticle than wild-type plants and this phenotype could be recapitulated by overexpressing other class III peroxidases. Such peroxidase overexpression was accompanied by the suppressed expression of cutin biosynthesis genes and the enhanced expression of genes associated with OG-signaling. Application of ABA completely removed ROS, restored the expression of genes associated with cuticle biosynthesis and led to decreased permeability of the leaf cuticle, and finally, abolished immunity to B. cinerea. Our work demonstrates that increased peroxidase activity increases permeability of the leaf cuticle. The loss of cuticle integrity primes plant defenses to necrotrophic pathogens via the activation of DAMP-responses.

ERD15--an attenuator of plant ABA responses and stomatal aperture.

Plants are continuously challenged by abiotic and biotic stress factors and need to mount appropriate responses to ensure optimal growth and survival. We have identified ERD15 as a central component in several stress responses in Arabidopsis thaliana. Comparative genomics demonstrates that ERD15 is a member of a small but highly conserved protein family ubiquitous but specific to the plant kingdom. The origin of ERD15 family of proteins can be traced to the time of emergence of land plants. The presence of the conserved PAM2 motif in ERD15 proteins is indicative of a possible interaction with poly(A) binding proteins and could suggest a role in posttranscriptional regulation of gene expression. The function of the other highly conserved motifs in ERD15 remains to be elucidated. The biological role of all ERD15 family members studied so far appears associated to stress responses and stress adaptation. Studies in Arabidopsis demonstrate a role in abiotic stress tolerance where ERD15 is a negative regulator of ABA signaling. The role in ABA signaling may also explain how ERD15 regulates stomatal aperture and consequently controls plant water relations.

Requirement of a homolog of glucosidase II beta-subunit for EFR-mediated defense signaling in Arabidopsis thaliana.

EFR is a plasma-membrane resident receptor responsible for recognition of microbial elongation factor Tu (EF-Tu) and thus triggering plant innate immunity to fend off phytopathogens. Functional EFR must be subject to the endoplasmic reticulum quality control (ERQC) machinery for the correct folding and proper assembly in order to reach its final destination. Genetic studies have demonstrated that ERD2b, a counterpart of the yeast or mammalian HDEL receptor ERD2 for retaining proteins in the endoplasmic reticulum (ER) lumen, is required for EFR function in plants (Li et al., 2009). In this study, we characterized the Arabidopsis glucosidase II beta-subunit via the HDEL motif against the non-redundant protein database. Data mining also revealed that the glucosidase II beta-subunit gene has a highly similar expression pattern to ERD2b and the other known ERQC components involved in EFR biogenesis. Importantly, the T-DNA insertion lines of the glucosidase II beta-subunit gene showed that EFR-controlled responses were substantially reduced or completely blocked in these mutants. The responses include seedling growth inhibition, induction of marker genes, MAP kinase activation, and callose deposition, triggered by peptide elf18, a full mimic of EF-Tu. Taken together, our data indicate a requirement of the glucosidase II beta-subunit for EFR function.

Blood pressure is lower in children and adolescents with a low-saturated-fat diet since infancy: the special turku coronary risk factor intervention project.

Blood pressure was measured in the prospective randomized Special Turku Coronary Risk Factor Intervention Project Study with an oscillometric method every year from 7 months to 15 years of age in 540 children receiving a low-saturated-fat, low-cholesterol diet and in 522 control children. Dietary intakes, family history of parental hypertension, and grandparental vascular disease were recorded. Systolic and diastolic blood pressures were 1.0 mm Hg lower (95% CI for systolic: -1.7 to -0.2 mm Hg; 95% CI for diastolic: -1.5 to -0.4 mm Hg) in children receiving low-saturated-fat counseling through childhood than in control children. Intakes of saturated fat were lower (P<0.001), those of polyunsaturated fat higher (P<0.001), and intakes of potassium slightly higher (P=0.002) in the intervention group, but sodium intakes were not influenced by the intervention (P=0.76). Children whose parents were hypertensive had 4- to 6-mm Hg higher systolic and 3- to 4-mm Hg higher diastolic blood pressures than children of normotensive parents (P<0.001). Diastolic blood pressure of children with grandparental vascular disease, ie, early cardiovascular or cerebrovascular disease, tended to be higher than that of children with no grandparental disease (P=0.051). We conclude that restriction of saturated fat from infancy until 15 years of age decreases childhood and adolescent blood pressure with a meaningful population-attributable amount. The importance of childhood lifestyle counseling and primary prevention of hypertension should be emphasized, especially in those children with a family history of hypertension or atherosclerotic vascular disease.

Wireless and inductively powered implant for measuring electrocardiogram.

The development of an active implantable device for measuring electrocardiogram (ECG) is presented. The study is a part of a project which aims at developing implantable ECG instrumentation with wireless data and power transfer ( http://www.ele.tut.fi/tule ). The developed implant presented here has all the measurement electronics as well as power and data communication instrumentation included. The implant itself contains no battery, while power for the implant is transferred electromagnetically from an external reader device. The results of testing the implant attached on the body surface and in vitro in a water container are also presented. The developed system was also successfully tested in in vivo measurements, which were conducted on four cows with an implantation time of 24 h. The in vivo testing of implant in cows was conducted by a veterinarian in supervised conditions under approved animal experiment licence.

Acceleration of ethanol and acetaldehyde oxidation by D-glycerate in rats.

The aim of the present study was to investigate whether d-glycerate (glycerate) could accelerate ethanol and acetaldehyde (AcH) oxidation in vivo in rats by circumventing the rate-limiting step, that is, the reoxidation of the reduced form of nicotinamide adenine dinucleotide. Male rats belonging to the ANA (Alko, nonalcohol) and AA (Alko, alcohol) rat lines were challenged with 1.2 g ethanol per kilogram with or without glycerate administration (0.1-1.0 g/kg). Blood ethanol, blood AcH, and liver free glycerol concentrations were determined during ethanol intoxication. Glycerate treatment, regardless of the dose, accelerated ethanol elimination by approximately 25% (P < .001) in the ANA animals. Glycerate also accelerated the AcH oxidation, but perhaps not as much as the ethanol oxidation, as indicated by a trend toward elevated AcH levels. In the experiments with the AA rats, glycerate treatment elevated hepatic free glycerol levels by about 50% (P < .05) during alcohol intoxication. The acceleration of ethanol and AcH oxidation in conjunction with elevated glycerol levels by the treatment with glycerate supports the hypothesis that the aldehyde dehydrogenase-mediated AcH oxidation can be coupled with the reduction of glycerate to d-glyceraldehyde catalyzed by the same enzyme. Such a coupling should increase the availability of the oxidized form of nicotinamide adenine dinucleotide and thus accelerate both ethanol and AcH oxidation. Further studies are needed to investigate how the AcH could be even more efficiently oxidized to reduce the harmful effects of ethanol-derived AcH.

EARLY RESPONSIVE TO DEHYDRATION 15, a negative regulator of abscisic acid responses in Arabidopsis.

EARLY RESPONSIVE TO DEHYDRATION 15 (ERD15) is rapidly induced in response to various abiotic and biotic stress stimuli in Arabidopsis (Arabidopsis thaliana). Modulation of ERD15 levels by overexpression or RNAi silencing altered the responsiveness of the transgenic plants to the phytohormone abscisic acid (ABA). Overexpression of ERD15 reduced the ABA sensitivity of Arabidopsis manifested in decreased drought tolerance and in impaired ability of the plants to increase their freezing tolerance in response to this hormone. In contrast, RNAi silencing of ERD15 resulted in plants that were hypersensitive to ABA and showed improved tolerance to both drought and freezing, as well as impaired seed germination in the presence of ABA. The modulation of ERD15 levels not only affected abiotic stress tolerance but also disease resistance: ERD15 overexpression plants showed improved resistance to the bacterial necrotroph Erwinia carotovora subsp. carotovora accompanied with enhanced induction of marker genes for systemic acquired resistance. We propose that ERD15 is a novel mediator of stress-related ABA signaling in Arabidopsis.

Differential responses of silver birch (Betula pendula) ecotypes to short-day photoperiod and low temperature.

We investigated interrelations of dormancy and freezing tolerance and the role of endogenous abscisic acid (ABA) in the development of silver birch (Betula pendula Roth) ecotypes in controlled environments. Short-day treatment induced growth cessation, bud set and dormancy development, as well as initiation of cold acclimation and an increase in freezing tolerance. Subsequent low temperature and short days (12-h photoperiod) resulted in a significant increase in freezing tolerance, whereas bud dormancy was gradually released. The concentration of ABA increased in response to short days and then remained high, but ABA concentrations fluctuated irregularly when the dormant plants were subsequently exposed to low temperature during short days. Although there was a parallel development of freezing tolerance and bud dormancy in response to short days, subsequent exposure to low temperature had opposite effects on these processes, enhancing freezing tolerance and releasing dormancy. Compared with the southern ecotype, the northern ecotype was more responsive to short days and low temperature, exhibiting earlier initiation of cold acclimation, growth cessation and an increase in ABA concentrations in short days, and higher freezing tolerance, faster dormancy release and greater alteration in ABA concentrations when subsequently exposed to low temperature during short days. The rates and extent of the increases in ABA concentration may be related to increases in freezing tolerance and dormancy development during short days, whereas the extent of the fluctuations in ABA concentration may play an important role in enhancing freezing tolerance and releasing dormancy during a subsequent exposure to low temperature during short days.

Short-day potentiation of low temperature-induced gene expression of a C-repeat-binding factor-controlled gene during cold acclimation in silver birch.

Development of winter hardiness in trees is a two-stage process involving sequential perception of distinct environmental cues, short-day (SD) photoperiod and low temperature (LT). We have shown that both SD and LT are recognized by leaves of silver birch (Betula pendula cv Roth) leading to increased freezing tolerance, and thus leaves can be used as an experimental model to study the physiological and molecular events taking place during cold acclimation. To obtain a molecular marker for the acclimation process in birch we cloned a gene, designated Bplti36, encoding a 36-kD acidic SK2 type of dehydrin. The gene was responsive to LT, drought, salt, and exogenous abscisic acid. This responsiveness to abiotic stresses and abscisic acid was retained when Bplti36 was introduced to Arabidopsis (Arabidopsis thaliana). The LT induction of the gene appeared to be under the control of the C-repeat-binding factor pathway as suggested by the presence of several C-repeat/dehydration-responsive element/LT-responsive elements in the Bplti36 promoter and its constitutive expression in C-repeat-binding factor overproducing Arabidopsis. In birch SD photoperiod at normal-growth temperature did not result in significant induction of Bplti36. However, preexposure to SD followed by LT treatment resulted in a remarkable increase in Bplti36 transcript accumulation as compared to LT-treated plants grown at long-day photoperiod. This suggests that SD photoperiod potentiates the LT response by conditioning the leaf tissue to be more responsive to the LT stimulus.

Overexpression of multiple dehydrin genes enhances tolerance to freezing stress in Arabidopsis.

To elucidate the contribution of dehydrins (DHNs) to freezing stress tolerance in Arabidopsis, transgenic plants overexpressing multiple DHN genes were generated. Chimeric double constructs for expression of RAB18 and COR47 (pTP9) or LTI29 and LTI30 (pTP10) were made by fusing the coding sequences of the respective DHN genes to the cauliflower mosaic virus 35S promoter. Overexpression of the chimeric genes in Arabidopsis resulted in accumulation of the corresponding dehydrins to levels similar or higher than in cold-acclimated wild-type plants. Transgenic plants exhibited lower LT50 values and improved survival when exposed to freezing stress compared to the control plants. Post-embedding immuno electron microscopy of high-pressure frozen, freeze-substituted samples revealed partial intracellular translocation from cytosol to the vicinity of the membranes of the acidic dehydrin LTI29 during cold acclimation in transgenic plants. This study provides evidence that dehydrins contribute to freezing stress tolerance in plants and suggests that this could be partly due to their protective effect on membranes.

Photoperiod and temperature differentially regulate the expression of two dehydrin genes during overwintering of birch (Betula pubescens Ehrh.).

The overwintering of trees in northern areas depends on processes regulated by photoperiod and temperature. To identify the physiological and genetic factors involved in this environmental control, three latitudinal ecotypes of pubescent birch (Betula pubescens Ehrh.) growing in a common garden experiment were used. Each ecotype responded to the shortening of the photoperiod according to its specific critical daylength, resulting in the induction of freezing tolerance and dehydration of buds first in the northern ecotype, followed by the central and southern ecotypes, respectively. By contrast, there was no clear difference in the timing of dormancy release, bud rehydration, and deacclimation in the spring, suggesting that these traits were controlled mainly by temperature. To elucidate the role of dehydrins (DHN) in the overwintering process, two DHN genomic clones were isolated from pubescent birch and expression of the corresponding genes, both in field and under controlled conditions, was characterized. BpuDhn1 was found to encode an Y(n)K(n)-type of basic DHN, while BpuDhn2 encoded an acidic, SK(n)-type of DHN. In field-grown trees the level of BpuDhn1 increased in buds during the autumn, while the level of BpuDhn2 was highest during the coldest winter months. Under controlled conditions BpuDhn1 increased in response to the combined effect of short daylength and low, non-freezing temperatures whereas the expression of BpuDhn2 was mainly controlled by low temperature while photoperiod had less effect on its expression. These results suggest that DHNs participate in the sensitive environmental regulation of the overwintering process in birch.

Different responses of northern and southern ecotypes of Betula pendula to exogenous ABA application.

We investigated responses of northern and southern ecotypes of silver birch (Betula pendula Roth) to exogenous abscisic acid (ABA) under controlled environmental conditions to determine the role of ABA in cold acclimation and dormancy development. Abscisic acid was sprayed on the leaves and changes in freezing tolerance, determined by the electrolyte leakage test, and bud dormancy were monitored. Applied ABA induced cold acclimation but had no effect on growth cessation in seedlings grown in long day conditions (LD, 24-h photoperiod at 18 degrees C). It enhanced freezing tolerance and accelerated growth cessation in seedlings grown in short day conditions (SD, 12-h photoperiod at 18 degrees C), and slightly enhanced freezing tolerance in seedlings grown at low temperature (LT, 24-h photoperiod at 4 degrees C) in both ecotypes. There were distinct ecotypic differences in ABA-induced cold acclimation and dormancy development. The northern ecotype was more responsive to applied ABA than the southern ecotype, resulting in more rapid development of freezing tolerance in all treatments, and earlier dormancy development in SD. When plants were grown in a photoperiod just above the critical photoperiod for the ecotype (defined as the longest photoperiod that induces growth cessation), applied ABA caused growth cessation and dormancy development. Compared with ABA-treated seedlings grown in SD, dormancy development was delayed in ABA-treated seedlings exposed to a near-critical photoperiod, but even in this treatment dormancy developed faster in the northern ecotype than in the southern ecotype.