Non-destructive insights into photosynthetic and photoprotective mechanisms in Arabidopsis thaliana grown under two light regimes.

Probing insights into understanding photosynthetic processes via non-invasive means has an added advantage when used in phenotyping or precision agriculture. We employed Raman spectroscopy and fluorescence-based methods to investigate both the changes in the photosynthetic processes and the underlying protective mechanisms on Arabidopsis thaliana wild-type (WT), and ros1, which is a mutant of a repressor of transcriptional gene silencing, both grown under low light (LL: 100 µmol m-2s-1) and high light (HL: 400 µmol m-2s-1) regimes. Raman imaging detected a lower carotenoid intensity after two weeks in those plants grown under HL, compared to those grown under the LL regime; we interpret this as the result of oxidative damage of ß-carotene molecules. Further, the data revealed a significant depletion in carotenoids with enhanced phenolics around the midrib and tip of the WT leaves, but not in the ros1. On the contrary, small necrotic zones appeared after two weeks of HL in the ros1 mutant, pointing to the starting oxidative damage. The lower maximum quantum yield of the photochemistry (Fv/Fm) in the WT as well as in the ros1 mutant grown in HL (compared to those in the LL two weeks post-exposure), indicates the HL partially inactivated photosystems. Chlorophyll a fluorescence imaging further showed high non-photochemical quenching (NPQ) in the plants grown under the HL regime for both the WT and the ros1 mutant, but the spatial heterogeneity of NPQ images was much higher in the HL-grown ros1 mutant. Fluorescence screening methods revealed significantly high values of chlorophyll proxies in the WT as well as in the ros1 mutant two weeks after in the HL compared to those under LL. The data generally revealed an increased accumulation of phenolics under HL in both the WT and ros1 mutant plants, but the proxies of anthocyanin and flavonols were significantly lower in the ros1 mutant than in the WT. The comparatively low accumulation of anthocyanin in the ros1 mutant compared to the WT supports the Raman data. We conclude that integrated use of these techniques can be efficiently applied for a better understanding of insights into photosynthetic mechanisms.

Interactive effects of nitrogen, UV and PAR on barley morphology and biochemistry are associated with the leaf C:N balance.

Environmental conditions to which plants acclimate prior exposure to abiotic or biotic stressors can greatly affect their subsequent resilience. This may have a significant impact on the response to ongoing climate change and can be useful for increasing the food security under adverse weather conditions associated with climate change.Within this study, we tested the hypothesis that plant morphological and biochemical acclimation to radiation conditions and nitrogen (N) availability is closely linked with carbon (C) and N balance. Four barley (Hordeum vulgare) varieties, differing in their morphological characteristics and sensitivity to photooxidative stress, were grown at two levels of N supply and four radiation regimes combining distinct levels of ultraviolet (UV) and photosynthetically active radiation (PAR). Changes in root and shoot morphology, accumulation of phenolic compounds, amino acids, and sugars were studied together with the analysis of C and N content in leaves. Both UV and PAR reduced leaf length and increased root-to-shoot ratio (R:S). Such effect was more pronounced under high N availability. High N supply reduced R:S, but this effect showed significant interactions with UV and PAR, and also with barley variety. Changes in R:S were positively related to C:N ratio in leaves that varied in response to both N availability and radiation treatments. UV radiation, particularly in combination with high PAR intensity, led to increases in most phenolic compounds (particularly flavones such as saponarin, homoorientin and isovitexin) which was also closely associated with changes in C:N ratio, while specifically phenolic acids (vanillic and syringic acids) decreased under high levels of UV and PAR, and hydroxycinnamic acids responded positively mainly to PAR. Although high N availability generally reduced the accumulation of phenolic compounds, this effect was genotype-specific and modulated by the radiation regime. A similar antagonistic effect of radiation treatment and N availability was also found for the accumulation of sugars (pentoses), resulting in a close relationship between the accumulation of pentoses and C:N ratio. The accumulation of most amino acids, in contrary to phenolic compounds, increases at high N and is also stimulated by high PAR and UV intensities. We conclude that radiation conditions and N availability have opposite effects on plant morphology and accumulation of most phenolic compounds and modulate the amino acid and sugar metabolism. Strong associations of these responses with changes in C:N ratio indicates that plant stoichiometry integrates acclimation processes and induction of relevant defence mechanisms.

Quality improvement project to improve providers' goal-setting activity for chronic disease self-management.

BACKGROUND: Patient-centered medical home (PCMH) places patients at the center of care and actively considers patients' goals, preferences and skill levels in caring for their disease while developing a care plan. We implemented a quality improvement (QI) project to improve internal medicine resident goal-setting activity with patients for chronic disease self-management. METHODS: The plan-do-study-act (PDSA) model was used and a root cause analysis was conducted with internal medicine residents (n=20) and faculty (n=7) to identify barriers to patient goal setting. Two main causes were identified - lack of awareness and lack of knowledge about where and how to set patient goals in the electronic medical record (EMR). Reminders during daily morning huddles, discussions with faculty, educational video presentation and organizational policy change interventions were implemented in four serial PDSA cycles. RESULTS: The goal setting activity documentation rate by resident providers increased to 14% following inclusion of patient goal setting reminders during daily morning huddles. There was notable increase in the goal setting documentation rate to 29% following discussion in faculty meeting. The goal setting documentation rate remained the same (29%) despite educational video and policy change intervention. However, goal setting documentation rate increased to 33% by the end of the study. CONCLUSION: Our QI project resulted in a measurable increase in the use of methods of healthcare delivery associated with improved outcomes. This model worked well in our setting, and sharing our success may benefit others seeking to achieve similar goals.

Interaction Detection with Depth Sensing and Body Tracking Cameras in Physical Rehabilitation.

INTRODUCTION: This article is part of the Focus Theme of Methods of Information in Medicine on "Methodologies, Models and Algorithms for Patients Rehabilitation". OBJECTIVES: This paper presents a camera based method for identifying the patient and detecting interactions between the patient and the therapist during therapy. Detecting interactions helps to discriminate between active and passive motion of the patient as well as to estimate the accuracy of the skeletal data. METHODS: Continuous face recognition is used to detect, recognize and track the patient with other people in the scene (e.g. the therapist, or a clinician). We use a method based on local binary patterns (LBP). After identifying users in the scene we identify interactions between the patient and other people. We use a depth map/point cloud for estimating the distance between two people. Our method uses the association of depth regions to user identities and computes the minimal distance between the regions. RESULTS: Our results show state-of-the-art performance of real-time face recognition using low-resolution images that is sufficient to use in adaptive systems. Our proposed approach for detecting interactions shows 91.9% overall recognition accuracy what is sufficient for applications in the context of serious games. We also discuss limitations of the proposed method as well as general limitations of using depth cameras for serious games. CONCLUSIONS: We introduced a new method for frame-by-frame automated identification of the patient and labeling reliable sequences of the patient's data recorded during rehabilitation (games). Our method improves automated rehabilitation systems by detecting the identity of the patient as well as of the therapist and by detecting the distance between both over time.

Similar local, but different systemic, metabolomic responses of closely related pine subspecies to folivory by caterpillars of the processionary moth.

Plants respond locally and systemically to herbivore attack. Most of the research conducted on plant-herbivore relationships at element and molecular levels have focused on the elemental composition or/and certain molecular compounds or specific families of defence metabolites showing that herbivores tend to select plant individuals or species with higher nutrient concentrations and avoid those with higher levels of defence compounds. We performed stoichiometric and metabolomics, both local and systemic, analyses in two subspecies of Pinus sylvestris under attack from caterpillars of the pine processionary moth, an important pest in the Mediterranean Basin. Both pine subspecies responded locally to folivory mainly by increasing relative concentrations of terpenes and some phenolics. Systemic responses differed between pine subspecies, and most of the metabolites presented intermediate concentrations between those of the affected parts and unattacked trees. Our results support the hypothesis that foliar nutrient concentrations are not a key factor for plant selection by adult female processionary moths for oviposition, since folivory was not associated with any of the elements analysed. Phenolic compounds generally did not increase in the attacked trees, questioning the suggestion of induction of phenolics following folivory attack and the anti-feeding properties of phenolics. Herbivory attack produced a general systemic shift in pines, in both primary and secondary metabolism, which was less intense and chemically different from the local responses. Local pine responses were similar between pine subspecies, while systemic responses were more distant.

Potassium ion channels and allergic asthma.

High-conductive calcium-sensitive potassium channels (BK+Ca) and ATP-sensitive potassium (K+ATP) channels play a significant role in the airway smooth muscle cell and goblet cell function, and cytokine production. The present study evaluated the therapeutic potential of BK+Ca and K+ATP openers, NS 1619 and pinacidil, respectively, in an experimental model of allergic inflammation. Airway allergic inflammation was induced with ovalbumine in guinea pigs during 21 days, which was followed by a 14-day treatment with BK+Ca and K+ATP openers. The outcome measures were airway smooth muscle cells reactivity in vivo and in vitro, cilia beating frequency and the level of exhaled NO (ENO), and the level of pro-inflammatory cytokines in the plasma and bronchoalveolar lavage fluid. The openers of both channels decreased airway smooth muscle cells reactivity, cilia beating frequency, and cytokine levels in the serum. Furthermore, NS1619 reduced ENO and inflammatory cells infiltration. The findings confirmed the presence of beneficial effects of BK+Ca and K+ATP openers on airway defence mechanisms. Although both openers dampened pro-inflammatory cytokines and mast cells infiltration, an evident anti-inflammatory effect was provided only by NS1619. Therefore, we conclude that particularly BK+Ca channels represent a promising new drug target in treatment of airway's allergic inflammation.

CRAC ion channels and airway defense reflexes in experimental allergic inflammation.

Calcium release-activated calcium channels (CRAC) play unambiguous role in secretory functions of mast cells, T cells, and eosinophils. Less knowledge exists about the role of CRAC, widely distributed in airway smooth muscle (ASM) cells, in airway contractility. The presented study seeks to determine the possible participation of CRAC in ASM-based inflammatory airway disorders in guinea pigs. The acute and long-term administration (14 days) of the CRAC antagonist 3-fluoropyridine-4-carboxylic acid was used to examine the ASM contractility and associated reflexes in the guinea pig model of allergic airway inflammation by the following methods: (i) evaluation of specific airway resistance in vivo; (ii) evaluation of the contractile response of isolated ASM strips in vitro; and (iii) citric acid-induced cough reflex; (iv) measurement of exhaled NO levels (E(NO)). Allergic airway inflammation was induced by repetitive exposure of guinea pigs to ovalbumin (10(-6) M). The CRAC antagonist administered in a single dose to guinea pigs with confirmed allergic inflammation significantly reduced the cough response and the airway resistance, which corresponded with the findings in vitro. Long-term application of the CRAC antagonist had more strongly expressed effects. The results confirm the role of CRAC in the pathophysiology of experimental animal asthma and have a potential meaning for anti-asthma therapy.

LC-MS analyses of microcystins in fish tissues overestimate toxin levels-critical comparison with LC-MS/MS.

Microcystins are cyclic peptide toxins with hepatotoxic and tumour-promoting properties which are produced in high quantities in freshwater cyanobacterial water blooms, and several studies have reported microcystin accumulation in fish with possible food transfer to humans. In this study, we provide the first comparison of liquid chromatography with single mass-spectrometric and with tandem mass-spectrometric detection for analyses of microcystins in complex fish tissue samples. Use of traditional single mass spectrometry (i.e. monitoring of ions with m/z 519.5 for microcystin-RR and m/z 995.5 for microcystin-LR) was found to provide false-positive responses, thus overestimating the concentrations of microcystins in the tissue samples. More selective tandem mass spectrometry seems to provide more reliable results. The concentrations of microcystins detected by tandem mass spectrometry in fish from controlled-exposure experiments were more than 50% lower in comparison with concentrations obtained by single mass spectrometry. Extensive analyses of edible fish parts-muscles (148 fish specimens from eight different species from five natural reservoirs with dense cyanobacterial water blooms)-showed negligible microcystin concentrations (all analyses below the limit of detection; limit of detection of 1.2-5.4 ng/g fresh weight for microcystin-RR, microcystin-YR and microcystin-LR in multiple reaction monitoring mode). Our findings have practical consequences for critical re-evaluation of the health risks of microcystins accumulated in fish.

Bismuth-based quadruple therapy with bismuth subcitrate, metronidazole, tetracycline and omeprazole in the eradication of Helicobacter pylori.

BACKGROUND: A previous study showed that 14 days of qid bismuth-based triple therapy with tetracycline 500 mg, metronidazole 250 mg and colloidal bismuth subcitrate 120 mg resulted in excellent Helicobacter pylori eradication rates (89.5%). The present study looked at a shorter treatment period by adding omeprazole and by reducing the dose of tetracycline. METHODS: One hundred sixty-one patients with H pylori confirmed by histology and (13)carbon urea breath test were included in the study. They were treated for seven days with bismuth subcitrate 120 mg plus metronidazole 250 mg plus tetracycline 250 mg qid plus omeprazole 20 mg bid (OBMT). Patients were 18 to 75 years of age and had dyspepsia with or without a history of peptic ulcer. Patients with irritable bowel syndrome, active ulcer or previous attempt at eradication, or those who had used antibiotics or antiulcer drugs in the previous 30 days were excluded. Eradication was determined by two (13)carbon urea breath tests done one and three months, respectively, after treatment. Strains with minimal inhibitory concentrations of 8 microg/mL or higher were considered to be resistant to metronidazole. RESULTS: The overall per protocol eradication rate was 84%-89.5% in metronidazole-sensitive and 70.8% in metronidazole-resistant strains. Modified intent-to-treat analysis resulted in a 80% eradication rate--82.5% in metronidazole-sensitive and 66.7% in metronidazole-resistant strains. Only one patient discontinued treatment because of adverse events. CONCLUSIONS: The OBMT regimen used in this study is safe and effective against metronidazole-sensitive H pylori strains.

[The little known Fay syndrome (benign carotidynia)]. / Málo známy Fayov syndróm (benígna karotidynia).

The authors describe the little known Fay syndrome (benign carotidynia, extracranial vascular cephalea). They emphasize the importance of a careful case-history (in particular sore throat--usually unilateral), with careful physical examination (palpation sensitivity of the carotid artery beneath the bifurcation). The authors assume that the syndrome is not as rare but that it is rarely correctly diagnosed. The drug of choice are non-steroid antirheumatic drugs. The authors describe in an epicritical manner eight of their own observations made during the last three years.

Hypothalamic-pituitary dopaminergic function in hepatic failure in man.

The growth hormone (GH) response to apomorphine HCl (Apo) (0.75 mg sc), a dopamine (DA) receptor agonist, was assessed in healthy chronic alcoholics without cirrhosis (N = 20) and in patients with alcoholic cirrhosis both with (N = 5) and without (N = 14) hepatic encephalopathy (HE). A significant number of cirrhotic patients with (P less than 0.004) and without (P less than 0.002) HE had an impaired GH response (peak increment less than 5 ng/ml) compared with non-cirrhotic individuals. An impaired GH response was independent of the presence of HE. The magnitude of the GH response was unrelated to plasma oestrone, oestradiol, or progesterone concentrations but was significantly correlated with plasma testosterone levels (P less than 0.01). None of the patients with an abnormally low testosterone concentration showed a normal GH response. None of the subjects with HE showed an arousal response to Apo. These results suggest that DA receptor sensitivity is decreased in liver cirrhosis and that this decrease is related to inadequate circulating levels of testosterone. The occurrence of HE is independent of impaired DA function. The present study only evaluates DA function in the hypothalamic-pituitary axis and therefore may not reflect changes in other regions of brain.

The effect of growth hormone on the metabolism of a tryptophan load in the liver and brain of hypophysectomized rats.

Growth hormone antagonizes the induction of tryptophan pyrrolase and tyrosine amino-transferase by cortisol. We have shown that contrary to previous reports, growth hormone is also capable of antagonizing the induction of these enzymes by tryptophan and alpha-methyl tryptophan. As alpha-methyl tryptophan is not metabolized appreciably in the rat, our data show that growth hormone does not act indirectly through changes in the liver tryptophan content as was suggested previously. Growth hormone decreases the rate of tryptophan catabolism in vivo after induction of tryptophan pyrrolase by tryptophan and alpha-methyl tryptophan. Because the rate of catabolism of a tryptophan is slower in animals treated with growth hormone, tissue tryptophan levels and the rate of synthesis of 5-hydroxyltryptamine in the brain are higher in these animals than in those receiving tryptophan alone. Thus, although tryptophan administration raises brain 5-hydroxytryptamine levels, induction of tryptophan pyrrolase in the liver, by the load, limits the extent and duration of the rise in brain 5-hydroxytryptamine synthesis. This has important implications for the clinical use of tryptophan in psychiatric disorders, where tryptophan is given to produce long-lasting elevations of brain 5-hydroxytryptamine.

Stimulation of tyrosine aminotransferase activity by dl-alpha-methyltryptophan.

DL-alpha-Methyltryptophan (alphaMeTrp), a synthetic analogue of tryptophan, has been found to be a potent inducer of hepatic tyrosine aminotransferase activity in the adrenalectomized rat. alphaMeTrp is inactive in vitro. Unlike the action of other known inducers (tryptophan, hydrocortisone, adenosine cyclic 3:5-monophosphate, and glucagon), maximal stimulation of enzyme activity occurs only 16 to 30 hours after alphaMeTrp administration and the activity is still elevated at 96 hours. Only the L isomer of alphaMeTrp is active, and addition of a hydroxyl group to position 5 of the indole ring renders an inactive compound. The induction can be prevented by actinomycin D or cycloheximide but not galactosamine. Administration of alphaMeTrp together with hydrocortisone produced an additive stimulation of enzyme activity. alphaMeTrp given along with glucagon or adenosine cyclic 3:5-monophosphate caused a further but not additive increase in enzyme activity. Tryptophan given along with alphaMeTrp promoted no extra stimulation whatsoever. These data indicate that alphaMeTrp and tryptophan may act via a common pathway which in part requires RNA synthesis. Other enzymes, namely alanine and aspartate aminotransferase, ornithine aminotransferase, ornithine carbamoyltransferase, serine dehydratase, and histidine ammonialyase, were not affected by treatment of rats with alphaMeTrp.