The expression of several barley (Hordeum vulgare) cold-regulated (cor) genes during cold acclimation was blocked in the albino mutant $a_{\text{n}}$, implying a chloroplast control on mRNAs ...accumulation. By using albino and xantha mutants ordered according to the step in chloroplast biogenesis affected, we show that the cold-dependent accumulation of cor14b, tmc-ap3, and blt14 mRNAs depends on plastid developmental stage. Plants acquire the ability to fully express cor genes only after the development of primary thylakoid membranes in their chloroplasts. To investigate the chloroplast-dependent mechanism involved in cor gene expression, the activity of a 643-bp cor14b promoter fragment was assayed in wild-type and albino mutant $a_{\text{n}}$ leaf explants using transient β-glucuronidase reporter expression assay. Deletion analysis identified a 27-bp region between nucleotides -274 and -247 with respect to the transcription start point, encompassing a boundary of some element that contributes to the cold-induced expression of cor14b. However, cor14b promoter was equally active in green and in albino $a_{\text{n}}$ leaves, suggesting that chloroplast controls cor14b expression by posttranscriptional mechanisms. Barley mutants lacking either photosystem I or II reaction center complexes were then used to evaluate the effects of redox state of electron transport chain components on COR14b accumulation. In the mutants analyzed, the amount of COR14b protein, but not the steady-state level of the corresponding mRNA, was dependent on the redox state of the electron transport chain. Treatments of the vir-zb63 mutant with electron transport chain inhibitors showed that oxidized plastoquinone promotes COR14b accumulation, thus suggesting a molecular relationship between plastoquinone/plastoquinol pool and COR14b.
We report on the results obtained by measuring the stoichiometry of antenna polypeptides in Photosystem I (PSI) from Arabidopsis thaliana. This analysis was performed by quantification of Coomassie ...blue binding to individual LHCI polypeptides, fractionation by SDS/PAGE, and by the use of recombinant light harvesting complex of Photosystem I (Lhca) holoproteins as a standard reference. Our results show that a single copy of each Lhca1–4 polypeptide is present in Photosystem I. This is in agreement with the recent structural data on PSI–LHCI complex Ben Shem, A., Frolow, F. and Nelson, N. (2003) Nature, 426, 630–635. The discrepancy from earlier estimations based on pigment binding and yielding two copies of each LHCI polypeptide per PSI, is explained by the presence of ‘gap’ and ‘linker’ chlorophylls bound at the interface between PSI core and LHCI. We showed that these chlorophylls are lost when LHCI is detached from the PSI core moiety by detergent treatment and that gap and linker chlorophylls are both Chl a and Chl b. Carotenoid molecules are also found at this interface between LHCI and PSI core. Similar experiments, performed on PSII supercomplexes, showed that dissociation into individual pigment‐proteins did not produce a significant loss of pigments, suggesting that gap and linker chlorophylls are a peculiar feature of Photosystem I.
FeFe-hydrogenases catalyze the reversible production of H2 in some bacteria and unicellular eukaryotes. These enzymes require ancillary proteins to assemble the unique active site H-cluster, a ...complex structure composed of a 2Fe center bridged to a 4Fe-4S cubane. The first crystal structure of a key factor in the maturation process, HydF, has been determined at 3 Å resolution. The protein monomer present in the asymmetric unit of the crystal comprises three domains: a GTP-binding domain, a dimerization domain, and a metal cluster-binding domain, all characterized by similar folding motifs. Two monomers dimerize, giving rise to a stable dimer, held together mainly by the formation of a continuous β-sheet comprising eight β-strands from two monomers. Moreover, in the structure presented, two dimers aggregate to form a supramolecular organization that represents an inactivated form of the HydF maturase. The crystal structure of the latter furnishes several clues about the events necessary for cluster generation/transfer and provides an excellent model to begin elucidating the structure/function of HydF in FeFe-hydrogenase maturation.
Background: HydF is a GTPase essential for maturation of FeFe-hydrogenase.
Results: The first crystal structure of HydF has been determined.
Conclusion: The protein monomer comprises a GTP-binding domain, a dimerization domain, and a metal-cluster binding domain. Two monomers dimerize, and two dimers can aggregate to a tetramer.
Significance: The crystal structure of the latter furnishes several clues about the events necessary for cluster generation.
BACKGROUND Tra le patologie disabilitanti che possono colpire i bambini fin dalla nascita, la Paralisi Cerebrale Infantile (PCI) occupa il primo posto, sia per la frequenza, sia per la molteplicità ...di disturbi intercorrenti. La cura della PCI richiede quindi un percorso riabilitativo lungo e complesso, che investe i servizi sanitari, le strutture educative, le agenzie sociali ma anche e soprattutto il contesto famigliare. Da circa trent’anni in Canada viene proposto un approccio centrato sulla famiglia definito Family Centered Care. Questo tipo di approccio presuppone una responsabilità condivisa tra i portatori di cure e le famiglie nella progettazione, pianificazione e applicazione delle cure rivolte al bambino. In Italia, Le Linee Guida prevedono la stesura del Piano Riabilitativo Individuale (PRI) che prevede il consenso informato della famiglia all’atto medico; la responsabilità del progetto terapeutico è assunta unicamente dal medico riabilitatore che informerà la famiglia e concordare con essa e gli altri operatori socio sanitari dell’equipe gli obiettivi riabilitativi, le modalità, le misure di esito attese e le tempistiche degli interventi attraverso la stesura di programmi terapeutici condivisi in un modello che si presenta così “child centered” ma con un coinvolgimento partecipativo costante della famiglia. Lo scopo di questo studio è confrontare la percezione delle famiglie Italiane e Canadesi riguardo ai due differenti modelli di assistenza sanitaria nella riabilitazione delle patologie neuromotorie infantili. Metodi Sono stati raccolti i dati di 219 questionari MPOC-20 e di 75 MPOC-SP, provenienti dai servizi territoriali infantili della regione Emilia Romagna, e comparati con i dati della provincia dell’Ontario pubblicati dal gruppo CanChild. Risultati Confrontando i risultati dei questionari MPOC-20 e MPOC-SP ottenuti in Emilia Romagna e in Ontario, abbiamo riscontrato che le valutazioni medie nei diversi domini presentano poche differenze. L’unico dominio che mostra un valore più basso nei dati italiani riguarda "Fornire le informazioni specifiche riguardanti il bambino" (in Emilia Romagna la media è 4.69, in Ontario 5.23). Al contrario, per tutti gli altri domini, in Emilia Romagna la media è più alta rispetto all'Ontario. Se invece consideriamo il questionario compilato dai Fisioterapisti, riscontriamo un più alto livello di soddisfazione per quanto riguarda il trattamento in Ontario. La media in Emilia Romagna, infatti, risulta 3.74, mentre in Ontario 4.68. La differenza maggiore viene riscontrata nel dominio “Fornire le Informazioni generali”. Al contrario, la media è più alta in Emila Romagna quando il fisioterapista valuta la sensibilità interpersonale. Nel dominio “Mostrare Sensibilità Interpersonale” la soddisfazione è alta in entrambe le nazioni: 5.76 in Emilia Romagna, 5.83 in Ontario. Discussione In Emilia Romagna, la comunicazione tra famiglie ed operatori sanitari riguardante le informazioni generali, la patologia e il trattamento dovrebbe essere migliorata per aumentare la soddisfazione e la collaborazione, limitare le problematiche sociali e psicologiche riducendo così la richiesta di trattamenti complementari o alternativi. Conclusioni I risultati dello studio ci permettono di concludere che la soddisfazione delle famiglie italiane e canadesi riguardo la qualità dell’assistenza è abbastanza simile e che, con alcune lievi modifiche, il modello italiano di riabilitazione nella PCI potrebbe rimanere competitivo. Un modello dei servizi che ponga al centro il bambino coinvolgendo in maniera costante la famiglia nel suo programma di cure che potremmo così definire “child and family centered” sembra essere la chiave di lettura per proporre servizi di qualità, efficacia ed efficienza.
BACKGROUND Among disabling pathologies, that affect children from birth, Cerebral Palsy (CP) is the most important for frequency and multiplicity of associated disorders. Care of CP requires a long ...and complex rehabilitation process that involves healthcare services, educational facilities, and social agencies, but above all family members (SCPE 2000). In Canada, families have decision-making power in childcare, which includes rehabilitation treatments and socio-educational interventions. This family-centered approach presupposes a shared responsibility between caregivers and family in planning and applying child rehabilitation therapies. In Italy, “Recommendations for cerebral palsy rehabilitation” provide for a drafting of an Individual Rehabilitation Plan (PRI), according to the ICF-CY model. Designing the therapeutic project (PRI) is the physician’s responsibility, who subsequently involves the family in reaching objectives, timing interventions, realizing setting modalities and measuring outcomes. This approach is child-centered, however with the participatory involvement of family. The aim of this study is to compare the perception of Italian and Canadian families regarding these two different healthcare models in CP rehabilitation. Methods Data from 219 MPOC-20 and 75 MPOC-SP questionnaires were collected from child healthcare services in Emilia Romagna Region and compared to Ontario province data published by CanChild. Results By comparing MPOC-20 and MPOC-SP results obtained in Emilia Romagna and Ontario, we found that average values of various domains reveal few differences. The only domain showing lower results for Emilia Romagna concerned child-specific information supply (Emilia Romagna average is 4.69, Ontario is 5.23). On the contrary, for all the remaining domains, Emilia Romagna had higher averages. Considering physiotherapist questionnaires, we found higher satisfaction levels regarding treatment in Ontario. The greatest difference related to the “Providing General Information” domain: parental perception; Emilia Romagna average was 3.74, while Ontario’s average was 4.68. For the domain “Showing Interpersonal Sensitivity”, satisfaction was high for both countries: 5.76 in Emilia Romagna, 5.83 in Ontario. Discussion Communication regarding general aspects, pathology and treatment information must be improved in Emilia Romagna in order to increase satisfaction and cooperation between families and healthcare professionals. Conclusions The study results allow us to conclude that Italian and Canadian family satisfaction of healthcare quality is quite similar, and that the Italian model of CP rehabilitation, with a few slight modifications, could be judged competitive. An organizational model focused on the child, constantly involving the family in care programs, which we could coin "Child and Family Centre approach", would seem to be the key to a higher quality, efficacy and efficiency service.
The expression of several barley (
Hordeum vulgare
) cold-regulated (
cor
) genes during cold acclimation was blocked in the
albino
mutant
a
n
, implying a chloroplast control on mRNAs accumulation. ...By using
albino
and
xantha
mutants ordered according to the step in chloroplast biogenesis affected, we show that the cold-dependent accumulation of
cor14b
,
tmc
-
ap3
, and
blt14
mRNAs depends on plastid developmental stage. Plants acquire the ability to fully express
cor
genes only after the development of primary thylakoid membranes in their chloroplasts. To investigate the chloroplast-dependent mechanism involved in
cor
gene expression, the activity of a 643-bp
cor14b
promoter fragment was assayed in wild-type and
albino
mutant
a
n
leaf explants using transient β-glucuronidase reporter expression assay. Deletion analysis identified a 27-bp region between nucleotides −274 and −247 with respect to the transcription start point, encompassing a boundary of some element that contributes to the cold-induced expression of
cor14b
. However,
cor14b
promoter was equally active in green and in albino
a
n
leaves, suggesting that chloroplast controls
cor14b
expression by posttranscriptional mechanisms. Barley mutants lacking either photosystem I or II reaction center complexes were then used to evaluate the effects of redox state of electron transport chain components on COR14b accumulation. In the mutants analyzed, the amount of COR14b protein, but not the steady-state level of the corresponding mRNA, was dependent on the redox state of the electron transport chain. Treatments of the
vir
-
zb63
mutant with electron transport chain inhibitors showed that oxidized plastoquinone promotes COR14b accumulation, thus suggesting a molecular relationship between plastoquinone/plastoquinol pool and COR14b.
Nine clones coding different components of the protein synthesis machinery were selected from an EST barley library prepared from cold exposed plants and tested for their expression at low ...temperature. Northern analysis revealed that expression of elongation factor (EF) 1Bβ and two ribosomal protein (RP) genes S7 and L7A was enhanced following exposure to 3 °C, but not during dehydration or exogenous ABA treatment. The mRNA levels of EF1Bβ, RPS7, and RPL7A did not vary between cultivars with different frost tolerance, but differences in expression were found between different species. Experiments with an
albino mutant and etiolated plants revealed that the cold-dependent regulation of EF1Bβ, RPS7, and RPL7A transcripts is controlled by a chloroplast-related pathway impaired in the mutant.
While smart metering applications have initially focused on energy and gas utility markets, water consumption has recently become the subject of increasing attention. Unfortunately, despite the large ...number of solutions available on the market, the lack of an open and widely accepted communication standard means that vendors typically propose proprietary data collection solutions whose adoption causes non-trivial problems to water utility companies in term of costs, vendor lock-in, and lack of control on the data collection infrastructure. There is the need for open and interoperable smart water metering solutions, capable of collecting data from the wide range of water meters on the market. This paper reports our experience in the development and field testing of a highly interoperable smart water metering solution, which we designed in collaboration with several water utility companies and which we deployed in Gorino Ferrarese, Italy, in collaboration with CADF (Consorzio Acque Delta Ferrarese), the water utility serving the city. At the core of our solution is SWaMM (Smart Water Metering Middleware), an interoperable wireless IoT middleware based on the Edge computing paradigm, which proved extremely effective in interfacing with several types of smart water meters operating with different protocols.
Abstract
Objective
To investigate risk factors for damage development in a prospective inception cohort of early diagnosed SLE patients.
Methods
The Early Lupus Project recruited an inception cohort ...of patients within 12 months of SLE classification (1997 ACR criteria). At enrolment and every 6 months thereafter, the SLICC/ACR Damage Index was recorded. The contribution of baseline and time-varying covariates to the development of damage, defined as any SLICC/ACR Damage Index increase from 0 to ≥1, was assessed using univariate analysis. Forward-backward Cox regression models were fitted with covariates with P < 0.05 to identify factors independently associated with the risk of damage development.
Results
Overall, 230 patients with a mean (s.d.) age of 36.5 (14.4) years were eligible for this study; the mean number of visits per patient was 5.3 (2.7). There were 51 (22.2%) patients with SLICC/ACR Damage Index ≥1 after 12 months, 59 (25.6%) after 24 months and 67 (29.1%) after 36 months. Dyslipidaemia P = 0.001; hazard ratio (HR) 2.9; 95% CI 1.5, 5.6, older age (P = 0.001; HR 3.0; 95% CI 1.6, 5.5), number of organs/systems involved (P = 0.002; HR 1.4; 95% CI 1.1, 1.8) and cardiorespiratory involvement (P = 0.041; HR 1.9; 95% CI 1.0, 3.7) were independently associated with an increased risk of developing damage. Risk profiles for damage development differed for glucocorticoid-related and -unrelated damage. HCQ use (P = 0.005; HR 0.4; 95% CI 0.2, 0.8) reduced the risk of glucocorticoid-unrelated damage.
Conclusion
We identified risk factors of damage development, but little effect of glucocorticoids, in this early SLE cohort. Addressing modifiable risk factors from the time of SLE diagnosis might improve patient outcomes.
•One-pot, low-cost and reproducible g-C3N4 grafting onto silica microparticles.•g-C3N4@silica novel application for SPE of Fluoroquinolones from natural water.•Quantitative recovery in tap and raw ...river waters (lowngL−1).•Good precision and sensitivity for g-C3N4@silica SPE coupled to HPLC-FD.•g-C3N4@silica advantageous in terms of preparation, cost, efficiency, reusability.
The analytical application of silica-supported graphitic carbon nitride (g-C3N4@silica) for solid-phase extraction (SPE) of fluoroquinolone (FQ) pollutants from water is presented for the first time. g-C3N4@silica was easily and quickly prepared by one-pot thermal condensation of dicyandiamide and characterized by powder X-ray diffraction, thermogravimetric analysis, scanning electron microscopy, Fourier transform infrared spectroscopy and surface area measurements. The novel composite was applied as sorbent for SPE of FQs from water prior high-performance liquid chromatography with fluorescence detection. The extraction efficiency of g-C3N4 was tested in tap and surface waters at actual concentrations (10–100ngL−1). Quantitative adsorption was achieved using 100mg sorbent (20wt% g-C3N4) for pre-concentration of 50–500mL sample, at the native pH (∼7.5–8). Elution was performed with 25mM H3PO4 aqueous solution-acetonitrile (80:20), obtaining recoveries in the range 70–114%, enrichment factors up to 500 and inter-day RSDs≤12%. The batch-to-batch reproducibility was assessed on three independently synthesized g-C3N4@silica preparations (RSD 6–12%). g-C3N4 supported on silica microparticles proved to be of easy preparation, inexpensive, reusable for at least 4 extractions of raw surface waters, and suitable for determination in real matrices.
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