Thermal inactivation and conformational lock studies on glucose oxidase

In this study, the dissociative thermal inactivation and conformational lock theories are applied for the homodimeric enzyme glucose oxidase (GOD) in order to analyze its structure. For this purpose, the rate of activity reduction of glucose oxidase is studied at various temperatures using b-D-glucose as the substrate by incubation of enzyme at various temperatures in the wide range between 40 and 70 �C using UV–Vis spectrophotometry. It was observed that in the two ranges of temperatures, the enzyme has two different forms. In relatively low temperatures, the enzyme is in its dimeric state and has normal activity. In high temperatures, the activity almost disappears and it aggregates. The above achievements are confirmed by dynamic light scattering. The experimental parameter ‘‘n’’ as the obvious number of conformational locks at the dimer interface of glucose oxidase is obtained by kinetic data, and the value is near to two. To confirm the above results, the X-ray crystallography structure of the enzyme, GOD (pdb, 1gal), was also studied. The secondary and tertiary structures of the enzyme to track the thermal inactivation were studied by circular dichroism and fluorescence spectroscopy, respectively. We proposed a mechanism model for thermal inactivation of GOD based on the absence of the monomeric form of the enzyme by circular dichroism and fluorescence spectroscopy

СТАН ТА ТЕНДЕНЦІЇ МІЖБЮДЖЕТНОГО РЕГУЛЮВАННЯ В УКРАЇНІ В УМОВАХ ФІСКАЛЬНОЇ ДЕЦЕНТРАЛІЗАЦІЇ

A balanced socio-economic development of any country is closely related to the efficiency of the system of intergovernmental relations, which should be aimed at optimizing the process of fiscal decentralization and clear definition of the powers of local government. Interbudgetary regulation in Ukraine reflects processes of the national and political importance and is of great importance in the composition of intergovernmental fiscal relations, which priority is growing in terms of decentralization reforms. Purpose. The main objectives of this study are: to summarize a complex task that is resolved in the process of budgetary control; to analyze the dynamics of revenues of the consolidated budget of Ukraine in 2007-2016 and the proportion of own local revenues in GDP; to distinguish formation features of Ukrainian local budgets by region; to analyze volumes of local budgets per capita in terms of area; to highlight the leverage rate of local budgets; to analyze the share of interbudget transfers in total local budgets. Results. The study is a synthesis of the complex challenges that must be resolved in the process of budgetary control, such as: balancing the budget; foster the interest of local governments in the maximum revenue mobilization; reallocation of existing budgetary resources, taking into account local features. The dynamics of the revenues of the consolidated budget of Ukraine in 2007-2016 was analyzed and concluded that the proportion of own local revenues in GDP tended to decrease with a simultaneous increase in the share of the interbudget transfers from the state budget. Abovementioned resulted in the ensuring of a stable share of local budgets in the GDP structure (at 14-15%). As a result of the intergovernmental fiscal decentralization reform by the results of 2016 the share of own revenues of local budgets Ukraine in GDP rose to 7.2%, or 1.1% compared to 2015 with a simultaneous decrease in the interbudget transfers share from the state budget to 8.2% (0.6% compared to 2015). Analyzed the share interbudget transfers in total local budgets, which for a long period tended to increase (from 47.2% in 2007 to 53.4% in 2016), which is contrary to the basic principles of fiscal decentralization. Conclusion. Grounded features of Ukrainian local budget revenues by region and proved that the unevenness of income between regions of Ukraine are often equivalent to the level of industrial production. Analyzed volumes of local budgets per capita in terms of area. Presented the list of influence instruments on local budgets indexes of incomes, including the increase in the minimum wage, inflation, continuing reform of the budget system in the direction of fiscal decentralization and transfer volumes.Представлен комплекс задач, который должен решаться в процессе межбюджетного регулирования, проанализирована динамика доходной части сводного бюджета Украины в 2007-2016 гг. и удельный вес собственных доходов местных бюджетов в структуре ВВП, выделены особенности формирования доходов местных бюджетов Украины по регионам. Проанализированы объемы доходов местных бюджетов на душу населения в разрезе территорий, освещены рычаги влияния на показатель доходов местных бюджетов, проведен анализ удельного веса межбюджетных трансфертов в общей структуре местных бюджетов.Узагальнено комплекс завдань, що має вирішуватись у процесі міжбюджетного регулювання, проаналізовано динаміку дохідної частини зведеного бюджету України у 2007–2016 рр. та питому вагу власних доходів місцевих бюджетів в структурі ВВП, виокремлено особливості формування доходів місцевих бюджетів України за регіонами. Проаналізовано обсяги доходів місцевих бюджетів на душу населення у розрізі територій, висвітлено важелі впливу на показник доходів місцевих бюджетів, здійснено аналіз питомої ваги міжбюджетних трансфертів у загальній структурі місцевих бюджетів

Solvent and thermal stability, and pH kinetics, of proline-specific dipeptidyl peptidase IV-like enzyme from bovine serum

Proline-specific dipeptidyl peptidase-like (DPP IV; EC 3.4.14.5) activity in bovine serum has attracted little attention despite its ready availability and the paucity of useful proline-cleaving enzymes. Bovine serum DPP IV-like peptidase is very tolerant of organic solvents, particularly acetonitrile: upon incubation for 1 h at room temperature in 70% acetonitrile, 47% dimethylformamide, 54% DMSO and 33% tetrahydrofuran (v/v concentrations) followed by dilution into the standard assay mixture, the enzyme retained half of its aqueous activity. As for thermal performance in aqueous buffer, its relative activity increased up to 50 ◦C. Upon thermoinactivation at 71 ◦C, pH 8.0 (samples removed periodically, cooled on ice, then assayed under optimal conditions), residual activities over short times fit a first-order decay with a k-value of 0.071±0.0034 min−1. Over longer times, residual activities fit to a double exponential decay with k1 and k2 values of 0.218±0.025 min−1 (46±4% of overall decay) and 0.040±0.002 min−1 (54±4% of overall decay), respectively. The enzyme’s solvent and thermal tolerances suggest that it may have potential for use as a biocatalyst in industry. Kinetic analysis with the fluorogenic substrate Gly-Pro-7-aminomethylcoumarin over a range of pH values indicated two pK values at 6.18±0.07 and at 9.70±0.50. We ascribe the lower value to the active site histidine; the higher may be due to the active site serine or to a free amino group in the substrate

Performance of the ABCN-25 readout chip for the ATLAS Inner Detector Upgrade

We present the test results of the ABCN-25 front end chip implemented in CMOS 0.25 μm technology and optimised for the short, 2.5 cm, silicon strips intended to be used in the upgrade of the ATLAS Inner Detector. We have obtained the full functionality of the readout part, the expected performance of the analogue front-end and the operation of the power control circuits. The performance is evaluated in view of the minimization of the power consumption, as the upgrade detector may contain up to 70 million of channels. System tests with different power distribution schemes proposed for the future tracker detectors are possible with this chip. The ABCN-25 ASIC is now serving as the prototype readout chip in the developments of the modules and staves for the upgrade of the ATLAS Inner Detector

MicroRNA-155 is induced during the macrophage inflammatory response

The mammalian inflammatory response to infection involves the induction of several hundred genes, a process that must be carefully regulated to achieve pathogen clearance and prevent the consequences of unregulated expression, such as cancer. Recently, microRNAs (miRNAs) have emerged as a class of gene expression regulators that has also been linked to cancer. However, the relationship between inflammation, innate immunity, and miRNA expression is just beginning to be explored. In the present study, we use microarray technology to identify miRNAs induced in primary murine macrophages after exposure to polyriboinosinic:polyribocytidylic acid or the cytokine IFN-{beta}. miR-155 was the only miRNA of those tested that was substantially up-regulated by both stimuli. It also was induced by several Toll-like receptor ligands through myeloid differentiation factor 88- or TRIF-dependent pathways, whereas up-regulation by IFNs was shown to involve TNF-{alpha} autocrine signaling. Pharmacological inhibition of the kinase JNK blocked induction of miR-155 in response to either polyriboinosinic:polyribocytidylic acid or TNF-{alpha}, suggesting that miR-155-inducing signals use the JNK pathway. Together, these findings characterize miR-155 as a common target of a broad range of inflammatory mediators. Importantly, because miR-155 is known to function as an oncogene, these observations identify a potential link between inflammation and cancer

Anthropology, Brokerage and Collaboration in the development of a Tongan Public Psychiatry: Local Lessons for Global Mental Health

The Global Mental Health (GMH) movement has revitalised questions of the translatability of psychiatric concepts and the challenges of community engagement in countries where knowledge of the biomedical basis for psychiatric diagnosis is limited or challenged by local cultural codes. In Tonga, the local psychiatrist Dr Puloka has successfully established a publicly accessible psychiatry that has raised admission rates for serious mental illness and addressed some of the stigma attached to diagnosis. On the basis of historical analysis and ethnographic fieldwork with healers, doctors and patients since 1998, this article offers an ethnographic contextualization of the development and reception of three key interventions during the 1990s inspired by traditional healing and reliant on the translation of psychiatric terms and diagnosis. Dr Puloka’s use of medical anthropological and transcultural psychiatry research informed a community engaged brokerage between the implications of psychiatric nosologies and local needs. As such it reveals deficiencies in current polarised positions on the GMH project and offers suggestions to address current challenges of the Global Mental Health movement

Expamers: a new technology to control T cell activation

T cell activation is a cornerstone in manufacturing of T cell-based therapies, and precise control over T cell activation is important in the development of the next generation T-cell based therapeutics. This need cannot be fulfilled by currently available methods for T cell stimulation, in particular not in a time dependent manner. Here, we describe a modular activation reagent called Expamers, which addresses these limitations. Expamers are versatile stimuli that are intended for research and clinical use. They are readily soluble and can be rapidly bound and removed from the cell surface, allowing nearly instantaneous initiation and termination of activation signal, respectively. Hence, Expamers enable precise regulation of T cell stimulation duration and provide promise of control over T cell profiles in future products. Expamers can be easily adopted to different T cell production formats and have the potential to increase efficacy of T cell immunotherapeutics

The challenges of communicating research evidence in practice: perspectives from UK health visitors and practice nurses

<p>Background: Health practitioners play a pivotal role in providing patients with up-to-date evidence and health information. Evidence-based practice and patient-centred care are transforming the delivery of healthcare in the UK. Health practitioners are increasingly balancing the need to provide evidence-based information against that of facilitating patient choice, which may not always concur with the evidence base. There is limited research exploring how health practitioners working in the UK, and particularly those more autonomous practitioners such as health visitors and practice nurses working in community practice settings, negotiate this challenge. This research provides a descriptive account of how health visitors and practice nurses negotiate the challenges of communicating health information and research evidence in practice.</p> <p>Methods: A total of eighteen in-depth telephone interviews were conducted in the UK between September 2008 and May 2009. The participants comprised nine health visitors and nine practice nurses, recruited via adverts on a nursing website, posters at a practitioner conference and through recommendation. Thematic analysis, with a focus on constant comparative method, was used to analyse the data.</p> <p>Results: The data were grouped into three main themes: communicating evidence to the critically-minded patient; confidence in communicating evidence; and maintaining the integrity of the patient-practitioner relationship. These findings highlight some of the daily challenges that health visitors and practice nurses face with regard to the complex and dynamic nature of evidence and the changing attitudes and expectations of patients. The findings also highlight the tensions that exist between differing philosophies of evidence-based practice and patient-centred care, which can make communicating about evidence a daunting task.</p> <p>Conclusions: If health practitioners are to be effective at communicating research evidence, we suggest that more research and resources need to be focused on contextual factors, such as how research evidence is negotiated, appraised and communicated within the dynamic patient-practitioner relationship.</p&gt