Structure-guided development of Pb2+-binding DNA aptamers

Owing to its great threat to human health and environment, Pb2+ pollution has been recognized as a major public problem by the World Health Organization (WHO). Many DNA aptamers have been utilized in the development of Pb2+-detection sensors, but the underlying mechanisms remain elusive. Here, we report three Pb2+-complexed structures of the thrombin binding aptamer (TBA). These high-resolution crystal structures showed that TBA forms intramolecular G-quadruplex and Pb2+ is bound by the two G-tetrads in the center. Compared to K+-stabilized G-quadruplexes, the coordinating distance between Pb2+ and the G-tetrads are much shorter. The T3T4 and T12T13 linkers play important roles in dimerization and crystallization of TBA, but they are changeable for Pb2+-binding. In combination with mutagenesis and CD spectra, the G8C mutant structure unraveled that the T7G8T9 linker of TBA is also variable. In addition to expansion of the Pb2+-binding aptamer sequences, our study also set up one great example for quick and rational development of other aptamers with similar or optimized binding activity.

Rational Development and Characterization of a Ubiquitin Variant with Selectivity for Ubiquitin C-Terminal Hydrolase L3

There is currently a lack of reliable methods and strategies to probe the deubiquitinating enzyme UCHL3. Current small molecules reported for this purpose display reduced potency and selectivity in cellular assays. To bridge this gap and provide an alternative approach to probe UCHL3, our group has carried out the rational design of ubiquitin-variant activity-based probes with selectivity for UCHL3 over the closely related UCHL1 and other DUBs. The approach successfully produced a triple-mutant ubiquitin variant activity-based probe, UbVQ40V/T66K/V70F-PRG, that was ultimately 20,000-fold more selective for UCHL3 over UCHL1 when assessed by rate of inactivation assays. This same variant was shown to selectively form covalent adducts with UCHL3 in MDA-MB-231 breast cancer cells and no reactivity toward other DUBs expressed. Overall, this study demonstrates the feasibility of the approach and also provides insight into how this approach may be applied to other DUB targets.

Evaluation and Scale Forecast of Underground Space Resources of Historical and Cultural Cities in China

Following economic growth in the past three decades, rapid urbanization has caused many pronounced issues, such as spatial scarcity and cultural discontinuity, in Chinese historical and cultural cities. In order to better deal with the diversification of underground space resources, data and information, this study introduces a random forest algorithm and proposes a multi-layer information superposition method. According to the characteristics of different information, starting from qualitative and quantitative aspects, we explore the effective performance of the rational development of underground space resources. Taking Yangzhou City, China, as an example, this paper evaluates the suitability and calculates the development volume of urban underground space. The development capacity, potential value, and comprehensive quality of underground space resources are explored in an attempt to demonstrate the practicality and scientificity of the evaluation method for achieving the developmental goals of urban space reconstruction and historic preservation. On this basis, an underground space scale forecast is carried out to provide decision support for relevant planners, managers, and construction personnel that is conducive to the orderly development of urban space, alleviation of increasing human–land conflicts, and coordination of the protection and development of underground space resources in historical and cultural cities, ultimately promoting sustainable development of cities.

Plant Protein-Based Delivery Systems: An Emerging Approach for Increasing the Efficacy of Lipophilic Bioactive Compounds

The development of plant protein-based delivery systems to protect and control lipophilic bioactive compound delivery (such as vitamins, polyphenols, carotenoids, polyunsaturated fatty acids) has increased interest in food, nutraceutical, and pharmaceutical fields. The quite significant ascension of plant proteins from legumes, oil/edible seeds, nuts, tuber, and cereals is motivated by their eco-friendly, sustainable, and healthy profile compared with other sources. However, many challenges need to be overcome before their widespread use as raw material for carriers. Thus, modification approaches have been used to improve their techno-functionality and address their limitations, aiming to produce a new generation of plant-based carriers (hydrogels, emulsions, self-assembled structures, films). This paper addresses the advantages and challenges of using plant proteins and the effects of modification methods on their nutritional quality, bioactivity, and techno-functionalities. Furthermore, we review the recent progress in designing plant protein-based delivery systems, their main applications as carriers for lipophilic bioactive compounds, and the contribution of protein-bioactive compound interactions to the dynamics and structure of delivery systems. Expressive advances have been made in the plant protein area; however, new extraction/purification technologies and protein sources need to be found Their functional properties must also be deeply studied for the rational development of effective delivery platforms.

Light-mediated discovery of surfaceome nanoscale organization and intercellular receptor interaction networks

The molecular nanoscale organization of the surfaceome is a fundamental regulator of cellular signaling in health and disease. Technologies for mapping the spatial relationships of cell surface receptors and their extracellular signaling synapses would unlock theranostic opportunities to target protein communities and the possibility to engineer extracellular signaling. Here, we develop an optoproteomic technology termed LUX-MS that enables the targeted elucidation of acute protein interactions on and in between living cells using light-controlled singlet oxygen generators (SOG). By using SOG-coupled antibodies, small molecule drugs, biologics and intact viral particles, we demonstrate the ability of LUX-MS to decode ligand receptor interactions across organisms and to discover surfaceome receptor nanoscale organization with direct implications for drug action. Furthermore, by coupling SOG to antigens we achieved light-controlled molecular mapping of intercellular signaling within functional immune synapses between antigen-presenting cells and CD8+ T cells providing insights into T cell activation with spatiotemporal specificity. LUX-MS based decoding of surfaceome signaling architectures thereby provides a molecular framework for the rational development of theranostic strategies.

An Exploratory Analysis of Changes in Circulating Plasma Protein Profiles Following Image-Guided Ablation of Renal Tumours Provides Evidence for Effects on Multiple Biological Processes

Further biological understanding of the immune and inflammatory responses following ablation is critical to the rational development of combination ablation-immunotherapies. Our pilot exploratory study evaluated the circulating plasma protein profiles after image-guided ablation (IGA) of small renal masses to determine the resultant systemic effects and provide insight into impact both on the tumour and immune system. Patients undergoing cryotherapy (CRYO), radiofrequency ablation (RFA) or microwave ablation (MWA) for small renal tumours were recruited. Blood samples were obtained at four timepoints; two baselines prior to IGA and at 24 h and 1–3 months post-IGA, and a panel of 164 proteins measured. Of 55 patients recruited, 35 underwent ablation (25 CRYO, 8 RFA, 2 MWA) and biomarker measurements. The most marked changes were 24 h post-CRYO, with 29 proteins increasing and 18 decreasing significantly, principally cytokines and proteins involved in regulating inflammation, danger-associated molecular patterns (DAMPs), cell proliferation, hypoxic response, apoptosis and migration. Intra-individual variation was low but inter-individual variation was apparent, for example all patients showed increases in IL-6 (1.7 to 29-fold) but only 50% in CD27. Functional annotation analysis highlighted immune/inflammation and cell proliferation/angiogenesis-related clusters, with interaction networks around IL-6, IL-10, VEGF-A and several chemokines. Increases in IL-8, IL-6, and CCL23 correlated with cryoprobe number (p = 0.01, rs = 0.546; p = 0.009, rs = 0.5515; p = 0.005, rs = 0.5873, respectively). This initial data provide further insights into ablation-induced biological changes of relevance in informing trial design of immunotherapies combined with ablation.

Rationality of internal control according to COSO-INTOSAI principles in public sector organizations

Subject. The article investigates the rationality of internal controls in accordance with COSO-INTOSAI recommendations in public sector organizations. Objectives. The aim is to classify the reasons for failures and the actual refusal to directly follow the COSO-INTOSAI recommendations in public sector organizations in Russia and the UK, as well as to determine the directions for the rational development of internal control in such organizations. Methods. The study employs the content analysis of domestic and foreign sources, the comparative analysis. Results. The study classifies the reasons for failures, i.e. excessive universalism and inanity of recommendations borrowed from professional participants of the financial market, hiring employees from financial markets, erroneous creation of additional units, procedures and documents, objective lack of authority, motivation and qualification of employees to implement the recommendations. The areas of rational development are determined on the basis of sector specifics, real powers of budget fund managers, professional duties of chief accountants as controllers, reasonable centralization and digitalization of public services. Conclusions. The findings may clarify the regulatory and departmental documents of the Ministry of Finance and managers of budget funds in internal control of public sector organizations, and improve the performance of such organizations, increase the efficiency of budget spending and off-budget operations.

Impact of Island Urbanization on Freshwater Lenses: A Case Study on a Small Coral Island

Freshwater resources on small coral islands mainly exist in the form of freshwater lenses. The freshwater lens is highly vulnerable to salinization due to natural recharge variations and urbanization construction. However, it is unclear how a freshwater lens evolves under the influence of urbanization construction and which factors control its evolution. Based on the hydrogeological data of a small coral island in China, a corresponding 3D numerical model was established by the Visual MODFLOW software to investigate the formation and evolution of freshwater lenses under natural conditions. Thereby, the island reclamation scenario and impermeable surface scenario were set up and the changes in morphology and volume of the freshwater lens were analyzed. The results show the following: (1) After island reclamation and island building, the freshwater lens would reach a stable state after 25 years and the freshwater lens would also appear in the newly added part of the island with a thickness of 9.5 m, while the volume of the total freshwater lens would increase to 1.22 times that of the original island. (2) When the impermeable surface is built at different positions of the island, the reduction in the volume of the freshwater lens, in the order from large to small, is Scenario B (northeast side), Scenario A (southwest side) and Scenario C (central); with the increase in the impermeable surface area, the volume of the freshwater lens would gradually decrease and the volume of the freshwater lens would decrease by more than 50% with the impermeable surface exceeding 30% of the island area. The study has important implications for the conservation and rational development of subsurface freshwater resources on islands.

ZAKAT WITHIN THE FRAME OF THE UNITARY STATE REPUBLIC OF INDONESIA: ADAPTABILITY OF SHARIA VALUES AND CULTURES

This paper aims to elaborate the dynamics of Islamic law assimilation with local culture. With the hope that in the future it will form a basic perspective in shaping the philanthropy of contemporary Islamic law based on the reality of Indonesian society. The basic questions which is going to be answered through this paper are, first, the extent of the adaptability of Islamic law in the midst of multicultural society conditions in Indonesia. Second, what are juridical, normative and sociological arguments in placing zakat as a support for the integrity of the Unitary State Republic of Indonesia. Third, How is the Formulation of the Concept of Zakat within the frame of Unitary State Republic of Indonesia? The conclusion from this study shows that, first, the characteristics of Islamic law indicate the ability of adaptability to the culture of the society in which it is accepted. Even in this case Islam has provided important principles regarding rational development in efforts to adapt to its new environment. Second, placing zakat as a support for the integrity of the Unitary State Republic of Indonesia is worth to be formulated. This is a logical consequence of the efforts of the Islamic ummah to always place al-Qur'an and al-Sunnah as limited texts. One of the most fundamental results of Indonesian social culture is the realization of the Unitary State of the Republic of Indonesia. Making the formulation of zakat in the frame of the Unitary State Republic of Indonesia is a clear proof that Islamic law contains universal values that are valid in any time and any place. Third, the methodological formulation of zakat in the frame of Unitary State of the Republic of Indonesia is in a dynamic and accommodating ijtihad towards change. This methodological framework is based on al-Mashlahah, ‘Urf, Sad Dzaria'ah and dialectics between Gama and the State.