Genetically encoded intracellular sensors based on fluorescent proteins

Green fluorescent protein from Aequorea victoria and its many homologs are now widely used in basic and applied research. These genetically encoded fluorescent markers can detect localization of cell proteins and organelles in living cells and also cells and tissues in living organisms. Unique instruments and methods for studies of molecular biology of a cell and high throughput drug screenings are based on fluorescent proteins. This review deals with the most intensively evolving directions in this field, the development of genetically encoded sensors. Changes in their spectral properties are used for monitoring of cell enzyme activities or changes in concentrations of particular molecules.     

Improved functional expression of recombinant human ether-a-go-go (hERG) K+ channels by cultivation at reduced temperature

Background

Genetically encoded sensors developed on the basis of green fluorescent protein (GFP)-like proteins are becoming more and more popular instruments for monitoring cellular analytes and enzyme activities in living cells and transgenic organisms. In particular, a number of Ca²⁺ sensors have been developed, either based on FRET (Fluorescence Resonance Energy Transfer) changes between two GFP-mutants or on the change in fluorescence intensity of a single circularly permuted fluorescent protein (cpFP).

Results

Here we report significant progress on the development of the latter type of Ca²⁺ sensors. Derived from the knowledge of previously reported cpFP-based sensors, we generated a set of cpFP-based indicators with different spectral properties and fluorescent responses to changes in Ca²⁺ concentration. Two variants, named Case12 and Case16, were characterized by particular high brightness and superior dynamic range, up to 12-fold and 16.5-fold increase in green fluorescence between Ca²⁺-free and Ca²⁺-saturated forms. We demonstrated the high potential of these sensors on various examples, including monitoring of Ca²⁺ response to a prolonged glutamate treatment in cortical neurons.

Conclusion

We believe that expanded dynamic range, high brightness and relatively high pH-stability should make Case12 and Case16 popular research tools both in scientific studies and high throughput screening assays.     

Centrale de mesure ambulatoire biomédicale sur PSoC

Improvement in quality and efficiency of health and medicine, at home and in hospital, has become of paramount importance. The solution to this problem would require the continuous monitoring of several key patient parameters, including the assessment of autonomic nervous system (ANS) activity using non-invasive sensors, providing information for emotional, sensorial, cognitive and physiological analysis of the patient. Recent advances in embedded systems, microelectronics, sensors and wireless networking enable the design of wearable systems capable of such advanced health monitoring. The subject of this article is an ambulatory system comprising of a small wrist device connected to several sensors for the detection of the autonomic nervous system activity. It affords monitoring of skin resistance, skin temperature and heart activity. It is also capable of recording the data on a removable media or sending it to computer via a wireless communication. The wrist device is based on a programmable system-on-chip (PSoC) from Cypress.     

Crack Detection in Fibre Reinforced Plastic Structures Using Embedded Fibre Bragg Grating Sensors: Theory,Model Development and Experimental Validation

In a fibre-reinforced polymer (FRP) structure designed using the emerging damage tolerance and structural health monitoring philosophy, sensors and models that describe crack propagation will enable a structure to operate despite the presence of damage by fully exploiting the material’s mechanical properties. When applying this concept to different structures, sensor systems and damage types, a combination of damage mechanics, monitoring technology, and modelling is required. The primary objective of this article is to demonstrate such a combination. This article is divided in three main topics: the damage mechanism (delamination of FRP), the structural health monitoring technology (fibre Bragg gratings to detect delamination), and the finite element method model of the structure that incorporates these concepts into a final and integrated damage-monitoring concept. A novel method for assessing a crack growth/damage event in fibre-reinforced polymer or structural adhesive-bonded structures using embedded fibre Bragg grating (FBG) sensors is presented by combining conventional measured parameters, such as wavelength shift, with parameters associated with measurement errors, typically ignored by the end-user. Conjointly, a novel model for sensor output prediction (virtual sensor) was developed using this FBG sensor crack monitoring concept and implemented in a finite element method code. The monitoring method was demonstrated and validated using glass fibre double cantilever beam specimens instrumented with an array of FBG sensors embedded in the material and tested using an experimental fracture procedure. The digital image correlation technique was used to validate the model prediction by correlating the specific sensor response caused by the crack with the developed model.     

Stream Restoration in the Upper Midwest, U.S.A.

Restoration activities intended to improve the condition of streams and rivers are widespread throughout the Upper Midwest, U.S.A. As with other regions, however, little information exists regarding types of activities and their effectiveness. We developed a database of 1,345 stream restoration projects implemented from the years 1970 to 2004 for the states of Michigan, Ohio, and Wisconsin in order to analyze regional trends in goals, presence of monitoring, spatial distribution, size, and cost of river restoration projects. We found that data on individual projects were fragmented across multiple federal, state, and county agencies, as well as nonprofit groups and consulting firms. The most common restoration goals reported for this region were in‐stream habitat improvement, bank stabilization, water‐quality management, and dam removal. The former two were most common in Michigan and Wisconsin, where salmonid fisheries enhancement appeared to be an important concern, whereas water‐quality management was most frequent in Ohio. The most common restoration activities were the use of sand traps and riprap, and other common activities were related to the improvement of fish habitat. The median cost was $12,957 for projects with cost data, and total expenditures since 1990 were estimated at $444 million. Over time, the cost of individual projects has increased, whereas the median size has decreased, suggesting that restoration resources are being spent on smaller, more localized, and more expensive projects. Only 11% of data records indicated that monitoring was performed, and more expensive projects were more likely to be monitored. Standardization of monitoring and record keeping and dissemination of findings are urgently needed to ensure that dollars are well spent and restoration effectiveness is maximized.     

Biosensors for process monitoring

Summary A short review about the biosensor research activities for bioprocess monitoring in the F.R.G. after its reunification is given. The principles of biosensor applications are presented. In situ sensors and sensors based on the principles of flow injection analysis are studied. Some applications of a four-channel enzyme thermistor, bio-field effect transistors, and immunoanalysis systems for real process monitoring are presented.     

The ecological integrity of the Lower Olifants River,Limpopo province,South Africa: 2009–2015 – Part A: Olifants River main stem

The major rivers of the South African ‘Lowveld’ (low-latitude savanna) suffer numerous impacts from upstream economic activities. Whereas monitoring these rivers is required to detect biodiversity losses, record pollution events and devise mitigation strategies, current monitoring programmes are inadequate. In 2009, the South African Earth Observation Network initiated an intensive long-term research programme on the Lowveld reaches of the Olifants River. Physico-chemical parameters, aquatic macroinvertebrates and fish abundances were recorded at four Lowveld sites in the Olifants River. We review six years of this programme. The results suggest deterioration in the ecological condition of the Olifants River with no discernible improvement through protected areas. Trends could not be detected. The parameters measured, sampling methods and/or sampling frequency might be responsible for the limited trends observed, or alternatively the results simply reflect stable conditions despite on-going pollution. Real time monitoring and an expansion in the parameters monitored would add value to the monitoring programme.     

Process analytical technologies to monitor the liquid phase of anaerobic cultures

Recent advances in sensor development and miniaturization offer new possibilities to monitor and control bioprocesses. Specific requirements for anaerobic processes in terms of low costs and high robustness against insoluble fractions and impurities in media led to a decelerated penetration of new technology in this field. Since no regulatory framework demands for process monitoring and documentation like in the pharmaceutical industry, the implementation of new sensors beyond long-established methods is not conducted as intensively.Nevertheless, many attempts have been made in recent years to adopt sensors and show their applicability in anaerobic fermentation processes. New possibilities arise for improved monitoring, control and faster process development through digitalization. This review aims to provide an overview over these recent attempts with the focus on the liquid phase in the upstream part of pure culture bioprocesses for anaerobic bulk compound, food, and beverage production. In particular, methods that monitor the viability, metabolic activity or related parameters are discussed, like electrochemical, impedance and spectroscopy probes, and methods related to fluid flow and gradient formation, like acoustic and mobile sensors.     

Human activity recognition based on Evolving Fuzzy Systems

Environments equipped with intelligent sensors can be of much help if they can recognize the actions or activities of their users. If this activity recognition is done automatically, it can be very useful for different tasks such as future action prediction, remote health monitoring, or interventions. Although there are several approaches for recognizing activities, most of them do not consider the changes in how a human performs a specific activity. We present an automated approach to recognize daily activities from the sensor readings of an intelligent home environment. However, as the way to perform an activity is usually not fixed but it changes and evolves, we propose an activity recognition method based on Evolving Fuzzy Systems.     

Academic network for nature conservation in Tandilia System,Buenos Aires,Argentina

The Tandilia System, located in the Southern Pampa region of Argentina, contains areas with conservation value scattered throughout its agricultural landscape. Unfortunately, efforts for their conservation and sustainable management are scarce and isolated, which may be explained by the lack of articulation and linkage between the different social actors in the territory. Based on 123 surveys of academics working in this field, we analyzed research lines, management and outreach experiences, and the synergies and interactions between them. The most frequent research topics were related to biodiversity, terrestrial ecosystems, landscapes, anthropic pressures, and environmental management. Twenty-six environmental management experiences were reported in which the creation and/or planning of protected areas were the most frequent. The analysis of academic-social networks allowed us to explore the relationship between actors. A high degree of network modularity was identified, with strong links within the research groups, but few connections between them. In general terms, these connections depend on a few actors with a high degree of intermediation. Additionally, some professionals presented a high degree of betweenness, which is an important potential for the efficient coordination of conservation and management projects. Our results suggest the relevance of fostering greater interaction between professionals and research groups, as well as of promoting joint management activities concerning nature care in the Tandilia System.     

Long-term <Emphasis Type="Italic">ex vivo</Emphasis> monitoring of <Emphasis Type="Italic">in vivo</Emphasis> microRNA activity in liver using a secreted luciferase sensor

Technology for monitoring in vivo microRNA (miRNA) activity is extremely important for elucidating miRNA biology. However, in vivo studies of miRNA have been hampered by the lack of a convenient approach to reliably reflect real-time functional changes in miRNAs. Sensors for miRNA were developed by adding miRNA target sequences to the 3′-untranslated region of Gaussia princeps luciferase (Gluc) mRNA. These sensors were then evaluated in vitro and in vivo by measuring Gluc activity in cell supernatants and in peripheral blood. Sensors driven by the CMV promoter were effective for monitoring miR-122 in living cells, but not for the long-term monitoring of miR-122 or miR-142 in mouse liver because of CMV-promoter silencing. Replacing the CMV promoter with a CAG promoter rendered these sensors effective for the long-term monitoring of relevant liver miRNA activities. We subsequently used the CAG-promoter-based sensor for the long-term monitoring of endogenous liver miR-122, miR142 and miR-34a activities, as well as for exogenous miR-34a activity. Our study demonstrates that real-time in vivo activities of miRNAs can be continuously and conveniently detected in mouse liver using the sensors that we have developed.     

无人机在生物多样性遥感监测中的应用现状与展望

近十年, 无人机平台由于其灵活机动、成本低等优势在植被生态调查、资源环境监测、生物多样性保护等领域逐渐兴起。本文从生物多样性遥感监测应用角度首先介绍了无人机分类系统, 为具体工作开展过程中如何选择合适的载体和传感器提供了参考; 继而总结了不同类型无人机的适用性及其可搭载传感器的用途与区别。在此基础上, 针对无人机平台的高精度遥感信息具体应用案例, 就反映生物多样性变化并揭示其驱动机制方面的无人机遥感直接和间接指标的相关研究进展展开阐述。最后, 就目前无人机遥感技术在生物多样性监测领域的应用中存在的限制, 如软硬件结合匹配程度不够、部分设备过于昂贵、法律法规不完善、与传统生物多样性监测手段结合较弱等问题进行探讨。我们认为: 无人机遥感技术可以很好地弥补地面监测与航天、卫星遥感之间的尺度空缺, 更好地将监测点上的结果以准确、可靠的推绎方法扩展到区域尺度供决策分析使用。今后迫切需要进一步加大生物多样性近地面遥感监测项目建设的实施力度, 从整体上提高生物多样性热点区域应对变化的分析预警能力。     

Patient Care Teams in treatment of diabetes and chronic heart failure in primary care: an observational networks study

Background

Patient care teams have an important role in providing medical care to patients with chronic disease, but insight into how to improve their performance is limited. Two potentially relevant determinants are the presence of a central care provider with a coordinating role and an active role of the patient in the network of care providers. In this study, we aimed to develop and test measures of these factors related to the network of care providers of an individual patient.

Methods

We performed an observational study in patients with type 2 diabetes or chronic heart failure, who were recruited from three primary care practices in The Netherlands. The study focused on medical treatment, advice on physical activity, and disease monitoring. We used patient questionnaires and chart review to measure connections between the patient and care providers, and a written survey among care providers to measure their connections. Data on clinical performance were extracted from the medical records. We used network analysis to compute degree centrality coefficients for the patient and to identify the most central health professional in each network. A range of other network characteristics were computed including network centralization, density, size, diversity of disciplines, and overlap among activity-specific networks. Differences across the two chronic conditions and associations with disease monitoring were explored.

Results

Approximately 50% of the invited patients participated. Participation rates of health professionals were close to 100%. We identified 63 networks of 25 patients: 22 for medical treatment, 16 for physical exercise advice, and 25 for disease monitoring. General practitioners (GPs) were the most central care providers for the three clinical activities in both chronic conditions. The GP's degree centrality coefficient varied substantially, and higher scores seemed to be associated with receiving more comprehensive disease monitoring. The degree centrality coefficient of patients also varied substantially but did not seem to be associated with disease monitoring.

Conclusions

Our method can be used to measure connections between care providers of an individual patient, and to examine the association between specific network parameters and healthcare received. Further research is needed to refine the measurement method and to test the association of specific network parameters with quality and outcomes of healthcare.

     

Biotechnology and the Third World: the missing link between research and applications

A United Nations University study investigated the activities of four major United Nations agencies that focussed on helping developing countries gain advanced capabilities in biotechnology. Relevant program and project documents were scrutinized at agency headquarters and managers were interviewed. Then, projects underway in three case countries (Egypt, Thailand, and Venezuela) were examined. The resulting information was used to assess whether United Nations projects were fulfilling these countries' needs and (or) advancing their capabilities in biotechnology. The minute, United Nations originated assistance available was directed solely at increasing capabilities in research and thus benefited bioscientists and their institutes. However, as virtually no linkage exists between the research establishment and the industrial--marketing sector, results from indigenous research does not reach industrialists or health workers. Consequently, biotechnology is neither advancing economic development in the case countries nor helping solve national problems. This situation is likely to persist because corrective systemic changes will be difficult to implement. Major implications of these findings are discussed, particularly as they bear on the United Nations system.     

Monitoring physical activities of COPD patients with a network of sensors

Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and the fourth leading cause of death worldwide with multiple comorbidities including cardiovascular diseases. Therefore it is interesting to monitor both cardiorespiratory parameters and physical activity of patients, during intensive physical exercises and also during normal daily activities. But to monitor the patient in his daily life, we need a system that can be used autonomously by the patient at home. Such monitoring can benefit from a network of collaborative sensors. We therefore propose a system composed of multiple medical sensors wirelessly interconnected to a Master device, in order to acquire medical data in daily life or in supervised medical exams.     

An integrated scheduling and control model for multi-mode projects

In today’s highly competitive uncertain project environments, it is of crucial importance to develop analytical models and algorithms to schedule and control project activities so that the deviations from the project objectives are minimized. This paper addresses the integrated scheduling and control in multi-mode project environments. We propose an optimization model that models the dynamic behavior of projects and integrates optimal control into a practically relevant project scheduling problem. From the scheduling perspective, we address the discrete time/cost trade-off problem, whereas an optimal control formulation is used to capture the effect of project control. Moreover, we develop a solution algorithm for two particular instances of the optimal project control. This algorithm combines a tabu search strategy and nonlinear programming. It is applied to a large scale test bed and its efficiency is tested by means of computational experiments. To the best of our knowledge, this research is the first application of optimal control theory to multi-mode project networks. The models and algorithms developed in this research are targeted as a support tool for project managers in both scheduling and deciding on the timing and quantity of control activities.     

CO2 and H2O: Understanding Different Stakeholder Perspectives on the Use of Carbon Credits to Finance Household Water Treatment Projects

Background

Carbon credits are an increasingly prevalent market-based mechanism used to subsidize household water treatment technologies (HWT). This involves generating credits through the reduction of carbon emissions from boiling water by providing a technology that reduces greenhouse gas emissions linked to climate change. Proponents claim this process delivers health and environmental benefits by providing clean drinking water and reducing greenhouse gases. Selling carbon credits associated with HWT projects requires rigorous monitoring to ensure households are using the HWT and achieving the desired benefits of the device. Critics have suggested that the technologies provide neither the benefits of clean water nor reduced emissions. This study explores the perspectives of carbon credit and water, sanitation and hygiene (WASH) experts on HWT carbon credit projects.

Methods

Thirteen semi-structured, in-depth interviews were conducted with key informants from the WASH and carbon credit development sectors. The interviews explored perceptions of the two groups with respect to the procedures applied in the Gold Standard methodology for trading Voluntary Emission Reduction (VER) credits.

Results

Agreement among the WASH and carbon credit experts existed for the concept of suppressed demand and parameters in the baseline water boiling test. Key differences, however, existed. WASH experts’ responses highlighted a focus on objectively verifiable data for monitoring carbon projects while carbon credit experts called for contextualizing observed data with the need for flexibility and balancing financial viability with quality assurance.

Conclusions

Carbon credit projects have the potential to become an important financing mechanism for clean energy in low- and middle-income countries. Based on this research we recommend that more effort be placed on building consensus on the underlying assumptions for obtaining carbon credits from HWT projects, as well as the approved methods for monitoring correct and consistent use of the HWT technologies in order to support public health impacts.     

Ostracoda (Crustacea) as indicators for surface water quality: a case study from the Ledra River basin (NE Italy)

Wastewater discharges associated with urbanisations, farming activities and industry may dramatically reduce the ecological health of river ecosystems. During the reconstruction of the Friuli Venezia Giulia region following the 1976 earthquake, a lot of resources were used to build large numbers of wastewater treatment plants to minimize the impact of human activities on lotic ecosystems. Their efficiency is usually assessed through monitoring of the physical and chemical environment near the discharge point. However, discontinuous monitoring of the abiotic environment may fail to detect periodic malfunctioning and do not recognize indirect effects on the ecosystem. We assessed the potential of an alternative approach to assess the impact of wastewater discharges, based on the monitoring of ostracod density, richness and community composition. We repeatedly measured physical, chemical and microbial parameters and collected ostracod samples at stations up- and downstream from wastewater discharge points scattered over a 21-km stretch of the Ledra River (NE Italy). The results indicate that monitoring ostracods is a potentially valuable approach, for two reasons. Communities appeared to be well differentiated even in the small spatial area of this study, indicating that they can provide sufficient resolution to pick up even minor impacts. Secondly, despite the seasonal succession in species composition, spatial differentiation was consistent over time, suggesting that ostracods provide a time-integrated picture of the water quality. The traditional approach failed to detect any consistent impact of wastewater discharges, apart from an ambiguous increase in nutrient levels. The density and/or richness of the ostracod communities was altered by some wastewater discharges, but not by others. We identified a general trend for wastewater discharges to systematically replace regionally rare ostracod species with common species. In particular, the species Ilyocypris inermis was very sensitive to discharges, and may be used as an indicator species for good ecosystem health.