Absolute pharmacokinetics of heparin in primates.

Heparin is a commonly used anticoagulant drug, derived from the tissues of animals including pigs, cows, and sheep. Measuring heparin concentration in plasma is challenging due to its complex molecular structure. Existing methods rely on measuring heparin's anticoagulant activity, which provides pharmacodynamic (PD) data but not pharmacokinetic (PK) data, measuring concentration over time. To overcome this limitation, we used liquid chromatography-mass spectrometry (LC-MS) and the multiple reaction monitoring (MRM) method to directly measure heparin's concentration in non-human primates after administering porcine, bovine, and ovine heparin. A protocol was developed to enable an MRM method for application to small plasma volumes without purification. The PK data obtained from LC-MS are then compared with the data obtained using the Heparin Red assay and the PD data determined using biochemical clinical assays. Results showed that LC-MS and Heparin Red assay measurements closely correlated with unfractionated heparin's biological activities, supporting the use of mass spectra and dye-binding assays to determine heparin levels in plasma. This study builds a way for the measurement of heparin concentration in plasma, which could lead to an improved understanding of heparin's metabolism and dosing safety.

Running a dense air temperature measurement field campaign at the urban neighbourhood level: Protocol and lessons learned.

The cooling capacity of urban green spaces constitutes a key measure for cities to mitigate heat events, which is gaining importance in climate change adaptation and mitigation. In this protocol article, we present details on two field campaigns aiming at collecting dense air temperature data in two urban inner city parks in Leipzig, Germany, under unprecedented heat and drought conditions. We introduce all the steps required to plan and conduct qualified fieldwork in environmental research, including study design, technical and logistical preparations, on-site work and data management steps from data acquisition, transfer into research outcomes to dissemination. We further share valuable lessons learned before, during and after fieldwork that helped us improve our work and that could support and improve similar future project campaigns.

Pathways linking biodiversity to human health: A conceptual framework.

Biodiversity is a cornerstone of human health and well-being. However, while evidence of the contributions of nature to human health is rapidly building, research into how biodiversity relates to human health remains limited in important respects. In particular, a better mechanistic understanding of the range of pathways through which biodiversity can influence human health is needed. These pathways relate to both psychological and social processes as well as biophysical processes. Building on evidence from across the natural, social and health sciences, we present a conceptual framework organizing the pathways linking biodiversity to human health. Four domains of pathways-both beneficial as well as harmful-link biodiversity with human health: (i) reducing harm (e.g. provision of medicines, decreasing exposure to air and noise pollution); (ii) restoring capacities (e.g. attention restoration, stress reduction); (iii) building capacities (e.g. promoting physical activity, transcendent experiences); and (iv) causing harm (e.g. dangerous wildlife, zoonotic diseases, allergens). We discuss how to test components of the biodiversity-health framework with available analytical approaches and existing datasets. In a world with accelerating declines in biodiversity, profound land-use change, and an increase in non-communicable and zoonotic diseases globally, greater understanding of these pathways can reinforce biodiversity conservation as a strategy for the promotion of health for both people and nature. We conclude by identifying research avenues and recommendations for policy and practice to foster biodiversity-focused public health actions.

Distant regions underpin interregional flows of cultural ecosystem services provided by birds and mammals.

Ecosystem service assessments rarely consider flows between distant regions. Hence, telecoupling effects such as conservation burdens in distant ecosystems are ignored. We identified service-providing species for two cultural ecosystem services (existence and bequest, and birdwatching) and two receiving, i.e. benefitting, regions (Germany, the Netherlands). We delineated and analysed sending, i.e. service-providing, regions on a global scale. The proportion of service-providing species with distant habitats was higher for birdwatching (Germany: 58.6%, Netherlands: 59.4%), than for existence and bequest (Germany: 49.3%, Netherlands: 57.1%). Hotspots of sending regions were predominantly situated in tropical and subtropical grasslands, savannas and shrublands and were significantly more threatened and poorer than the global mean. Hotspot protection levels for flows to Germany were higher than the global mean, and lower for the Dutch hotspots. Our findings increase understanding on how distant regions underpin ecosystem services and necessitate interregional assessment as well as conservation efforts.

Using Semistructured Surveys to Improve Citizen Science Data for Monitoring Biodiversity.

Biodiversity is being lost at an unprecedented rate, and monitoring is crucial for understanding the causal drivers and assessing solutions. Most biodiversity monitoring data are collected by volunteers through citizen science projects, and often crucial information is lacking to account for the inevitable biases that observers introduce during data collection. We contend that citizen science projects intended to support biodiversity monitoring must gather information about the observation process as well as species occurrence. We illustrate this using eBird, a global citizen science project that collects information on bird occurrences as well as vital contextual information on the observation process while maintaining broad participation. Our fundamental argument is that regardless of what species are being monitored, when citizen science projects collect a small set of basic information about how participants make their observations, the scientific value of the data collected will be dramatically improved.

Development of a Semiglobal Reaction Mechanism for the Thermal Decomposition of a Polymer Containing Reactive Flame Retardants: Application to Glass-Fiber-Reinforced Polybutylene Terephthalate Blended with Aluminum Diethyl Phosphinate and Melamine Polyphosphate.

This work details a methodology for parameterization of the kinetics and thermodynamics of the thermal decomposition of polymers blended with reactive additives. This methodology employs Thermogravimetric Analysis, Differential Scanning Calorimetry, Microscale Combustion Calorimetry, and inverse numerical modeling of these experiments. Blends of glass-fiber-reinforced polybutylene terephthalate (PBT) with aluminum diethyl phosphinate and melamine polyphosphate were used to demonstrate this methodology. These additives represent a potent solution for imparting flame retardancy to PBT. The resulting lumped-species reaction model consisted of a set of first- and second-order (two-component) reactions that defined the rate of gaseous pyrolyzate production. The heats of reaction, heat capacities of the condensed-phase reactants and products, and heats of combustion of the gaseous products were also determined. The model was shown to reproduce all aforementioned experiments with a high degree of detail. The model also captured changes in the material behavior with changes in the additive concentrations. Second-order reactions between the material constituents were found to be necessary to reproduce these changes successfully. The development of such models is an essential milestone toward the intelligent design of flame retardant materials and solid fuels.

Incorporating threat in hotspots and coldspots of biodiversity and ecosystem services.

Spatial prioritization could help target conservation actions directed to maintain both biodiversity and ecosystem services. We delineate hotspots and coldspots of two biodiversity conservation features and five regulating and cultural services by incorporating an indicator of 'threat', i.e. timber harvest profitability for forest areas in Telemark (Norway). We found hotspots, where high values of biodiversity, ecosystem services and threat coincide, ranging from 0.1 to 7.1% of the area, depending on varying threshold levels. Targeting of these areas for conservation follows reactive conservation approaches. In coldspots, high biodiversity and ecosystem service values coincide with low levels of threat, and cover 0.1-3.4% of the forest area. These areas might serve proactive conservation approaches at lower opportunity cost (foregone timber harvest profits). We conclude that a combination of indicators of biodiversity, ecosystem services and potential threat is an appropriate approach for spatial prioritization of proactive and reactive conservation strategies.

Determination of direct oral anticoagulants from human serum samples.

Dabigatran, rivaroxaban, and apixaban are direct oral anticoagulants (DOAC) inhibiting thrombin or factor Xa and effectively preventing thromboembolic complications using fixed doses without need for laboratory-guided dose adjustment. Plasma samples are needed to determine the actual concentration or activity of DOACs, which may be required for special patient populations such as those with acute deterioration of renal function due to any disease, before surgical interventions, during bleeding or thrombotic episodes while on therapy with DOACs, the elderly and youngest populations, unexpected pregnancy, suspicion of overdose and toxication, and to control adherence to therapy. Serum samples have several advantages over plasma samples such as no need of sampling with a specific coagulation tube, reduced pre-analytical errors, and longer storage stability. Determination of rivaroxaban and apixaban from serum samples of patients on treatment performed well and better than samples of patients treated with dabigatran compared with plasma samples. Specific adaption to automated coagulation platforms may improve the performance of the assays from serum samples.

Measurement of the new anticoagulants.

New oral anticoagulants are given at fixed daily doses without laboratory dose adjustment for prevention of venous thromboembolism following elective total knee- and hip replacement, for treatment and prevention of recurrent events of acute venous thromboembolism, and for prevention of embolic events in atrial fibrillation. However, it may be necessary to determine the anticoagulant effect of new oral anticoagulants in special patient populations such as in elderly, for renal impairment, before operation, bleeding or thrombotic episodes and to monitor self-compliance. Oral factor Xa and oral thrombin inhibitors influence dose dependently global and specific coagulation assays. Standardization of assays is currently undertaken. Determination of the new oral anticoagulants in serum samples would facilitate blood sampling and analysis from samples taken and stored for creatinine or other biochemical parameters. Point of care methods from plasma or urine for the new oral anticoagulants would improve patient care. First data demonstrate the feasibility of such assays in urine.

Public access resuscitation program including defibrillator training for laypersons: a randomized trial to evaluate the impact of training course duration.

BACKGROUND: Time to cardiopulmonary resuscitation (CPR) is a main determinant of survival after out-of-hospital cardiac arrest. Only widespread implementation of training courses for laypersons can decrease response time. METHODS AND RESULTS: In this prospective randomized trial, we evaluated how laypersons retained CPR skills and skills in using the automated external defibrillator (AED). A total of 1095 volunteers were randomly assigned to receive CPR/AED-training courses of 2h (375 persons), 4h (378 persons) or 7h (342 persons) duration. Courses were held in accordance with the guidelines for CPR. All trainees were tested immediately after the initial class in a standardized test scenario using an AED and a manikin. Either at 6 or at 12 months, retests were given to 164 and 206 volunteers, respectively. In 479 volunteers, retesting was completed at both 6- and 12-month intervals. At the immediate tests, the 7-h training group showed a slightly higher rate of correct responses (7h: 96%, 4h: 94%, 2h: 92%) (p<0.001). Skill retention decreased significantly in the three groups and was lowest after 12 months if no 6-month retests were done. In trainees who did undergo retesting at 6 months, skills did not deteriorate at 12 months. There were no significant differences between the three groups (overall correct responses: 2h: 72%, 4h: 73%, 7h: 74%) (ns). CONCLUSIONS: A 2-h class is sufficient to acquire and retain CPR and AED skills for an extended time period provided that a brief re-evaluation is performed after 6 months.

Fluorescent sensor for Cu2+ with a tunable emission wavelength.

A concept of fluorescent metal ion sensing with an easily tunable emission wavelength is presented and its principle demonstrated by detection of Cu(2+). A fluorescein dye was chemically modified with a metal chelating group and then attached to the terminus of ss-DNA. This was combined with a complementary ss-DNA modified with another fluorescent dye (ATTO 590), emitting at a longer wavelength. In the assembled duplex, fluorescence resonance energy transfer (FRET) between the fluorescein donor (excited at 470 nm) and the ATTO 590 acceptor (emitting at 624 nm) is observed. Proper positioning within the rigid DNA double helix prevents intramolecular contact quenching of the two dyes. Coordination of paramagnetic Cu(2+) ions by the chelating unit of the sensor results in direct fluorescence quenching of the fluorescein dye and indirect (by loss of FRET) quenching of the ATTO 590 emission at 624 nm. As a result, emission of the acceptor dye can be used for monitoring of the concentration of Cu(2+), with a 20 nM detection limit. The emission wavelength is readily tuned by replacement of ATTO-DNA by other commercially available DNA-acceptor dye conjugates. Fluorescent metal ion sensors emitting at >600 nm are very rare. The possibility of tuning the emission wavelength is important with respect to the optimization of this sensor type for application to biological samples, which usually show broad autofluorescence at <550 nm.

Biphenyl derived oxovanadium(IV) and copper(II) salen-type complexes--structure and redox tuning.

A series of vanadyl(IV) salen (N,N'-bis(salicylidene)ethylenediaminato)-type complexes (1-4) bearing phenyl or 2-hydroxyphenyl moieties have been prepared and characterized by means of mass spectrometry, infra-red, electron paramagnetic resonance (EPR), UV/Vis spectroscopy, cyclovoltammetry and X-ray crystallography. Their structures have been compared to their copper(II) analogs 5-8. Hydrogen intralinkages have been observed in the crystal structure of 5. The pendant hydroxy groups fine-tune the redox properties of the complexes. The catalytic activity in the oxygenation of ethyl phenyl sulfide to the corresponding sulfoxide was investigated. Results indicate that complex 1 bearing hydroxyphenyl subunits and a phenylene bridge is the most selective under these reaction conditions, with the smallest amount of the over-oxidized product, sulfone.

Copper(II)-quenched oligonucleotide probes for fluorescent DNA sensing.

A copper(II)-quenched molecular beacon was prepared by attaching fluorescein to the 3'-end and a copper(II) complex to the 5'-end of DNA. In the presence of complementary DNA, copper(II) and dye are spatially separated in the duplex and fluorescence increases up to 15-fold, with excellent discrimination of single base mismatches.

DNA-templated metal catalysis.

Hydrolysis of an ester substrate by a CuII complex catalyst, both attached to oligo-peptide nucleic acids (PNA), is accelerated up to 485-fold in the presence of a complementary DNA template. The approach combines the sequence selectivity of DNA-templated reactions with signal amplification by multiple turnover and the versatility of metal catalysis.

Conjugates of PNA with naphthalene diimide derivatives having a broad range of DNA affinities.

Peptide nucleic acids (PNAs) are neutral DNA analogues, which bind single-stranded DNA (ssDNA) strongly and with high sequence specificity. However, binding efficiency is dependent on the purine content of the PNA strand. This property make more difficult application of PNA as hybridization probes in, e.g., PNA chips, since at a set temperature the hybridization of a fraction of the DNA targets to the PNA probes does not obey Watson-Crick binding rules. The polypurine PNAs, for example, bind the mismatch containing DNA targets stronger, than the pyrimidine rich PNAs their fully complementary targets. Herein we show that PNA-DNA binding efficiency can be finely tuned by the conjugation of derivatives of naphthalene diimide (NADI) to the N-terminus of PNA using polyamide linkers of different lengths. This approach can potentially be used for the design of PNA probes, which bind their DNA targets with similar affinity independently of the PNA sequence.

Synthesis and DNA binding properties of terminally modified peptide nucleic acids.

PNAs with terminal modifications of varying structure and charge were synthesized and their binding to DNA was studied. A variation in thermal stability of 19. 8 degrees C has been observed between the least and the most stable PNA-DNA duplexes. The most stable duplex melts 7.7 degrees C higher than the duplex of the corresponding non-modified PNA and complementary DNA. It has been shown that sequence fidelity of the PNA conjugate having the highest DNA affinity is significantly better than that of non-modified PNA. The results obtained can be used for the design of PNA probes, whose binding to DNA is sequence independent.

Peptide nucleic acid-metal complex conjugates: facile modulation of PNA-DNA duplex stability.

Conjugates of peptide nucleic acids (PNA) and metal binding ligands were prepared using solid-phase synthesis. Stability of duplexes of bis-picolylamine-PNA conjugates and DNA was found to be modulated by equimolar concentrations of bioavailable metal ions: Ni(2+), Zn(2+)>Cu(2+). Sequence specificity of PNA was not compromised in the presence of these metal ions.