Towards enhanced functionality of vagus neuroprostheses through in silico optimized stimulation.

Bioelectronic therapies modulating the vagus nerve are promising for cardiovascular, inflammatory, and mental disorders. Clinical applications are however limited by side-effects such as breathing obstruction and headache caused by non-specific stimulation. To design selective and functional stimulation, we engineered VaStim, a realistic and efficient in-silico model. We developed a protocol to personalize VaStim in-vivo using simple muscle responses, successfully reproducing experimental observations, by combining models with trials conducted on five pigs. Through optimized algorithms, VaStim simulated the complete fiber population in minutes, including often omitted unmyelinated fibers which constitute 80% of the nerve. The model suggested that all Aα-fibers across the nerve affect laryngeal muscle, while heart rate changes were caused by B-efferents in specific fascicles. It predicted that tripolar paradigms could reduce laryngeal activity by 70% compared to typically used protocols. VaStim may serve as a model for developing neuromodulation therapies by maximizing efficacy and specificity, reducing animal experimentation.

Nanomaterials as a Service (NaaS) concept: on-demand protocols for volume synthesis of nanomaterials.

Establishing scalable nanomaterials synthesis protocols remains a bottleneck towards their commercialisation and, thus, a topic of intense research and development. Herein, we present an automated machine-learning microfluidic platform capable of synthesising optically active nanomaterials from target spectra originating from prior experience, theorised or published. Implementing unsupervised Bayesian optimisation with Gaussian processes reduces the optimisation time and the need for prior knowledge to initiate the process. Using PTFE tubing and connectors enables facile change in reactor design. Ultimately, the platform substitutes the labour-intensive trial-and-error synthesis and provides a pathway to standardisation and volume synthesis, slowing down the translation and commercialisation of high-quality nanomaterials. As a proof-of-concept, Ag nanoplates and Prussian-blue nanoparticle protocols were optimised and validated for volume production.

Vertical optical complexity shaped by submerged macrophytes.

The influence of macrophytes on the optical environment of the littoral zone was assessed by studying the effect of monospecific Potamogeton perfoliatus on the quantitative and qualitative properties of light and the response of plants to this altered environment. P. perfoliatus was shown to alter the optical environment and consequently its own architecture: in high-density pondweed patches, 67 percent of incident light was absorbed in the top 10 cm, while spectral properties of light was significantly altered. Leaf morphology and photophysiology adapted to these changes, with photosynthetically active biomass concentrated in the upper water layer and stem biomass increasing in the basal parts due to self-shading. This study highlights the importance of submerged macrophytes in shaping the optical environment and ecological dynamics of littoral zones. Not only do pondweed plants from different sites show very similar vertical patterns of morphological and physiological parameters, but they also contribute to similar vertical spatial variability in water optics, thus increasing habitat complexity. This added optical heterogeneity not only increases the diversity of the littoral zone, but also enriches the entire aquatic ecosystem of shallow lakes by providing additional optical ecological niches.

Organ- and function-specific anatomical organization of vagal fibers supports fascicular vagus nerve stimulation.

Vagal fibers travel inside fascicles and form branches to innervate organs and regulate organ functions. Existing vagus nerve stimulation (VNS) therapies activate vagal fibers non-selectively, often resulting in reduced efficacy and side effects from non-targeted organs. The transverse and longitudinal arrangement of fibers inside the vagal trunk with respect to the functions they mediate and organs they innervate is unknown, however it is crucial for selective VNS. Using micro-computed tomography imaging, we tracked fascicular trajectories and found that, in swine, sensory and motor fascicles are spatially separated cephalad, close to the nodose ganglion, and merge caudad, towards the lower cervical and upper thoracic region; larynx-, heart- and lung-specific fascicles are separated caudad and progressively merge cephalad. Using quantified immunohistochemistry at single fiber level, we identified and characterized all vagal fibers and found that fibers of different morphological types are differentially distributed in fascicles: myelinated afferents and efferents occupy separate fascicles, myelinated and unmyelinated efferents also occupy separate fascicles, and small unmyelinated afferents are widely distributed within most fascicles. We developed a multi-contact cuff electrode to accommodate the fascicular structure of the vagal trunk and used it to deliver fascicle-selective cervical VNS in anesthetized and awake swine. Compound action potentials from distinct fiber types, and physiological responses from different organs, including laryngeal muscle, cough, breathing, and heart rate responses are elicited in a radially asymmetric manner, with consistent angular separations that agree with the documented fascicular organization. These results indicate that fibers in the trunk of the vagus nerve are anatomically organized according to functions they mediate and organs they innervate and can be asymmetrically activated by fascicular cervical VNS.

Scientists without borders: lessons from Ukraine.

Conflicts and natural disasters affect entire populations of the countries involved and, in addition to the thousands of lives destroyed, have a substantial negative impact on the scientific advances these countries provide. The unprovoked invasion of Ukraine by Russia, the devastating earthquake in Turkey and Syria, and the ongoing conflicts in the Middle East are just a few examples. Millions of people have been killed or displaced, their futures uncertain. These events have resulted in extensive infrastructure collapse, with loss of electricity, transportation, and access to services. Schools, universities, and research centers have been destroyed along with decades' worth of data, samples, and findings. Scholars in disaster areas face short- and long-term problems in terms of what they can accomplish now for obtaining grants and for employment in the long run. In our interconnected world, conflicts and disasters are no longer a local problem but have wide-ranging impacts on the entire world, both now and in the future. Here, we focus on the current and ongoing impact of war on the scientific community within Ukraine and from this draw lessons that can be applied to all affected countries where scientists at risk are facing hardship. We present and classify examples of effective and feasible mechanisms used to support researchers in countries facing hardship and discuss how these can be implemented with help from the international scientific community and what more is desperately needed. Reaching out, providing accessible training opportunities, and developing collaborations should increase inclusion and connectivity, support scientific advancements within affected communities, and expedite postwar and disaster recovery.

Leaf reflectance can surrogate foliar economics better than physiological traits across macrophyte species.

BACKGROUND: Macrophytes are key players in aquatic ecosystems diversity, but knowledge on variability of their functional traits, among and within species, is still limited. Remote sensing is a high-throughput, feasible option for characterizing plant traits at different scales, provided that reliable spectroscopy models are calibrated with congruous empirical data, but existing applications are biased towards terrestrial plants. We sampled leaves from six floating and emergent macrophyte species common in temperate areas, covering different phenological stages, seasons, and environmental conditions, and measured leaf reflectance (400-2500 nm) and leaf traits (dealing with photophysiology, pigments, and structure). We explored optimal spectral band combinations and established non-parametric reflectance-based models for selected traits, eventually showing how airborne hyperspectral data could capture spatial-temporal macrophyte variability. RESULTS: Our key finding is that structural-leaf dry matter content, leaf mass per area-and biochemical-chlorophyll-a content and chlorophylls to carotenoids ratio-traits can be surrogated by leaf reflectance with normalized error under 17% across macrophyte species. On the other hand, the performance of reflectance-based models for photophysiological traits substantively varies, depending on macrophyte species and target parameters. CONCLUSIONS: Our main results show the link between leaf reflectance and leaf economics (structure and biochemistry) for aquatic plants, thus envisioning a crucial role for remote sensing in enhancing the level of detail of macrophyte functional diversity analysis to intra-site and intra-species scales. At the same time, we highlighted some difficulties in establishing a general link between reflectance and photosynthetic performance under high environmental heterogeneity, potentially opening further investigation directions.

Diastereoselective Double C-H Functionalization of Chiral Ferrocenes with Heteroaromatics.

Diastereoselective double C-H heteroarylation of chiral ferrocenes provides valuable compounds with multiple functionalities using mild reaction conditions and simple reagents. Pd-Complexes with chiral mono-protected amino acids afforded corresponding heteroarylated ferrocenyl amines in good yields and high diastereomeric purities. In this way, a variety of indole, thiophene, pyrrole, or furan substituents were introduced to the ferrocene moiety. Furthermore, a range of relevant functional groups, for example ketone, ester, chloro, nitro, or silyl, are tolerated by this method. An alternative combination of amino acid and ferrocenyl amine configurations was leveraged to provide the complementary diastereomeric products. The products of C-H heteroarylation can be transformed into corresponding phosphines. Absolute configurations of CH-activation products were confirmed by the combination of X-ray crystallographic analysis and CD spectroscopy. 19 F NMR kinetic study and DFT calculations provided insights into the reaction mechanism and reasons governing stereoinduction.

Photomorphogenesis in the Picocyanobacterium Cyanobium gracile Includes Increased Phycobilisome Abundance Under Blue Light, Phycobilisome Decoupling Under Near Far-Red Light, and Wavelength-Specific Photoprotective Strategies.

Photomorphogenesis is a process by which photosynthetic organisms perceive external light parameters, including light quality (color), and adjust cellular metabolism, growth rates and other parameters, in order to survive in a changing light environment. In this study we comprehensively explored the light color acclimation of Cyanobium gracile, a common cyanobacterium in turbid freshwater shallow lakes, using nine different monochromatic growth lights covering the whole visible spectrum from 435 to 687 nm. According to incident light wavelength, C. gracile cells performed great plasticity in terms of pigment composition, antenna size, and photosystem stoichiometry, to optimize their photosynthetic performance and to redox poise their intersystem electron transport chain. In spite of such compensatory strategies, C. gracile, like other cyanobacteria, uses blue and near far-red light less efficiently than orange or red light, which involves moderate growth rates, reduced cell volumes and lower electron transport rates. Unfavorable light conditions, where neither chlorophyll nor phycobilisomes absorb light sufficiently, are compensated by an enhanced antenna size. Increasing the wavelength of the growth light is accompanied by increasing photosystem II to photosystem I ratios, which involve better light utilization in the red spectral region. This is surprisingly accompanied by a partial excitonic antenna decoupling, which was the highest in the cells grown under 687 nm light. So far, a similar phenomenon is known to be induced only by strong light; here we demonstrate that under certain physiological conditions such decoupling is also possible to be induced by weak light. This suggests that suboptimal photosynthetic performance of the near far-red light grown C. gracile cells is due to a solid redox- and/or signal-imbalance, which leads to the activation of this short-term light acclimation process. Using a variety of photo-biophysical methods, we also demonstrate that under blue wavelengths, excessive light is quenched through orange carotenoid protein mediated non-photochemical quenching, whereas under orange/red wavelengths state transitions are involved in photoprotection.

Let Sleeping Patients Lie, avoiding unnecessary overnight vitals monitoring using a clinically based deep-learning model.

Impaired sleep for hospital patients is an all too common reality. Sleep disruptions due to unnecessary overnight vital sign monitoring are associated with delirium, cognitive impairment, weakened immunity, hypertension, increased stress, and mortality. It is also one of the most common complaints of hospital patients while imposing additional burdens on healthcare providers. Previous efforts to forgo overnight vital sign measurements and improve patient sleep used providers' subjective stability assessment or utilized an expanded, thus harder to retrieve, set of vitals and laboratory results to predict overnight clinical risk. Here, we present a model that incorporates past values of a small set of vital signs and predicts overnight stability for any given patient-night. Using data obtained from a multi-hospital health system between 2012 and 2019, a recurrent deep neural network was trained and evaluated using ~2.3 million admissions and 26 million vital sign assessments. The algorithm is agnostic to patient location, condition, and demographics, and relies only on sequences of five vital sign measurements, a calculated Modified Early Warning Score, and patient age. We achieved an area under the receiver operating characteristic curve of 0.966 (95% confidence interval [CI] 0.956-0.967) on the retrospective testing set, and 0.971 (95% CI 0.965-0.974) on the prospective set to predict overnight patient stability. The model enables safe avoidance of overnight monitoring for ~50% of patient-nights, while only misclassifying 2 out of 10,000 patient-nights as stable. Our approach is straightforward to deploy, only requires regularly obtained vital signs, and delivers easily actionable clinical predictions for a peaceful sleep in hospitals.

Single-axon level automatic segmentation and feature extraction from immuhistochemical images of peripheral nerves.

Quantitative descriptions of the morphology and structure of peripheral nerves is central in the development of bioelectronic devices interfacing the nerves. While histological procedures and microscopy techniques yield high-resolution detailed images of individual axons, automated methods to extract relevant information at the single-axon level are not widely available. We implemented a segmentation algorithm that allows for subsequent feature extraction in immunohistochemistry (IHC) images of peripheral nerves at the single fiber scale. These features include short and long cross-sectional diameters, area, perimeter, thickness of surrounding myelin and polar coordinates of single axons within a nerve or nerve fascicle. We evaluated the performance of our algorithm using manually annotated IHC images of 27 fascicles of the swine cervical vagus; the accuracy of single-axon detection was 82%, and of the classification of fiber myelination was 89%.

Machine learning to assist clinical decision-making during the COVID-19 pandemic.

BACKGROUND: The number of cases from the coronavirus disease 2019 (COVID-19) global pandemic has overwhelmed existing medical facilities and forced clinicians, patients, and families to make pivotal decisions with limited time and information. MAIN BODY: While machine learning (ML) methods have been previously used to augment clinical decisions, there is now a demand for "Emergency ML." Throughout the patient care pathway, there are opportunities for ML-supported decisions based on collected vitals, laboratory results, medication orders, and comorbidities. With rapidly growing datasets, there also remain important considerations when developing and validating ML models. CONCLUSION: This perspective highlights the utility of evidence-based prediction tools in a number of clinical settings, and how similar models can be deployed during the COVID-19 pandemic to guide hospital frontlines and healthcare administrators to make informed decisions about patient care and managing hospital volume.

Aspects of Invasiveness of Ludwigia and Nelumbo in Shallow Temperate Fluvial Lakes.

The relationship between invasive plant functional traits and their invasiveness is still the subject of scientific investigation, and the backgrounds of transition from non-native to invasive species in ecosystems are therefore poorly understood. Furthermore, our current knowledge on species invasiveness is heavily biased toward terrestrial species; we know much less about the influence of allochthonous plant traits on their invasiveness in aquatic ecosystems. In this paper, we present the results of a study on physiological and ecological traits of two introduced and three native macrophyte species in the Mantua lakes system (northern Italy). We compared their photophysiology, pigment content, leaf reflectance, and phenology in order to assess how the invasive Nelumbo nucifera and Ludwigia hexapetala perform compared to native species, Nuphar lutea, Nymphaea alba, and Trapa natans. We found L. hexapetala to have higher photosynthetic efficiency and to tolerate higher light intensities than N. nucifera and the native species especially at extreme weather conditions (prolonged exposure to high light and higher temperatures). Chlorophyll a and b, and carotenoids content of both allochthonous species were substantially higher than those of native plants, suggesting adaptive response to the ecosystem of Mantua lakes system. Higher variability of recorded data in invasive species was also observed. These observations suggest advanced photosynthetic efficiency of the invasive species, especially L. hexapetala, resulting in faster growth rates and higher productivity. This was supported by the evaluation of seasonal dynamics mapped from satellite remote sensing data. This study provides empirical evidence for the relationship between specific plant physiological traits and invasiveness of aquatic plant species, highlighting the importance of trait studies in predicting ecosystem-level impacts of invasive plant species.

Exocyclic Sulfur and Selenoorganic Compounds Towards Their Anticancer Effects: Crystallographic and Biological Studies.

BACKGROUND/AIM: Multidrug resistance leads to therapeutic difficulties. There is great interest in experimental chemotherapy regarding multidrug resistance inhibitors and new anticancer agents. The aim of this study was to evaluate the anticancer activity of exocyclic sulfur and selenoorganic compounds on mouse T-lymphoma cell lines. MATERIALS AND METHODS: A series of eighteen sulfur and selenium analogues of 2[1H]-pyrimidinone and hydantoin derivatives were evaluated towards their efflux modulating, cytotoxic and antiproliferative effects in mouse T-lymphoma cells. The combination assay with doxorubicin on multidrug resistant mouse T-lymphoma cells was performed in order to see the nature of drug interactions. Crystal structures were determined for two selected compounds with the highest efflux-modulating activity. RESULTS: The sulfur analogues with aromatic rings almost perpendicular to pyrimidinethione ring at positions 1 and 6 showed the highest efflux inhibitory action, while all selenium analogues showed good antiproliferative and cytotoxic activities. CONCLUSION: The sulfur analogues can be modified towards improving their efflux inhibitory activity, whereas the selenium towards antiproliferative and cytotoxic activities.

Monitoring Spatial Variability and Temporal Dynamics of Phragmites Using Unmanned Aerial Vehicles.

Littoral zones of freshwater lakes are exposed to environmental impacts from both terrestrial and aquatic sides, while substantial anthropogenic pressure also affects the high spatial, and temporal variability of the ecotone. In this study, the possibility of monitoring seasonal and spatial changes in reed (Phragmites australis) stands using an unmanned aerial vehicle (UAV) based remote sensing technique was examined. Stands in eutrophic and mesotrophic parts of Lake Balaton including not deteriorating (stable) and deteriorating (die-back) patches, were tracked throughout the growing season using a UAV equipped with a Normalized Difference Vegetation Index (NDVI) camera. Photophysiological parameters of P. australis were also measured with amplitude modulated fluorescence. Parameters characterizing the dynamics of seasonal changes in NDVI data were used for phenological comparison of eutrophic and mesotrophic, stable and die-back, terrestrial and aquatic, mowed and not-mowed patches of reed. It was shown that stable Phragmites plants from the eutrophic part of the lake reached specific phenological stages up to 3.5 days earlier than plants from the mesotrophic part of the lake. The phenological changes correlated with trophic (total and nitrate-nitrite nitrogen) and physical (organic C and clay content) properties of the sediment, while only minor relationships with air and water temperature were found. Phenological differences between the stable and die-back stands were even more pronounced, with ~34% higher rates of NDVI increase in stable than die-back patches, while the period of NDVI increase was 16 days longer. Aquatic and terrestrial parts of reed stands showed no phenological differences, although intermediate areas (shallow water parts of stands) were found to be less vigorous. Winter mowing of dried Phragmites sped up sprouting and growth of reed in the spring. This study showed that remote sensing-derived photophysiological and phenological variability within and between reed stands may provide valuable early indicators of environmental stress. The flexibility of the method makes it usable for mapping fine-scale temporal variability and spatial zonation within a stand, revealing ecophysiological hotspots that might require particular attention, and obtaining information vital for conservation and management of plants in the littoral zones.

The effects of interspecific interactions between bloom forming cyanobacteria and Scenedesmus quadricauda (chlorophyta) on their photophysiology.

Eutrophication and enhanced external nutrient loading of lakes and seas are most clearly reflected by increased cyanobacterial blooms, which are often toxic. Freshwater cyanobacteria produce a number of bioactive secondary metabolites, some of which have allelopathic properties, significantly influencing the biological processes of other algae, thereby affecting species composition and succession of the phytoplankton. The goal of this work was to investigate the influence of bloom-forming cyanobacterial exudates on the photophysiology of the green alga Scenedesmus quadricauda by chlorophyll fluorescence analysis. We were able to prove the effect of algal cell-free filtrates on the performance of S. quadricauda and demonstrate for the first time that the freshwater picocyanobacterium Cyanobium gracile has strong negative impact on the coexisting green alga. Neither the cyanotoxin (MYC, CYN and ATX) producing, nor the non-toxic strains showed any systematic effect on the production of S. quadricauda. Various strains of the cyanobacterium Cylindrospermopsis raciborskii inhibited the performance of the green alga independently of their origin. Our results urge further studies for a better understanding of the factors affecting the release of allelopathic compounds and the mechanisms of their effects on target organisms.

Evaluating Sentinel-2 for Lakeshore Habitat Mapping Based on Airborne Hyperspectral Data.

Monitoring of lakeshore ecosystems requires fine-scale information to account for the high biodiversity typically encountered in the land-water ecotone. Sentinel-2 is a satellite with high spatial and spectral resolution and improved revisiting frequency and is expected to have significant potential for habitat mapping and classification of complex lakeshore ecosystems. In this context, investigations of the capabilities of Sentinel-2 in regard to the spatial and spectral dimensions are needed to assess its potential and the quality of the expected output. This study presents the first simulation of the high spatial resolution (i.e., 10 m and 20 m) bands of Sentinel-2 for lakeshore mapping, based on the satellite's Spectral Response Function and hyperspectral airborne data collected over Lake Balaton, Hungary in August 2010. A comparison of supervised classifications of the simulated products is presented and the information loss from spectral aggregation and spatial upscaling in the context of lakeshore vegetation classification is discussed. We conclude that Sentinel-2 imagery has a strong potential for monitoring fine-scale habitats, such as reed beds.

Tuning the aggregation of titanate nanowires in aqueous dispersions.

Electrophoretic and dynamic light scattering (DLS) measurements revealed that aggregation in aqueous dispersion of titanate nanowires (TiONWs) can be tuned by poly(diallyldimethylammonium) chloride (PDADMAC) polyelectrolyte. The nanowires possessed negative charge under alkaline conditions which was compensated by the oppositely charged PDADMAC adsorbed on the surface. Such adsorption led to charge neutralization and subsequent charge reversal at the appropriate polyelectrolyte doses. The dispersions were stable at low PDADMAC concentration where the TiONWs possessed negative charge. However, fast aggregation of the nanowires occurred close to the charge neutralization point where the overall charge of the particles was zero. Charge inversion at high polyelectrolyte doses gave rise to restabilization of the samples and slow aggregation of the TiONWs even at higher ionic strengths where the original bare TiONW dispersions were unstable. The colloid stability of the bare nanowires can be explained well qualitatively by the Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory; however, polyelectrolyte adsorption led to additional patch-charge attractions and osmotic repulsion between the particles. On the basis of the knowledge generated by the present work, experimental conditions (e.g., salt level, polyelectrolyte, and particle concentrations) can be adjusted in order to design stable and processable aqueous dispersions of TiONWs for further applications.

Support for the thermal origin of the pioneer anomaly.

We investigate the possibility that the anomalous acceleration of the Pioneer 10 and 11 spacecraft is due to the recoil force associated with an anisotropic emission of thermal radiation off the vehicles. To this end, relying on the project and spacecraft design documentation, we constructed a comprehensive finite-element thermal model of the two spacecraft. Then, we numerically solve thermal conduction and radiation equations using the actual flight telemetry as boundary conditions. We use the results of this model to evaluate the effect of the thermal recoil force on the Pioneer 10 spacecraft at various heliocentric distances. We found that the magnitude, temporal behavior, and direction of the resulting thermal acceleration are all similar to the properties of the observed anomaly. As a novel element of our investigation, we develop a parametrized model for the thermal recoil force and estimate the coefficients of this model independently from navigational Doppler data. We find no statistically significant difference between the two estimates and conclude that, once the thermal recoil force is properly accounted for, no anomalous acceleration remains.

Support for temporally varying behavior of the Pioneer anomaly from the extended Pioneer 10 and 11 Doppler data sets.

The Pioneer anomaly is a small sunward anomalous acceleration found in the trajectory analysis of the Pioneer 10 and 11 spacecraft. As part of the investigation of the effect, the analysis of recently recovered Doppler data for both spacecraft has been completed. The presence of a small anomalous acceleration is confirmed by using data spans more than twice as long as those that were previously analyzed. We examine the constancy and direction of the Pioneer anomaly and conclude that (i) the data favor a temporally decaying anomalous acceleration (~2×10(-11) m/s(2)/yr) with an over 10% improvement in the residuals compared to a constant acceleration model, (ii) although the direction of the acceleration remains imprecisely determined, we find no support in favor of a Sun-pointing direction over the Earth-pointing or along the spin-axis directions, and (iii) support for an early "onset" of the acceleration remains weak in the pre-Saturn Pioneer 11 tracking data. We present these new findings and discuss their implications for the nature of the Pioneer anomaly.

The Pioneer Anomaly.

Radio-metric Doppler tracking data received from the Pioneer 10 and 11 spacecraft from heliocentric distances of 20-70 AU has consistently indicated the presence of a small, anomalous, blue-shifted frequency drift uniformly changing with a rate of ∼ 6 × 10-9 Hz/s. Ultimately, the drift was interpreted as a constant sunward deceleration of each particular spacecraft at the level of aP = (8.74 ± 1.33) × 10-10 m/s2. This apparent violation of the Newton's gravitational inverse-square law has become known as the Pioneer anomaly; the nature of this anomaly remains unexplained. In this review, we summarize the current knowledge of the physical properties of the anomaly and the conditions that led to its detection and characterization. We review various mechanisms proposed to explain the anomaly and discuss the current state of efforts to determine its nature. A comprehensive new investigation of the anomalous behavior of the two Pioneers has begun recently. The new efforts rely on the much-extended set of radio-metric Doppler data for both spacecraft in conjunction with the newly available complete record of their telemetry files and a large archive of original project documentation. As the new study is yet to report its findings, this review provides the necessary background for the new results to appear in the near future. In particular, we provide a significant amount of information on the design, operations and behavior of the two Pioneers during their entire missions, including descriptions of various data formats and techniques used for their navigation and radio-science data analysis. As most of this information was recovered relatively recently, it was not used in the previous studies of the Pioneer anomaly, but it is critical for the new investigation.