The Role of EU Trade Secrets Law in the Data Economy: An Empirical Analysis.

This article draws on a recently completed study for the European Commission on trade secrets in the data economy. It distils the main findings of that Study and advances it by reflecting on and analyzing these findings in the context of existing legal, management and economics literature, as well as their implications for EU legal policymaking when it comes to trade secrets law. In order to facilitate data sharing, the article argues for a cautious approach, with very modest legislative reforms to the EU Trade Secrets Directive, instead preferring soft law and practical steps to be taken. There is, however, greater scope to reform legal regimes that are complementary to EU trade secrets law, such as the sui generis database right.

When things get MESI: The Manipulation Experiments Synthesis Initiative-A coordinated effort to synthesize terrestrial global change experiments.

Responses of the terrestrial biosphere to rapidly changing environmental conditions are a major source of uncertainty in climate projections. In an effort to reduce this uncertainty, a wide range of global change experiments have been conducted that mimic future conditions in terrestrial ecosystems, manipulating CO2 , temperature, and nutrient and water availability. Syntheses of results across experiments provide a more general sense of ecosystem responses to global change, and help to discern the influence of background conditions such as climate and vegetation type in determining global change responses. Several independent syntheses of published data have yielded distinct databases for specific objectives. Such parallel, uncoordinated initiatives carry the risk of producing redundant data collection efforts and have led to contrasting outcomes without clarifying the underlying reason for divergence. These problems could be avoided by creating a publicly available, updatable, curated database. Here, we report on a global effort to collect and curate 57,089 treatment responses across 3644 manipulation experiments at 1145 sites, simulating elevated CO2 , warming, nutrient addition, and precipitation changes. In the resulting Manipulation Experiments Synthesis Initiative (MESI) database, effects of experimental global change drivers on carbon and nutrient cycles are included, as well as ancillary data such as background climate, vegetation type, treatment magnitude, duration, and, unique to our database, measured soil properties. Our analysis of the database indicates that most experiments are short term (one or few growing seasons), conducted in the USA, Europe, or China, and that the most abundantly reported variable is aboveground biomass. We provide the most comprehensive multifactor global change database to date, enabling the research community to tackle open research questions, vital to global policymaking. The MESI database, freely accessible at doi.org/10.5281/zenodo.7153253, opens new avenues for model evaluation and synthesis-based understanding of how global change affects terrestrial biomes. We welcome contributions to the database on GitHub.

End-to-end approach for the characterization and control of product-related impurities in T cell bispecific antibody preparations.

Antibody-based T cell-activating biologics are promising therapeutic medicines being developed for a number of indications, mainly in the oncology field. Among those, T cell bispecific antibodies are designed to bind one tumor-specific antigen and the T cell receptor at the same time, leading to a robust T cell response against the tumor. Although their unique format and the versatility of the CrossMab technology allows for the generation of safer molecules in an efficient manner, product-related variants cannot be completely avoided. Therefore, it is of extreme importance that both a manufacturing process that limits or depletes product-related impurities, as well as a thorough analytical characterization are in place, starting from the development of the manufacturing cell line until the assessment of potential toxicities. Here, we describe such an end-to-end approach to minimize, quantify and control impurities and -upon their functional characterization- derive specifications that allow for the release of clinical material.

Optimal stomatal theory predicts CO2 responses of stomatal conductance in both gymnosperm and angiosperm trees.

Optimal stomatal theory predicts that stomata operate to maximise photosynthesis (Anet ) and minimise transpirational water loss to achieve optimal intrinsic water-use efficiency (iWUE). We tested whether this theory can predict stomatal responses to elevated atmospheric CO2 (eCO2 ), and whether it can capture differences in responsiveness among woody plant functional types (PFTs). We conducted a meta-analysis of tree studies of the effect of eCO2 on iWUE and its components Anet and stomatal conductance (gs ). We compared three PFTs, using the unified stomatal optimisation (USO) model to account for confounding effects of leaf-air vapour pressure difference (D). We expected smaller gs , but greater Anet , responses to eCO2 in gymnosperms compared with angiosperm PFTs. We found that iWUE increased in proportion to increasing eCO2 in all PFTs, and that increases in Anet had stronger effects than reductions in gs . The USO model correctly captured stomatal behaviour with eCO2 across most datasets. The chief difference among PFTs was a lower stomatal slope parameter (g1 ) for the gymnosperm, compared with angiosperm, species. Land surface models can use the USO model to describe stomatal behaviour under changing atmospheric CO2 conditions.

Temperature Effects on the Survival and Development of Two Pest Bark Beetles Hylurgus ligniperda F. (Coleoptera: Curculionidae) and Hylastes ater Paykull (Coleoptera: Curculionidae).

Hylurgus ligniperda (F.) and Hylastes ater (Paykull) are secondary bark beetles that have successfully spread beyond their native range, particularly into Pinus spp. plantations in the Southern Hemisphere. They feed on the phloem and cambial regions of highly stressed and recently dead Pinus spp. Here H. ligniperda and H. ater egg, larval, and pupal survival and development rates were modeled. Survival was variably influenced by temperatures depending on the life stage, but general trends were for H. ligniperda to tolerate warmer temperatures in comparison to H. ater. Nonlinear models showed 26, 29, and 34°C are the optimal temperature (maximum development rates) for the development of eggs, larvae, and pupae of H. ligniperda. In contrast, optimal temperature predictions were lower for H. ater, with estimates of 26, 22, and 23°C for the development of eggs, larvae, and pupae, respectively. H. ligniperda pre-imaginal stages were more tolerant to high temperatures, and H. ater pre-imaginal stages were more tolerant to low temperatures. Understanding the thermal requirements and limits for development for these two pests can assist in modeling emergence times, their current and potential species distribution and have potential phytosanitary applications.

Soil Inoculation With Beneficial Microbes Buffers Negative Drought Effects on Biomass, Nutrients, and Water Relations of Common Myrtle.

Common myrtle (Myrtus communis L.) occurs in (semi-)arid areas of the Palearctic region where climate change, over-exploitation, and habitat destruction imperil its existence. The evergreen shrub is of great economic and ecological importance due to its pharmaceutical value, ornamental use, and its role in urban greening and habitat restoration initiatives. Under greenhouse conditions, we investigated the effect of soil inoculation with arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) on biomass allocation, water relations, and nutritional status of drought-stressed myrtle seedlings. Single and dual AMF (Funneliformis mosseae and Rhizophagus irregularis) and PGPR (Pseudomonas fluorescens and P. putida) soil inoculations were applied to myrtle seedlings growing under different soil water regimes (100, 60, and 30% of field capacity) for 6 months using a full factorial, completely randomized design. AMF and PGPR treatments, especially dual inoculations, alleviated negative drought effects on biomass and morpho-physiological traits, except for water-use efficiency, which peaked under severe drought conditions. Under the greatest soil water deficit, dual inoculations promoted leaf biomass (104%-108%), root biomass (56%-73%), mesophyll conductance (58%), and relative water content (1.4-fold) compared to non-inoculated controls. Particularly, dual AMF and PGPR inoculations stimulated nutrient dynamics in roots (N: 138%-151%, P: 176%-181%, K: 112%-114%, Ca: 124%-136%, and Mg: 130%-140%) and leaves (N: 101%-107%, P: 143%-149%, K: 83%-84%, Ca: 98%-107%, and Mg: 102%-106%). Our findings highlight soil inoculations with beneficial microbes as a cost-effective way to produce highly drought resistant seedling stock which is vital for restoring natural myrtle habitats and for future-proofing myrtle crop systems.

High light-induced photoinhibition is not limiting seedling establishment at abrupt treeline ecotones in New Zealand.

Seedlings of New Zealand's treeline-forming Fuscospora cliffortioides (Hook.f.) perform poorly beyond the established canopy, limiting treeline advance. To test the long-standing assumption that photoinhibition impairs regeneration in the subalpine belt of New Zealand's Southern Alps, we assessed photosystem II (PSII) performance of seedling-sized individuals and microclimate variation. We performed diurnal, non-invasive chlorophyll-a-fluorescence measurements on exposed and canopy-sheltered individuals at two sites in New Zealand's Southern Alps during summer and winter. Diurnal recordings of the effective (ΦPSII) and optimal (Fv/Fm) photosynthetic quantum yield were supplemented with light response curves and micro-temperature recordings. ΦPSII returned to near-optimal values around 0.8 after 30 min of shading, which rules out accumulative or long-term photoinhibition effects. The maximum electron transport rate derived from rapid light curves was significantly higher (+12%) in exposed compared with canopy-shaded individuals. Summer temperature fluctuated widely on the scree (-0.5 to 60.5 °C) and near seedlings (-2 to 26.5 °C). Our results revealed a remarkable level of light adaptation and contradict previous studies hinting at high light-induced photoinhibition as a treeline-limiting factor in the Southern Alps. By linking low ΦPSII on winter mornings, and large, sudden temperature drops in summer, we suspect that cold-induced photoinhibition might occur but the rapid recovery of ΦPSII seen across a wide temperature range makes lethal photo-oxidative damage rather unlikely. Given the demonstrably low summer frost tolerance of F. cliffortioides, cold-related damage resulting from frost events during the growing season or embolism induced by frost drought may offer more plausible explanations for the poor seedling establishment. Duration and frequency of these events could diminish with global warming, which may promote treeline advance.

Author Correction: Behaviour reactions of bottlenose dolphins (Tursiops truncatus) to multirotor Unmanned Aerial Vehicles (UAVs).

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

No carbon limitation after lower crown loss in Pinus radiata.

BACKGROUND AND AIMS: Biotic and abiotic stressors can cause different defoliation patterns within trees. Foliar pathogens of conifers commonly prefer older needles and infection with defoliation that progresses from the bottom crown to the top. The functional role of the lower crown of trees is a key question to address the impact of defoliation caused by foliar pathogens. METHODS: A 2 year artificial defoliation experiment was performed using two genotypes of grafted Pinus radiata to investigate the effects of lower-crown defoliation on carbon (C) assimilation and allocation. Grafts received one of the following treatments in consecutive years: control-control, control-defoliated, defoliated-control and defoliated-defoliated. RESULTS: No upregulation of photosynthesis either biochemically or through stomatal control was observed in response to defoliation. The root:shoot ratio and leaf mass were not affected by any treatment, suggesting prioritization of crown regrowth following defoliation. In genotype B, defoliation appeared to impose C shortage and caused reduced above-ground growth and sugar storage in roots, while in genotype A, neither growth nor storage was altered. Root C storage in genotype B decreased only transiently and recovered over the second growing season. CONCLUSIONS: In genotype A, the contribution of the lower crown to the whole-tree C uptake appears to be negligible, presumably conferring resilience to foliar pathogens affecting the lower crown. Our results suggest that there is no C limitation after lower-crown defoliation in P. radiata grafts. Further, our findings imply genotype-specific defoliation tolerance in P. radiata.

Contrasting the Pathogen Loads in Co-Existing Populations of Phytophthora pluvialis and Nothophaeocryptopus gaeumannii in Douglas Fir Plantations in New Zealand and the Pacific Northwest United States.

The emergence of Phytophthora pluvialis as a foliar pathogen of Douglas fir in New Zealand and the Pacific Northwest United States has raised questions about its interaction with the widespread Swiss needle cast (SNC) disease. During Spring 2017, we repeatedly sampled 30 trees along an environmental gradient in each region and 292 additional trees in a longitudinal transect to assess the P. pluvialis epidemic and the association between P. pluvialis and Nothophaeocryptopus gaeumannii, which are causal agents of SNC. Both pathogens were consistently more abundant in the host's exotic environment in New Zealand. In both areas, the two pathogens co-exist in different spatial scales for regions and needles. The relative abundance of both pathogens was negatively correlated in the Pacific Northwest, where both presumably have co-existed for longer. Our findings confirmed the interaction of P. pluvialis and N. gaeumannii as foliar pathogens of Douglas fir and suggest a within-site spatial variation in the Pacific Northwest.

Behaviour reactions of bottlenose dolphins (Tursiops truncatus) to multirotor Unmanned Aerial Vehicles (UAVs).

Unmanned aerial vehicles (UAVs) represent a novel and cost effective research tool to investigate cetacean behaviour, as conventional aircraft are expensive, limited in the altitude they can fly at and potentially disturb sensitive wildlife. In addition, the aerial observation from the UAVs allows assessment of cetacean behaviour from an advantageous perspective and can collect high spatial and temporal resolution data, providing the opportunity to gather accurate data about group size, age class and subsurface behaviour. However, concerns have been raised about the potential risks of disturbance to animals caused by the UAV's visual and acoustic stimuli. Boat-based surveys were conducted to assess the short-term behavioural responses of resting bottlenose dolphins (Tursiops truncatus) to a lightweight Vertical take-off and landing (VTOL) UAV flown at 10, 25, and 40 m altitude. Changes in group swim direction and frequencies of surface and aerial behavioural events were recorded from an anchored research vessel before (control) and during the aerial survey. The number of reorientation and tail slap events increased significantly between controls and flights when the UAV was flown at 10 m over the animals. In contrast, no significant differences were detected when the aircraft was flown at 25 and 40 m altitude. However, a precautionary approach is recommended for research applications requiring lower flight altitudes, with further research recommended to assess how different cetacean species and age class may respond to the UAV presence.

Can Copper Be Used to Treat Foliar Phytophthora Infections in Pinus radiata?

Red needle cast is a significant foliar disease of commercial stands of Pinus radiata caused by Phytophthora pluvialis in New Zealand. The effect of copper, applied as a foliar spray of cuprous oxide at a range of doses between 0 and 1.72 kg ha-1, was investigated in two controlled trials with potted plants and in an operational trial with mature P. radiata. In all trials, lesions formed on needles after artificial exposure to the infecting propagules (zoospores) of P. pluvialis were used to determine treatment efficacy, with the number and/or length of lesions as the dependent variable. Results across all trials indicated that cuprous oxide was highly effective at reducing infection of P. radiata with P. pluvialis. Application rates equivalent to ≥0.65 kg ha-1 significantly reduced infection levels relative to a control treatment, with foliar surface copper levels as low as 13 to 26 mg kg-1 of needle tissue preventing infection. Greater copper content was associated with a reduction in the proportion of needles with P. pluvialis lesions, with the probability of lesions developing decreasing approximately 1% for every 1 unit (in milligrams per kilogram) increase in copper content. Over a 90-day period, surface copper content declined to 30% of that originally applied, indicating an approximate period of treatment efficacy of 3 months. Our findings highlight the potential of cuprous oxide for the control of red needle cast in P. radiata stands. Further information about the optimal field dose, timing, and the frequency of foliar cuprous oxide application is key to prevent infection and also reduce the build up of inoculum during severe outbreaks of this pathogen.

Modelling the key drivers of an aerial Phytophthora foliar disease epidemic, from the needles to the whole plant.

Understanding the epidemiology of infectious diseases in a host population is a major challenge in forestry. Radiata pine plantations in New Zealand are impacted by a foliar disease, red needle cast (RNC), caused by Phytophthora pluvialis. This pathogen is dispersed by water splash with polycyclic infection affecting the lower part of the tree canopy. In this study, we extended an SI (Susceptible-Infectious) model presented for RNC to analyse the key epidemiological drivers. We conducted two experiments to empirically fit the extended model: a detached-needle assay and an in vivo inoculation. We used the detached-needle assay data to compare resistant and susceptible genotypes, and the in vivo inoculation data was used to inform sustained infection of the whole plant. We also compared isolations and real-time quantitative PCR (qPCR) to assess P. pluvialis infection. The primary infection rate and the incubation time were similar for susceptible and resistant genotypes. The pathogen death rate was 2.5 times higher for resistant than susceptible genotypes. Further, external proliferation of mycelium and sporangia were only observed on 28% of the resistant ramets compared to 90% of the susceptible ones. Detection methods were the single most important factor influencing parameter estimates of the model, giving qualitatively different epidemic outputs. In the early stages of infection, qPCR proved to be more efficient than isolations but the reverse was true at later points in time. Isolations were not influenced by the presence of lesions in the needles, while 19% of lesioned needle maximized qPCR detection. A primary infection peak identified via qPCR occurred at 4 days after inoculation (dai) with a secondary peak observed 22 dai. Our results have important implications to the management of RNC, by highlighting the main differences in the response of susceptible and resistant genotypes, and comparing the most common assessment methods to detect RNC epidemics.

Temperature Effects on Biomass and Regeneration of Vegetation in a Geothermal Area.

Understanding the effects of increasing temperature is central in explaining the effects of climate change on vegetation. Here, we investigate how warming affects vegetation regeneration and root biomass and if there is an interactive effect of warming with other environmental variables. We also examine if geothermal warming effects on vegetation regeneration and root biomass can be used in climate change experiments. Monitoring plots were arranged in a grid across the study area to cover a range of soil temperatures. The plots were cleared of vegetation and root-free ingrowth cores were installed to assess above and below-ground regeneration rates. Temperature sensors were buried in the plots for continued soil temperature monitoring. Soil moisture, pH, and soil chemistry of the plots were also recorded. Data were analyzed using least absolute shrinkage and selection operator and linear regression to identify the environmental variable with the greatest influence on vegetation regeneration and root biomass. There was lower root biomass and slower vegetation regeneration in high temperature plots. Soil temperature was positively correlated with soil moisture and negatively correlated with soil pH. Iron and sulfate were present in the soil in the highest quantities compared to other measured soil chemicals and had a strong positive relationship with soil temperature. Our findings suggest that soil temperature had a major impact on root biomass and vegetation regeneration. In geothermal fields, vegetation establishment and growth can be restricted by low soil moisture, low soil pH, and an imbalance in soil chemistry. The correlation between soil moisture, pH, chemistry, and plant regeneration was chiefly driven by soil temperature. Soil temperature was negatively correlated to the distance from the geothermal features. Apart from characterizing plant regeneration on geothermal soils, this study further demonstrates a novel approach to global warming experiments, which could be particularly useful in low heat flow geothermal systems that more realistically mimic soil warming.

Phytophthora pluvialis Studies on Douglas-fir Require Swiss Needle Cast Suppression.

Phytophthora pluvialis is associated with early defoliation and shoot dieback in Douglas-fir in Oregon and New Zealand. In 2013, P. pluvialis was described from mixed tanoak-Douglas-fir forests in the Pacific Northwest and concurrently recognized as the main causal agent of red needle cast (RNC) in New Zealand radiata pine plantations. Little is known about its infection cycle and impact on host physiology. P. pluvialis studies in Douglas-fir are challenging due to the ubiquitous presence of the endophyte Phaeocryptopus gaeumannii, which produces similar symptoms and premature defoliation with persistent needle wetness, known as Swiss needle cast (SNC). Nonetheless, our study showed P. pluvialis infection in the presence of SNC. Exclusive expression of P. pluvialis is difficult to achieve as both diseases are promoted by high humidity. Here we established a 'dry leaf' strategy to suppress SNC when inoculating Douglas-fir needles for RNC studies. Sheltering plants along with drip irrigation to avoid needle wetness during the P. gaeumannii sporulation period suppressed its development in the new season flush. The diminished endophyte inoculum enabled bias-reduced studies of P. pluvialis impacts on Douglas-fir without the confounding effects of stomatal blockage and premature defoliation caused by P. gaeumannii.

Olfactory Cues, Visual Cues, and Semiochemical Diversity Interact During Host Location by Invasive Forest Beetles.

Plant-feeding insects use visual and olfactory cues (shape, color, plant volatiles) for host location, but the relative importance of different cues and interactions with non-host-plant volatiles in ecosystems of varying plant biodiversity is unclear for most species. We studied invasive bark beetles and wood borers associated with pine trees to characterize interactions among color, host and non-host volatiles, by employing traps that mimic tree trunks. Cross-vane flight intercept traps (black, green, red, white, yellow, clear) and black funnel traps were used with and without attractants (α-pinene + ethanol), repellents (non-host green leaf volatiles, 'GLV'), and attractant/repellent combinations in four pine forests in New Zealand. We trapped 274,594 Hylurgus ligniperda, 7842 Hylastes ater, and 16,301 Arhopalus ferus. Trap color, attractant, and color × attractant effects were highly significant. Overall, black and red traps had the highest catches, irrespective of the presence of attractants. Alpha-pinene plus ethanol increased trap catch of H. ligniperda 200-fold but only 6-fold for H. ater and 2-fold for A. ferus. Green leaf volatiles had a substantial repellent effect on trap catch of H. ligniperda but less on H. ater and A. ferus. Attack by H. ligniperda was halved when logs were treated with GLV, and a similar effect was observed when logs were placed among broadleaved understory shrubs emitting GLV. Overall, H. ligniperda was most strongly affected by the olfactory cues used, whereas H. ater and A. ferus were more strongly affected by visual cues. Collectively, the results support the semiochemical diversity hypothesis, indicating that non-host plant volatiles from diverse plant communities or artificial dispensers can contribute to resistance against herbivores by partly disrupting host location.

Foliar phosphite application has minor phytotoxic impacts across a diverse range of conifers and woody angiosperms.

Phytophthora plant pathogens cause tremendous damage in planted and natural systems worldwide. Phosphite is one of the only effective chemicals to control broad-scale Phytophthora disease. Little work has been done on the phytotoxic effects of phosphite application on plant communities especially in combination with plant physiological impacts. Here, we tested the phytotoxic impact of phosphite applied as foliar spray at 0, 12, 24 and 48 kg a.i. ha(-1) . Eighteen-month-old saplings of 13 conifer and angiosperm species native to New Zealand, and two exotic coniferous species were treated and the development of necrotic tissue and chlorophyll-a-fluorescence parameters (optimal quantum yield, Fv /Fm ; effective quantum yield of photosystem II, ΦPSII ) were assessed. In addition, stomatal conductance (gs ) was measured on a subset of six species. Significant necrosis assessed by digital image analysis occurred in only three species: in the lauraceous canopy tree Beilschmiedia tawa (8-14%) and the understory shrub Dodonaea viscosa (5-7%) across phosphite concentrations and solely at the highest concentration in the myrtaceous pioneer shrub Leptospermum scoparium (66%). In non-necrotic tissue, Fv /Fm , ΦPSII and gs remained unaffected by the phosphite treatment. Overall, our findings suggest minor phytotoxic effects resulting from foliar phosphite application across diverse taxa and regardless of concentration. This study supports the large-scale use of phosphite as a management tool to control plant diseases caused by Phytophthora pathogens in plantations and natural ecosystems. Long-term studies are required to ascertain potential ecological impacts of repeated phosphite applications.

A dynamic leaf gas-exchange strategy is conserved in woody plants under changing ambient CO2 : evidence from carbon isotope discrimination in paleo and CO2 enrichment studies.

Rising atmospheric [CO2 ], ca , is expected to affect stomatal regulation of leaf gas-exchange of woody plants, thus influencing energy fluxes as well as carbon (C), water, and nutrient cycling of forests. Researchers have proposed various strategies for stomatal regulation of leaf gas-exchange that include maintaining a constant leaf internal [CO2 ], ci , a constant drawdown in CO2 (ca  - ci ), and a constant ci /ca . These strategies can result in drastically different consequences for leaf gas-exchange. The accuracy of Earth systems models depends in part on assumptions about generalizable patterns in leaf gas-exchange responses to varying ca . The concept of optimal stomatal behavior, exemplified by woody plants shifting along a continuum of these strategies, provides a unifying framework for understanding leaf gas-exchange responses to ca . To assess leaf gas-exchange regulation strategies, we analyzed patterns in ci inferred from studies reporting C stable isotope ratios (δ(13) C) or photosynthetic discrimination (∆) in woody angiosperms and gymnosperms that grew across a range of ca spanning at least 100 ppm. Our results suggest that much of the ca -induced changes in ci /ca occurred across ca spanning 200 to 400 ppm. These patterns imply that ca  - ci will eventually approach a constant level at high ca because assimilation rates will reach a maximum and stomatal conductance of each species should be constrained to some minimum level. These analyses are not consistent with canalization toward any single strategy, particularly maintaining a constant ci . Rather, the results are consistent with the existence of a broadly conserved pattern of stomatal optimization in woody angiosperms and gymnosperms. This results in trees being profligate water users at low ca , when additional water loss is small for each unit of C gain, and increasingly water-conservative at high ca , when photosystems are saturated and water loss is large for each unit C gain.

Developability assessment during the selection of novel therapeutic antibodies.

Therapeutic antibodies and antibody derivatives comprise the majority of today's biotherapeutics. Routine methods to generate novel antibodies, such as immunization and phage-display, often give rise to several candidates with desired functional properties. On the contrary, resource-intense steps such as the development of a cell line, a manufacturing process, or a formulation, are typically carried out for only one candidate. Therefore, "developability," that is, the likelihood for the successful development of a lead candidate into a stable, manufacturable, safe, and efficacious drug, may be used as an additional selection criterion. Employing a set of small-scale, fast, and predictive tests addressing biochemical and biophysical features, as well as in vivo fate can help to identify a clinical candidate molecule with promising properties at an early stage of drug development. This article gives an overview of existing methods for developability testing and shows how these assays can be interlaced in the lead selection process.

Combined inhibition of tumor necrosis factor α and interleukin-17 as a therapeutic opportunity in rheumatoid arthritis: development and characterization of a novel bispecific antibody.

OBJECTIVE: Rheumatoid arthritis therapies that are based on inhibition of a single cytokine, e.g., tumor necrosis factor α (TNFα) or interleukin-6 (IL-6), produce clinically meaningful responses in only about half of the treated patients. This study was undertaken to investigate whether combined inhibition of TNFα and IL-17 has additive or synergistic effects in the suppression of mesenchymal cell activation in vitro and inflammation and tissue destruction in arthritis in vivo. METHODS: Cultures of human fibroblast-like synoviocytes (FLS) were stimulated with TNFα, IL-17, or a combination of both. Single/combined neutralizing antibodies against TNFα and IL-17 were used to examine in vitro cytokine responses and in vivo development of arthritis and bone and cartilage destruction in TNFα-transgenic mice. Bispecific anti-TNFα/IL-17 antibodies were designed, and their potential to block cytokine responses in human FLS was tested. RESULTS: TNFα and IL-17 had additive/synergistic effects in promoting production of IL-6, IL-8, and granulocyte colony-stimulating factor, as well as matrix metalloproteinases, in FLS. Bispecific anti-TNFα/IL-17 antibodies showed superior efficacy in blocking cytokine and chemokine responses in vitro. Furthermore, dual versus single inhibition of both cytokines using neutralizing antibodies was more effective in inhibiting the development of inflammation and bone and cartilage destruction in arthritic mice. CONCLUSION: Combined blockade of TNFα and IL-17 was more effective than single blockade in inhibiting cytokine, chemokine, and matrix enzyme responses from human mesenchymal cells and in blocking tissue destruction associated with arthritis, and additionally showed a positive impact on rebalance of bone homeostasis. Bispecific anti-TNFα/IL-17 antibodies may have superior efficacy in the treatment of arthritis and may overcome the limited therapeutic responses obtained with single cytokine neutralization.