DLX1008 (brolucizumab), a single-chain anti-VEGF-A antibody fragment with low picomolar affinity, leads to tumor involution in an in vivo model of Kaposi Sarcoma.

Kaposi Sarcoma (KS) is among the most angiogenic cancers in humans and an AIDS-defining condition. KS-associated herpesvirus (KSHV) is necessary for KS development, as is vascular endothelial growth factor (VEGF-A). DLX1008 is a novel anti-VEGF-A antibody single-chain variable fragment (scFv) with low picomolar affinity for VEGF-A. In vivo imaging techniques were used to establish the efficacy of DLX1008 and to establish the mechanism of action; this included non-invasive imaging by ultrasound and optical fluorescence, verified by post-mortem histochemistry. The results showed that DLX1008 was efficacious in a KS mouse model. The NSG mouse xenografts suffered massive internal necrosis or involution, consistent with a lack of blood supply. We found that imaging by ultrasound was superior to external caliper measurements in the validation of the angiogenesis inhibitor DLX1008. Further development of DLX1008 against VEGF-dependent sarcomas is warranted.

Correction: Soot and the city: Evaluating the impacts of Clean Heat policies on indoor/outdoor air quality in New York City apartments.

[This corrects the article DOI: 10.1371/journal.pone.0199783.].

Soot and the city: Evaluating the impacts of Clean Heat policies on indoor/outdoor air quality in New York City apartments.

New York City has had a long history of implementing local policies to reduce air pollution. Enacted as a part of PlaNYC, the Clean Heat policies aim to lower wintertime ambient air pollution by phasing out dirty No. 6 heating fuel oil and transitioning to comparatively cleaner No. 4, No. 2, or natural gas. This study evaluates the impacts of policies on ambient air pollution and, given that people spend the majority of their time inside, importantly, indoor air pollution. Using a natural experiment, we evaluate the effects of the policies by measuring average two-week levels of indoor and outdoor black carbon (BC) and fine particulate matter (PM2.5) in 48 upper Manhattan apartments in successive winter heating seasons before and after mandated fuel transition. We failed to observe systematic improvements in indoor BC and PM2.5 concentrations in follow-up. However, outdoor levels of PM2.5 did improve, with statistical differences observed among buildings converting to the cleanest fuels. Non-statistical improvements were observed for outdoor BC. However, when accounting for meteorological differences, apartment characteristics, and behavioral patterns that may have influenced air pollution measurements, these differences were not significant. The study results have important policy and equity implications considering the differential improvements in air quality by conversion to No. 4 oil as compared to the cleaner No. 2 oil and natural gas.

Antitumor Activity of DLX1008, an Anti-VEGFA Antibody Fragment with Low Picomolar Affinity, in Human Glioma Models.

Angiogenesis mediated by vascular endothelial growth factor (VEGF) is a hallmark of glioblastoma. Based on the response rate and improved progression-free survival, although not on overall survival, the 149-kDa anti-VEGF-A IgG antibody bevacizumab (Avastin) has been approved in the United States and Japan for recurrent glioblastoma and in Japan for newly diagnosed glioblastoma; however, it is not approved in the EU. Here we characterize the biologic activity of DLX1008, a 26-kDa anti-VEGF-A single-chain antibody fragment that shows 30-fold stronger affinity to human VEGF-A than bevacizumab. The small molecular size of DLX1008 is predicted to result in improved target coverage over bevacizumab. DLX1008 showed superiority to bevacizumab in the inhibition of VEGF-A binding to VEGF receptor (VEGFR) 1 in enzyme-linked immunosorbent assay by a factor of around 10 and comparable efficacy for the inhibition of VEGF-A-stimulated VEGFR2 dimerization. In a tube-formation assay with human cerebral microvascular endothelial cells, DLX1008 was at least as active as bevacizumab. In vivo, DLX1008 delayed growth in a mouse subcutaneous U87 xenograft model (P = 0.0021) and improved survival in a mouse orthotopic U87 xenograft model (P = 0.00026). Given the exceptionally high affinity and small molecular size of DLX1008, these data warrant further clinical development of DLX1008 as an antiangiogenic agent in glioblastoma.

Brugada Syndrome Induced by an Interscalene Block: A Case Report.

CASE: A 57-year-old woman with no noteworthy medical or surgical history underwent an interscalene block with bupivacaine in preparation for an arthroscopic rotator cuff repair. Following administration of the bupivacaine, the patient sustained a ventricular fibrillation arrest. After successful cardiopulmonary resuscitation, she was diagnosed with Brugada syndrome. An implantable cardioverter-defibrillator was placed, and the rotator cuff repair was performed 1 month later. CONCLUSION: Brugada syndrome is an abnormality of the cardiac conduction system that leads to cardiac arrhythmias. Several anesthetic agents trigger Brugada-like electrocardiographic abnormalities. To our knowledge, this is the first report of an interscalene block inducing Brugada syndrome.

Half-life extension using serum albumin-binding DARPin® domains.

A long systemic half-life is key for therapeutic proteins. To that end we have generated serum albumin-binding designed ankyrin repeat domains. These domains bind serum albumin of different species with nanomolar affinities, and have significantly improved pharmacokinetic properties both in mouse and cynomolgus monkey compared to non-serum albumin-binding DARPin® domains. In addition, they exhibit high thermal stability and long storage stability, which is an essential feature for their use in drug development. Covalently linking a serum albumin-binding DARPin® domain to domains with other target specificities results in improvements of multiple orders of magnitude in exposure and terminal half-life, both in mouse and cynomolgus monkey. Pharmacokinetic assessment of such constructs revealed terminal half-life values ranging from 27 h to 80 h in mouse, and from 2.6 days to 20 days in cynomolgus monkey. Extrapolation by allometric scaling on these findings suggests terminal half-life values of 5-50 days in human, indicating that pharmacokinetic properties in the range of monoclonal antibodies can be achieved with DARPin® drug candidates. Such serum albumin-binding DARPin® domains are thus valuable tools for the generation of multi-functional drugs with an extended in vivo half-life.

Design and characterization of MP0250, a tri-specific anti-HGF/anti-VEGF DARPin® drug candidate.

MP0250 is a multi-domain drug candidate currently being tested in clinical trials for the treatment of cancer. It comprises one anti-vascular endothelial growth factor-A (VEGF-A), one anti-hepatocyte growth factor (HGF), and two anti-human serum albumin (HSA) DARPin® domains within a single polypeptide chain. While there is first clinical validation of a single-domain DARPin® drug candidate, little is known about DARPin® drug candidates comprising multiple domains. Here, we show that MP0250 can be expressed at 15 g/L in soluble form in E. coli high cell-density fermentation, it is stable in soluble/frozen formulation for 2 years as assessed by reverse phase HPLC, it has picomolar potency in inhibiting VEGF-A and HGF in ELISA and cellular assays, and its domains are simultaneously active as shown by surface plasmon resonance. The inclusion of HSA-binding DARPin® domains leads to a favorable pharmacokinetic profile in mouse and cynomolgus monkey, with terminal half-lives of ∼ 30 hours in mouse and ∼ 5 days in cynomolgus monkey. MP0250 is thus a highly potent drug candidate that could be particularly useful in oncology. Beyond MP0250, the properties of MP0250 indicate that multi-domain DARPin® proteins can be valuable next-generation drug candidates.

Housing hardship and energy insecurity among native-born and immigrant low-income families with children in the United States.

The costs for rent and utilities account for the largest share of living expenses, yet these two critical dimensions of material hardship have seldom been examined concurrently in population-based studies. This paper employs multivariate statistical analysis using American Community Survey data to demonstrate the relative risk ratio of low-income renter-occupied households with children experiencing "rent burden," "energy insecurity," or a "double burden" as opposed to no burden. Findings suggest that low-income households are more likely to experience these economic hardships in general but that specific groups are disproportionately burdened in different ways. For instance, whereas immigrants are more likely to experience rental burden, they are less likely to experience energy insecurity and are also spared from the double burden. In contrast, native-born African Americans are more likely than all other groups to experience the double burden. These results may be driven by the housing stock available to certain groups due to racial residential segregation, decisions regarding the quality of housing low-income householders are able to afford, as well as home-country values, such as modest living and energy conservation practices, among immigrant families. This paper also points to important policy gaps in safety net benefits related to housing and energy targeting low-income households.

Exploring the Housing and Household Energy Pathways to Stress: A Mixed Methods Study.

Chronic stress, known to contribute to negative physical and mental health outcomes, is closely associated with broader issues of material hardship, poor neighborhood conditions, residential instability, and inadequate housing conditions. However, few studies have comprehensively explored pathways to stress in a low-income housing environment. A mixed-methods pilot study investigated the concept of energy insecurity by looking at the impacts of weatherization and energy efficiency interventions on low-income households in the South Bronx neighborhood of New York City. In-depth interviews were conducted with 20 low-income heads of household; participants also completed health, housing and budget assessments. Physical deficiencies, economic hardship, and health issues all interacted to directly and indirectly produce living conditions that contribute to chronic stress. Households with higher stress reported more health problems. Poor quality housing led to coping responses that increased expenses, which in turn increased stress around housing and energy affordability. This study provides further support for the connections between both health and the built environment and between low socio-economic status populations and net negative health outcomes. Energy insecurity is an important contributor to chronic stress in low-income households, and isolating pathways to stress where there is potential for interventions is important for future policy and housing-based strategies.

Benefit or burden? Perceptions of energy efficiency efforts among low-income housing residents in New York City.

Low-income households contend with high energy costs and poor thermal comfort due to poor structural conditions and energy inefficiencies in their homes. Energy efficiency upgrades can potentially reduce energy expenses and improve thermal comfort, while also addressing problematic issues in the home environment. The present mixed method pilot study explored the impacts of energy efficiency upgrades in 20 households in a low-income community in New York City. Surveys and interviews were administered to the heads of household in a variety of housing types. Interviews were also conducted with landlords of buildings that had recently undergone upgrades. Findings indicate that energy efficiency measures resulted in improved thermal comfort, enhanced health and safety and reduced energy costs. Participants reported largely positive experiences with the upgrades, resulting in direct and indirect benefits. However, results also indicate negative consequences associated with the upgrades and further illustrate that weatherization alone was insufficient to address all of the issues facing low-income households. Moreover, qualitative results revealed differing experiences of low-income renters compared to homeowners. Overall, energy efficiency upgrades are a promising intervention to mitigate the energy and structurally related challenges facing low-income households, but larger scale research is needed to capture the long-term implications of these upgrades.

Detection and tracking of a novel genetically tagged biological simulant in the environment.

A variant of Bacillus thuringiensis subsp. kurstaki containing a single, stable copy of a uniquely amplifiable DNA oligomer integrated into the genome for tracking the fate of biological agents in the environment was developed. The use of genetically tagged spores overcomes the ambiguity of discerning the test material from pre-existing environmental microflora or from previously released background material. In this study, we demonstrate the utility of the genetically "barcoded" simulant in a controlled indoor setting and in an outdoor release. In an ambient breeze tunnel test, spores deposited on tiles were reaerosolized and detected by real-time PCR at distances of 30 m from the point of deposition. Real-time PCR signals were inversely correlated with distance from the seeded tiles. An outdoor release of powdered spore simulant at Aberdeen Proving Ground, Edgewood, MD, was monitored from a distance by a light detection and ranging (LIDAR) laser. Over a 2-week period, an array of air sampling units collected samples were analyzed for the presence of viable spores and using barcode-specific real-time PCR assays. Barcoded B. thuringiensis subsp. kurstaki spores were unambiguously identified on the day of the release, and viable material was recovered in a pattern consistent with the cloud track predicted by prevailing winds and by data tracks provided by the LIDAR system. Finally, the real-time PCR assays successfully differentiated barcoded B. thuringiensis subsp. kurstaki spores from wild-type spores under field conditions.