Innovative Detection and Segmentation of Mobility Activities in Patients Living with Parkinson's Disease Using a Single Ankle-Positioned Smartwatch.

BACKGROUND: The automatic detection of activities of daily living (ADL) is necessary to improve long-term home-based monitoring of Parkinson's disease (PD) symptoms. While most body-worn sensor algorithms for ADL detection were developed using laboratory research systems covering full-body kinematics, it is now crucial to achieve ADL detection using a single body-worn sensor that remains commercially available and affordable for ecological use. AIM: to detect and segment Walking, Turning, Sitting-down, and Standing-up activities of patients with PD using a Smartwatch positioned at the ankle. METHOD: Twenty-two patients living with PD performed a Timed Up and Go (TUG) task three times before engaging in cleaning ADL in a simulated free-living environment during a 3 min trial. Accelerations and angular velocities of the right or left ankle were recorded in three dimensions using a Smartwatch. The TUG task was used to develop detection algorithms for Walking, Turning, Sitting-down, and Standing-up, while the 3 min trial in the free-living environment was used to test and validate these algorithms. Sensitivity, specificity, and F-scores were calculated based on a manual segmentation of ADL. RESULTS: Sensitivity, specificity, and F-scores were 96.5%, 94.7%, and 96.0% for Walking; 90.0%, 93.6%, and 91.7% for Turning; 57.5%, 70.5%, and 52.3% for Sitting-down; and 57.5%, 72.9%, and 54.1% for Standing-up. The median of time difference between the manual and automatic segmentation was 1.31 s for Walking, 0.71 s for Turning, 2.75 s for Sitting-down, and 2.35 s for Standing-up. CONCLUSION: The results of this study demonstrate that segmenting ADL to characterize the mobility of people with PD based on a single Smartwatch can be comparable to manual segmentation while requiring significantly less time. While Walking and Turning were well detected, Sitting-down and Standing-up will require further investigation to develop better algorithms. Nonetheless, these achievements increase the odds of success in implementing wearable technologies for PD monitoring in ecological environments.

Using cephalopod-inspired chemistry to extend long-wavelength ultraviolet and visible light protection of mineral sunscreens.

OBJECTIVE: The emergence of new human and environmental-related toxicity data associated with some common UV filters has catalysed growing interest in the inclusion of boosters and stabilizing ingredients in sunscreens. One approach is to incorporate alternative materials inspired by or mimetic of systems in biology, which offer a notable evolutionary advantage of multifunctionality and stability with increased biocompatibility. We describe the use of a natural product, Xanthochrome® (INCI: Ammonium Xanthommatin), in a series of studies designed to not only assess its safety with marine systems but also its formulation compatibility and function in water-in-oil mineral sunscreens. Xanthochrome is the synthetic form of the naturally occurring chromophore xanthommatin (XA) present in cephalopod skin, which doubles as a photostable antioxidant; however, it has never been explored in combination with mineral UV filters in finished formulations. METHODS: Given the recent controversies associated with the environmental toxicological effects of some chemicals used in sunscreens, the safety of XA with coral cuttings was first validated at concentrations 5× above those used in our formulations. Next, a particle-based delivery of XA was designed and incorporated into a zinc oxide (ZnO)-based water-in-oil sunscreen, where the SPF, critical wavelength, and visible light (VL) blocking potential were measured. RESULTS: We observed no adverse effects of XA at 100 mg/L when tested with coral cuttings, demonstrating its safety at concentrations exceeding those used in our sunscreens. When formulated with ZnO-based sunscreens, the inclusion of XA increased the total UV absorbance profile by 28% and the total blocking potential of VL by 45%. The formulations also elicited no dermal irritation or sensitization in a human insult repeat patch test (N = 100 subjects). CONCLUSIONS: XA is differentiated as a photostable, water-soluble compound that is a VL booster proven safe for skin and coral cuttings. To the best of our knowledge, there are no other boosters that can be classified as such, despite a growing body of literature highlighting the need in the industry.

OBJECTIF: L'émergence de nouvelles données de toxicité liées à l'environnement et à l'homme associées à certains filtres UV courants a suscité un intérêt croissant pour l'inclusion d'agents rehausseurs et d'ingrédients stabilisants dans les écrans solaires. Une approche consiste à intégrer des matériaux alternatifs inspirés ou mimétiques des systèmes en biologie, ce qui offre une biocompatibilité accrue et un avantage évolutif notable en matière de multifonctionnalité et de stabilité. Nous décrivons l'utilization d'un produit naturel, le Xanthochrome® (INCI: ammonium de xanthommatine), dans une série d'études conçues non seulement pour évaluer sa sécurité d'emploi avec les systèmes marins, mais également sa compatibilité et sa fonction de formulation dans les écrans solaires minéraux à base d'eau en huile. Le xanthochrome est la forme synthétique de la xanthommatine chromophore (XA) naturellement présente dans la peau des céphalopodes, qui fait office d'antioxydant photostable; cependant, il n'a jamais été étudié en association avec des filtres minéraux UV dans des formulations finies. MÉTHODES: Compte tenu des controverses récentes associées aux effets toxicologiques environnementaux de certains produits chimiques utilisés dans les écrans solaires, la sécurité d'emploi de l'XA sur les coupes de corail a été d'abord validée à des concentrations 5 fois supérieures à celles utilisées dans nos formulations. Ensuite, une application d'XA à base de particules a été conçue et incorporée dans un écran solaire à base d'oxyde de zinc (ZnO), dans lequel le facteur SPF, la longueur d'onde critique et le potentiel bloquant de la lumière visible (LV) ont été mesurés. RÉSULTATS: Nous n'avons pas observé d'effets indésirables de l'XA à raison de 100 mg/L lors de tests sur des coupes de corail, ce qui démontre sa sécurité d'emploi à des concentrations supérieures à celles utilisées dans nos écrans solaires. Lorsqu'il est formulé avec des écrans solaires à base de ZnO, l'inclusion de XA a augmenté le profil d'absorbance totale des UV de 28% et le potentiel bloquant total de la LV de 45%. Les formulations n'ont également entraîné aucune irritation ou sensibilisation cutanée lors d'un test épicutané d'exposition répétée chez l'homme (N = 100 sujets). CONCLUSIONS: Le XA se différencie par un composé soluble dans l'eau et photostable qui est un amplificateur de la LV éprouvé sans danger pour les coupes de peau et de corail. À notre connaissance, il n'existe aucun autre agent rehausseur pouvant être classé comme tel, malgré un corpus croissant de littérature soulignant le besoin dans l'industrie.

Instrumented Pre-Hospital Care Simulation Mannequin for Use in Spinal Motion Restrictions Scenarios: Validation of Cervical and Lumbar Motion Assessment.

BACKGROUND: A mid-fidelity simulation mannequin, equipped with an instrumented cervical and lumbar spine, was developed to investigate best practices and train healthcare professionals in applying spinal motion restrictions (SMRs) during the early mobilization and transfer of accident victims with suspected spine injury. The study objectives are to (1) examine accuracy of the cervical and lumbar motions measured with the mannequin; and (2) confirm that the speed of motion has no bearing on this accuracy. METHODS: Accuracy was evaluated by concurrently comparing the orientation data obtained with the mannequin with that from an optoelectronic system. The mannequin's head and pelvis were moved in all anatomical planes of motion at different speeds. RESULTS: Accuracy, assessed by root-mean-square error, varied between 0.7° and 1.5° in all anatomical planes of motion. Bland-Altman analysis revealed a bias ranging from -0.7° to 0.6°, with the absolute limit of agreement remaining below 3.5°. The minimal detectable change varied between 1.3° and 2.6°. Motion speed demonstrated no impact on accuracy. CONCLUSIONS: The results of this validation study confirm the mannequin's potential to provide accurate measurements of cervical and lumbar motion during simulation scenarios for training and research on the application of SMR.

Removal of the cervical collar from alpine rescue protocols? A biomechanical non-inferiority trial in real-life mountain conditions.

BACKGROUND: Alpine skiing rescues are challenging because of the mountainous environment and risks of cervical spine motion (CSM) induced during victims' extrications (EXs) and downhill evacuations (DEs). The benefits of applying a cervical collar (CC) over manual in-line stabilization without CC (MILS) in terms of spinal motion restriction during simulated alpine rescues are undocumented. Our hypothesis was that CSM recorded using MILS alone is non-inferior to CSM recorded with a CC according to a 10 degrees margin. METHODS: A total of 32 alpine extrications and 4 downhill evacuations on different slope conditions were performed using a high fidelity mannequin designed with a motion sensors instrumented cervical spine. The primary outcome was the peak extrication 3D excursion angle (Peak 3D θEX,) of the mannequin's head. The secondary objectives were to describe the time to extrication completion (tEX) and to highlight which extrication manipulation is more likely to induce CSM. RESULTS: The median Peak 3D θEX recorded during flat terrain extrications using CC was 10.77° (95% CI 7.31°-16.45°) compared to 13.06° (95% CI 10.20°-30.36°) using MILS, and 16.09° (95% CI 9.07°-37.43°) for CC versus 16.65° (95% CI 13.80°-23.40°) using MILS on a steep slope. Peak 3D θEX with CC or using MILS during extrications were equivalent according to a 10 degrees non-inferiority hypothesis testing (p < 0.05). Time to extrication completion (tEX) was significantly reduced using MILS without CC on a flat terrain with a median duration of 237,3 s (95% CI 197.8 s, 272.2 s) compared to 358.7 s (95% CI 324.1 s, 472.4 s). During downhill evacuations, CSM with and without CC across all terrain conditions were negligible (< 5°). When CC is used; its installation manipulation induces the highest CSM. When EXs are done using MILS without CC, the logroll initiation is the manipulation inducing the highest risk of CSM. CONCLUSION: For experienced ski patrollers, the biomechanical benefits of spinal motion restriction provided by CC over MILS during alpine skiing rescues appear to be marginal and CC use negatively affects rescue time.

Evaluation of Biologically Inspired Ammonium Xanthommatin as a Multifunctional Cosmetic Ingredient.

We describe the investigation of an organic natural product, ammonium xanthommatin (Xanthochrome), in a series of studies designed to not only assess its impact on endocrine receptor function in vitro but also interrogate its mutagenic potential using bacterial reverse mutation assays. As a multifunctional raw material, ammonium xanthommatin functions as an antioxidant with a broad absorption profile spanning the UV through the visible spectrum, making it an interesting target for cosmetic applications. In solution, ammonium xanthommatin contributes to <30% inhibition of hormonal activities, indicating that it is not an endocrine disruptor. Furthermore, the compound does not cause gene mutations in the bacterial strains used, indicating that it is nonmutagenic. Applications are also described, highlighting xanthommatin's ability to boost the UVA and UVB absorptive properties of traditional chemical UV filters by >50% across all filters tested. In addition to these features, xanthommatin exhibited no phototoxic hazards in vitro when irradiated with UVA and visible light, demonstrating its utility as a multifunctional cosmetic ingredient. Although these findings encourage the use of xanthommatin in cosmetics, they represent only the beginning of the complete in vitro and in vivo data package needed to support safety and efficacy claims for future applications in skin health.

Therapeutic Potential of MF-TTZ-MMAE, a Site-Specifically Conjugated Antibody-Drug Conjugate, for the Treatment of HER2-Overexpressing Breast Cancer.

With three clinically approved antibody-drug conjugates targeting HER2, this target is clearly identified to be of interest in oncology. Moreover, the advent of new bioconjugation technologies producing site-specific homogenous conjugates led to the opportunity of developing new medicines linking antibodies and payloads. Here, a new relevant HER2-targeting ADC was obtained by the conjugation of monomethyl auristatin E onto trastuzumab using McSAF Inside bioconjugation technology. The antibody-drug conjugate formed presented an average drug-to-antibody ratio of 4 with a high homogeneity and an excellent stability especially when incubated with human serum albumin or in human plasma. Moreover, it demonstrated a strong efficacy in an HER2 xenograft tumor model in mice, superior to the clinically approved antibody-drug conjugate ado-trastuzumab emtansine, with a complete tumor regression observed both macroscopically and microscopically demonstrating its therapeutic potential.

Antibody-Drug Conjugates as an Emerging Therapy in Oncodermatology.

Antibody-drug conjugates (ADCs) are an emerging class of therapeutics, with twelve FDA- and EMA-approved drugs for hematological and solid cancers. Such drugs consist in a monoclonal antibody linked to a cytotoxic agent, allowing a specific cytotoxicity to tumor cells. In recent years, tremendous progress has been observed in therapeutic approaches for advanced skin cancer patients. In this regard, targeted therapies (e.g., kinase inhibitors) or immune checkpoint-blocking antibodies outperformed conventional chemotherapy, with proven benefit to survival. Nevertheless, primary and acquired resistances as well as adverse events remain limitations of these therapies. Therefore, ADCs appear as an emerging therapeutic option in oncodermatology. After providing an overview of ADC design and development, the goal of this article is to review the potential ADC indications in the field of oncodermatology.

Innovative Bioconjugation Technology for Antibody-Drug Conjugates: Proof of Concept in a CD30-Positive Lymphoma Mouse Model.

To overcome stability and heterogeneity issues of antibody-drug conjugates (ADCs) produced with existing bioconjugation technologies incorporating a maleimide motif, we developed McSAF Inside, a new technology based on a trifunctionalized di(bromomethyl)pyridine scaffold. Our solution allows the conjugation of a linker-payload to previously reduced interchain cysteines of a native antibody, resulting in disulfide rebridging. This leads to highly stable and homogeneous ADCs with control over the drug-to-antibody ratio (DAR) and the linker-payload position. Using our technology, we synthesized an ADC, MF-BTX-MMAE, built from anti-CD30 antibody cAC10 (brentuximab), and compared it to Adcetris, the first line treatment against CD30-positive lymphoma, in a CD30-positive lymphoma model. MF-BTX-MMAE displayed improved DAR homogeneity, with a solid batch-to-batch reproducibility, as well as enhanced stability in thermal stress conditions or in the presence of a free thiol-containing protein, such as human serum albumin (HSA). MF-BTX-MMAE showed antigen-binding, in vitro cytotoxicity, in vivo efficacy, and tolerability similar to Adcetris. Therefore, in accordance with current regulatory expectations for the development of new ADCs, McSAF Inside technology gives access to relevant ADCs with improved characteristics and stability.

Biomimetic Colorants and Coatings Designed with Cephalopod-Inspired Nanocomposites.

Brilliant and dynamic colors in nature have stimulated the design of dyes and pigments with broad applications ranging from electronic displays to apparel. Inspired by the nanostructured pigment granules present in cephalopod chromatophore organs, we describe the design and fabrication of biohybrid colorants containing the cephalopod-specific pigment, xanthommatin (Xa), encased within silica-based nanostructures. We employed a biomimetic approach to encapsulate Xa with amine-terminated polyamidoamine (PAMAM) dendrimer templates, which helped stabilize the pigment during encapsulation. Depending on the concentration of Xa used in the reaction, the resultant biohybrid nanomaterials generated a range of neutral colors of differing hues. When applied as coatings, these colorants can be triggered to change color from yellow/gold to red in the presence of a chemical reducing agent, as we leverage the natural redox-dependent color change of Xa. Altogether, these capabilities demonstrated the ability to process biochromes like Xa as nanomaterials that can be applied as coatings with a tunable and dynamic range.

Alcohol consumption among French physicians: A cross-sectional study.

BACKGROUND: Data regarding French physicians' alcohol behaviours are scarce and most studies address this issue within the population of either medical students or residents. We aim to describe and assess the prevalence of hazardous alcohol consumption among French physicians. METHODS: A regional, cross-sectional, survey was conducted in 2018 using an online questionnaire among Parisian general practitioners and hospital doctors. Hazardous alcohol consumption was defined by an Alcohol Use Disorders Identification Test (AUDIT) score ≥ 8. Data were analysed in 2020. RESULTS: Five hundred fifteen physicians completed the survey: 108 general practitioners and 407 hospital physicians. The median age was 40 years [32-55] and 59 % were women. They considered their physical and mental health as average or bad in 10 % and 8% of cases, respectively. The prevalence of hazardous alcohol consumption was 12.6 %. Among the 65 physicians with hazardous alcohol consumption, 27 (41.5 %) did not considered it as risky and four (6.2 %) mentioned a potentially negative impact on patients' care. Factors independently associated with hazardous alcohol consumption were illegal drugs consumption (OR 4.62 [2.05-10.37]) and fixed term contract for hospital doctors (OR 2.69 [1.14-6.36]). CONCLUSIONS: The prevalence of hazardous alcohol consumption among French physicians was 12.6 %. Illegal substance users and fix-termed contract hospital doctors were more likely to have risky alcohol consumption. A large-scale national study would confirm the factors associated with hazardous alcohol consumption and could explore the efficacy of preventive measures to insure the safety and health of physicians and their patients.

Food Anticipatory Activity on Circadian Time Scales Is Not Dependent on Central Serotonin: Evidence From Tryptophan Hydroxylase-2 and Serotonin Transporter Knockout Mice.

A number of studies implicate biogenic amines in regulating circadian rhythms. In particular, dopamine and serotonin influence the entrainment of circadian rhythms to daily food availability. To study circadian entrainment to feeding, food availability is typically restricted to a short period within the light cycle daily. This results in a notable increase in pre-meal activity, termed "food anticipatory activity" (FAA), which typically develops within about 1 week of scheduled feeding. Several studies have implicated serotonin as a negative regulator of FAA: (1) aged rats treated with serotonin 5-HT2 and 3 receptor antagonists showed enhanced FAA, (2) mice lacking for the 2C serotonin receptor demonstrate enhanced FAA, and (3) pharmacologically increased serotonin levels suppressed FAA while decreased serotonin levels enhanced FAA in mice. We sought to confirm and extend these findings using genetic models with impairments in central serotonin production or re-uptake, but were surprised to find that both serotonin transporter (Slc6a4) and tryptophan hydroxylase-2 knockout mice demonstrated a normal behavioral response to timed, calorie restricted feeding. Our data suggest that FAA is largely independent of central serotonin and/or serotonin reuptake and that serotonin may not be a robust negative regulator of FAA.

Impact of Maleimide Disubstitution on Chemical and Biological Characteristics of HER2 Antibody-Drug Conjugates.

Antibody-drug conjugates (ADCs) are the spearhead of targeted therapies. According to the technology used, the conjugation of a cytotoxic drug to an antibody can produce suboptimal heterogeneous species, impacting the overall efficacy. Herein, we describe the synthesis of HER2-targeting ADCs with three disulfide rebridging heads, allowing homogeneous and site-specific bioconjugation: dibromomaleimide (DBM), dithiomaleimide (DTM), and hybrid thio-bromomaleimide (TBM) chemical bricks to combine the properties of both previously used heads. The primary purpose of this study was to compare the reactivity of these three chemical bricks in the bioconjugation process. Then, the resulting ADCs were evaluated in terms of physicochemical stability, binding, and biological activity. We have demonstrated that the higher percentage of a drug-to-antibody ratio of 4 was obtained with TBM. Additionally, the reaction time was drastically reduced with TBM in comparison to DTM. The three ADCs showed good binding to HER2 and in vitro cytotoxicity, which validate the TBM structure as an attractive alternative scaffold for rebridging bioconjugation.

In Vitro Characterization and Stability Profiles of Antibody-Fluorophore Conjugates Derived from Interchain Cysteine Cross-Linking or Lysine Bioconjugation.

Fluorescent labelling of monoclonal antibodies (mAbs) is classically performed by chemical bioconjugation methods. The most frequent labelling technique to generate antibody-fluorophore conjugates (AFCs) involves the bioconjugation onto the mAb lysines of a dye bearing an N-hydroxysuccinimide ester or an isothiocyanate group. However, discrepancies between labelling experiments or kits can be observed, related to reproducibility issues, alteration of antigen binding, or mAb properties. The lack of information on labelling kits and the incomplete characterization of the obtained labelled mAbs largely contribute to these issues. In this work, we generated eight AFCs through either lysine or interchain cysteine cross-linking bioconjugation of green-emitting fluorophores (fluorescein or BODIPY) onto either trastuzumab or rituximab. This strategy allowed us to study the influence of fluorophore solubility, bioconjugation technology, and antibody nature on two known labelling procedures. The structures of these AFCs were thoroughly analyzed by mass spectroscopy, and their antigen binding properties were studied. We then compared these AFCs in vitro by studying their respective spectral properties and stabilities. The shelf stability profiles and sensibility to pH variation of these AFCs prove to be dye-, antibody- and labelling-technology-dependent. Fluorescence emission in AFCs was higher when lysine labelling was used, but cross-linked AFCs were revealed to be more stable. This must be taken into account for the design of any biological study involving antibody labelling.

A bioinspired, photostable UV-filter that protects mammalian cells against UV-induced cellular damage.

While commercially available suncare products are effective at absorbing ultraviolet (UV)-light, recent studies indicate systemic toxicities associated with many traditional chemical and physical UV-filters. We demonstrate the application of xanthommatin, a biochrome present in arthropods and cephalopods, as an alternative chemical UV-filter that is cytocompatible while maintaining its photostability and photoprotective properties.

Glycoside Hydrolase Genes Are Required for Virulence of Agrobacterium tumefaciens on Bryophyllum daigremontiana and Tomato.

Agrobacterium tumefaciens is a rhizosphere bacterium that can infect wound sites on plants. The bacterium transfers a segment of DNA (T-DNA) from the Ti plasmid to the plant host cell via a type IV secretion system where the DNA becomes integrated into the host cell chromosomes. The expression of T-DNA in the plant results in tumor formation. Although the binding of the bacteria to plant surfaces has been studied previously, there is little work on possible interactions of the bacteria with the plant cell wall. Seven of the 48 genes encoding putative glycoside hydrolases (Atu2295, Atu2371, Atu3104, Atu3129, Atu4560, Atu4561, and Atu4665) in the genome of A. tumefaciens C58 were found to play a role in virulence on tomato and Bryophyllum daigremontiana Two of these genes (pglA and pglB; Atu3129 and Atu4560) encode enzymes capable of digesting polygalacturonic acid and, thus, may play a role in the digestion of pectin. One gene (arfA; Atu3104) encodes an arabinosylfuranosidase, which could remove arabinose from the ends of polysaccharide chains. Two genes (bglA and bglB; Atu2295 and Atu4561) encode proteins with ß-glycosidase activity and could digest a variety of plant cell wall oligosaccharides and polysaccharides. One gene (xynA; Atu2371) encodes a putative xylanase, which may play a role in the digestion of xylan. Another gene (melA; Atu4665) encodes a protein with α-galactosidase activity and may be involved in the breakdown of arabinogalactans. Limited digestion of the plant cell wall by A. tumefaciens may be involved in tumor formation on tomato and B. daigremontianaIMPORTANCEA. tumefaciens is used in the construction of genetically engineered plants, as it is able to transfer DNA to plant hosts. Knowledge of the mechanisms of DNA transfer and the genes required will aid in the understanding of this process. Manipulation of glycoside hydrolases may increase transformation and widen the host range of the bacterium. A. tumefaciens also causes disease (crown gall tumors) on a variety of plants, including stone fruit trees, grapes, and grafted ornamentals such as roses. It is possible that compounds that inhibit glycoside hydrolases could be used to control crown gall disease caused by A. tumefaciens.

Dynamic pigmentary and structural coloration within cephalopod chromatophore organs.

Chromatophore organs in cephalopod skin are known to produce ultra-fast changes in appearance for camouflage and communication. Light-scattering pigment granules within chromatocytes have been presumed to be the sole source of coloration in these complex organs. We report the discovery of structural coloration emanating in precise register with expanded pigmented chromatocytes. Concurrently, using an annotated squid chromatophore proteome together with microscopy, we identify a likely biochemical component of this reflective coloration as reflectin proteins distributed in sheath cells that envelop each chromatocyte. Additionally, within the chromatocytes, where the pigment resides in nanostructured granules, we find the lens protein Ω- crystallin interfacing tightly with pigment molecules. These findings offer fresh perspectives on the intricate biophotonic interplay between pigmentary and structural coloration elements tightly co-located within the same dynamic flexible organ - a feature that may help inspire the development of new classes of engineered materials that change color and pattern.

Xanthommatin-Based Electrochromic Displays Inspired by Nature.

Color is a signature visual feature in nature; however, the ability to trigger color change in the presence of different environmental stimuli is unique to only a handful of species in the animal kingdom. We exploit the natural color-changing properties of the predominant pigment in arthropods and cephalopods-xanthommatin (Xa)-and describe its utility as a new broad-spectrum electrochromic material. To accomplish this goal, we explored the spectroelectrochemical properties of Xa adsorbed to an indium-doped tin oxide-coated substrate chemically modified with poly(3,4-ethylene dioxythiophene) doped with poly(styrenesulfonate) (PEDOT:PSS). We identified a synergistic role between PEDOT:PSS and Xa that contributed to its absorption profile, which could be modulated across multiple cycles. By varying the ratio of the two electroactive components, we also altered the perceived visible color of Xa-based devices, which cycled from different shades of red to yellow under reducing and oxidizing potentials, respectively. Together, our data illustrate the utility of Xa-based devices as new broad-spectrum electrochromic materials.

Site-Specific Conjugation of Auristatins onto Engineered scFv Using Second Generation Maleimide to Target HER2-positive Breast Cancer in Vitro.

Antibody-drug conjugates (ADC) are spearheading vectorized chemotherapy against cancer, with 4 FDA-approved ADCs and 79 in clinical trials. However, most ADCs are produced using a stochastic bioconjugation method, target hematological cancers, and are derived from a full immunoglobulin-G (IgG). These factors limit their efficacy, especially against solid tumors which remain difficult to treat. Here we report the site-specific conjugation of a single auristatin derivative onto an engineered anti-HER2 single chain fragment variable (scFv) of the trastuzumab antibody, generating new scFv-drug conjugates (SDC). Two cysteines were judiciously incorporated at the beginning of the scFv hexahistidine tag, in order to allow controlled bioconjugation of a heterobifunctional linker including a second generation maleimide (SGM), either cleavable (for monomethyl auristatin E) or noncleavable (for monomethyl auristatin F). Our data indicated that both SDCs conserved their affinity to HER2 in comparison to the native scFv, and were efficiently able to kill in vitro HER2-positive SK-BR-3 cells at subnanomolar concentrations (EC50 of 0.68 nM and 0.32 nM). No effect was observed on HER2-negative MCF-7 cells. Ours results showed efficient targeting of site-specific SDCs against HER2-positive breast cancer cells. This work represents a first important step in the design of more effective small conjugates, paving the way for future in vivo translation to evaluate their full potential.

Impact of Site-Specific Conjugation of ScFv to Multifunctional Nanomedicines Using Second Generation Maleimide.

Biocompatible multifunctional nanomedicines (NMs) are known to be an attractive platform for targeted anticancer theranosis. However, these nanomedicines are of interest only if they efficiently target diseased cells and accumulate in tumors. Here we report the synthesis of a new generation of immunotargeted nanomedicines composed of a superparamagnetic iron oxide nanoparticle (SPION) core, polyethylene glycol coating and the anti-HER2 single chain fragment variable (scFv) of Trastuzumab antibody. We developed two novel bioengineered scFv carrying two cysteines located (i) at the end (4D5.1-cys2) or (ii) at the beginning (4D5.2-cys2) of its hexahistidine tag. The scFv bioconjugation was controlled via heterobifunctional linkers including a second generation maleimide (SGM). Our data indicated that the insertion of cysteines at the beginning of the hexahistidine tag was allowed to obtain nearly 2-fold conjugation efficiency (13 scFv/NP) compared to NMs using classical maleimide. As a result, the NMs-4D5.2 built using the optimal 4D5-cys2 and linkers equipped with SGM showed the enhanced recognition of HER2 in an ELISA format and on the surface of SK-BR-3 breast cancer cells in vitro. Their stability in serum was also significantly improved compared to the NMs-4D5. Our results showed the fundamental importance of the controlled ligand conjugation in the perspective of rational design of NMs with tailored physicochemical and biological properties.

Sex differences in circadian food anticipatory activity are not altered by individual manipulations of sex hormones or sex chromosome copy number in mice.

Recent studies in mice have demonstrated a sexual dimorphism in circadian entrainment to scheduled feeding. On a time restricted diet, males tend to develop food anticipatory activity (FAA) sooner than females and with a higher amplitude of activity. The underlying cause of this sex difference remains unknown. One study suggests that sex hormones, both androgens and estrogens, modulate food anticipatory activity in mice. Here we present results suggesting that the sex difference in FAA is unrelated to gonadal sex hormones. While a sex difference between males and females in FAA on a timed, calorie restricted diet was observed there were no differences between intact and gonadectomized mice in the onset or magnitude of FAA. To test other sources of the sex difference in circadian entrainment to scheduled feeding, we used sex chromosome copy number mutants, but there was no difference in FAA when comparing XX, XY-, XY-;Sry Tg, and XX;Sry Tg mice, demonstrating that gene dosage of sex chromosomes does not mediate the sex difference in FAA. Next, we masculinized female mice by treating them with 17-beta estradiol during the neonatal period; yet again, we saw no difference in FAA between control and masculinized females. Finally, we observed that there was no longer a sex difference in FAA for older mice, suggesting that the sex difference in FAA is age-dependent. Thus, our study demonstrates that singular manipulations of gonadal hormones, sex chromosomes, or developmental patterning are not able to explain the difference in FAA between young male and female mice.