Ultrasound-Responsive Nrf2-Targeting siRNA-Loaded Nanobubbles for Enhancing the Treatment of Melanoma

The siRNA-mediated inhibition of nuclear factor E2-related factor 2 (Nrf2) can be an attractive approach to overcome chemoresistance in various malignant tumors, including melanoma. This work aims at designing a new type of chitosan-shelled nanobubble for the delivery of siRNA against Nrf2 in combination with an ultrasound. A new preparation method based on a water–oil–water (W/O/W) double-emulsion was purposely developed for siRNA encapsulation in aqueous droplets within a nanobubble core. Stable, very small NB formulations were obtained, with sizes of about 100 nm and a positive surface charge. siRNA was efficiently loaded in NBs, reaching an encapsulation efficiency of about 90%. siNrf2-NBs downregulated the target gene in M14 cells, sensitizing the resistant melanoma cells to the cisplatin treatment. The combination with US favored NB cell uptake and transfection efficiency. Based on the results, nanobubbles have shown to be a promising US responsive tool for siRNA delivery, able to overcome chemoresistance in melanoma cancer cells

RoboCrane: a system for providing a power and a communication link between lunar surface and lunar caves for exploring robots

Lava caves are the result of a geological process related to the cooling of basaltic lava flows. On the Moon, this process may lead to caves several kilometers long and diameters of hundreds of meters. Access to lava tubes can be granted through skylights, a vertical pit between the lava tube and the lunar surface. This represents an outstanding opportunity for long-term missions, for future permanent human settlements, and for accessing pristine samples of lava, secondary minerals and volatiles. Given this, the ESA launched a campaign through the Open Space Innovation Platform calling for ideas that would tackle the many challenges of exploring lava pits. Five projects, including Robocrane, were selected. Solar light and direct line of sight (for communications) with the lunar surface are not available inside lava tubes. This is a problem for any robot (or swarm of robots) exploring the lava tubes. Robocrane tackles both problems by deploying an element (called the Charging head, or CH) at the bottom of the skylight by means of a crane. This CH behaves as a battery charger and a communication relay for the exploring robots. The required energy is extracted from the crane’s solar panel (on the surface) and driven to the bottom of the skylight through an electrical wire running in parallel to the crane hoisting wire. Using a crane allows the system to deal with unstable terrain around the skylight rim and protect the wires from abrasion from the rocky surface and the pit rim. The charger in the CH is wireless so that the charging process can begin as soon as any of the robots get close enough to the CH. This avoids complex and time-consuming docking operations, aggravated by the skylight floor orography. The crane infrastructure can also be used to deploy the exploring robots inside the pit, reducing their design constraints and mass budget, as the robots do not need to implement their own self-deployment system. Finally, RoboCrane includes all the sensors and actuators for remote operation from a ground station. RoboCrane has been designed in a parametric tool so it can be dynamically and rapidly adjusted to input-variable changes, such as the number of exploring robots, their electrical characteristics, and crane reach, etc.Agencia Estatal de Investigación | Ref. RTI2018-099682-A-I0

Evaluation of the effectiveness of some drugs for the treatment of canine hepatozoonosis

La hepatozoonosis canina es una enfermedad parasitaria, adquirida por la ingestión de garrapatas infectadas con protozoarios del género Hepatozoon (H). Hepatozoon canis y Hepatozoon americanum son las dos especies que pueden infectar al perro. La presentación clínica de la infección con Hepatozoon canis es muy variable, pudiendo ser asintomática o manifestarse con signos de enfermedad leves a severos. Los exámenes hematológicos revelan ligera anemia no regenerativa, marcada leucocitosis neutrofílica con desvío a la izquierda y monocitosis en los casos de hepatozoonosis clínica. El diagnóstico se realiza mediante la visualización mediante el microscopio óptico de los gamontes en neutrófilos y monocitos en frotis de sangre coloreados. No existe, hasta el momento, un tratamiento eficaz para esta parasitosis. El fármaco más utilizado es el dipropionato de imidocarb, aunque con resultados variables. El objetivo de este trabajo fue evaluar tres opciones terapéuticas para la erradicación de Hepatozoon spp. de la sangre de perros infectados. Se utilizaron 18 perros parasitados naturalmente y se evaluaron tres fármacos: dipropionato de imidocarb, toltrazuril y espiramicina. En todos los perros tratados disminuyó el promedio de infección, presentando similares valores medios de leucocitos infectados. El análisis estadístico arrojó una diferencia significativa solo en el tratamiento de la parasitosis con dipropionato de imidocarb. En virtud de los resultados obtenidos con los tratamientos llevados a cabo con los diferentes fármacos, se concluye que ninguno es totalmente efectivo para la desaparición del parásito de la sangre, siendo el dipropionato de imidocarb el de mejor comportamiento.Canine hepatozoonosis is a parasitic disease, acquired by ingestion of ticks infected with protozoa of the genus Hepatozoon (H). Hepatozoon canis and Hepatozoon americanum are the two species that can infect dogs. The clinical presentation of Hepatozoon canis infection is highly variable and may be asymptomatic or show signs of mild to severe disease. In clinical hepatozoonosis, haematological tests reveal slight non regenerative anemia, marked neutrophilic leukocytosis with left shift and monocytosis. Diagnosis is made by the visualization of gamonts in neutrophils and monocytes in stained blood smears using an optical microscope. To date, there is no effective treatment for hepatozoonsis; imidocarb propionate is the most commonly used drug, although results are variable. The aim of this study was to evaluate three therapeutic options for the eradication of Hepatozoon spp. from blood of infected dogs. Three drugs (imidocarb dipropionate, toltrazuril and spiramycin) were evaluated in 18 naturally infected dogs. In all treated dogs the infection averages decreased, showing similar mean values of infected leukocytes. The statistical analysis showed a significant difference only in the treatment of this parasitosis with imidocarb dipropionate. It is concluded that none of the drugs are fully effective in removing the parasite from the blood, but imidocarb dipropionate showed the best results.Facultad de Ciencias Veterinaria

Discovery of novel herpes simplexviruses in wild gorillas, bonobos, and chimpanzees supports zoonotic origin of HSV-2

Viruses closely related to human pathogens can reveal the origins of human infectious diseases. Human herpes simplexvirus type 1 (HSV-1) and type 2 (HSV-2) are hypothesized to have arisen via host-virus codivergence and cross-species transmission. We report the discovery of novel herpes simplexviruses during a large-scale screening of fecal samples from wild gorillas, bonobos, and chimpanzees. Phylogenetic analysis indicates that, contrary to expectation, simplexviruses from these African apes are all more closely related to HSV-2 than to HSV-1. Molecular clock-based hypothesis testing suggests the divergence between HSV-1 and the African great ape simplexviruses likely represents a codivergence event between humans and gorillas. The simplexviruses infecting African great apes subsequently experienced multiple cross-species transmission events over the past 3 My, the most recent of which occurred between humans and bonobos around 1 Ma. These findings revise our understanding of the origins of human herpes simplexviruses and suggest that HSV-2 is one of the earliest zoonotic pathogens

A novel patient-derived tumorgraft model with TRAF1-ALK anaplastic large-cell lymphoma translocation.

Although anaplastic large-cell lymphomas (ALCL) carrying anaplastic lymphoma kinase (ALK) have a relatively good prognosis, aggressive forms exist. We have identified a novel translocation, causing the fusion of the TRAF1 and ALK genes, in one patient who presented with a leukemic ALK+ ALCL (ALCL-11). To uncover the mechanisms leading to high-grade ALCL, we developed a human patient-derived tumorgraft (hPDT) line. Molecular characterization of primary and PDT cells demonstrated the activation of ALK and nuclear factor kB (NFkB) pathways. Genomic studies of ALCL-11 showed the TP53 loss and the in vivo subclonal expansion of lymphoma cells, lacking PRDM1/Blimp1 and carrying c-MYC gene amplification. The treatment with proteasome inhibitors of TRAF1-ALK cells led to the downregulation of p50/p52 and lymphoma growth inhibition. Moreover, a NFkB gene set classifier stratified ALCL in distinct subsets with different clinical outcome. Although a selective ALK inhibitor (CEP28122) resulted in a significant clinical response of hPDT mice, nevertheless the disease could not be eradicated. These data indicate that the activation of NFkB signaling contributes to the neoplastic phenotype of TRAF1-ALK ALCL. ALCL hPDTs are invaluable tools to validate the role of druggable molecules, predict therapeutic responses and implement patient specific therapies

Regulation of Glucose Homeostasis by KSR1 and MARK2

Protein scaffolds control the intensity and duration of signaling and dictate the specificity of signaling through MAP kinase pathways. KSR1 is a molecular scaffold of the Raf/MEK/ERK MAP kinase cascade that regulates the intensity and duration of ERK activation. Relative to wild-type mice, ksr1-/- mice are modestly glucose intolerant, but show a normal response to exogenous insulin. However, ksr1-/- mice also demonstrate a three-fold increase in serum insulin levels in response to a glucose challenge, suggesting a role for KSR1 in insulin secretion. The kinase MARK2 is closely related to C-TAK1, a known regulator of KSR1. Mice lacking MARK2 have an increased rate of glucose disposal in response to exogenous insulin, increased glucose tolerance, and are resistant to diet-induced obesity. mark2-/-ksr1-/- (DKO) mice were compared to wild type, mark2-/-, and ksr1-/- mice for their ability to regulate glucose homeostasis. Here we show that disruption of KSR1 in mark2-/- mice reverses the increased sensitivity to exogenous insulin resulting from MARK2 deletion. DKO mice respond to exogenous insulin similarly to wild type and ksr1-/- mice. These data suggest a model whereby MARK2 negatively regulates insulin sensitivity in peripheral tissue through inhibition of KSR1. Consistent with this model, we found that MARK2 binds and phosphorylates KSR1 on Ser392. Phosphorylation of Ser392 is a critical regulator of KSR1 stability, subcellular location, and ERK activation. These data reveal an unexpected role for the molecular scaffold KSR1 in insulin-regulated glucose metabolism

A Missense Variant in PTPN22 is a Risk Factor for Drug-induced Liver Injury

Background & Aims We performed genetic analyses of a multiethnic cohort of patients with idiosyncratic drug-induced liver injury (DILI) to identify variants associated with susceptibility. Methods We performed a genome-wide association study of 2048 individuals with DILI (cases) and 12,429 individuals without (controls). Our analysis included subjects of European (1806 cases and 10,397 controls), African American (133 cases and 1,314 controls), and Hispanic (109 cases and 718 controls) ancestry. We analyzed DNA from 113 Icelandic cases and 239,304 controls to validate our findings. Results We associated idiosyncratic DILI with rs2476601, a nonsynonymous polymorphism that encodes a substitution of tryptophan with arginine in the protein tyrosine phosphatase, nonreceptor type 22 gene (PTPN22) (odds ratio [OR] 1.44; 95% confidence interval [CI] 1.28–1.62; P = 1.2 × 10–9 and replicated the finding in the validation set (OR 1.48; 95% CI 1.09–1.99; P = .01). The minor allele frequency showed the same effect size (OR > 1) among ethnic groups. The strongest association was with amoxicillin and clavulanate-associated DILI in persons of European ancestry (OR 1.62; 95% CI 1.32–1.98; P = 4.0 × 10–6; allele frequency = 13.3%), but the polymorphism was associated with DILI of other causes (OR 1.37; 95% CI 1.21–1.56; P = 1.5 × 10–6; allele frequency = 11.5%). Among amoxicillin- and clavulanate-associated cases of European ancestry, rs2476601 doubled the risk for DILI among those with the HLA risk alleles A*02:01 and DRB1*15:01. Conclusions In a genome-wide association study, we identified rs2476601 in PTPN22 as a non-HLA variant that associates with risk of liver injury caused by multiple drugs and validated our finding in a separate cohort. This variant has been associated with increased risk of autoimmune diseases, providing support for the concept that alterations in immune regulation contribute to idiosyncratic DILI