Anti-hyperalgesic and anti-inflammatory effects of 4R-tobacco cembranoid in a mouse model of inflammatory pain.

4R is a tobacco cembranoid that binds to and modulates cholinergic receptors and exhibits neuroprotective and anti-inflammatory activity. Given the established function of the cholinergic system in pain and inflammation, we propose that 4R is also analgesic. Here, we tested the hypothesis that systemic 4R treatment decreases pain-related behaviors and peripheral inflammation via modulation of the alpha 7 nicotinic acetylcholine receptors (α7 nAChRs) in a mouse model of inflammatory pain. We elicited inflammation by injecting Complete Freund's Adjuvant (CFA) into the hind paw of male and female mice. We then assessed inflammation-induced hypersensitivity to cold, heat, and tactile stimulation using the Acetone, Hargreaves, and von Frey tests, respectively, before and at different time points (2.5 h - 8d) after a single systemic 4R (or vehicle) administration. We evaluated the contribution of α7 nAChRs 4R-mediated analgesia by pre-treating mice with a selective antagonist of α7 nAChRs followed by 4R (or vehicle) administration prior to behavioral tests. We assessed CFA-induced paw edema and inflammation by measuring paw thickness and quantifying immune cell infiltration in the injected hind paw using hematoxylin and eosin staining. Lastly, we performed immunohistochemical and flow cytometric analyses of paw skin in α7 nAChR-cre::Ai9 mice to measure the expression of α7 nAChRs on immune subsets. Our experiments show that systemic administration of 4R decreases inflammation-induced peripheral hypersensitivity in male and female mice and inflammation-induced paw edema in male but not female mice. Notably, 4R-mediated analgesia and anti-inflammatory effects lasted up to 8d after a single systemic administration on day 1. Pretreatment with an α7 nAChR-selective antagonist prevented 4R-mediated analgesia and anti-inflammatory effects, demonstrating that 4R effects are via modulation of α7 nAChRs. We further show that a subset of immune cells in the hind paw expresses α7 nAChRs. However, the number of α7 nAChR-expressing immune cells is unaltered by CFA or 4R treatment, suggesting that 4R effects are independent of α7 nAChR-expressing immune cells. Together, our findings identify a novel function of the 4R tobacco cembranoid as an analgesic agent in both male and female mice that reduces peripheral inflammation in a sex-dependent manner, further supporting the pharmacological targeting of the cholinergic system for pain treatment.

Sex differences in pain-related behaviors and clinical progression of disease in mouse models of colonic pain.

ABSTRACT: Previous studies have reported sex differences in patients with irritable bowel syndrome and inflammatory bowel disease, including differences in visceral pain perception. Despite this, sex differences in behavioral manifestations of visceral pain and underlying pathology of the gastrointestinal tract have been largely understudied in preclinical research. In this study, we evaluated potential sex differences in spontaneous nociceptive responses, referred abdominal hypersensitivity, disease progression, and bowel pathology in mouse models of acute and persistent colon inflammation. Our experiments show that females exhibit more nociceptive responses and referred abdominal hypersensitivity than males in the context of acute but not persistent colon inflammation. We further demonstrate that, after acute and persistent colon inflammation, pain-related behavioral responses in females and males are distinct, with increases in licking of the abdomen only observed in females and increases in abdominal contractions only seen in males. During persistent colon inflammation, males exhibit worse disease progression than females, which is manifested as worse physical appearance and higher weight loss. However, no measurable sex differences were observed in persistent inflammation-induced bowel pathology, stool consistency, or fecal blood. Overall, our findings demonstrate sex differences in pain-related behaviors and disease progression in the context of acute and persistent colon inflammation, highlighting the importance of considering sex as a biological variable in future mechanistic studies of visceral pain as well as in the development of diagnostics and therapeutic options for chronic gastrointestinal diseases.

Diseño de un dispositivo de asistencia ventilatoria temporal de lazo cerrado basado en bolsa válvula-mascarilla / Design of a closed-loop temporary ventilation assistance device based on bag valve mask

Desde inicios del 2020, el mundo se ha visto afectado por la COVID-19 causada por el SARS-CoV-2, que en agosto lo padecen más de 31 millones de pacientes, algunos de los cuales presentan el síndrome de distrés respiratorio, que requiere de ventilación mecánica. Por el alto número de contagios, la disponibilidad de ventiladores para el tratamiento es escasa. Se presenta la descripción de un prototipo de un dispositivo de asistencia ventilatoria temporal de lazo cerrado de bajo costo; el AR_CODEX, basado en una bolsa válvula-mascarilla (BVM), que contribuye al mantenimiento ventilatorio mínimo del paciente durante un tiempo corto en casos donde no hay disponibilidad de ventiladores mecánicos. Para esto, se diseñó y construyó un sistema mecánico ajustable que compresiona la bolsa de ventilación, el cual cuenta con sensores de flujo y presión. Además, se elaboró una interfaz gráfica para un adecuado monitoreo del paciente y un sistema de control para variables como volumen, presión máxima, frecuencia respiratoria y relación inspiración: espiración. Por otro lado, existe un problema de sensibilidad en el sensor de flujo debido a varios factores, como la variación del voltaje en los motores. Adicionalmente, la implementación de un lazo cerrado es importante para compensar variaciones aleatorias en el funcionamiento del dispositivo. Es necesario realizar pruebas en animales para evaluar el correcto funcionamiento de AR_CODEX en seres vivos.

In early 2020, the world has been affected from Covid-19 caused by SARS-CoV-2. By August there were more than 31 million patients, some of them suffering from respiratory distress that requires mechanical ventilation. Due to the rise of infection rates there is no ventilator availability for the treatment. In this work we describe a reduced cost closed loop temporal assisted ventilation device prototype, AR_CODEX. It is based on mask valve bag (BVM from its Spanish initials), contributing to the minimum ventilation maintenance for the patient du-ring a short period of time when there is no mechanical ventilation availability. For this purpose an adjustable mechanical system was designed and built to pressurize the ventilation bag that is equipped with flux and pres-sure sensors. Additionally a graphical interface was developed to include adequate monitoring and controlling system for volume, maximum pressure, respiratory frequency and inhalation/exhalation rate. In addition there is a sensibility issue on the flux sensor due to engine voltage variation. A closed loop implementation is important to overcome aleatory variations during the device operation. It is needed to run AR_CODEX device performance test on animals to evaluate prior to use it directly on human patients.

A UAV-Based Framework for Semi-Automated Thermographic Inspection of Belt Conveyors in the Mining Industry.

Frequent and accurate inspections of industrial components and equipment are essential because failures can cause unscheduled downtimes, massive material, and financial losses or even endanger workers. In the mining industry, belt idlers or rollers are examples of such critical components. Although there are many precise laboratory techniques to assess the condition of a roller, companies still have trouble implementing a reliable and scalable procedure to inspect their field assets. This article enumerates and discusses the existing roller inspection techniques and presents a novel approach based on an Unmanned Aerial Vehicle (UAV) integrated with a thermal imaging camera. Our preliminary results indicate that using a signal processing technique, we are able to identify roller failures automatically. We also proposed and implemented a back-end platform that enables field and cloud connectivity with enterprise systems. Finally, we have also cataloged the anomalies detected during the extensive field tests in order to build a structured dataset that will allow for future experimentation.

Autonomous Aeromagnetic Surveys Using a Fluxgate Magnetometer.

Recent advances in the research of autonomous vehicles have showed a vast range of applications, such as exploration, surveillance and environmental monitoring. Considering the mining industry, it is possible to use such vehicles in the prospection of minerals of commercial interest beneath the ground. However, tasks such as geophysical surveys are highly dependent on specific sensors, which mostly are not designed to be used in these new range of autonomous vehicles. In this work, we propose a novel magnetic survey pipeline that aims to increase versatility, speed and robustness by using autonomous rotary-wing Unmanned Aerial Vehicles (UAVs). We also discuss the development of a state-of-the-art three-axis fluxgate, where our goal in this work was to refine and adjust the sensor topology and coupled electronics specifically for this type of vehicle and application. The sensor was built with two ring-cores using a specially developed stress-annealed CoFeSiB amorphous ribbon, in order to get sufficient resolution to detect concentrations of small ferrous minerals. Finally, we report on the results of experiments performed with a real UAV in an outdoor environment, showing the efficacy of the methodology in detecting an artificial ferrous anomaly.

Four monoclonal antibodies against capsular polysaccharides of Neisseria meningitidis serogroups A, C, Y and W135: its application in identity tests.

Murine hybridoma monoclonal antibodies (MAbs) were produced against the capsular polysaccharide (CPs) of serogroups A, C, W135 and Y meningococci (MenA, MenC, MenW, MenY) in order to develop immunological reagents for the identification of meningococcal polysaccharides. Each serogroup-specific MAb reacted with the CPs from its homologous serogroup only and did not react with CPs from the other three serogroups. The affinity constant (Ka) of the four MAbs measured by non-competitive ELISA was 6.62 × 10(9), 2.76 × 10(9), 1.48 × 10(9) and 3.8 × 10(9) M(-1) for MenA, MenC, MenW and MenY MAbs respectively. The application of these MAbs for identity tests was demonstrated by their abilities to correctly identify the CPs from serogroups A, C, W135 and Y in meningococcal CPs-based vaccines through ELISA. The MAbs obtained in this work are a very valuable set of tools for study meningococcal polysaccharides vaccines.

A proteoliposome formulation derived from Bordetella pertussis induces protection in two murine challenge models.

Whooping cough remains a health problem despite high vaccination coverage. It has been recommended that development of new strategies provide long-lasting immunity. The aim of this work was to evaluate the potential of proteoliposomes (PL) extracted from Bordetella pertussis as a vaccine candidate against whooping cough. The size of the B. pertussis PL was estimated to be 96.7 ± 50.9 nm by Scanning Correlation Spectroscopy and the polydispersity index was 0.268. Western blots using monoclonal antibodies revealed the presence of pertussis toxin, pertactin, and fimbriae 3. The Limulus Amebocyte Lisate (LAL) assay showed endotoxin levels lower than those reported for whole cell pertussis licensed vaccines, while the Pyrogen Test indicated 75 ng/mL/Kg. The PL showed high protection capacity in mouse challenge models. There was 89.7% survival in the intracerebral challenge and total reduction of the number of CFU in the intranasal challenge. No significant differences (p > 0.05) were observed between mice immunized with B. pertussis PL and the Cuban DTwP vaccine, whichever challenge model used. These results encouraged us to continue the development of the B. pertussis PL as a component of a new combined vaccine formulated with tetanus and diphtheria toxoids or as a booster dose for adolescents and adults.

Evaluation of enteric-coated tablets as a whole cell inactivated vaccine candidate against Vibrio cholerae.

A vaccine candidate against cholera was developed in the form of oral tablets to avoid difficulties during application exhibited by current whole cell inactivated cholera vaccines. In this study, enteric-coated tablets were used to improve the protection of the active compound from gastric acidity. Tablets containing heat-killed whole cells of Vibrio cholerae strain C7258 as the active pharmaceutical compound was enteric-coated with the polymer Kollicoat(®) MAE-100P, which protected them efficiently from acidity when a disintegration test was carried out. Enzyme-linked immunosorbent assay (ELISA) anti-lipopolysaccharide (LPS) inhibition test and Western blot assay revealed the presence of V. cholerae antigens as LPS, mannose-sensitive haemagglutinin (MSHA) and outer membrane protein U (Omp U) in enteric-coated tablets. Immunogenicity studies (ELISA and vibriocidal test) carried out by intraduodenal administration in rabbits showed that the coating process of tablets did not affect the immunogenicity of V. cholerae-inactivated cells. In addition, no differences were observed in the immune response elicited by enteric-coated or uncoated tablets, particularly because the animal model and immunization route used did not allow discriminating between acid resistances of both tablets formulations in vivo. Clinical studies with volunteers will be required to elucidate this aspect, but the results suggest the possibility of using enteric-coated tablets as a final pharmaceutical product for a cholera vaccine.