Recent advances in the use of CRISPR/Cas for understanding the early development of molecular gaps in glial cells.

Glial cells are non-neuronal elements of the nervous system (NS) and play a central role in its development, maturation, and homeostasis. Glial cell interest has increased, leading to the discovery of novel study fields. The CRISPR/Cas system has been widely employed for NS understanding. Its use to study glial cells gives crucial information about their mechanisms and role in the central nervous system (CNS) and neurodegenerative disorders. Furthermore, the increasingly accelerated discovery of genes associated with the multiple implications of glial cells could be studied and complemented with the novel screening methods of high-content and single-cell screens at the genome-scale as Perturb-Seq, CRISP-seq, and CROPseq. Besides, the emerging methods, GESTALT, and LINNAEUS, employed to generate large-scale cell lineage maps have yielded invaluable information about processes involved in neurogenesis. These advances offer new therapeutic approaches to finding critical unanswered questions about glial cells and their fundamental role in the nervous system. Furthermore, they help to better understanding the significance of glial cells and their role in developmental biology.

Immunoprotection evaluation of the recombinant N-terminal domain of Cys-loop receptors against Rhipicephalus (Boophilus) microplus tick infestation.

Rhipicephalus (Boophilus) microplus ticks are obligatory hematophagous ectoparasites of cattle and act as vectors for disease-causing microorganisms. Conventional tick control is based on the application of chemical acaricides; however, their uncontrolled use has increased resistant tick populations, as well as food and environmental contamination. Alternative immunological tick control has shown to be partially effective. Therefore, there is a need to characterize novel antigens in order to improve immunological protection. The aim of this work was to evaluate Cys-loop receptors as vaccine candidates. N-terminal domains of a glutamate receptor and of a glycine-like receptor were recombinantly produced in Escherichia coli. Groups of BALB/c mice were independently immunized with four doses of each recombinant protein emulsified with Freund's adjuvant. Both vaccine candidates were immunogenic in mice as demonstrated by western blot analysis. Next, recombinant proteins were independently formulated with the adjuvant Montanide ISA 50 V2 and evaluated in cattle infested with Rhipicephalus microplus tick larvae. Groups of three European crossbred calves were immunized with three doses of each adjuvanted protein. ELISA test was used to evaluate the IgG immune response elicited against the recombinant proteins. Results showed that vaccine candidates generated a moderate humoral response on vaccinated cattle. Vaccination significantly affected the number of engorged adult female ticks, having no significant effects on tick weight, egg weight and egg fertility values. Vaccine efficacies of 33% and 25% were calculated for the glutamate receptor and the glycine-like receptor, respectively.

TITLE: Évaluation de l'immunoprotection du domaine N-terminal recombinant des récepteurs Cys-loop contre l'infestation par les tiques Rhipicephalus (Boophilus) microplus. ABSTRACT: Les tiques Rhipicephalus (Boophilus) microplus sont des ectoparasites hématophages obligatoires des bovins et agissent comme vecteurs de micro-organismes pathogènes. Le contrôle conventionnel des tiques est basé sur l'application d'acaricides chimiques, mais leur utilisation incontrôlée a augmenté les populations de tiques résistantes ainsi que la contamination des aliments et de l'environnement. Le contrôle immunologique alternatif des tiques s'est avéré partiellement efficace. Par conséquent, il est nécessaire de caractériser de nouveaux antigènes afin d'améliorer la protection immunologique. Le but de ce travail était d'évaluer les récepteurs Cys-loop comme candidats vaccins. Les domaines N-terminaux d'un récepteur du glutamate et d'un récepteur de type glycine ont été produits par recombinaison chez Escherichia coli. Des groupes de souris BALB/c ont été immunisés indépendamment avec quatre doses de chaque protéine recombinante émulsionnée avec l'adjuvant de Freund. Les deux vaccins candidats étaient immunogènes chez la souris, comme l'a démontré l'analyse par transfert Western. Ensuite, des protéines recombinantes ont été formulées indépendamment avec l'adjuvant Montanide ISA 50 V2 et évaluées chez des bovins infestés de larves de tiques Rhipicephalus microplus. Des groupes de trois veaux croisés européens ont été immunisés avec trois doses de chaque protéine avec adjuvant. Le test ELISA a été utilisé pour évaluer la réponse immunitaire IgG induite contre les protéines recombinantes. Les résultats ont montré que les candidats vaccins généraient une réponse humorale modérée sur les bovins vaccinés. La vaccination a affecté de manière significative le nombre de tiques femelles adultes engorgées mais n'a eu aucun effet significatif sur le poids des tiques, le poids des œufs et les valeurs de fertilité des œufs. Des efficacités vaccinales de 33 % et 25 % ont été calculées pour le récepteur du glutamate et le récepteur de type glycine, respectivement.

Gastroprotective activity and pharmacological safety evaluation of Eupatorium aschenbornianum.

Eupatorium aschenbornianum has been widely used in traditional Mexican and folk medicine for the treatment of wounds, skin lesions, hemorrhages and gastric ulcers in humans. Phytochemical studies have indicated that hexane extracts of E. aschenbornianum have anti-microbial and anti-fungal activities. In the present study, an accurate and reliable approach using a murine model was pursued to evaluate the anti-ulcer activity, lipid peroxidation properties and acute toxicity of powdered dried stem of E. aschenbornianum. The results indicated that administration of E. aschenbornianum exerted an anti-ulcerative effect and decreased lipid peroxidation in gastric ulcers induced by acetylsalicylic acid. An acute toxicity assay indicated normal behavior and no significant variations in the weight and food consumption of animals. In addition, quantitative analysis of biochemical parameters did not indicate any liver or kidney damage. The results indicated that E. aschenbornianum may be a safe therapeutic agent for the prevention of gastric ulcers.

In vitro and In vivo Postprandial Glycemic Activity of Citrus limetta Peel Flour.

BACKGROUND: Previous studies of Citrus spp. peel have shown hypoglycemic and antioxidant activities. Citrus limetta has been studied for its therapeutic properties. Diabetes mellitus (DM) is a health problem in Mexico and worldwide, that takes a vital importance due to its high incidence. Recently, scientists have searched natural sources to control the disease. MATERIALS AND METHODS: In this study, we evaluated the in vitro hypoglycemic activity and in vivo postprandial glycemic effect of C. limetta peel flour by glucose adsorption and retardation assays as well as postprandial serum glucose levels using a group of female Balb-c mice, respectively. RESULTS: C. limetta peel flour showed a glucose adsorption capacity of 16.58 mM, having a similar effect regarding the positive control. The glucose diffusion in the dialysate was elevated, with a glucose dialysis retardation index of 33.79% in a period of 3 h, showing similar results to positive control. Postprandial serum glucose levels in the animal group treated with C. limetta peel flour showed a glucose level of 41.4 mg/dL, being this value significantly lower than negative control group and similar to positive control. Toxicity tests showed good tolerance to the dose of 2000 mg/kg. CONCLUSION: C. limetta peel flour could act as a source of functional compounds for the control of DM. SUMMARY: Citrus limetta peel flour showed a glucose adsorption capacity similar to the positive controlThe glucose diffusion in the dialysate was elevated, showing similar results to positive controlPostprandial serum glucose levels in the animal group treated with C. limetta peel flour showed a glucose level significantly lower than negative control group and similar to positive controlToxicity tests showed good toleranceC. limetta peel flour could act as a source of functional compounds for the control of diabetes mellitus. Abbreviations used: CIATEJ: Center for Research and Assistance in Technology and Design of Jalisco; DM: Diabetes mellitus; FGC: Final glucose concentration; GDRI: Glucose dialysis retardation index; IGC: Initial glucose concentration; OECD: Organization for Economic Cooperation and Development.

Molecular characterization and expression analysis of three novel autophagy-related genes from the cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The cattle tick Rhipicephalus (Boophilus) microplus is a hematophagous ectoparasite of major importance for the livestock industry. It shows a remarkable ability to survive over long periods without feeding. However, the mechanisms used to endure long-term starvation are poorly understood. It is believed that autophagy, a process of intracellular protein degradation, may play a significant role to confront adverse environmental conditions. To advance our understanding of autophagy in R. microplus, in the present study we report the molecular characterization of three autophagy-related (ATG) genes, namely, RmATG3, RmATG4 and RmATG6, as well as their expression profiles in different developmental stages and organs of the parasite. The deduced amino acid sequences derived from the characterized gene sequences were subjected to Basic Local Alignment Search Tool analysis. The testing produced significant alignments with respective ATG proteins from Haemaphysalis longicornis and Ixodes scapularis ticks. Real-time polymerase chain reaction assays revealed that RmATG4 and RmATG6 transcripts were elevated in egg and ovary tissue, when compared with larva and midgut samples, while RmATG3 expression in midgut was 2-fold higher than in egg, larva and ovary samples.