Cytotoxicity analysis of oxazine 4-perchlorate fluorescence nerve potential clinical biomarker for guided surgery

Biological tissue discrimination is relevant in guided surgery. Nerve identification is critical to avoid potentially severe collateral damage. Fluorescence imaging by oxazine 4-perchlorate (O4P) has been recently proposed. In this work, the cytotoxicity of O4P on U87 human-derived glioma cells has been investigated as a function of concentration and operating room irradiation modes. A custom-built optical irradiation device was employed for controlled optical dosimetry. DNA damage and O4P intracellular localization was also investigated by immunofluorescence and confocal microscopy. The results show that concentration below 100 µM can be considered safe. These results contribute to the assessment of the feasibility of O4P as a nerve biomarker.Spanish Ministry of Science, Research and Universities, cofunded by FEDER funds (PGC2018-101464-BI00); San Cándido Foundation

Genetic markers associated with divergent selection against the parasite Marteilia cochillia in common cockle (Cerastoderma edule) using transcriptomics and population genomics data

The common cockle (Cerastoderma edule) plays an important role in marine ecosystems and represents a valuable socioeconomic resource for coastal communities. In 2012, the cockle beds from Rı́a de Arousa (Galicia, NW Spain) were seriously decimated by the protozoan Marteilia cochillia responsible for marteiliosis. We aimed to identify single nucleotide polymorphisms (SNP) markers potentially associated with resilience to marteiliosis to be used in marker-assisted selection programs for restoring affected cockle beds and recovering their production. For this, we carried out a population genomics approach using 2b-RADseq, where 38 naive samples (before the first detection of M. cochillia in 2012) from two beds of Rı́a de Arousa were compared with 39 affected samples collected in 2018/2019 (after several years of marteiliosis occurring in the area), collected either before (15 non-exposed samples) or during (24 exposed samples) the marteiliosis outbreak. Additionally, 767 differentially expressed genes (DEG) from a previous transcriptomic study addressed during the aforementioned 2018/19 marteiliosis outbreak, were evaluated to identify SNPs showing signals of selection. Using 2b-RADseq, 9,154 SNPs were genotyped and among them, 110 consistent outliers for divergent selection were identified. This set of SNPs was able to discriminate the samples according to their marteiliosis status (naive vs affected; exposed vs non-exposed), while another 123 SNPs were identified linked to DEGs associated with the level of infection across a temporal series. Finally, combining the population genomics and transcriptomics information, we selected the 60 most reliable SNPs associated with marteiliosis resilience. These SNPs were close to or within DEGs, and many of them were related to immune response (phagocytosis and cell adhesion), defence, such as apoptosis, stress, and cellular cycle, among other functions. This set of SNPs will eventually be validated to develop a cost-effective genotyping tool for their application for obtaining cockle-resilient strains for marteiliosis.This study was possible due to funds granted by the European Union through the project COCKLES within the INTERREG-AA programme (EAPA_458/2016) and the scholarship granted to M.R.M.Coimbra from CNPq (202015/2020-3). This research has been carried out under the framework of the Spain's Recovery and Resilience Plan, and more specifically under the investment line no.1 of its component number 17, where the complementary RTDI plan with the autonomous regions of Spain are foreseen, with one of those being the Complementary RTDI Plan for Marine Science which includes the Marine Science Programme for Galicia. This research in this paper corresponds to the Programme Work Package nº 6 and activity no. 6.3.A.2 about "Genetic architecture of marteiliosis resistance in common cockle" and has been funded by the Resilience and Recovery Funds).S

Clinical Outcomes and Cost Analysis of Fibula Free Flaps: A Retrospective Comparison of CAD/CAM versus Conventional Technique

(1) Background: A decrease in operative time can not only improve patient outcomes through a reduction in the risk of developing complications but can also result in cost savings. The aim of this study is to determine whether there an intraoperative time gain can be achieved by using the preoperative virtual planning of mandibular reconstruction using a free fibula flap compared with freehand plate bending and osteotomies. (2) Methods: A retrospective comparative study was carried out in the Oral and Maxillofacial Department of La Paz University Hospital, Madrid, Spain. The study compared 18 patients in the CAD/CAM group with 19 patients in the conventional freehand group. A comparison was made between the total surgical time, the comorbidities, and the hospital stay. The resource consumption was estimated using a cost analysis. (3) Results: Although CAD/CAM was a statistically more expensive procedure in the perioperative phase, no significant differences were observed in total health care costs between the two groups. There was a non-significant trend towards an increase in complications with conventional reconstruction plates compared to patient-specific plates (PSI). (4) Conclusions: CAD/CAM technology and a 3D printed cutting guide offer a significantly shorter surgical time, which is associated with a reduction in hospital days, PACU days, and complications. The cost of CAD/CAM technology is comparable to that of the conventional freehand technique

Exploring Biginelli-based scaffolds as A2B adenosine receptor antagonists: Unveiling novel structure-activity relationship trends, lead compounds, and potent colorectal anticancer agents

Antagonists of the A(2B) adenosine receptor have recently emerged as targeted anticancer agents and immune checkpoint inhibitors within the realm of cancer immunotherapy. This study presents a comprehensive evaluation of novel Biginelli-assembled pyrimidine chemotypes, including mono-, bi-, and tricyclic derivatives, as A(2B)AR antagonists. We conducted a comprehensive examination of the adenosinergic profile (both binding and functional) of a large compound library consisting of 168 compounds. This approach unveiled original lead compounds and enabled the identification of novel structure-activity relationship (SAR) trends, which were supported by extensive computational studies, including quantum mechanical calculations and free energy perturbation (FEP) analysis. In total, 25 molecules showed attractive affinity (K-i < 100 nM) and outstanding selectivity for A(2B)AR. From these, five molecules corresponding to the new benzothiazole scaffold were below the K-i < 10 nM threshold, in addition to a novel dual A(2A)/A(2B) antagonist. The most potent compounds, and the dual antagonist, showed enantiospecific recognition in the A(2B)AR. Two A(2B)AR selective antagonists and the dual A(2A)AR/A(2B)AR antagonist reported in this study were assessed for their impact on colorectal cancer cell lines. The results revealed a significant and dose-dependent reduction in cell proliferation. Notably, the A(2B)AR antagonists exhibited remarkable specificity, as they did not impede the proliferation of non-tumoral cell lines. These findings support the efficacy and potential that A(2B)AR antagonists as valuable candidates for cancer therapy, but also that they can effectively complement strategies involving A(2A)AR antagonism in the context of immune checkpoint inhibition

Vocabulario de química [galego-español-inglés]

Dirección Xeral de Política Lingüística. Consellería de Educación e Ordenación Universitaria. Xunta de Galici

Reactivity of a recombinant esterase from Thermus thermophilus HB27 in aqueous and organic media

The thermoalkalophilic membrane-associated esterase E34Tt from Thermus thermophilus HB27 was cloned and expressed in Kluyveromyces lactis (KLEST-3S esterase). The recombinant enzyme was tested as a biocatalyst in aqueous and organic media. It displayed a high thermal stability and was active in the presence of 10% (v/v) organic solvents and 1% (w/v) detergents. KLEST-3S hydrolysed triglycerides of various acyl chains, which is a rare characteristic among carboxylic ester hydrolases from extreme thermophiles, with maximum activity on tributyrin. It also displayed interfacial activation towards triacetin. KLEST-3S was also tested as a biocatalyst in organic media. The esterase provided high yields for the acetylation of alcohols. In addition, KLEST-3S catalyzed the stereoselective hydrolysis of (R,S)-ibuprofen methyl ester (87% ee). Our results indicate that KLEST-3S may be a robust and efficient biocatalyst for application in industrial bioconversions.Xunta de Galicia | Ref. 10MDS373027PRXunta de Galicia | Ref. GRC ED431C 2020/08European Commision | Ref. FP7-PEOPLE 2012-IAPP, n. 32443