PRPF8 increases the aggressiveness of hepatocellular carcinoma by regulating FAK/AKT pathway via fibronectin 1 splicing

López-Cánovas Juan L.,Hermán-Sánchez Natalia,del Rio-Moreno Mercedes,Fuentes-Fayos Antonio C.,Lara-López Araceli,Sánchez-Frias Marina E.,Amado Víctor,Ciria Rubén,Briceño Javier,de la Mata Manuel,Casta 생화학분자생물학회 2023 Experimental and molecular medicine Vol.55 No.-

Hepatocellular carcinoma (HCC) pathogenesis is associated with alterations in splicing machinery components (spliceosome and splicing factors) and aberrant expression of oncogenic splice variants. We aimed to analyze the expression and potential role of the spliceosome component PRPF8 (pre-mRNA processing factor 8) in HCC. PRPF8 expression (mRNA/protein) was analyzed in a retrospective cohort of HCC patients (n = 172 HCC and nontumor tissues) and validated in two in silico cohorts (TCGA and CPTAC). PRPF8 expression was silenced in liver cancer cell lines and in xenograft tumors to understand the functional and mechanistic consequences. In silico RNAseq and CLIPseq data were also analyzed. Our results indicate that PRPF8 is overexpressed in HCC and associated with increased tumor aggressiveness (patient survival, etc.), expression of HCC-related splice variants, and modulation of critical genes implicated in cancer-related pathways. PRPF8 silencing ameliorated aggressiveness in vitro and decreased tumor growth in vivo. Analysis of in silico CLIPseq data in HepG2 cells demonstrated that PRPF8 binds preferentially to exons of protein-coding genes, and RNAseq analysis showed that PRPF8 silencing alters splicing events in multiple genes. Integrated and in vitro analyses revealed that PRPF8 silencing modulates fibronectin (FN1) splicing, promoting the exclusion of exon 40.2, which is paramount for binding to integrins. Consistent with this finding, PRPF8 silencing reduced FAK/AKT phosphorylation and blunted stress fiber formation. Indeed, HepG2 and Hep3B cells exhibited a lower invasive capacity in membranes treated with conditioned medium from PRPF8-silenced cells compared to medium from scramble-treated cells. This study demonstrates that PRPF8 is overexpressed and associated with aggressiveness in HCC and plays important roles in hepatocarcinogenesis by altering FN1 splicing, FAK/AKT activation and stress fiber formation.

Revision of Infected Total Knee Arthroplasty: Two-Stage Reimplantation Using an Antibiotic-Impregnated Static Spacer

Antonio Silvestre,Fernando Almeida,Pablo Renovell,Elena Morante,Raúl López 대한정형외과학회 2013 Clinics in Orthopedic Surgery Vol.5 No.3

Background: A two-stage revision remains as the “gold standard” treatment for chronically infected total knee arthroplasties. Methods: Forty-five septic knee prostheses were revised with a minimum follow-up of 5 years. Static antibiotic-impregnated cement spacers were used in all cases. Intravenous antibiotics according to sensitivity test of the culture were applied during patients' hospital stay. Oral antibiotics were given for another 5 weeks. Second-stage surgery was undertaken after control of infection with normal erythrocyte sedimentation rate and C-reactive protein values. Extensile techniques were used if needed and metallic augments were employed for bone loss in 32 femoral and 29 tibial revisions. Results: The average interval between the first-stage resection and reimplantation was 4.4 months. Significant improvement was obtained with respect to visual analog scale pain and clinical and functional scores, and infection was eradicated in 95.6% of cases following a two-stage revision total knee arthroplasty. Radiographic evaluation showed suitable alignment without signs of mechanical loosening. Conclusions: This technique is a reasonable procedure to eradicate chronic infection in knee arthroplasty and provides proper functional and clinical results. However, it sometimes requires extensile surgical approaches that could imply arduous surgeries. Metallic augments with cementless stems available in most of the knee revision systems are a suitable alternative to handle bone deficiencies, avoiding the use of bone allografts with its complications.

Oxidative Refolding of Lysozyme Assisted by DsbA, DsbC and the GroEL Apical Domain Immobilized in Cellulose

Aurora Antonio-Pérez,Tania Rivera-Hernández,Luz María Aldaz-Martínez,Jaime Ortega-López 한국생물공학회 2012 Biotechnology and Bioprocess Engineering Vol.17 No.4

Expression of recombinant proteins in Escherichia coli often leads to formation of inclusion bodies (IB). If a recombinant protein contains one or more disulfide bonds,protein refolding and thiol oxidation reactions are required to recover its biological activity. Previous studies have demonstrated that molecular chaperones and foldases assist with the in vitro protein refolding. However, their use has been limited by the stoichiometric amount required for the refolding reaction. In search of alternatives to facilitate the use of these folding biocatalysts in this study, DsbA, DsbC,and the apical domain of GroEL (AD) were fused to the carbohydrate-binding module CBDCex of Cellulomonas fimi. The recombinant proteins were purified and immobilized in cellulose and used to assist the oxidative refolding of denatured and reduced lysozyme. The assisted refolding yields obtained with immobilized folding biocatalysts were at least twice of those obtained in the spontaneous refolding,suggesting that the AD, DsbA, and DsbC immobilized in cellulose might be useful for the oxidative refolding of recombinant proteins that are expressed as inclusion bodies. In addition, the spontaneous or assisted refolding kinetics data fitted well (r2 > 0.9) to a previously reported lysozyme refolding model. The estimated refolding (kN)and aggregation (kA) constants were consistent with the hypothesis that foldases assisted the oxidative refolding of lysozyme by decreasing protein aggregation rather than increasing the refolding rate.

Modeling for the strap combined footings Part II: Mathematical model for design

Juan Antonio Yáñez-Palafox,Arnulfo Luévanos-Rojas,Sandra López-Chavarría,Manuel Medina-Elizondo 국제구조공학회 2019 Steel and Composite Structures, An International J Vol.30 No.2

This paper presents the second part of the modeling for the strap combined footings, this part shows a mathematical model for design of strap combined footings subject to axial load and moments in two directions to each column considering the soil real pressure acting on the contact surface of the footing for one and/or two property lines of sides opposite restricted, the pressure is presented in terms of an axial load, moment around the axis “X” and moment around the axis “Y” to each column, and the methodology is developed using the principle that the derived of the moment is the shear force. The first part shows the optimal contact surface for the strap combined footings to obtain the most economical dimensioning on the soil (optimal area). The classic model considers an axial load and a moment around the axis “X” (transverse axis) applied to each column, i.e., the resultant force from the applied loads is located on the axis “Y” (longitudinal axis), and its position must match with the geometric center of the footing, and when the axial load and moments in two directions are presented, the maximum pressure and uniform applied throughout the contact surface of the footing is considered the same. A numerical example is presented to obtain the design of strap combined footings subject to an axial load and moments in two directions applied to each column. The mathematical approach suggested in this paper produces results that have a tangible accuracy for all problems and it can also be used for rectangular and T-shaped combined footings.

Thermo and pH-Responsive Poly(DEGMA-co-OEGMA)-b-Poly(DEAEM) Synthesized by RAFT Polymerization and Its Self-Assembly Study

Ángela M. López,Antonio Tirado-Guizar,Angel Licea-Claverie,Alejandro Ramírez-Jiménez 한국고분자학회 2022 Macromolecular Research Vol.30 No.12

Drug delivery and release systems using stimuli-responsive nanostructures based on amphiphilic copolymers have gained great importance. In this work copolymers from oligo(ethylene glycol) methyl ether methacrylate (OEGMA) and di(ethylene glycol) methyl ether methacrylate (DEGMA) as thermo-responsive moiety and block copolymers incorporating N,N′-diethylaminoethyl methacrylate (DEAEM) as pH-responsive one, were synthesized by reversible addition-fragmentation chain transfer polymerization (RAFT) using 5-amino-4-methyl-4-(propylthiocarbonothioylthio)- 5-oxo pentanoic acid as chain transfer agent (CTA). The number average molecular weights (Mn), and dispersity Đ confirmed a good RAFT control. Polymers were characterized by Fourier transform infrared spectroscopy using a attenuated total reflection (FTIR-ATR), nuclear magnetic resonance (NMR), gel permeation chromatography (GPC), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic light scattering (DLS), transmission electron microscope (TEM), and Turbidimetry. Their self-assembly behavior is described by its hydrodynamic diameter (Dh), its ζ-potential and as well as its cloud point temperature (Tcp). The results showed thermo-responsive and pH-responsive systems with Tcp in phosphate-buffered saline (PBS) pH=7.4 around 37 and 42℃ the isoelectric point was found in a pH range between 7.0 and 7.5, further the critical micelle concentration (CMC) indicated very low values between 5.85 to 4.20 × 10-4 mg mL-1. The developed block copolymeric systems have potential to be used for drug transport and delivery.

Characterization of hypotensive and vasorelaxant effects of PHAR-DBH-Me a new cannabinoid receptor agonist

Oscar Alberto López-Canales,Natalia Pavón,Laura Matilde Ubaldo-Reyes,Marco Antonio Juárez-Oropeza,Patricia Victoria Torres-Durán,Ignacio Regla,María Cristina Paredes-Carbajal 대한약리학회 2022 The Korean Journal of Physiology & Pharmacology Vol.26 No.2

The effect of PHAR-DBH-Me, a cannabinoid receptor agonist, on different cardiovascular responses in adult male rats was analyzed. The blood pressure was measured directly and indirectly. The coronary flow was measured by Langendorff preparation, and vasomotor responses induced by PHAR-DBH-Me in aortic rings precontracted with phenylephrine (PHEN) were analyzed. The intravenous injection of the compound PHAR-DBH-Me (0.018–185 μg/kg) resulted in decreased blood pressure; maximum effect was observed at the dose of 1,850 μg/kg. A concentrationdependent increase in the coronary flow was observed in a Langendorff preparation. In the aortic rings, with and without endothelium, pre-contracted with PHEN (10–6 M), the addition of PHAR-DBH-Me to the superfusion solution (10–12–10–5 M), produced a vasodilator response, which depends on the concentration and presence of the endothelium. L-NAME inhibited these effects. Addition of CB1 receptor antagonist (AM 251) did not modify the response, while CB2 receptor antagonist (AM630) decreased the potency of relaxation elicited by PHAR-DBH-Me. Indomethacin shifted the curve concentration-response to the left and produced an increase in the magnitude of the maximum endothelium dependent response to this compound. The maximum effect of PHAR-DBH-Me was observed with the concentration of 10–5 M. These results show that PHAR-DBH-Me has a concentration-dependent and endothelium-dependent vasodilator effect through CB2 receptor. This vasodilation is probably mediated by the synthesis/release of NO. On the other hand, it is suggested that PHAR-DBH-Me also induces the release of a vasoconstrictor prostanoid.

Molecular epidemiology of Aleutian mink disease virus causing outbreaks in mink farms from Southwestern Europe: a retrospective study from 2012 to 2019

Alberto Prieto,Ricardo Fernández-Antonio,Gonzalo López-Lorenzo,José Manuel Díaz-Cao,Cynthia López-Novo,Susana Remesar,Rosario Panadero,Pablo Díaz,Patrocinio Morrondo,Pablo Díez-Baños,Gonzalo Fernández 대한수의학회 2020 Journal of Veterinary Science Vol.21 No.4

Background: Aleutian mink disease virus (AMDV) causes major economic losses in fur-bearing animal production. The control of most AMDV outbreaks is complex due to the difficulties of establishing the source of infection based only on the available on-farm epidemiological data. In this sense, phylogenetic analysis of the strains present in a farm may help elucidate the origin of the infection and improve the control and biosecurity measures. Objectives: This study had the following aims: characterize the AMDV strains from most outbreaks produced at Spanish farms between 2012–2019 at the molecular level, and assess the utility of the combined use of molecular and epidemiological data to track the possible routes of infection. Methods: Thirty-seven strains from 17 farms were partially sequenced for the NS1 and VP2 genes and analyzed phylogenetically with other strains described worldwide. Results: Spanish AMDV strains are clustered in four major clades that generally show a good geographical correlation, confirming that most had been established in Spain a long time ago. The combined study of phylogenetic results and epidemiological information of each farm suggests that most of the AMDV outbreaks since 2012 had been produced by within-farm reservoirs, while a few of them may have been due to the introduction of the virus through international trade. Conclusions: The combination of phylogenetic inference, together with epidemiological data, helps assess the possible origin of AMDV infections in mink farms and improving the control and prevention of this disease.

Zn-22Al-2Cu Alloy Phase Transformations at Different Homogenizing Temperatures

Héctor Javier Dorantes-Rosales,Víctor Manuel López-Hirata,Ramón Esquivel-González,Jorge Luis González-Velazquez,Joel Moreno-Palmerin,Antonio Torres-Castillo 대한금속·재료학회 2012 METALS AND MATERIALS International Vol.18 No.3

The effect of the homogenizing temperature on phase transformations and hardening behavior was studied in a Zn-22Al-2Cu (wt%) alloy homogenized for 7 days at 315, 350, and 390°C. The homogenized alloy was subsequently quenched and aged naturally at room temperature or artificially at 200°C for different time periods. The β phase was unstable at room temperature and it decomposed through the β→α+ηreaction during natural aging. An increase in the homogenizing temperature caused a decrease in the kinet-ics of the decomposition of the β phase in the alloy. Natural aging also caused an increase in the hardness due to the formation of nanometric grains with α and η phases and the presence of an intermetallic εphase. This result agrees with the time exponents (n) between 1.5 and 2.6, which were obtained using the Johnson-Mehl-Avrami-Kolmogorov equation for the decomposition of the β phase. The transformed frac-tion curves exhibited the fastest and slowest growth kinetics of the â phase decomposition at homogeniz-ing temperatures of 350 and 390°C, respectively. The artificial aging induced the formation of equilibrium phases and began a decrease in hardness due to the coarsening process of the equilibrium α, η, and τ'phases.

Osteoblastic cells biomineralized on bioactive glass and glass-ceramics of the SiO2.Na2O.CaO.K2O.MgO.P2O5 system modified with Al2O3 and B2O3

Irene Barrios de Arenas,Karem Noris-Suárez,Maritza Vasquez,Yotselys López,Thalia San Antonio 한양대학교 세라믹연구소 2010 Journal of Ceramic Processing Research Vol.11 No.2

A treatment study evaluates glass and glass-ceramics of the Na2O·CaO·K2O·MgO·SiO2·P2O5 system, modified with B2O3, Al2O3or B2O3/Al2O3 up to 0.66. Both, bioactivity using simulated body fluid (SBF) immersion and mineral nodule formation using calvaria derived osteoblasts, were tested. The surface structure changes were examined by SEM-EDX line scan X-ray microanalysis and calcium deposition by the alizarin-red test. It was found that all compositions of glasses as well as glassceramics reacted with the SBF, allowing the development of a SiO2 rich layer. In some cases the presence of a rich calcium and phosphorous layer was observed, that according to other studies, corresponds to the apatite layer identified in previous studies. This result suggests a bioactivity behavior, with the exception of the glass containing B2O3/Al2O3 = 0.66 and the glassceramic of the same composition with a 2nd heat treatment. The composition containing B2O3/Al2O3 = 0.66 showed the best biocompatibility with hydroxyapatite nodule formation on its surface, especially on the glass-ceramics.

Mediterranean Diet and Physical Activity Protect from Silent Brain Infarcts in a Cohort of Patients with Atrial Fibrillation

Irene Escudero-Martínez,Fernando Mancha,Ángela Vega-Salvatierra,María Irene Ayuso,Rafael F. Ocete,Pilar Algaba,Antonio López-Rueda,Pilar Piñero,Elena Fajardo,José Román Fernández-Engo,Eva María Martín 대한뇌졸중학회 2019 Journal of stroke Vol.21 No.3

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