Development and evaluation of different electroactive poly(vinylidene fluoride) architectures for endothelial cell culture

Tissue engineering (TE) aims to develop structures that improve or even replace the biological functions of tissues and organs. Mechanical properties, physical-chemical characteristics, biocompatibility, and biological performance of the materials are essential factors for their applicability in TE. Poly(vinylidene fluoride) (PVDF) is a thermoplastic polymer that exhibits good mechanical properties, high biocompatibility and excellent thermal properties. However, PVDF structuring, and the corresponding processing methods used for its preparation are known to significantly influence these characteristics. In this study, doctor blade, salt-leaching, and electrospinning processing methods were used to produce PVDF-based structures in the form of films, porous membranes, and fiber scaffolds, respectively. These PVDF scaffolds were subjected to a variety of characterizations and analyses, including physicochemical analysis, contact angle measurement, cytotoxicity assessment and cell proliferation. All prepared PVDF scaffolds are characterized by a mechanical response typical of ductile materials. PVDF films displayed mostly vibration modes for the a-phase, while the remaining PVDF samples were characterized by a higher content of electroactive beta-phase due the low temperature solvent evaporation during processing. No significant variations have been observed between the different PVDF membranes with respect to the melting transition. In addition, all analysed PVDF samples present a hydrophobic behavior. On the other hand, cytotoxicity assays confirm that cell viability is maintained independently of the architecture and processing method. Finally, all the PVDF samples promote human umbilical vein endothelial cells (HUVECs) proliferation, being higher on the PVDF film and electrospun randomly-oriented membranes. These findings demonstrated the importance of PVDF topography on HUVEC behavior, which can be used for the design of vascular implants.This work has been partially funded by the Junta de Extremadura (Spain), the Spanish Ministry of Science and Innovation, the European Social Fund, the European Regional Development Fund, and the European Next Generation Funds (Grant Numbers PD18077, TA18023, and GR21201). The authors also thanks to Portuguese Foundation for Science and Technology (FCT) for financial support under grants SFRH/BD/140698/2018 (RP), 2020.04163. CEECIND (CR). The also authors acknowledge funding by Spanish State Research Agency (AEI) and the European Regional Development Fund (ERFD) through the project PID 2019-106099RB-C43/AEI/10.13039/501100011033 and from the Basque Government Industry Departments under the ELKARTEK program

Phonon structure, infra-red and raman spectra of Li2MnO3 by first-principles calculations

The layer-structured monoclinic Li2MnO3 is a key material, mainly due to its role in Li-ion batteries and as a precursor for adsorbent used in lithium recovery from aqueous solutions. In the present work, we used first-principles calculations based on density functional theory (DFT) to study the crystal structure, optical phonon frequencies, infra-red (IR), and Raman active modes and compared the results with experimental data. First, Li2MnO3 powder was synthesized by the hydrothermal method and successively characterized by XRD, TEM, FTIR, and Raman spectroscopy. Secondly, by using Local Density Approximation (LDA), we carried out a DFT study of the crystal structure and electronic properties of Li2MnO3. Finally, we calculated the vibrational properties using Density Functional Perturbation Theory (DFPT). Our results show that simulated IR and Raman spectra agree well with the observed phonon structure. Additionally, the IR and Raman theoretical spectra show similar features compared to the experimental ones. This research is useful in investigations involving the physicochemical characterization of Li2MnO3 materia

CSF SERPINA3 Levels Are Elevated in Patients With Progressive MS

Multiple sclerosis; SERPINA3Esclerosi múltiple; SERPINA3Esclerosis múltiple; SERPINA3Objective To identify biomarkers associated with progressive phases of MS and with neuroprotective potential. Methods Combined analysis of the transcriptional and proteomic profiles obtained in CNS tissue during chronic progressive phases of experimental autoimmune encephalomyelitis (EAE) with the transcriptional profile obtained during the differentiation of murine neural stem cells into neurons. Candidate biomarkers were measured by ELISA in the CSF of 65 patients with MS (29 with relapsing-remitting MS [RRMS], 20 with secondary progressive MS, and 16 with primary progressive MS [PPMS]) and 30 noninflammatory neurologic controls (NINCs). Results Integrative analysis of gene and protein expression data identified 2 biomarkers, the serine protease inhibitor Serpina3n and the calcium-binding protein S100A4, which were upregulated in chronic progressive EAE and whose expression was induced during neuronal differentiation. Immunofluorescence studies revealed a primarily neuronal expression of S100A4 and Serpina3n during EAE. CSF levels of SERPINA3, the human ortholog of murine Serpina3n, and S100A4 were increased in patients with MS compared with NINCs (SERPINA3: 1,320 vs 838.6 ng/mL, p = 0.0001; S100A4: 1.6 vs 0.8 ng/mL, p = 0.02). Within the MS group, CSF SERPINA3 levels were significantly elevated in patients with progressive forms, mainly patients with PPMS compared with patients with RRMS (1,617 vs 1,129 ng/mL, p = 0.02) and NINCs (1,617 vs 838.6 ng/mL, p = 0.0001). Of interest, CSF SERPINA3 levels significantly correlated with CSF neurofilament light chain levels only in the PPMS group (r = 0.62, p = 0.01). Conclusion These results point to a role of SERPINA3 as a biomarker associated with the progressive forms of MS, particularly PPMS.This work was supported by grants from the Fondo de Investigación Sanitaria (FIS; grant number PI17/00596), Ministry of Science and Innovation, Spain; Generalitat de Catalunya Suport Grups de Recerca (2017 SGR 0527); and the Red Española de Esclerosis Múltiple (RD16/0015/0004) funded by the FIS

Unveiling the Synergistic Interplay of Neuropeptides for Novel Therapeutic Approaches in Neurodegenerative and Depressive Disorders.

The intricate relationship between hippocampal neurogenesis dysregulation and neurodegenerative diseases such as Alzheimer's, as well as depression, has sparked an urgent call for innovative therapeutic strategies. Our groundbreaking study delves into the interaction of Neuropeptide Y (NPY) and galanin (GAL) agonists, two neuromodulatory systems with a substantial presence in the limbic system, and their potential neurogenic impact on both the dorsal and ventral hippocampus. Through meticulous examination of the subchronic e"ects of NPY Y1 (Y1R) and GAL2 (GALR2) agonists on hippocampal cell proliferation, survival, and neuroprotective factor expression, we reveal a fascinating cascade of cellular responses. These include increased cell proliferation (PCNA), enhanced hippocampal cell survival (BrdU), and induction of neuroprotective factors (BDNF). Our functional assessment showcases the resulting improvements in spatial memory performance in the object-in-place task and antidepressant-like e"ects in the forced swimming test. These outcomes are attributed to the synergistic interaction between Y1R and GALR2 receptors, which promote neuronal survival and neurite outgrowth in hippocampal cells. This pioneering research paves the way for the development of heterobivalent agonist pharmacophores that target Y1R-GALR2 heterocomplexes. By acting on neuronal precursor cells in the dentate gyrus of the dorsal hippocampus, these novel compounds hold immense promise as transformative therapies for cognitive and a"ective impairments in neurodegenerative and depressive diseases.Supported by the Proy_Excel_2021_0613 (Junta de Andalucía) and Proyecto Puente (B4-2021) (UMA), Spain to MN. Swedish Medical Research Council, Sweden (62X-00715-50-3), to KF, by Stiftelsen Olle Engkvist Byggmästare to KF, and by Hjärnfonden, Sweden (F02018-0286), Hjärnfonden, Sweden (F02019-0296) and EMERGIA2020 (Junta de Andalucía), to DOBE. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

How hepatitis C virus modifies the immunological profile of Sjögren syndrome: analysis of 783 patients.

Introduction: We conducted a study to analyze how infection by hepatitis C virus (HCV) may influence the immunological serum pattern of patients with Sjögren syndrome (SS). Methods: Since 1994, we have tested serum HCV-IgG antibodies in 783 patients with SS diagnosed according to the 1993 European classification criteria. The immunological profile at diagnosis was compared according to the presence or absence of HCV. Results: Of the 783 patients with SS, 105 (13.4 %) tested positive for HCV-IgG antibodies (88 females, 17 males,mean age at SS diagnosis: 62.9 years). Multivariate analysis showed that patients with SS-HCV had a higher mean age and a higher frequency of low C3/C4 levels, cryoglobulins, and hematological neoplasia compared with patients without HCV. The frequency of anti-La antibodies compared with anti-Ro antibodies was higher in patients with SS-HCV (17 % vs. 15 %) and lower in patients without HCV infection (30 % vs. 43 %). The frequency of concomitant detection of the three main cryoglobulin-related markers (cryoglobulins, rheumatoid factor activity, and C4 consumption) was threefold higher in patients with SS-HCV compared with patients without HCV. SS-HCV patients with genotype 1b showed the highest frequencies of immunological abnormalities related to cryoglobulins and the lowest frequencies of anti-Ro/La antibodies. Conclusions: We found HCV infection in 13 % of a large series of Spanish patients with SS. The HCV-driven autoimmune response was characterized by a lower frequency of anti-Ro/La antibodies, an abnormal predominance of anti-La among anti-Ro antibodies, and a higher frequency of cryoglobulinemic-related immunological markers in comparison with patients without HCV infection. This immunological pattern may contribute to the poor outcomes found in patients with SS-HCV

Pruning height and frequency of Moringa oleifera and Leucaena leucocephala in a silvopastoral system

Objetive: Evaluate two heights (50 and 70 cm) and pruning frequencies (60 and 135 d) in Moringa oleifera and Leucaena leucocephala in a developing silvopastoral system. Desing/methodology/approach: Four treatments were established, T1: Moringa oleifera with pruning at 50 cm, T2: Moringa oleifera with pruning at 70 cm, T3: Leucaena leucocephala with pruning at 50 cm and T4: Leucaena leucocephala with pruning at 70 cm and pruning frequencies at the 60 and 135 d. The number of sprouts, diameter and length of the largest sprout, and biomass were recorded. The data were analyzed using a mixed model, analysis of variance and Tukey's tests. Results: The number of regrowths in L. leucocephala was higher than M. oleifera. The T2 had the highest behavior at 60 d, for L. leucocephala it was similar between pruning and frequencies. The diameter and length were greater for T2 at both frequencies, at 60 d for T3 and at 135 d for T4. Biomass was higher in M. oleífera than L. leucocephala, pruning at 70 cm was better than at 50 cm. Pruning at 135 days obtained higher biomass than at 60 days. T2 obtained higher biomass at 60 d, T1 and T4 at 135 d. Study limitations/implications: Forage species, frequencies and pruning heights determine the growth potential and amount of biomass available to the animals. Findings/conclusions:Leucaena leucocephala has better agronomic performance in response to pruning without influencing the height and frequency of pruning in a silvopastoral system.Objective: To evaluate two different heights (50 and 70 cm) and pruning frequencies (60 and 135 days) in Moringa oleifera and Leucaena leucocephala in a developing silvopastoral system. Design/methodology/approach: Four treatments were established: T1=Moringa oleifera with pruning at 50 cm; T2=Moringa oleifera with pruning at 70 cm; T3=Leucaena leucocephala with pruning at 50 cm; and T4=Leucaena leucocephala with pruning at 70 cm. In all cases, pruning frequencies were 60 and 135 days. The number of sprouts, diameter, and length of the largest sprout, as well as the amount of biomass, were recorded. A mixed model, variance analysis, and Tukey's test were used to analyze the data. Results: The number of regrowths for L. leucocephala was higher than for M. oleifera. Treatment T2 had the highest performance at day 60; in the case of L. leucocephala, performance was similar between pruning frequencies. For T2, diameter and length were larger at both frequencies; for T3, at 60 days; and for T4, at 135 days. Biomass was higher for M. oleifera than for L. leucocephala, while pruning at 70 cm was better than at 50 cm. Pruning at 135 days produced a higher biomass than at 60 days. For T2, biomass was higher at 60 days, while for T1 and T4, it was higher at 135 days. Study limitations/implications: Forage species, as well as pruning frequencies and heights, determine tree growth potential and the amount of biomass available for animals. Findings/conclusions: The agronomic performance of Leucaena leucocephala has a better response to pruning, regardless of pruning height and frequency, in a silvopastoral system

JKST6, a novel multikinase modulator of the BCR-ABL1/STAT5 signaling pathway that potentiates direct BCR-ABL1 inhibition and overcomes imatinib resistance in chronic myelogenous leukemia

Chronic myelogenous leukemia (CML) is a hematological malignancy that highly depends on the BCR-ABL1/STAT5 signaling pathway for cell survival. First-line treatments for CML consist of tyrosine kinase inhibitors that efficiently target BCR-ABL1 activity. However, drug resistance and intolerance are still therapeutic limitations in Ph+ cells. Therefore, the development of new anti-CML drugs that exhibit alternative mechanisms to overcome these limitations is a desirable goal. In this work, the antitumoral activity of JKST6, a naphthoquinone-pyrone hybrid, was assessed in imatinib-sensitive and imatinib-resistant human CML cells. Live-cell imaging analysis revealed JKST6 potent antiproliferative activity in 2D and 3D CML cultures. JKST6 provoked cell increase in the subG1 phase along with a reduction in the G0/G1 phase and altered the expression of key proteins involved in the control of mitosis and DNA damage. Rapid increases in Annexin V staining and activation/cleavage of caspases 8, 9 and 3 were observed after JKST6 treatment in CML cells. Of interest, JKST6 inhibited BCR-ABL1/STAT5 signaling through oncokinase downregulation that was preceded by rapid polyubiquitination. In addition, JKST6 caused a transient increase in JNK and AKT phosphorylation, whereas the phosphorylation of P38-MAPK and Src was reduced. Combinatory treatment unveiled synergistic effects between imatinib and JKST6. Notably, JKST6 maintained its antitumor efficacy in BCR-ABL1-T315I-positive cells and CML cells that overexpress BCR-ABL and even restored imatinib efficacy after a short exposure time. These findings, together with the observed low toxicity of JKST6, reveal a novel multikinase modulator that might overcome the limitations of BCR-ABL1 inhibitors in CML therapy.This research has been funded by Spanish Ministry of Economy and Competitiveness - MINECO - (SAF 2015–65113-C2–1-R and RTI2018–094356-B-C21 to AEB, SAF2015–65113-C2–2 to LFP, SAF2017–88026-R to JL) with the co-funding of European Regional Development Fund (EU-ERDF), Canary Islands Government (CEI2018–23/ACIISI to BG, CEI2019–08/ACIISI to BG and LFP, ProID2021010037 to AEB, LFP and BG) and "Juan de la Cierva Incorporacion" Grant Program from the Ministry of Science, Innovation and Universities (IJC2018-035193-I to CR). This project has been also supported by Alfredo Martin-Reyes Foundation (Arehucas)-Canary Islands Foundation for Cancer Research (FICIC). HAT is recipient of a predoctoral program grant from ULPGC (2016). JCM was funded by the Instituto de Salud Carlos III through a Miguel Servet program (CPII17/ 00015)

How hepatitis C virus modifies the immunological profile of Sjögren syndrome: analysis of 783 patients

Abstract Introduction: We conducted a study to analyze how infection by hepatitis C virus (HCV) may influence the immunological serum pattern of patients with Sjögren syndrome (SS). Methods: Since 1994, we have tested serum HCV-IgG antibodies in 783 patients with SS diagnosed according to the 1993 European classification criteria. The immunological profile at diagnosis was compared according to the presence or absence of HCV. Results: Of the 783 patients with SS, 105 (13.4 %) tested positive for HCV-IgG antibodies (88 females, 17 males, mean age at SS diagnosis: 62.9 years). Multivariate analysis showed that patients with SS-HCV had a higher mean age and a higher frequency of low C3/C4 levels, cryoglobulins, and hematological neoplasia compared with patients without HCV. The frequency of anti-La antibodies compared with anti-Ro antibodies was higher in patients with SS-HCV (17 % vs. 15 %) and lower in patients without HCV infection (30 % vs. 43 %). The frequency of concomitant detection of the three main cryoglobulin-related markers (cryoglobulins, rheumatoid factor activity, and C4 consumption) was threefold higher in patients with SS-HCV compared with patients without HCV. SS-HCV patients with genotype 1b showed the highest frequencies of immunological abnormalities related to cryoglobulins and the lowest frequencies of anti-Ro/La antibodies. Conclusions: We found HCV infection in 13 % of a large series of Spanish patients with SS. The HCV-driven autoimmune response was characterized by a lower frequency of anti-Ro/La antibodies, an abnormal predominance of anti-La among anti-Ro antibodies, and a higher frequency of cryoglobulinemic-related immunological markers in comparison with patients without HCV infection. This immunological pattern may contribute to the poor outcomes found in patients with SS-HCV