Deciphering Distinct Genetic Risk Factors for FTLD-TDP Pathological Subtypes via Whole-Genome Sequencing.

Frontotemporal lobar degeneration with neuronal inclusions of the TAR DNA-binding protein 43 (FTLD-TDP) is a fatal neurodegenerative disorder with only a limited number of risk loci identified. We report our comprehensive genome-wide association study as part of the International FTLD-TDP Whole-Genome Sequencing Consortium, including 985 cases and 3,153 controls, and meta-analysis with the Dementia-seq cohort, compiled from 26 institutions/brain banks in the United States, Europe and Australia. We confirm UNC13A as the strongest overall FTLD-TDP risk factor and identify TNIP1 as a novel FTLD-TDP risk factor. In subgroup analyses, we further identify for the first time genome-wide significant loci specific to each of the three main FTLD-TDP pathological subtypes (A, B and C), as well as enrichment of risk loci in distinct tissues, brain regions, and neuronal subtypes, suggesting distinct disease aetiologies in each of the subtypes. Rare variant analysis confirmed TBK1 and identified VIPR1 , RBPJL , and L3MBTL1 as novel subtype specific FTLD-TDP risk genes, further highlighting the role of innate and adaptive immunity and notch signalling pathway in FTLD-TDP, with potential diagnostic and novel therapeutic implications.

The role of artificial intelligence in scientific publishing: Perspectives from Hospital Pharmacy. / El rol de la inteligencia artificial en la publicación científica: perspectivas desde la farmacia hospitalaria.

The article examines the impact of artificial intelligence on scientific writing, with a particular focus on its application in hospital pharmacy. It analyzes artificial intelligence tools that enhance information retrieval, literature analysis, writing quality, and manuscript drafting. Chatbots like Consensus, along with platforms such as Scite and SciSpace, enable precise searches in scientific databases, providing evidence-based responses and references. SciSpace facilitates the generation of comparative tables and the formulation of queries regarding studies, while ResearchRabbit maps the scientific literature to identify trends. Tools like DeepL and ProWritingAid improve writing quality by correcting grammatical, stylistic, and plagiarism errors. A.R.I.A. enhances reference management, and Jenny AI assists in overcoming writer's block. Python libraries such as LangChain enable advanced semantic searches and the creation of agents. Despite their benefits, artificial intelligence raises ethical concerns including biases, misinformation, and plagiarism. The importance of responsible use and critical review by experts is emphasized. In hospital pharmacy, artificial intelligence can enhance efficiency and precision in research and scientific communication. Pharmacists can use these tools to stay updated, enhance the quality of their publications, optimize information management, and facilitate clinical decision-making. In conclusion, artificial intelligence is a powerful tool for hospital pharmacy, provided it is used responsibly and ethically.

DLK1-DIO3 region as a source of tumor suppressor miRNAs in papillary thyroid carcinoma.

BACKGROUND: In previous studies, we demonstrated the downregulation of several miRNAs from the DLK1-DIO3 genomic region in papillary thyroid carcinoma (PTC). Due to the large number of miRNAs within this region, the individual contribution of these molecules to PTC development and progression remains unclear. OBJECTIVE: In this study, we aimed to clarify the contribution of DLK1-DIO3-derived miRNAs to PTC. METHODS: We used different computational approaches and in vitro resources to assess the biological processes and signaling pathways potentially modulated by these miRNAs. RESULTS: Our analysis suggests that, out of more than 100 mature miRNAs originated from the DLK1-DIO3 region, a set of 12 miRNAs accounts for most of the impact on PTC development and progression, cooperating to modulate distinct cancer-relevant biological processes, such as cell migration, extracellular matrix remodeling, and signal transduction. The restoration of the expression of one of these miRNAs (miR-485-5p) in a BRAFT199A-positive PTC cell line impaired proliferation and migration, suppressing the expression of GAB2 and RAC1, validated miR-485-5p targets. CONCLUSIONS: Overall, our results shed light on the role of the DLK1-DIO3 region, which harbors promising tumor suppressor miRNAs in thyroid cancer, and open prospects for the functional exploration of these miRNAs as therapeutic targets for PTC.

C-terminal TMEM106B fragments in human brain correlate with disease-associated TMEM106B haplotypes.

Transmembrane protein 106B (TMEM106B) is a tightly regulated glycoprotein predominantly localized to endosomes and lysosomes. Genetic studies have implicated TMEM106B haplotypes in the development of multiple neurodegenerative diseases with the strongest effect in frontotemporal lobar degeneration with TDP-43 pathology (FTLD-TDP), especially in progranulin (GRN) mutation carriers. Recently, cryo-electron microscopy studies showed that a C-terminal fragment (CTF) of TMEM106B (amino acid residues 120-254) forms amyloid fibrils in the brain of patients with FTLD-TDP, but also in brains with other neurodegenerative conditions and normal ageing brain. The functional implication of these fibrils and their relationship to the disease-associated TMEM106B haplotype remain unknown. We performed immunoblotting using a newly developed antibody to detect TMEM106B CTFs in the sarkosyl-insoluble fraction of post-mortem human brain tissue from patients with different proteinopathies (n = 64) as well as neuropathologically normal individuals (n = 10) and correlated the results with age and TMEM106B haplotype. We further compared the immunoblot results with immunohistochemical analyses performed in the same study population. Immunoblot analysis showed the expected ∼30 kDa band in the sarkosyl-insoluble fraction of frontal cortex tissue in at least some individuals with each of the conditions evaluated. Most patients with GRN mutations showed an intense band representing TMEM106B CTF, whereas in most neurologically normal individuals it was absent or much weaker. In the overall cohort, the presence of TMEM106B CTFs correlated strongly with both age (rs = 0.539, P < 0.001) and the presence of the TMEM106B risk haplotype (rs = 0.469, P < 0.001). Although there was a strong overall correlation between the results of immunoblot and immunohistochemistry (rs = 0.662, P < 0.001), 27 cases (37%) were found to have higher amounts of TMEM106B CTFs detected by immunohistochemistry, including most of the older individuals who were neuropathologically normal and individuals who carried two protective TMEM106B haplotypes. Our findings suggest that the formation of sarkosyl-insoluble TMEM106B CTFs is an age-related feature which is modified by TMEM106B haplotype, potentially underlying its disease-modifying effect. The discrepancies between immunoblot and immunohistochemistry in detecting TMEM106B pathology suggests the existence of multiple species of TMEM106B CTFs with possible biological relevance and disease implications.

Accumulation of TMEM106B C-terminal fragments in neurodegenerative disease and aging.

Several studies using cryogenic electron microscopy (cryo-EM) techniques recently reported the isolation and characterization of novel protein filaments, composed of a C-terminal fragment (CTF) of the endolysosomal transmembrane protein 106B (TMEM106B), from human post-mortem brain tissue with various neurodegenerative conditions and normal aging. Genetic variation in TMEM106B is known to influence the risk and presentation of several neurodegenerative diseases, especially frontotemporal dementia (FTD) caused by mutations in the progranulin gene (GRN). To further elucidate the significance of TMEM106B CTF, we performed immunohistochemistry with antibodies directed against epitopes within the filament-forming C-terminal region of TMEM106B. Accumulation of TMEM106B C-terminal immunoreactive (TMEM-ir) material was a common finding in all the conditions evaluated, including frontotemporal lobar degeneration with TDP-43 pathology (FTLD-TDP), Alzheimer's disease, tauopathies, synucleinopathies and neurologically normal aging. TMEM-ir material was present in a wide range of brain cell types and in a broad neuroanatomical distribution; however, there was no co-localization of TMEM-ir material with other neurodegenerative proteins in cellular inclusions. In most conditions, the presence and abundance of TMEM-ir aggregates correlated strongly with patient age and showed only a weak correlation with the TMEM106B haplotype or the primary pathological diagnosis. However, all patients with FTD caused by GRN mutations were found to have high levels of TMEM-ir material, including several who were relatively young (< 60 years). These findings suggest that the accumulation of TMEM106B CTF is a common age-related phenomenon, which may reflect lysosomal dysfunction. Although its significance in most neurodegenerative conditions remains uncertain, the consistent finding of extensive TMEM-ir material in cases of FTLD-TDP with GRN mutations further supports a pathomechanistic role of TMEM106B and lysosomal dysfunction in this specific disease population.

Architectural Ultrasound Pennation Angle Measurement of Lumbar Multifidus Muscles: A Reliability Study.

The pennation angle has been shown to be a relevant parameter of muscle architecture. This parameter has not previously been measured in the lumbar multifidus musculature, and it is for this reason that it has been considered of great interest to establish an assessment protocol to generate new lines of research in the future. OBJECTIVE: The objective of this study was to establish a protocol for measuring the pennation angle of the multifidus muscles, with a study of intra-rater and interrater reliability values. DESIGN: This was a reliability study following the recommendations of the Guidelines for Reporting Reliability and Agreement Studies (GRRAS). SETTING: The study was carried out at University of Alcalá, Department of Physiotherapy. SUBJECTS: Twenty-seven subjects aged between 18 and 55 years were recruited for this study. METHODS: Different ultrasound images of the lumbar multifidus musculature were captured. Subsequently, with the help of ImageJ software, the pennation angle of this musculature was measured. Finally, a complex statistical analysis determined the intra- and interrater reliability. RESULTS: The intra-rater reliability of the pennation angle measurement protocol was excellent for observer 1 in the measurement of the left-sided superficial multifidus 0.851 (0.74, 0.923), and for observer 2 in the measurement of the right-sided superficial 0.711 (0.535, 0.843) and deep multifidus 0.886 (0.798, 0.942). Interrater reliability was moderate to poor, and correlation analysis results were high for thickness vs. pennation angle. CONCLUSIONS: The designed protocol for ultrasound measurement of the pennation angle of the lumbar multifidus musculature has excellent intra-rater reliability values, supporting the main conclusions and interpretations. Normative ranges of pennation angles are reported. High correlation between variables is described.

The role of platelet-rich plasma in the mild and severe stages of atherosclerotic disease in mice.

Background: Platelet-rich plasma (PRP) has a high concentration of growth factors (GFs), which present a therapeutic wound healing effect. Despite having been correlated with an immunomodulatory function, the administration of PRP has not yet been investigated in atherosclerosis models. Aim: Evaluate the effect of lyophilized PRP on atherosclerosis in mice models through serum analysis. Methods: Animals received a high-fat diet for disease induction and a weekly PRP retro-orbital application. Effectiveness was evaluated by measuring inflammatory markers in plasma following the treatment of mice with either PRP or saline solution. Results: PRP was well characterized for platelet and GF concentrations; the atherosclerotic profile was established. Cytokine concentrations were altered after PRP applications. Conclusion: PRP could modulate the inflammatory pattern in the early stages of atherosclerosis.

Platelet-rich plasma (PRP) contains growth factors, which stimulate normal wound healing. This product seems to be a good modulator of white blood cells and has not been investigated in atherosclerosis. This study aimed to evaluate PRP in atherosclerosis using mice models. The PRP was produced from animals and preserved using the lyophilization technique; the product was then applied weekly in the vein. For atherosclerosis induction, genetically modified animals were fed a high-fat diet. The effectiveness was evaluated by measuring plasma inflammatory markers after treatment, and PRP seemed to alter cell-signaling molecules (cytokines). This study concluded that PRP was capable of modulating the inflammatory pattern during the early stages of atherosclerosis.

A collagen I derived matricryptin increases aorta vascular wall remodeling after induced thrombosis in mouse.

Matricryptins are collagen fragments proteolytically released from the extracellular matrix (ECM) with biological activity that can regulate several processes involved in ECM remodeling. Vessel wall matrix reorganization after lesion is important to the recovery of vascular function. This study aimed to analyze the effect of the peptide p1158/59 (Lindsey, 2015) on thrombosis, neointimal formation, and vascular remodeling of C57BL6 mice abdominal aorta. We used a FeCl3 induced vascular injury mice model and analyzed thrombus size, neointima formation, gelatinase activities in situ, re-endothelization, and collagen fibers organization on the arterial wall using polarization microscopy. As result, we observed that 2 days after injury the treatment with p1158/59 increased thrombus size and gelatinase activity, vascular lesion and it did not recover the endothelium loss induced by the chemical injury. We also observed that the peptide increased neointima growth and collagen birefringence, indicating collagen fibers reorganization. It also promoted increased re-endothelization and decreased activity of gelatinases 14 days after injury. Thus, we conclude that the peptide p1158/59 impaired the initial thrombosis recovery 2 days after injury but was able to induce vascular ECM remodeling after 14 days, improving vessel re-endothelization, collagen fibers deposition, and organization.

Shared brain transcriptomic signature in TDP-43 type A FTLD patients with or without GRN mutations.

Frontotemporal lobar degeneration with TDP-43 inclusions (FTLD-TDP) is a complex heterogeneous neurodegenerative disorder for which mechanisms are poorly understood. To explore transcriptional changes underlying FTLD-TDP, we performed RNA-sequencing on 66 genetically unexplained FTLD-TDP patients, 24 FTLD-TDP patients with GRN mutations and 24 control participants. Using principal component analysis, hierarchical clustering, differential expression and coexpression network analyses, we showed that GRN mutation carriers and FTLD-TDP-A patients without a known mutation shared a common transcriptional signature that is independent of GRN loss-of-function. After combining both groups, differential expression as compared to the control group and coexpression analyses revealed alteration of processes related to immune response, synaptic transmission, RNA metabolism, angiogenesis and vesicle-mediated transport. Deconvolution of the data highlighted strong cellular alterations that were similar in FTLD-TDP-A and GRN mutation carriers with NSF as a potentially important player in both groups. We propose several potentially druggable pathways such as the GABAergic, GDNF and sphingolipid pathways. Our findings underline new disease mechanisms and strongly suggest that affected pathways in GRN mutation carriers extend beyond GRN and contribute to genetically unexplained forms of FTLD-TDP-A.

The Bisaccia and Meccariello technique in pediatric femoral shaft fractures with intramedullary titanium nail osteosynthesis linked external-fixator (IOLE): validity and reliability.

BACKGROUND: Over the last 20 years, the incidence of pediatric femoral shaft fractures was increased, due to changes in the children's daily activities. The healing times are different according to the chosen treatment and to other factors such as age, type of fracture, the involvement of the soft tissues, and concomitance with other injuries. The Bisaccia and Meccariello technique ( Intramedullary titanium nail Osteosynthesis Linked External-fixator -IOLE) was born to prevent rotationally and lengthening malunion or nonunion in the treatment of pediatric femoral shaft fractures. The aim or the objective of this paper is to compare the IOLE with the two most used methods for the treatment of femoral fractures in children. METHODS: From 2000 to 2016, 58 pediatric patients with femoral shaft fractures were surgically treated and enrolled in the study. The ranged age of the patients was between 3 and 15 years. Twenty-two patients were treated with endomedullary titanium nails (TEN), 16 with external axial or modular external fixators and 14 patients treated with IOLE technique. The IOLE technique is the hybridization of titanium intramedullary nails with a modular external fixator. It is divided into three phases, the first revenue given the length of the femur with the external fixator; the second, the rotations are dominated by the elastic nails; and the third finally they are hybridized on the external fixator. Comparing the three groups, radiographic images were taken to assess fracture reduction and consolidation.  RESULTS: At the final follow-up, there were no differences between three groups in terms of significant rotation defects, angulation, growth, and/or nonunion but there was a statistical in IOLE groups for the early weight-bearing. CONCLUSIONS: The Bisaccia- Meccariello technique (IOLE) showed to lead to healing the pediatric femoral shaft fracture of the femur but allows an early weight-bearing to these patients and normal life like that.

An Internal Promoter Drives the Expression of a Truncated Form of CCC1 Capable of Protecting Yeast from Iron Toxicity.

In yeast, iron storage and detoxification depend on the Ccc1 transporter that mediates iron accumulation in vacuoles. While deletion of the CCC1 gene renders cells unable to survive under iron overload conditions, the deletion of its previously identified regulators only partially affects survival, indicating that the mechanisms controlling iron storage and detoxification in yeast are still far from well understood. This work reveals that CCC1 is equipped with a complex transcriptional structure comprising several regulatory regions. One of these is located inside the coding sequence of the gene and drives the expression of a short transcript encoding an N-terminally truncated protein, designated as s-Ccc1. s-Ccc1, though less efficiently than Ccc1, is able to promote metal accumulation in the vacuole, protecting cells against iron toxicity. While the expression of the s-Ccc1 appears to be repressed in the normal genomic context, our current data clearly demonstrates that it is functional and has the capacity to play a role under iron overload conditions.

[Tobacco use in Spain during COVID-19 lockdown: an evaluation through social media.] / Consumo de tabaco en España durante el estado de alarma por COVID-19: resultados de una evaluación a través de redes sociales.

OBJECTIVE: During the state of alarm and once the confinement decreed by the COVID-19 pandemic ended, a cross-sectorial study was carried out in Spain between May 4th and 22nd, 2020 by volunteers who completed a self-administered online survey. The objective of this study was to know how the confinement period affected the consumption of tobacco and other related products in the adult Spanish population. METHODS: The survey consisted of 18 questions concerning sociodemographic characteristics, the consumption of tobacco and other related products, exposure to secondhand smoke and perception of COVID-19 risk associated with consumption. Questions about tobacco and other related products were posed in order to compare consumption prior to and during confinement. The survey was completed by 17,017 people. The analysis of association of variables was carried out with T-student. Variable frequency analysis was performed with χ2. RESULTS: There was a reduction in the prevalence of daily tobacco smoking and no changes were observed in the products consumed in either period (6.73%). The prevalence of exposure to secondhand smoke at home during confinement among non-smokers decreased (61.83%). Most of survey respondents reported that tobacco and e-cigarette consumption increased the risk of contracting COVID-19 and suffering severe complications (39.09% and 31.80% respectively). CONCLUSIONS: During the COVID-19 lockdown in Spain, the tobacco consumption decreased. Also, secondhand smoke exposition reduces in Spain during this period.

OBJETIVO: Durante el estado de alarma y una vez finalizado el confinamiento decretado por la pandemia por COVID-19, en España se realizó, entre el 4 y el 22 de mayo de 2020, un estudio transversal en voluntarios aplicando una encuesta autocumplimentada online. El objetivo de este estudio fue conocer cómo afectó el periodo de confinamiento al consumo de tabaco y relacionados en la población adulta española. METODOS: El cuestionario constaba de 18 preguntas e incluía características sociodemográficas, el consumo de tabaco y otros productos relacionados, exposición al humo ambiental de tabaco y percepción del riesgo de enfermedad por COVID-19 asociada a su consumo. La encuesta fue completada por 17.017 personas. El análisis de la asociación entre variables cuantitativas, fue realizado mediante el test de la T de Student y el de frecuencias de las variables categóricas mediante el test de χ2. RESULTADOS: Se observó reducción en la prevalencia de fumadores diarios de tabaco (6,73%) y no se observaron cambios en los productos consumidos. La prevalencia de exposición al humo ambiental en casa durante el confinamiento entre personas no fumadoras disminuyó (61,83%). Los encuestados declararon que el consumo de tabaco y de cigarrillos electrónicos aumentaba el riesgo de contraer la enfermedad del COVID-19 y sufrir complicaciones (39,09% y 31,80% respectivamente). CONCLUSIONES: Durante el periodo de confinamiento en España debido al COVID-19, se produjo una reducción en el consumo de tabaco y similares. Además de observó una reducción a la exposición al humo ambiental.

Current status of localized submental fat treatment with sodium deoxicolate (ATX-101).

Aim Facial aesthetics is at present a concept intricately linked to the degree of self-esteem. Unwanted submental fat (SMF) leads to an unattractive submental profile. Sodium deoxicolate (ATX) -101 is the only injectable drug approved to decrease submental fat of moderate to severe intensity. Methods We carried out a bibliographic review in PubMed using the key words: deoxycholic acid, ATX-101, and submental fat. Only complete articles published between 2009 and 2019, and focused on submental fat were reviewed, excluding those articles relating to that spoke of deoxycholate in the treatment of fat in other locations or in which deoxycholate was associated with other drugs. Results In several phase III clinical trials, injection of 2 mg/cm2 deoxycholic acid in SMF has reduced moderate-severe fullness compared to the placebo group. These results were maintained in most cases during a long follow-up period. Injections of deoxycholic acid are generally well tolerated, with limited adverse effects in the treatment area, with a mild and complete resolution without sequelae. However, not all patients with SMF are suitable for deoxycholic acid therapy, and therefore a proper selection is very important to achieve the desired aesthetic results. Conclusion Deoxycholic acid injections are effective and are a generally well-tolerated, minimally invasive option for the treatment of moderate to severe intensity SMF in selected adults.

Dacomitinib in first-line treatment of advanced EGFR-mutated non-small-cell lung cancer: a cost-effectiveness analysis.

Aim: To assess the cost-effectiveness of first-line treatment with dacomitinib compared with gefitinib in patients newly diagnosed with advanced NSCLC EGFR-positive in the context of Spain. Materials & methods: A partitioned survival model was developed including costs, utilities and disutilities to estimate quality-adjusted life-year (QALY) and incremental cost-effectiveness ratio when treating with dacomitinib versus gefitinib. Results: Dacomitinib presented higher QALYs (0.51) compared with gefitinib (0.45). Dacomitinib costs were €33,061 in comparison with €26,692 for gefitinib arm. An incremental cost-effectiveness ratio of €111,048 was obtained for dacomitinib. Conclusion: Dacomitinib was more effective in terms of QALYs gained than gefitinib. However, to obtain a cost-effectiveness alternative, a discount greater than 25% in dacomitinib acquisition cost is required.

Electrospun Nanofibrous Architectures of Thrombin-Loaded Poly(ethylene oxide) for Faster in Vivo Wound Clotting.

Wound healing materials to prevent blood loss are crucial during emergency medical treatment because uncontrolled bleeding can lead to patient death. Herein, bioabsorbable fibrous architectures of thrombin-loaded poly(ethylene oxide)-PEO/thrombin-are conceptualized and accomplished via electrospinning for faster wound clotting. Membranes with average fiber diameters ranging from 188 to 264 nm are achieved, where the active thrombin is entrapped within the nanofibers. The results of in vitro and in vivo wound healing activity tests revealed that when the nanofibers with thrombin-loaded capacity are in contact with the wound, the presence of water in the skin or blood catalyzes the degradation of the membranes, thus releasing thrombin. Thrombin then accelerates the wound clotting process. In contrast to other hemostatic materials, PEO/thrombin nanofibers do not require mechanical removal after application, and the viscoelastic nature of such biomaterials enables their conformation to a variety of wound topographies. Remarkably, PEO/thrombin membranes are promising functional materials and their use is a powerful strategy for hemostatic treatment, ranging from simple first aid and sealing to a wound to small surgical procedures.

Cost-effectiveness analysis of infliximab, adalimumab, golimumab, vedolizumab and tofacitinib for moderate to severe ulcerative colitis in Spain.

OBJECTIVE: Despite the biological drugs, the treatment of moderate to severe ulcerative colitis is still a challenge, particularly in resource-limited settings. The aim of this study was to assess the efficiency of biological drugs and tofacitinib for moderate to severe ulcerative colitis in the Spanish context. METHODS: A Markov model was built to simulate the progression of moderate to severe ulcerative colitis in a cohort of patients. The model used a time horizon of 10 years. The perspective chosen was the National Health Service, with a discount rate of 3%, and a threshold of €30,000/quality adjusted life-year (QALY). It carried out a one-way sensitivity analysis and probabilistic sensitivity analysis. RESULTS: The comparison of infliximab with adalimumab and golimumab estimated an incremental cost-effectiveness ratio (ICER) of €43,928.07/QALY and €31,340.69/QALY, with a difference of - 0.43 and - 0.82 QALY, respectively. Vedolizumab vs infliximab achieved an ICER of €122,890.19/QALY with a gain of 0.46 QALY. The comparison of infliximab with tofacitinib yielded an estimated ICER of €270,503.19/QALY, with a slight gain in QALY (0.16). The one-way sensitivity analysis showed a robust study. CONCLUSION: For a threshold of €30,000/QALY, adalimumab was the most cost-effective treatment versus infliximab for moderate to severe ulcerative colitis in Spain.

The sodium-glucose cotransporter-2 (SGLT2) inhibitors synergize with nitric oxide and prostacyclin to reduce human platelet activation.

Gliflozins (canagliflozin, dapagliflozin and empagliflozin) are the newest anti-hyperglycemic class and have offered cardiovascular and renal benefits. Because platelets are involved in the atherothrombosis process, this study is aimed to evaluate the direct effect of gliflozins on platelet reactivity. Platelet-rich plasma (PRP) or washed platelets (WP) were obtained from healthy volunteers. Aggregation, flow cytometry for glycoprotein IIb/IIIa, cyclic nucleotides and intracellular calcium levels, Western blot, thromboxane B2 (TXB2) measurement and COX-1 activity were performed in the presence of gliflozins (1-30 µM) alone or in combination with sodium nitroprusside (SNP, 10 or 100 nM) + iloprost (ILO, 0.1 or 1 nM). SGLT2 protein is not expressed on human platelets. Gliflozins produced little inhibitory effect in agonists-induced aggregation and this effect was greatly potentiated by ~10-fold in the presence of SNP + ILO, accompanied by lower levels of TXB2 (58.1 ± 5.1%, 47.1 ± 7.2% and 43.4 ± 9.2% inhibition for canagliflozin, dapagliflozin and empagliflozin, respectively). The activity of COX-1 was not affected by gliflozins. Collagen increased Ca2+ levels and α(IIb)ß(3) activation, both of which were significantly reduced by gliflozins + SNP + ILO. The intracellular levels of cAMP and cGMP and the protein expression of p-VASPSer157 and p-VASPSer239 were not increased by gliflozins while the expression of the serine-threonine kinase, AktSer473 was markedly reduced. Our results showed that the antiplatelet activity of gliflozins were greatly enhanced by nitric oxide and prostacyclin, thus suggesting that the cardiovascular protection seen by this class of drugs could be in part due to platelet inhibition.

Microcurrent and adipose-derived stem cells modulate genes expression involved in the structural recovery of transected tendon of rats.

Tendon injuries are common and have a high incidence of re-rupture that can cause loss of functionality. Therapies with adipose-derived stem cells (ASC) and the microcurrent (low-intensity electrical stimulation) application present promising effects on the tissue repair. We analyzed the expression of genes and the participation of some molecules potentially involved in the structural recovery of the Achilles tendon of rats, in response to the application of both therapies, isolated and combined. The tendons were distributed in five groups: normal (N), transected (T), transected and ASC (C) or microcurrent (M) or with ASC, and microcurrent (MC). Microcurrent therapy was beneficial for tendon repair, as it was observed a statistically significant increase in the organization of the collagen fibers, with involvement of the TNC, CTGF, FN, FMDO, and COL3A1 genes as well as PCNA, IL-10, and TNF-α. ASC therapy significantly increased the TNC and FMDO genes expression with no changes in the molecular organization of collagen. With the association of therapies, a significant greater collagen fibers organization was observed with involvement of the FMOD gene. The therapies did not affect the expression of COL1A1, SMAD2, SMAD3, MKX, and EGR1 genes, nor did they influence the amount of collagen I and III, caspase-3, tenomodulin (Tnmd), and hydroxyproline. In conclusion, the application of the microcurrent isolated or associated with ASC increased the organization of the collagen fibers, which can result in a greater biomechanical resistance in relation to the tendons treated only with ASC. Future studies will be needed to demonstrate the biological effects of these therapies on the functional recovery of injured tendons.