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1.
J Biomed Sci ; 31(1): 71, 2024 Jul 15.
Article in English | MEDLINE | ID: mdl-39004727

ABSTRACT

BACKGROUND: Critical limb-threatening ischemia (CLTI) constitutes the most severe manifestation of peripheral artery disease, usually induced by atherosclerosis. CLTI patients suffer from high risk of amputation of the lower extremities and elevated mortality rates, while they have low options for surgical revascularization due to associated comorbidities. Alternatively, cell-based therapeutic strategies represent an effective and safe approach to promote revascularization. However, the variability seen in several factors such as cell combinations or doses applied, have limited their success in clinical trials, being necessary to reach a consensus regarding the optimal "cellular-cocktail" prior further application into the clinic. To achieve so, it is essential to understand the mechanisms by which these cells exert their regenerative properties. Herein, we have evaluated, for the first time, the regenerative and vasculogenic potential of a combination of endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs) isolated from adipose-tissue (AT), compared with ECFCs from umbilical cord blood (CB-ECFCs) and AT-MSCs, in a murine model of CLTI. METHODS: Balb-c nude mice (n:32) were distributed in four different groups (n:8/group): control shams, and ischemic mice (after femoral ligation) that received 50 µl of physiological serum alone or a cellular combination of AT-MSCs with either CB-ECFCs or AT-ECFCs. Follow-up of blood flow reperfusion and ischemic symptoms was carried out for 21 days, when mice were sacrificed to evaluate vascular density formation. Moreover, the long-term molecular changes in response to CLTI and both cell combinations were analyzed in a proteomic quantitative approach. RESULTS: AT-MSCs with either AT- or CB-ECFCs, promoted a significant recovery of blood flow in CLTI mice 21 days post-ischemia. Besides, they modulated the inflammatory and necrotic related processes, although the CB group presented the slowest ischemic progression along the assay. Moreover, many proteins involved in the repairing mechanisms promoted by cell treatments were identified. CONCLUSIONS: The combination of AT-MSCs with AT-ECFCs or with CB-ECFCs promoted similar revascularization in CLTI mice, by restoring blood flow levels, together with the modulation of the inflammatory and necrotic processes, and reduction of muscle damage. The protein changes identified are representative of the molecular mechanisms involved in ECFCs and MSCs-induced revascularization (immune response, vascular repair, muscle regeneration, etc.).


Subject(s)
Adipose Tissue , Disease Models, Animal , Ischemia , Mesenchymal Stem Cells , Mice, Inbred BALB C , Mice, Nude , Animals , Mice , Ischemia/therapy , Ischemia/physiopathology , Umbilical Cord/cytology , Male , Mesenchymal Stem Cell Transplantation/methods , Neovascularization, Physiologic , Endothelial Cells , Humans
4.
Int Forum Allergy Rhinol ; 14(7): 1245-1248, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38268107

ABSTRACT

KEY POINTS: T-cell activation in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) is enriched by late cytotoxic T cells. The proportion of early and intermediate activated cytotoxic T cells decreases in nasal polyps of patients with CRSwNP. Our results identify late activated cytotoxic T cells as potential biomarkers or therapeutic targets for patients with CRSwNP.


Subject(s)
Immunophenotyping , Lymphocyte Activation , Nasal Polyps , Rhinitis , Sinusitis , Humans , Nasal Polyps/immunology , Sinusitis/immunology , Rhinitis/immunology , Chronic Disease , Lymphocyte Activation/immunology , Male , Adult , Middle Aged , Female , T-Lymphocytes, Cytotoxic/immunology , Aged , Rhinosinusitis
5.
Int J Biol Sci ; 19(6): 1664-1680, 2023.
Article in English | MEDLINE | ID: mdl-37063416

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection significantly affects the cardiovascular system, causing vascular damage and thromboembolic events in critical patients. Endothelial dysfunction represents one of the first steps in response to COVID-19 that might lead to cardiovascular complications and long-term sequelae. However, despite the enormous efforts in the last two years, the molecular mechanisms involved in such processes remain poorly understood. Herein, we analyzed the protein changes taking place in endothelial colony forming cells (ECFCs) after the incubation with the serum from individuals infected with COVID-19, whether asymptomatic or critical patients, by application of a label free-quantitative proteomics approach. Specifically, ECFCs from healthy individuals were incubated ex-vivo with the serum of either COVID-19 negative donors (PCR-/IgG-, n:8), COVID-19 asymptomatic donors at different infective stages (PCR+/ IgG-, n:8and PCR-/IgG+, n:8), or hospitalized critical COVID-19 patients (n:8), followed by proteomics analysis. In total, 590 proteins were differentially expressed in ECFCs in response to all infected serums. Predictive analysis highlighted several proteins like CAPN5, SURF4, LAMP2 or MT-ND1, as highly discriminating features between the groups compared. Protein changes correlated with viral infection, RNA metabolism or autophagy, among others. Remarkably, the angiogenic potential of ECFCs in response to the infected serums was impaired, and many of the protein alterations in response to the serum of critical patients were associated with cardiovascular-related pathologies.


Subject(s)
COVID-19 , Cardiovascular System , Humans , Proteomics , SARS-CoV-2 , Immunoglobulin G , Cells, Cultured , Membrane Proteins , Calpain
6.
Glia ; 71(1): 91-102, 2023 01.
Article in English | MEDLINE | ID: mdl-35411970

ABSTRACT

In the central nervous system, oligodendrocytes synthesize the myelin, a specialized membrane to wrap axons in a discontinuous way allowing a rapid saltatory nerve impulse conduction. Oligodendrocytes express a number of growth factors and neurotransmitters receptors that allow them to sense the environment and interact with neurons and other glial cells. Depending on the cell cycle stage, oligodendrocytes may respond to these signals by regulating their survival, proliferation, migration, and differentiation. Among these signals are the endocannabinoids, lipidic molecules synthesized from phospholipids in the plasma membrane in response to cell activation. Here, we discuss the evidence showing that oligodendrocytes express a full endocannabinoid signaling machinery involved in physiological oligodendrocyte functions that can be therapeutically exploited to promote remyelination in central nervous system pathologies.


Subject(s)
Endocannabinoids , Oligodendroglia , Endocannabinoids/metabolism , Oligodendroglia/metabolism , Myelin Sheath/metabolism , Axons/metabolism , Central Nervous System/metabolism , Cell Differentiation/physiology
7.
Polymers (Basel) ; 14(13)2022 Jul 01.
Article in English | MEDLINE | ID: mdl-35808751

ABSTRACT

The addition of natural substances with pharmacoactive properties to polymeric biomedical devices would provide beneficial regarding the assimilation of these endoprostheses when implanted into a patient's body. The added drug would facilitate endothelization by regulating the inflammatory processes that such interventions entail, preventing contamination hazards and favoring the angiogenesis or formation of blood vessels in the tissue. The present work used mango leaf extract (MLE) obtained through pressurized ethanol for this purpose. Polylactic acid (PLA) in the form of filaments or 3D-printed disks was impregnated by means of supercritical technology with MLE for the culture essays. The release kinetics has been studied and the polymer matrices have been examined by scanning electron microscopy (SEM). The impregnated devices were subjected to in vitro culture of colony-forming endothelial cells. The influence of the different impregnation conditions used for the production of the MLE impregnated polymeric devices on the development of the cell culture was determined by fluorescence microscopy. The best results were obtained from the calcein cultures on 35 °C MLE impregnated into 3D-printed polymer disks.

8.
Stem Cell Res Ther ; 13(1): 266, 2022 06 21.
Article in English | MEDLINE | ID: mdl-35729651

ABSTRACT

BACKGROUND: Endothelial colony forming cells (ECFCs), alone or in combination with mesenchymal stem cells, have been selected as potential therapeutic candidates for critical limb-threatening ischemia (CLTI), mainly for those patients considered as "no-option," due to their capability to enhance revascularization and perfusion recovery of ischemic tissues. Nevertheless, prior to translating cell therapy to the clinic, biodistribution assays are required by regulatory guidelines to ensure biosafety as well as to discard undesired systemic translocations. Different approaches, from imaging technologies to qPCR-based methods, are currently applied. METHODS: In the current study, we have optimized a cell-tracking assay based on DiR fluorescent cell labeling and near-infrared detection for in vivo and ex vivo assays. Briefly, an improved protocol for DiR staining was set up, by incubation of ECFCs with 6.67 µM DiR and intensive washing steps prior cell administration. The minimal signal detected for the residual DiR, remaining after these washes, was considered as a baseline signal to estimate cell amounts correlated to the DiR intensity values registered in vivo. Besides, several assays were also performed to determine any potential effect of DiR over ECFCs functionality. Furthermore, the optimized protocol was applied in combination with qPCR amplification of specific human Alu sequences to assess the final distribution of ECFCs after intramuscular or intravenous administration to a murine model of CLTI. RESULTS: The optimized DiR labeling protocol indicated that ECFCs administered intramuscularly remained mainly within the hind limb muscle while cells injected intravenously were found in the spleen, liver and lungs. CONCLUSION: Overall, the combination of DiR labeling and qPCR analysis in biodistribution assays constitutes a highly sensitive approach to systemically track cells in vivo. Thereby, human ECFCs administered intramuscularly to CLTI mice remained locally within the ischemic tissues, while intravenously injected cells were found in several organs. Our data corroborate the need to perform biodistribution assays in order to define specific parameters such as the optimal delivery route for ECFCs before their application into the clinic.


Subject(s)
Cell Tracking , Neovascularization, Physiologic , Animals , Cells, Cultured , Disease Models, Animal , Humans , Ischemia/therapy , Mice , Tissue Distribution
9.
Mol Ther Nucleic Acids ; 29: 76-87, 2022 Sep 13.
Article in English | MEDLINE | ID: mdl-35721225

ABSTRACT

Despite the extraordinary advances achieved to beat COVID-19 disease, many questions remain unsolved, including the mechanisms of action of SARS-CoV-2 and which factors determine why individuals respond so differently to the viral infection. Herein, we performed an in silico analysis to identify host microRNA targeting ACE2, TMPRSS2, and/or RAB14, all genes known to participate in viral entry and replication. Next, the levels of six microRNA candidates previously linked to viral and respiratory-related pathologies were measured in the serum of COVID-19-negative controls (n = 16), IgG-positive COVID-19 asymptomatic individuals (n = 16), and critical COVID-19 patients (n = 17). Four of the peripheral microRNAs analyzed (hsa-miR-32-5p, hsa-miR-98-3p, hsa-miR-423-3p, and hsa-miR-1246) were upregulated in COVID-19 critical patients compared with COVID-19-negative controls. Moreover, hsa-miR-32-5p and hsa-miR-1246 levels were also altered in critical versus asymptomatic individuals. Furthermore, these microRNA target genes were related to viral infection, inflammatory response, and coagulation-related processes. In conclusion, SARS-CoV-2 promotes the alteration of microRNAs targeting the expression of key proteins for viral entry and replication, and these changes are associated with disease severity. The microRNAs identified could be taken as potential biomarkers of COVID-19 progression as well as candidates for future therapeutic approaches against this disease.

10.
Antioxidants (Basel) ; 11(5)2022 Apr 27.
Article in English | MEDLINE | ID: mdl-35624715

ABSTRACT

Cardiovascular diseases remain the leading cause of death worldwide, mainly triggered by the formation of atherosclerotic plaques that reduce blood flow. Angiogenic cell therapy based on endothelial colony forming cells (ECFCs) constitutes a promising alternative to promote vascular revascularization; however, under the oxidative environment that prevails in ischemic areas, these cells become impaired. Thus, it is necessary to investigate strategies to enhance their regenerative properties. Antioxidant substances, such as polyphenols, have been shown to be useful for this purpose. In the current study we evaluated the potential of mango leaves, olive leaves and red grape pomace extracts, rich in polyphenols, to promote ECFC reparative effects. For this, aqueous and ethanolic extracts of the aforementioned raw materials were obtained by pressurized liquid extraction (PLE). After evaluating the polyphenol content and the antioxidant activity, in vitro assays were carried out, and we found that ethanolic extracts at low concentrations improved angiogenic capacities of ECFCs and reduced proliferation, apoptosis, and the inflammatory response of these cells. Overall, mango leaves ethanolic extract provided the most promising results, but all three extracts ameliorated the functionality of ECFCs.

11.
Cells ; 11(8)2022 04 09.
Article in English | MEDLINE | ID: mdl-35455962

ABSTRACT

The risk of complications following surgical procedures is significantly increased in patients with SARS-CoV-2 infection. However, the mechanisms underlying these correlations are not fully known. Spinal cord injury (SCI) patients who underwent reconstructive surgery for pressure ulcers (PUs) before and during the COVID-19 pandemic were included in this study. The patient's postoperative progression was registered, and the subcutaneous white adipose tissue (s-WAT) surrounding the ulcers was analyzed by proteomic and immunohistochemical assays to identify the molecular/cellular signatures of impaired recovery. Patients with SCI and a COVID-19-positive diagnosis showed worse recovery and severe postoperative complications, requiring reintervention. Several proteins were upregulated in the adipose tissue of these patients. Among them, CKMT2 and CKM stood out, and CKM increased for up to 60 days after the COVID-19 diagnosis. Moreover, CKMT2 and CKM were largely found in MGCs within the s-WAT of COVID patients. Some of these proteins presented post-translational modifications and were targeted by autoantibodies in the serum of COVID patients. Overall, our results indicate that CKMT2, CKM, and the presence of MGCs in the adipose tissue surrounding PUs in post-COVID patients could be predictive biomarkers of postsurgical complications. These results suggest that the inflammatory response in adipose tissue may underlie the defective repair seen after surgery.


Subject(s)
COVID-19 , Pressure Ulcer , Spinal Cord Injuries , Adipose Tissue/metabolism , COVID-19/complications , COVID-19 Testing , Creatine Kinase/metabolism , Creatine Kinase, Mitochondrial Form/metabolism , Humans , Pandemics , Pressure Ulcer/epidemiology , Pressure Ulcer/etiology , Pressure Ulcer/surgery , Proteomics , SARS-CoV-2 , Spinal Cord Injuries/complications , Spinal Cord Injuries/surgery , Suppuration/complications , Up-Regulation
12.
Mol Med ; 28(1): 40, 2022 04 09.
Article in English | MEDLINE | ID: mdl-35397534

ABSTRACT

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has already caused 6 million deaths worldwide. While asymptomatic individuals are responsible of many potential transmissions, the difficulty to identify and isolate them at the high peak of infection constitutes still a real challenge. Moreover, SARS-CoV-2 provokes severe vascular damage and thromboembolic events in critical COVID-19 patients, deriving in many related deaths and long-hauler symptoms. Understanding how these processes are triggered as well as the potential long-term sequelae, even in asymptomatic individuals, becomes essential. METHODS: We have evaluated, by application of a proteomics-based quantitative approach, the effect of serum from COVID-19 asymptomatic individuals over circulating angiogenic cells (CACs). Healthy CACs were incubated ex-vivo with the serum of either COVID-19 negative (PCR -/IgG -, n:8) or COVID-19 positive asymptomatic donors, at different infective stages: PCR +/IgG - (n:8) and PCR -/IgG + (n:8). Also, a label free quantitative approach was applied to identify and quantify protein differences between these serums. Finally, machine learning algorithms were applied to validate the differential protein patterns in CACs. RESULTS: Our results confirmed that SARS-CoV-2 promotes changes at the protein level in the serum of infected asymptomatic individuals, mainly correlated with altered coagulation and inflammatory processes (Fibrinogen, Von Willebrand Factor, Thrombospondin-1). At the cellular level, proteins like ICAM-1, TLR2 or Ezrin/Radixin were only up-regulated in CACs treated with the serum of asymptomatic patients at the highest peak of infection (PCR + /IgG -), but not with the serum of PCR -/IgG + individuals. Several proteins stood out as significantly discriminating markers in CACs in response to PCR or IgG + serums. Many of these proteins particiArticle title: Kindly check and confirm the edit made in the article title.pate in the initial endothelial response against the virus. CONCLUSIONS: The ex vivo incubation of CACs with the serum of asymptomatic COVID-19 donors at different stages of infection promoted protein changes representative of the endothelial dysfunction and inflammatory response after viral infection, together with activation of the coagulation process. The current approach constitutes an optimal model to study the response of vascular cells to SARS-CoV-2 infection, and an alternative platform to test potential inhibitors targeting either the virus entry pathway or the immune responses following SARS-CoV-2 infection.


Subject(s)
COVID-19 , Humans , Immunoglobulin G , Nucleic Acid Amplification Techniques , SARS-CoV-2
13.
Philos Ethics Humanit Med ; 17(1): 5, 2022 03 16.
Article in English | MEDLINE | ID: mdl-35292071

ABSTRACT

BACKGROUND: During the first weeks of March 2020 in Spain, the cases of severe respiratory failure progressively increased, generating an imbalance between the clinical needs for advanced life support (ALS) measures and the effective availability of ALS resources. To address this problem, the creation of triage committees (TC) was proposed, whose main function is to select the best candidates to receive ALS. The main objective of our study is to describe the clinical characteristics of the patients evaluated by the TC of the Alcorcón Foundation University Hospital (AFUH) during the first wave of SARS CoV-2. Other objectives are to determine if there are differences between the patients considered candidates / not candidates for ALS and to analyze the functioning of the TC. METHODS: Retrospective observational study of all patients assessed by the AFUH TC. RESULTS: There were 19 meetings, in which 181 patients were evaluated, 65.4% male and with a mean age of 70.1 years. 31% had some degree of functional dependence, the Barthel median was 100 and Charlson 4. 58.5% were not considered a candidate for ALS at that time. The patients considered candidates to receive ALS were younger (72 vs 66; p < 0.001), had less comorbidity (Charlson 4 vs 3; p < 0.001) and had a better previous functional situation. A median of 5 physicians participated in each meeting and, after being assessed by the TC, 13.6% received ALS: 29.3% of those considered candidates for ALS and 2% of the non-candidates. CONCLUSIONS: The patients evaluated by the TC had a mean age of 70 years, high comorbidity and almost a third had some degree of functional dependence. More than half were not considered candidates for ALS at that time, these patients being older, with more comorbidity and a worse previous functional situation. TC decisions, based on objective clinical criteria, were almost always respected. Public institutions must get involved in triage procedures, which should and in our opinion must include the creation of TC in health centers. The implementation of Anticipated Decision programs (ADP) would help enable patients affected by triage decisions to participate in them.


Subject(s)
COVID-19 , Triage , Advanced Cardiac Life Support , Aged , Female , Humans , Male , Pandemics , Retrospective Studies
14.
Int J Antimicrob Agents ; 59(3): 106536, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35091054

ABSTRACT

This study aimed to assess the efficacy of ceftazidime/avibactam (C/A) in the treatment of infections due to Gram-negative bacteria (GNB) in critically ill patients. A multicentre, retrospective, observational study was conducted in critically ill patients receiving C/A for GNB infections. We evaluated demographic data, localisation and severity of infection, clinical and microbiological outcomes, and mortality. A total of 68 patients received C/A for serious GNB infections. The main infections were respiratory (33.8%), intra-abdominal (22.1%) and urinary tract infections (10.3%); bacteraemia was found in 22 cases (32.4%). Most infections were complicated by septic shock (58.8%) or sepsis (36.8%) and most of them required life-supporting therapies. Enterobacterales (79.4%) and Pseudomonas aeruginosa (19.1%) were the most frequently isolated bacteria; 84.2% of isolates were carbapenem-resistant. Thirty-four patients (50.0%) received C/A in combination with other antimicrobials. Fifty patients (73.5%) presented a favourable clinical response. Microbiological eradication was documented in 25 cases (36.8%). No significant differences were found in clinical response between patients treated with monotherapy or combined therapy (79.4% vs. 67.6%; P = 0.27). Overall intensive care unit (ICU) mortality was 41.2%. Univariate analysis showed that 30-day all-cause mortality was significantly (P < 0.05) associated with bacteraemia, previous corticosteroid use and the need of life-supporting therapies. C/A appears to be an effective therapy for severe infections due to GNB, including carbapenem-resistant isolates, in critically ill patients. C/A combination therapy was not associated with a higher clinical response. Mortality correlated significantly with the presence of bacteraemia, previous corticosteroid use and the need for life-supporting therapies.


Subject(s)
Ceftazidime , Gram-Negative Bacterial Infections , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Azabicyclo Compounds/therapeutic use , Ceftazidime/therapeutic use , Critical Illness , Drug Combinations , Drug Resistance, Multiple, Bacterial , Gram-Negative Bacteria , Gram-Negative Bacterial Infections/drug therapy , Gram-Negative Bacterial Infections/microbiology , Humans , Microbial Sensitivity Tests , Retrospective Studies
15.
J Strength Cond Res ; 36(7): 1936-1943, 2022 Jul 01.
Article in English | MEDLINE | ID: mdl-32740285

ABSTRACT

ABSTRACT: Neto, FR, Dorneles, JR, Luna, RM, Spina, MA, Gonçalves, CW, and Gomes Costa, RR. Performance differences between the arched and flat bench press in beginner and experienced Paralympic powerlifters. J Strength Cond Res 36(7): 1936-1943, 2022-The present study aimed to verify the differences of the total load, trajectory of the barbell in the sagittal plane, and mean velocity of the barbell between the arched and flat techniques of the bench press in beginner (BG) and experienced (EG) Paralympic powerlifters. Twenty beginners (age: 34.4 years; experience: 3.3 months) and 23 experienced (age: 35.5 years; experience: 9.8 months) Paralympic powerlifters were selected from a Rehabilitation Hospital Network and a Paralympic sports center. Subjects were assessed in the one-maximum repetition test of the bench press exercise using the flat and arched bench press techniques (48-72-hour interval between sessions). Maximum strength, trajectory of the barbell in the sagittal plane, and mean velocity of the barbell were measured to compare the techniques and the groups. The total load corrected with the Haleczko formula was significantly higher in EG compared with BG (∆ = 21.1%; effect sizes [ES] = 0.39, p ≤ 0.05). There were no significant differences for all analyzed outcomes comparing the arched and flat techniques. During the eccentric phase of the bench press, all assessed differences ranged from -16.6 to 23.1% and presented ES of trivial to moderate. On the concentric phase, the assessed differences ranged from -20.7 to 13.9% and presented ES of trivial to moderate. The total load, trajectory of the barbell in the sagittal plane, and mean velocity of the barbell were not significantly different between the arched and flat techniques for experienced and beginner powerlifting athletes during both the eccentric and concentric phase of the movement. However, further analyses are essential to determine the best technique for athletes.


Subject(s)
Resistance Training , Weight Lifting , Adult , Exercise Therapy , Humans , Movement , Muscle Strength , Resistance Training/methods
16.
Appl Opt ; 60(17): 5153-5160, 2021 Jun 10.
Article in English | MEDLINE | ID: mdl-34143087

ABSTRACT

We present a method for calibration and data extraction for a Stokes polarimeter working with three different wavelengths simultaneously. In the Stokes polarimeter considered in this work, we use two liquid crystal variable retarders (LCVRs) combined with a Glan-Thompson linear polarizer. A recently developed fitting calibration procedure is used. We use the same calibration samples and LCVR voltages for all three wavelengths, giving simultaneous measurement and calibration. We compare the performance of the polarimeter, after calibration, using four or six calibration samples in our experiment. To generate the four known calibration beams, we use a linear polarizer oriented at 130° and 30° with respect to the horizontal, a horizontal linear polarizer followed by a half-wave plate (at 632 nm) with its fast axis at 30°, and a horizontal linear polarizer followed by a quarter-wave plate (at 632 nm) with its fast axis at 30°. For calibration with six reference beams, we add two known calibration beams by setting the fast axis of the half- and quarter-wave plates at 130°. Experimental results show good agreement with the expected results, with the fitting calibration procedure giving an approximately 50% reduction in total RMS error with four calibration samples. There is a negligible reduction in the error when six calibration samples are used compared to the case with four samples.

17.
J Neurochem ; 158(3): 640-656, 2021 08.
Article in English | MEDLINE | ID: mdl-33942314

ABSTRACT

CB1 cannabinoid receptor is widely expressed in the central nervous system of animals from late prenatal development to adulthood. Appropriate activation and signaling of CB1 cannabinoid receptors in cortical interneurons are crucial during perinatal/postnatal ages and adolescence, when long-lasting changes in brain activity may elicit subsequent appearance of disorders in the adult brain. Here we used an optimized immunoprecipitation protocol based on specific antibodies followed by shot-gun proteomics to find CB1 interacting partners in postnatal rat GABAergic cortical neurons in vitro at two different stages of maturation. Besides describing new proteins associated with CB1 like dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex (DLAT), fatty acid synthase (FASN), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ), voltage-dependent anion channel 1 (VDAC1), myosin phosphatase Rho-interacting protein (MPRIP) or usher syndrome type-1C protein-binding protein 1 (USHBP1), we show that the signaling complex of CB1 is different between maturational stages. Interestingly, the CB1 signaling complex is enriched at the more immature stage in mitochondrial associated proteins and metabolic molecular functions, whereas at more mature stage, CB1 complex is increased in maturation and synaptic-associated proteins. We describe also interacting partners specifically immunoprecipitated with either N-terminal or C-terminal CB1 directed antibodies. Our results highlight new players that may be affected by altered cannabinoid signaling at this critical window of postnatal cortical development.


Subject(s)
Cerebral Cortex/embryology , Cerebral Cortex/metabolism , GABAergic Neurons/physiology , Receptor, Cannabinoid, CB1/genetics , Receptor, Cannabinoid, CB1/metabolism , Animals , Cells, Cultured , Female , Pregnancy , Rats , Rats, Wistar , Signal Transduction/physiology
18.
Int J Antimicrob Agents ; 57(3): 106270, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33347991

ABSTRACT

BACKGROUND: This study aimed to assess the efficacy of ceftolozane-tazobactam (C/T) for treating infections due to Pseudomonas aeruginosa (P. aeruginosa) in critically ill patients. PATIENTS AND METHODS: A multicenter, retrospective and observational study was conducted in critically ill patients receiving different C/T dosages and antibiotic combinations for P. aeruginosa infections. Demographic data, localisation and severity of infection, clinical and microbiological outcome, and mortality were evaluated. RESULTS: Ninety-five patients received C/T for P. aeruginosa serious infections. The main infections were nosocomial pneumonia (56.2%), intra-abdominal infection (10.5%), tracheobronchitis (8.4%), and urinary tract infection (6.3%). Most infections were complicated with sepsis (49.5%) or septic shock (45.3%), and bacteraemia (10.5%). Forty-six episodes were treated with high-dose C/T (3 g every 8 hours) and 38 episodes were treated with standard dosage (1.5 g every 8 hours). Almost half (44.2%) of the patients were treated with C/T monotherapy, and the remaining group received combination therapy with other antibiotics. Sixty-eight (71.6%) patients presented a favourable clinical response. Microbiological eradication was documented in 42.1% (40/95) of the episodes. The global ICU mortality was 36.5%. Univariate analysis showed that 30-day mortality was significantly associated (P < 0.05) with Charlson Index at ICU admission and the need of life-supporting therapies. CONCLUSIONS: C/T appeared to be an effective therapy for severe infections due to P. aeruginosa in critically ill patients. Mortality was mainly related to the severity of the infection. No benefit was observed with high-dose C/T or combination therapy with other antibiotics.


Subject(s)
Cephalosporins/therapeutic use , Pseudomonas Infections/drug therapy , Pseudomonas Infections/mortality , Pseudomonas aeruginosa/drug effects , Tazobactam/therapeutic use , Adult , Aged , Aged, 80 and over , Anti-Bacterial Agents/therapeutic use , Critical Illness , Cross Infection/drug therapy , Cross Infection/mortality , Dose-Response Relationship, Drug , Drug Resistance, Multiple, Bacterial , Female , Humans , Intensive Care Units , Male , Microbial Sensitivity Tests , Middle Aged , Pseudomonas aeruginosa/isolation & purification , Retrospective Studies , Spain , Treatment Outcome
19.
Cir Cir ; 88(6): 679-683, 2020.
Article in English | MEDLINE | ID: mdl-33254181

ABSTRACT

BACKGROUND: At present benign and malignant pancreatobiliar diseases, required a multidisciplinary management, and the different techniques to solve this pathology has been improve with new technology and surgeons training. OBJECTIVE: The purpose of this article is to describe that de minimally invasive surgery on benign and malignant pancreatobiliar diseases is feasible and offer good results. METHOD: In Hospital regional ISSSTE Puebla from July 2013 to July 2018, 30 consecutive patients with different pancreas and biliary pathologies have been operated by totally laparoscopic, by the same surgeon and surgical team, and following for up to 4 years. RESULTS: None all the patients presented sever complications during surgery or after immediate surgery and no reoperations. Surgical time and blood loss was acceptable. CONCLUSION: The results we obtained are positive and encouraging because the morbidity and mortality are similar to the different sources we reviewed, nevertheless, this study is our initial experience and we required more cases to obtain a significant sample.


ANTECEDENTES: En la actualidad las enfermedades benignas y malignas de las vías biliares y del páncreas requieren un manejo multidisciplinario, y las técnicas para resolver este tipo de padecimientos han avanzado gracias al mejoramiento en la tecnología y la capacidad del personal humano. OBJETIVO: El propósito de este trabajo es presentar varios casos y demostrar que la cirugía de mínima invasión en patología benigna y maligna pancreatobiliar es viable y se pueden obtener adecuados resultados. MÉTODO: De julio de 2013 a julio de 2018, 30 pacientes consecutivos han sido intervenidos quirúrgicamente por vía laparoscópica por diversas enfermedades que involucran páncreas y vías biliares, por el mismo cirujano y equipo quirúrgico, con un seguimiento de hasta 4 años. RESULTADOS: Ninguno de los pacientes intervenidos presentó complicaciones graves en el transoperatorio ni en el posquirúrgico inmediato, ninguno fue sometido a una segunda cirugía y los tiempos quirúrgicos y el sangrado fueron aceptables. CONCLUSIÓN: Los resultados son positivos y muy alentadores, ya que la morbilidad fue muy similar a la de las series que se revisaron; sin embargo, este estudio es nuestra experiencia inicial y requerimos aumentar el número de casos para obtener una muestra significativa.


Subject(s)
Laparoscopy , Surgeons , Hospitals , Humans , Pica , Retrospective Studies
20.
J Pers Med ; 10(4)2020 Oct 21.
Article in English | MEDLINE | ID: mdl-33096745

ABSTRACT

Despite promising advances in the medical management of spinal cord injury (SCI), there is still no available effective therapy to repair the neurological damage in patients who experience this life-transforming condition. Recently, we performed a phase II/III placebo-controlled randomized trial of safety and efficacy of growth hormone (GH) treatment in incomplete chronic traumatic spinal cord injury. The main findings were that the combined treatment of GH plus rehabilitation treatment is feasible and safe, and that GH but not placebo slightly improves the SCI individual motor score. Moreover, we found that an intensive and long-lasting rehabilitation program per se increases the functional outcome of SCI individuals. To understand the possible mechanisms of the improvement due to GH treatment (motor score) and due to rehabilitation (functional outcome), we used a proteomic approach. Here, we used a multiple proteomic strategy to search for recovery biomarkers in blood plasma with the potential to predict response to somatropin treatment and to delayed intensive rehabilitation. Forty-six patients were recruited and followed for a minimum period of 1 year. Patients were classified into two groups based on their treatment: recombinant somatropin (0.4 mg) or placebo. Both groups received rehabilitation treatment. Our strategy allowed us to perform one of the deepest plasma proteomic analyses thus far, which revealed two proteomic signatures with predictive value: (i) response to recombinant somatropin treatment and (ii) response to rehabilitation. The proteins implicated in these signatures are related to homeostasis, inflammation, and coagulation functions. These findings open novel possibilities to assess and therapeutically manage patients with SCI, which could have a positive impact on their clinical response.

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