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2.
Nat Struct Mol Biol ; 2024 May 02.
Article in English | MEDLINE | ID: mdl-38698207

ABSTRACT

Vortioxetine (VTX) is a recently approved antidepressant that targets a variety of serotonin receptors. Here, we investigate the drug's molecular mechanism of operation at the serotonin 5-HT3 receptor (5-HT3R), which features two properties: VTX acts differently on rodent and human 5-HT3R, and VTX appears to suppress any subsequent response to agonists. Using a combination of cryo-EM, electrophysiology, voltage-clamp fluorometry and molecular dynamics, we show that VTX stabilizes a resting inhibited state of the mouse 5-HT3R and an agonist-bound-like state of human 5-HT3R, in line with the functional profile of the drug. We report four human 5-HT3R structures and show that the human receptor transmembrane domain is intrinsically fragile. We also explain the lack of recovery after VTX administration via a membrane partition mechanism.

3.
Cell ; 187(12): 2990-3005.e17, 2024 Jun 06.
Article in English | MEDLINE | ID: mdl-38772370

ABSTRACT

Integrins link the extracellular environment to the actin cytoskeleton in cell migration and adhesiveness. Rapid coordination between events outside and inside the cell is essential. Single-molecule fluorescence dynamics show that ligand binding to the bent-closed integrin conformation, which predominates on cell surfaces, is followed within milliseconds by two concerted changes, leg extension and headpiece opening, to give the high-affinity integrin conformation. The extended-closed integrin conformation is not an intermediate but can be directly accessed from the extended-open conformation and provides a pathway for ligand dissociation. In contrast to ligand, talin, which links the integrin ß-subunit cytoplasmic domain to the actin cytoskeleton, modestly stabilizes but does not induce extension or opening. Integrin activation is thus initiated by outside-in signaling and followed by inside-out signaling. Our results further imply that talin binding is insufficient for inside-out integrin activation and that tensile force transmission through the ligand-integrin-talin-actin cytoskeleton complex is required.


Subject(s)
Integrins , Talin , Animals , Humans , Mice , Actin Cytoskeleton/metabolism , Actin Cytoskeleton/chemistry , Cell Adhesion , CHO Cells , Cricetulus , Integrins/metabolism , Integrins/chemistry , Ligands , Protein Binding , Protein Conformation , Signal Transduction , Single Molecule Imaging , Talin/metabolism , Talin/chemistry
4.
J Med Chem ; 66(21): 14928-14947, 2023 11 09.
Article in English | MEDLINE | ID: mdl-37797083

ABSTRACT

The multifactorial origin and neurochemistry of Alzheimer's disease (AD) call for the development of multitarget treatment strategies. We report a first-in-class triple acting compound that targets serotonin type 6 and 3 receptors (5-HT-Rs) and monoamine oxidase type B (MAO-B) as an approach for treating AD. The key structural features required for MAO-B inhibition and 5-HT6R antagonism and interaction with 5-HT3R were determined using molecular dynamic simulations and cryo-electron microscopy, respectively. Bioavailable PZ-1922 reversed scopolamine-induced cognitive deficits in the novel object recognition test. Furthermore, it displayed superior pro-cognitive properties compared to intepirdine (a 5-HT6R antagonist) in the AD model, which involved intracerebroventricular injection of an oligomeric solution of amyloid-ß peptide (oAß) in the T-maze test in rats. PZ-1922, but not intepirdine, restored levels of biomarkers characteristic of the debilitating effects of oAß. These data support the potential of a multitarget approach involving the joint modulation of 5-HT6R/5-HT3R/MAO-B in AD.


Subject(s)
Alzheimer Disease , Serotonin , Rats , Animals , Serotonin/adverse effects , Cryoelectron Microscopy , Receptors, Serotonin , Serotonin Antagonists/pharmacology , Serotonin Antagonists/therapeutic use , Alzheimer Disease/drug therapy , Alzheimer Disease/chemically induced , Monoamine Oxidase , Cognition , Monoamine Oxidase Inhibitors/pharmacology , Monoamine Oxidase Inhibitors/therapeutic use
5.
J Phys Chem C Nanomater Interfaces ; 127(29): 14146-14154, 2023 Jul 27.
Article in English | MEDLINE | ID: mdl-37529663

ABSTRACT

Transition-metal nanocatalysis represents a novel alternative currently experiencing flourishing progress to tackle the tumor microenvironment (TME) in cancer therapy. These nanomaterials aim at attacking tumor cells using the intrinsic selectivity of inorganic catalysts. In addition, special attention to tune and control the release of these transition metals is also required. Understanding the chemical reactions behind the catalytic action of the transition-metal nanocatalysts and preventing potential undesired side reactions caused by acute cytotoxicity of the released ionic species represent another important field of research. Specifically, copper-based oxides may suffer from acute leaching that potentially may induce toxicity not only to target cancer cells but also to nearby cells and tissues. In this work, we propose the synthesis of chalcopyrite (CuFeS2) nanostructures capable of triggering two key reactions for an effective chemodynamic therapy (CDT) in the heterogeneous phase: (i) glutathione (GSH) oxidation and (ii) oxidation of organic substrates using H2O2, with negligible leaching of metals under TME-like conditions. This represents an appealing alternative toward the development of safer copper-iron-based nanocatalytic materials with an active catalytic response without incurring leaching side phenomena.

6.
Biophys J ; 122(10): 1846-1857, 2023 05 16.
Article in English | MEDLINE | ID: mdl-37077048

ABSTRACT

Single-particle electron cryo-microscopy (cryo-EM) has become an effective and straightforward approach to determine the structure of membrane proteins. However, obtaining cryo-EM grids of sufficient quality for high-resolution structural analysis remains a major bottleneck. One of the difficulties arises from the presence of detergents, which often leads to a lack of control of the ice thickness. Amphipathic polymers such as amphipols (APols) are detergent substitutes, which have proven to be valuable tools for cryo-EM studies. In this work, we investigate the physico-chemical behavior of APol- and detergent-containing solutions and show a correlation with the properties of vitreous thin films in cryo-EM grids. This study provides new insight on the potential of APols, allowing a better control of ice thickness while limiting protein adsorption at the air-water interface, as shown with the full-length mouse serotonin 5-HT3A receptor whose structure has been solved in APol. These findings may speed up the process of grid optimization to obtain high-resolution structures of membrane proteins.


Subject(s)
Detergents , Surface-Active Agents , Animals , Mice , Surface-Active Agents/chemistry , Cryoelectron Microscopy , Electrons , Ice , Membrane Proteins
7.
Biotechnol Appl Biochem ; 70(3): 1176-1188, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36575961

ABSTRACT

This work describes the genetic transformation of a strain of Aspergillus niger with five different constructs containing 16 different heterologous genes, coding for four oxidoreductases, two cellobiohydrolases, one endoglucanase, one ß-glucosidase, six enzymes involved in xylose metabolism, and two enzymes involved in fermentation. The aim was to try and engineer a consolidated bioprocessing in A. niger. The fungus already contains most of these enzymes and we only enhanced endogenous activities. We recovered nine transformants containing all genes, as indicated by polymerase chain reaction (PCR). To confirm that the products of the genes were functional, we measured the activity of five different enzymes in all the strains, and they all showed enhanced activity over the wild-type (wt) strain. The strains were grown on carboxymethyl cellulose (CMC) and xylan as substrates, and they produced considerably more ethanol than the wt. The levels of ethanol production were comparable to those reported in the literature.


Subject(s)
Aspergillus niger , Cellulase , Ethanol/metabolism , Metabolic Engineering , Cellulase/metabolism , Fermentation
8.
Brain Sci ; 11(11)2021 Oct 29.
Article in English | MEDLINE | ID: mdl-34827445

ABSTRACT

BACKGROUND: Previous systematic reviews report that arterial hypertension (AHT) is associated with lower performance in cognition in the elderly. However, some studies show that with higher blood pressure, a better cognitive performance is obtained. OBJECTIVE: The aim of this study was to determine the relationship between AHT with cognitive performance in the elderly. METHODS: the review involved a search on PubMed, Scopus and PsycINFO databases from January 1990 to March, 2020 to identify the relationship among AHT and cognitive performance in older people. RESULTS: 1170 articles were identified, 136 complete papers were reviewed, a qualitative analysis of 26 studies and a quantitative analysis of eight studies were carried out. It was found that people with AHT have a lower performance in processing speed SMD = 0.40 (95% CI: 0.25, 0.54), working memory SMD = 0.28 (95% CI: 0.15, 0.41) in short-term memory and learning SMD = -0.27 (95% CI: -0.37, -0.17) and delayed recall SMD = -0.20 (95% CI: -0.35, -0.05). Only one study found that higher blood pressure was associated with better memory performance. CONCLUSION: Our results suggest that high blood pressure primarily affects processing speed, working memory, short-term memory and learning and delayed recall.

9.
Nat Methods ; 18(1): 60-68, 2021 01.
Article in English | MEDLINE | ID: mdl-33408403

ABSTRACT

Nanobodies are popular and versatile tools for structural biology. They have a compact single immunoglobulin domain organization, bind target proteins with high affinities while reducing their conformational heterogeneity and stabilize multi-protein complexes. Here we demonstrate that engineered nanobodies can also help overcome two major obstacles that limit the resolution of single-particle cryo-electron microscopy reconstructions: particle size and preferential orientation at the water-air interfaces. We have developed and characterized constructs, termed megabodies, by grafting nanobodies onto selected protein scaffolds to increase their molecular weight while retaining the full antigen-binding specificity and affinity. We show that the megabody design principles are applicable to different scaffold proteins and recognition domains of compatible geometries and are amenable for efficient selection from yeast display libraries. Moreover, we demonstrate that megabodies can be used to obtain three-dimensional reconstructions for membrane proteins that suffer from severe preferential orientation or are otherwise too small to allow accurate particle alignment.


Subject(s)
Cryoelectron Microscopy/methods , Lipids/chemistry , Multiprotein Complexes/chemistry , Receptors, GABA-A/chemistry , Single Molecule Imaging/methods , Single-Cell Analysis/methods , Single-Domain Antibodies/chemistry , Humans , Models, Molecular , Molecular Structure , Protein Conformation
10.
Int J Qual Health Care ; 32(10): 658-662, 2020 Dec 15.
Article in English | MEDLINE | ID: mdl-32986101

ABSTRACT

OBJECTIVE: Identifying when and how often decisions are made based on high-quality evidence can inform the development of evidence-based treatment plans and care pathways, which have been shown to improve quality of care and patient safety. Evidence to guide decision-making, national guidelines and clinical pathways for many conditions in pediatric orthopedic surgery are limited. This study investigated decision-making rationale and quantified the evidence supporting decisions made by pediatric orthopedic surgeons in an outpatient clinic. DESIGN/SETTING/PARTICIPANTS/INTERVENTION(S)/MAIN OUTCOME MEASURE(S): We recorded decisions made by eight pediatric orthopedic surgeons in an outpatient clinic and the surgeon's reported rationale behind the decisions. Surgeons categorized the rationale for each decision as one or a combination of 12 possibilities (e.g. 'Experience/anecdote,' 'First principles,' 'Trained to do it,' 'Arbitrary/instinct,' 'General study,' 'Specific study'). RESULTS: Out of 1150 total decisions, the most frequent decisions were follow-up scheduling, followed by bracing prescription/removal. The most common decision rationales were 'First principles' (n = 310, 27.0%) and 'Experience/anecdote' (n = 253, 22.0%). Only 17.8% of decisions were attributed to scientific studies, with 7.3% based on studies specific to the decision. As high as 34.6% of surgical intervention decisions were based on scientific studies, while only 10.4% of follow-up scheduling decisions were made with studies in mind. Decision category was significantly associated with a basis in scientific studies: surgical intervention and medication prescription decisions were more likely to be based on scientific studies than all other decisions. CONCLUSIONS: With increasing emphasis on high value, evidence-based care, understanding the rationale behind physician decision-making can educate physicians, identify common decisions without supporting evidence and help create clinical care pathways in pediatric orthopedic surgery. Decisions based on evidence or consensus between surgeons can inform pathways and national guidelines that minimize unwarranted variation in care and waste. Decision support tools and aids could also be implemented to guide these decisions.


Subject(s)
Orthopedic Procedures , Orthopedics , Surgeons , Child , Clinical Decision-Making , Humans
11.
Front Immunol ; 11: 1562, 2020.
Article in English | MEDLINE | ID: mdl-32793218

ABSTRACT

Thymocyte differentiation is dependent on the availability and transport of metabolites in the thymus niche. As expression of metabolite transporters is a rate-limiting step in nutrient utilization, cell surface transporter levels generally reflect the cell's metabolic state. The GLUT1 glucose transporter is upregulated on actively dividing thymocytes, identifying thymocytes with an increased metabolism. However, it is not clear whether transporters of essential elements such as phosphate are modulated during thymocyte differentiation. While PiT1 and PiT2 are both phosphate transporters in the SLC20 family, we show here that they exhibit distinct expression profiles on both murine and human thymocytes. PiT2 expression distinguishes thymocytes with high metabolic activity, identifying immature murine double negative (CD4-CD8-) DN3b and DN4 thymocyte blasts as well as immature single positive (ISP) CD8 thymocytes. Notably, the absence of PiT2 expression on RAG2-deficient thymocytes, blocked at the DN3a stage, strongly suggests that high PiT2 expression is restricted to thymocytes having undergone a productive TCRß rearrangement at the DN3a/DN3b transition. Similarly, in the human thymus, PiT2 was upregulated on early post-ß selection CD4+ISP and TCRαß-CD4hiDP thymocytes co-expressing the CD71 transferrin receptor, a marker of metabolic activity. In marked contrast, expression of the PiT1 phosphate importer was detected on mature CD3+ murine and human thymocytes. Notably, PiT1 expression on CD3+DN thymocytes was identified as a biomarker of an aging thymus, increasing from 8.4 ± 1.5% to 42.4 ± 9.4% by 1 year of age (p < 0.0001). We identified these cells as TCRγδ and, most significantly, NKT, representing 77 ± 9% of PiT1+DN thymocytes by 1 year of age (p < 0.001). Thus, metabolic activity and thymic aging are associated with distinct expression profiles of the PiT1 and PiT2 phosphate transporters.


Subject(s)
Cell Differentiation , Phosphate Transport Proteins/metabolism , Thymocytes/metabolism , Animals , Biomarkers , Cell Differentiation/genetics , Gene Expression Profiling , Glucose Transporter Type 1/genetics , Glucose Transporter Type 1/metabolism , Humans , Immunophenotyping , Lymphocyte Subsets/immunology , Lymphocyte Subsets/metabolism , Mice , Mice, Knockout , Phosphate Transport Proteins/genetics , Sodium-Phosphate Cotransporter Proteins, Type III/genetics , Sodium-Phosphate Cotransporter Proteins, Type III/metabolism , Thymocytes/cytology , Thymocytes/immunology , Thymus Gland/cytology , Thymus Gland/immunology , Thymus Gland/metabolism , Transcriptome
12.
J Biol Chem ; 295(28): 9366-9378, 2020 07 10.
Article in English | MEDLINE | ID: mdl-32393577

ABSTRACT

Solute carrier family 20 member 2 (SLC20A2) and xenotropic and polytropic retrovirus receptor 1 (XPR1) are transporters with phosphate uptake and efflux functions, respectively. Both are associated with primary familial brain calcification (PFBC), a genetic disease characterized by cerebral calcium-phosphate deposition and associated with neuropsychiatric symptoms. The association of the two transporters with the same disease suggests that they jointly regulate phosphate fluxes and cellular homeostasis, but direct evidence is missing. Here, we found that cross-talk between SLC20A2 and XPR1 regulates phosphate homeostasis, and we identified XPR1 as a key inositol polyphosphate (IP)-dependent regulator of this process. We found that overexpression of WT SLC20A2 increased phosphate uptake, as expected, but also unexpectedly increased phosphate efflux, whereas PFBC-associated SLC20A2 variants did not. Conversely, SLC20A2 depletion decreased phosphate uptake only slightly, most likely compensated for by the related SLC20A1 transporter, but strongly decreased XPR1-mediated phosphate efflux. The SLC20A2-XPR1 axis maintained constant intracellular phosphate and ATP levels, which both increased in XPR1 KO cells. Elevated ATP levels are a hallmark of altered inositol pyrophosphate (PP-IP) synthesis, and basal ATP levels were restored after phosphate efflux rescue with WT XPR1 but not with XPR1 harboring a mutated PP-IP-binding pocket. Accordingly, inositol hexakisphosphate kinase 1-2 (IP6K1-2) gene inactivation or IP6K inhibitor treatment abolished XPR1-mediated phosphate efflux regulation and homeostasis. Our findings unveil an SLC20A2-XPR1 interplay that depends on IPs such as PP-IPs and controls cellular phosphate homeostasis via the efflux route, and alteration of this interplay likely contributes to PFBC.


Subject(s)
Homeostasis , Inositol Phosphates/metabolism , Receptors, G-Protein-Coupled/metabolism , Receptors, Virus/metabolism , Sodium-Phosphate Cotransporter Proteins, Type III/metabolism , Adenosine Triphosphate/genetics , Adenosine Triphosphate/metabolism , Cell Line , Humans , Inositol Phosphates/genetics , Phosphotransferases (Phosphate Group Acceptor)/genetics , Phosphotransferases (Phosphate Group Acceptor)/metabolism , Receptors, G-Protein-Coupled/genetics , Receptors, Virus/genetics , Sodium-Phosphate Cotransporter Proteins, Type III/genetics , Xenotropic and Polytropic Retrovirus Receptor
13.
Protein Expr Purif ; 168: 105570, 2020 04.
Article in English | MEDLINE | ID: mdl-31953182

ABSTRACT

Aspergillus niger has been employed to produce heterologous proteins due to its high capacity for expression and secretion; nevertheless, expression levels of human proteins have been modest. We were interested in investigating whether A. niger can express and secret human erythropoietin (HuEPO) at high yields. Our strategy was to combine the presence of introns with CRISPR-Cas9 to increase the yield of the recombinant protein. The epo gene was codon-optimized and its expression driven by the PmbfA promoter. Another version of epo contained introns from the fructose-1,6-bisphosphatase (fbp) gene. Two recombinant clones, uME12 (no introns) and uME23 (with introns), were selected based on the resistance to the antibiotic and because they showed a protein profile different from that of the parental strain, as shown by SDS-PAGE. Expression of epo was confirmed by RT-PCR in both colonies but the recombinant EPO protein (rHUEPO) was detected by Western blot only in uME23. The rHuEPO yield from uME23 was estimated at about 1.8 mg L-1 by ELISA, demonstrating that the presence of introns resulted in higher yield, possibly by conferring more stability to mRNA. On the other hand, as part of our strategy we decided to inactivate in the strain uME23 the following genes vps, prtT, algC and och1 which are involved in protein secretion, regulating of protease expression and protein glycosylation in A. niger, with CRISPR-Cas9, yielding the muPS20 transformant. muPS20 is a protease-free strain and its rHuEPO production level was increased 41.1-fold. Moreover, its molecular weight was ≈27 kDa showing that mutations in the above mentioned genes improved secretion, prevented proteolytic degradation and hyperglycosylation of heterologous protein.


Subject(s)
Aspergillus niger/genetics , Erythropoietin/biosynthesis , Genes, Fungal , Introns , Plasmids/metabolism , RNA, Messenger/genetics , Aspergillus niger/metabolism , CRISPR-Cas Systems , Cloning, Molecular , Erythropoietin/genetics , Fructose-Bisphosphatase/chemistry , Fructose-Bisphosphatase/genetics , Gene Expression , Gene Knockdown Techniques , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Glycosylation , Humans , Plasmids/chemistry , Promoter Regions, Genetic , Protein Stability , Proteolysis , RNA, Messenger/metabolism , Recombinant Proteins/biosynthesis , Recombinant Proteins/genetics
14.
JCI Insight ; 4(19)2019 10 03.
Article in English | MEDLINE | ID: mdl-31578306

ABSTRACT

BACKGROUNDFecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridioides difficile infection (rCDI) in adults and children, but donor stool samples are currently screened for only a limited number of potential pathogens. We sought to determine whether putative procarcinogenic bacteria (enterotoxigenic Bacteroides fragilis, Fusobacterium nucleatum, and Escherichia coli harboring the colibactin toxin) could be durably transmitted from donors to patients during FMT.METHODSStool samples were collected from 11 pediatric rCDI patients and their respective FMT donors prior to FMT as well as from the patients at 2-10 weeks, 10-20 weeks, and 6 months after FMT. Bacterial virulence factors in stool DNA extracts and stool cultures were measured by quantitative PCR: Bacteroides fragilis toxin (bft), Fusobacterium adhesin A (fadA), and Escherichia coli colibactin (clbB).RESULTSFour of 11 patients demonstrated sustained acquisition of a procarcinogenic bacteria. Whole genome sequencing was performed on colony isolates from one of these donor/recipient pairs and demonstrated that clbB+ E. coli strains present in the recipient after FMT were identical to a strain present in the donor, confirming strain transmission. Conversely, 2 patients exhibited clearance of procarcinogenic bacteria following FMT from a negative donor.CONCLUSIONBoth durable transmission and clearance of procarcinogenic bacteria occurred following FMT, suggesting that additional studies on appropriate screening measures for FMT donors and the long-term consequences and/or benefits of FMT are warranted.FUNDINGCrohn's & Colitis Foundation, the Bloomberg~Kimmel Institute for Cancer Immunotherapy at Johns Hopkins University School of Medicine, the National Cancer Institute, and the Canadian Institutes of Health Research.


Subject(s)
Bacteria/classification , Clostridium Infections/microbiology , Clostridium Infections/therapy , Fecal Microbiota Transplantation/methods , Adolescent , Bacteria/genetics , Child , Child, Preschool , Clostridioides difficile , Cohort Studies , DNA, Bacterial , Feces/microbiology , Female , Humans , Male , RNA, Ribosomal, 16S/genetics , Virulence Factors , Whole Genome Sequencing
15.
Sci Rep ; 9(1): 6776, 2019 05 01.
Article in English | MEDLINE | ID: mdl-31043717

ABSTRACT

Primary familial brain calcification (PFBC) is a rare neurological disease characterized by deposits of calcium phosphate in the basal ganglia and other regions of the brain. Pathogenic variants in the XPR1/SLC53A1 gene, which encodes the only known inorganic phosphate exporter, cause an autosomal dominant form of PFBC. These variants are typically located in the SPX N-terminal domain of the protein. Here, we characterize three XPR1 variants outside of SPX in three PFBC patients with an apparently sporadic presentation: c.1375C > T p.(R459C), c.1855A > G p.(N619D) and c.1886T > G p.(I629S), with the latter identified as the first XPR1/SLC53A1 de novo mutation to occur in a PFBC proband. When tested in an in vitro physiological complementation assay, the three XPR1 variants were impaired in phosphate export function, although they were normally expressed at the cell surface and could serve as functional receptors for retrovirus entry. Moreover, peripheral blood cells from the p.N619D patient could be assayed ex vivo and displayed significantly impaired phosphate export. Our results establish for the first time the clinical and molecular characteristics of XPR1 variants located outside the SPX domain and assert a direct link between these variants, deficient phosphate export, and PFBC. Moreover, we unveiled new structural features in XPR1 C-terminal domain that play a role in phosphate export and disease.


Subject(s)
Brain Diseases/pathology , Calcinosis/pathology , Mutation , Peptide Hormones/genetics , Phosphates/metabolism , Receptors, G-Protein-Coupled/genetics , Receptors, Virus/genetics , Brain Diseases/genetics , Calcinosis/genetics , Female , Genetic Predisposition to Disease , Humans , Male , Pedigree , Protein Domains , Xenotropic and Polytropic Retrovirus Receptor
16.
Repert. med. cir ; 27(2): 100-104, 2018. ilus., tab.
Article in English, Spanish | LILACS, COLNAL | ID: biblio-981827

ABSTRACT

Objetivo: determinar la concordancia entre el cateterismo cardiaco y el ecocardiograma en pacientes con diagnóstico de síndrome coronario en las primeras 48 horas de su ingreso al servicio de urgencias. Métodos: estudio descriptivo de concordancia, se incluyeron pacientes con diagnóstico de angina inestable e infarto agudo del miocardio (IAM) con y sin elevación del segmento ST y se obtuvo la información de las dos pruebas en las historias clínicas. Se calculó el grado de acuerdo entre los hallazgos de contractibilidad y el compromiso de obstrucción por medio del índice kappa. Resultados: se incluyeron 129 pacientes con edad promedio de 65.2 años (DE 12.15), 65.1% fueron mujeres. La frecuencia de oclusión del flujo coronario en el cateterismo fue 74.4%, la alteración de contractibilidad en el ecocardiograma 69.8%. Se observó una baja concordancia entre los hallazgos de estas pruebas diagnósticas (acuerdo: 70.5%; kappa: 0.27, p=0.001). El principal trastorno de contractilidad fue la hipoquinesia (42.6%) y en la mayoría se presentó obstrucción de un único vaso (44.2%). Conclusiones: aunque la concordancia entre el ecocardiograma y el cateterismo es baja en este estudio, el primero junto con la valoración clínica es un procedimiento importante en el diagnóstico y seguimiento de los pacientes con patología cardiaca en el servicio de urgencias, para proceder a una intervención temprana y disminuir el riesgo de lesión miocárdica, así como la mortalidad y la morbilidad.


Objective: to determine agreement between findings on cardiac catheterization and echocardiogram in patients diagnosed with coronary syndrome within 48 hours of admission to the emergency department (ED). Methods: a descriptive study on agreement. Patients diagnosed with unstable angina and ST-segment elevation or non-ST-segment elevation acute myocardial infarction (AMI), were included, obtaining data on the two tests from clinical records. The level of agreement between contractility and coronary flow obstructive findings was reported using the kappa coefficient. Results: the study included 129 patients with mean age 65.2 years (SD 12.15), of which 65.1% were women. The coronary occlusion rate on catheterization was 74.4%, and contractility alteration on echocardiogram was 69.8%. Low agreement between findings on these two tests was observed (agreement: 70.5%; kappa: 0.27, p=0.001). The main contractility alteration was hypokinesia (42.6%) and most presented obstruction of only one artery (44.2%). Conclusion: although agreement is low between echocardiogram and catheterization in this study, echocardiogram coupled with clinical examination constitutes an important diagnostic and follow-up procedure for patients with cardiovascular disease at the ED, in order to permit an early intervention minimizing the risk of myocardial damage, as well as, mortality and morbidity.


Subject(s)
Humans , Female , Aged , Cardiac Catheterization , Echocardiography , Acute Coronary Syndrome
17.
J Neurol ; 263(8): 1559-64, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27230854

ABSTRACT

Mutations in XPR1, a gene encoding an inorganic phosphate exporter, have recently been identified in patients with primary familial brain calcification (PFBC). Using Sanger sequencing, we screened XPR1 in 18 unrelated patients with PFBC and no SLC20A2, PDGFB, or PDGFRB mutation. XPR1 variants were tested in an in vitro physiological complementation assay and patient blood cells were assessed ex vivo for phosphate export. We identified a novel c.260T > C, p.(Leu87Pro) XPR1 variant in a 41-year-old man complaining of micrographia and dysarthria and demonstrating mild parkinsonism, cerebellar ataxia and executive dysfunction. Brain (123)I-Ioflupane scintigraphy showed marked dopaminergic neuron loss. Peripheral blood cells from the patient exhibited decreased phosphate export. XPR1 in which we introduced the mutation was not detectable at the cell surface and did not lead to phosphate export. These results confirm that loss of XPR1-mediated phosphate export function causes PFBC, occurring in less than 8 % of cases negative for the other genes, and may be responsible for parkinsonism.


Subject(s)
Brain Diseases/genetics , Calcinosis/genetics , Family Health , Mutation/genetics , Receptors, G-Protein-Coupled/genetics , Receptors, Virus/genetics , Adult , Brain Diseases/diagnostic imaging , Calcinosis/diagnostic imaging , Female , Genetic Association Studies , Humans , Magnetic Resonance Imaging , Male , Nortropanes/pharmacokinetics , RNA, Small Interfering/genetics , RNA, Small Interfering/metabolism , Radionuclide Imaging , Transfection , Xenotropic and Polytropic Retrovirus Receptor
18.
Intervirology ; 59(5-6): 235-242, 2016.
Article in English | MEDLINE | ID: mdl-28329739

ABSTRACT

AIMS: The aim of this study was to design peptides derived from glycoproteins H (gH) and B (gB) of herpes simplex viruses type 1 (HSV-1) and type 2 (HSV-2) with the potential to block herpetic infection and to evaluate their ability to inhibit HSV-1 and HSV-2 infection in vitro. METHODS: A library of continuous 15-25 residue stretches (CRSs) located at the surface of gH and gB from HSV-1 and HSV-2 was created. These CRSs were analyzed, and only those that were highly flexible and rich in charged residues were selected for the design of the antiviral peptides (AVPs). The toxicity of the AVPs was evaluated by MTT reduction assays. Virucidal activity of the AVPs was determined by a plaque reduction assay, and their antiviral effect was measured by cell viability assays. RESULTS AND CONCLUSION: Four AVPs (CB-1, CB-2, U-1, and U-2) derived from gB and gH were designed and synthetized, none of which showed high levels of toxicity in Vero cells. The U-1 and U-2 gB-derived AVPs showed high virucidal and antiviral activities against both HSV-1 and HSV-2. The gH-derived peptide CB-1 showed high virucidal and antiviral activities against HSV-2, while CB-2 showed similar results against HSV-1. The peptides CB-1 and CB-2 showed higher IC50 values than the U-1 and U-2 peptides.

19.
Nat Genet ; 47(6): 579-81, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25938945

ABSTRACT

Primary familial brain calcification (PFBC) is a neurological disease characterized by calcium phosphate deposits in the basal ganglia and other brain regions and has thus far been associated with SLC20A2, PDGFB or PDGFRB mutations. We identified in multiple families with PFBC mutations in XPR1, a gene encoding a retroviral receptor with phosphate export function. These mutations alter phosphate export, implicating XPR1 and phosphate homeostasis in PFBC.


Subject(s)
Brain Diseases, Metabolic, Inborn/genetics , Calcinosis/genetics , Receptors, G-Protein-Coupled/genetics , Receptors, Virus/genetics , DNA Mutational Analysis , Female , Genetic Association Studies , Genetic Predisposition to Disease , HEK293 Cells , Humans , Lod Score , Male , Middle Aged , Mutation, Missense , Neurodegenerative Diseases/genetics , Pedigree , Xenotropic and Polytropic Retrovirus Receptor
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