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BACKGROUND Platelets are now recognized as immunological sentries in the first line of defense that participate in the detection and response to pathogens. This frequently results in a decrease in the number of circulating platelets. Different mechanisms have been hypothesized to explain the thrombocytopenia in patients with severe dengue, one of them is the participation of the non-structural protein 1 (NS1) of dengue virus (DENV), which can be secreted into circulation during DENV infection and promotes a more efficient infection. OBJECTIVE The present study aimed to investigate the ability of platelet response to stimulation with full-length DENV NS1 protein and its domains. METHODS DENV NS1 plasmid was transfected into HEK-293T. Proteins were purified by Niquel Sepharose affinity chromatography. Secreted proteins were assessed by sodium dodecylsulfate polyacrylamide gel electrophoresis, Coomassie staining and western blot. Platelet-rich plasma was directly incubated with DENV NS1 proteins. Platelet activation was confirmed by expression of αIIbβIII and P-selectin by flow cytometry. Platelet aggregation was also assessed using DENV NS1 protein and its individual domains as agonists. RESULTS DENV NS1 protein and its domains induce P-selectin and αIIbβ3 complex expression on platelet surfaces. DENV NS1 induce a stable platelet aggregation after the addition of a minimal dose of adenosine diphosphate (ADP), epinephrine (EPI), or collagen. Interestingly, only EPI could induce the formation of platelet aggregates after incubation with the protein domains of NS1. CONCLUSION Our results suggest that the full DENV NS1 protein and also its domains promote platelet recognition, activation, and aggregation.

Background Zika (ZIKV) is a mosquito-borne virus that has caused multiple outbreaks in Americas. The early and accurate diagnosis of ZIKV is the key to minimize the health burden imposed on the infected patients. Many commercial ZIKV diagnosis kit are available based on ZIKV envelope antigen with varying sensitivity and specificity. Objective The aim of this study was to develop sensitive ELISA methodology based on recombinant ZIKV NS1 and NS1 β-ladder antigens for seroprevalence detection in a cohort of patients in an arbovirus endemic Mexican region in comparison to a commercial kit. Study design We obtained serum samples from 60 patients presenting with febrile illness and from 10 healthy donors in Michoacan state in 2016–2017. These samples were screened for ZIKV by RT-PCR and used to develop NS1 based ELISA and compared to a commercial kit. Results Our results indicate that both ZIKV sNS1 and β-ladder-based ELISA can reliably detect anti-ZIKV NS1 antibodies in infected patients, relevant for the serodiagnosis of ZIKV. Determination of antibody titers showed that it offered higher sensitivity than a commercially available ZIKV ELISA. Over 90% seropositivity against ZIKV for the febrile patients was detected in Lazaro Cardenas which is an arbovirus endemic region while lower seropositivity was observed in the healthy volunteers in Morelia (non-endemic area). Conclusion We conclude that our simple and sensitive in-house NS1 based ELISA used in this study has excellent sensitivity, is easy to use and can provide fast results suitable for larger population-based seroprevalence studies in the future.

Among COVID-19 hospitalized patients, high incidence of alterations in inflammatory and coagulation biomarkers correlates with a poor prognosis. Comorbidities such as chronic degenerative diseases are frequently associated with complications in COVID-19 patients. The aim of this study was to evaluate inflammatory and procoagulant biomarkers in COVID-19 patients from a public hospital in Mexico. Blood was sampled within the first 48 h after admission in 119 confirmed COVID-19 patients that were classified in 3 groups according to oxygen demand, evolution and the severity of the disease as follows: 1) Non severe: nasal cannula or oxygen mask; 2) Severe: high flow nasal cannula and 3) Death: mechanical ventilation eventually leading to fatal outcome. Blood samples from 20 healthy donors were included as a Control Group. Analysis of inflammatory and coagulation biomarkers including D-dimer, interleukin 6, interleukin 8, PAI-1, P-selectin and VWF was performed in plasma. Routine laboratory and clinical biomarkers were also included and compared among groups. Concentrations of D-dimer (14.5 ± 13.8 µg/ml) and PAI-1 (1223 ± 889.6 ng/ml) were significantly elevated in severe COVID-19 patients (P < 0.0001). A significant difference was found in interleukin-6, PAI-1 and P-selectin in non-severe and healthy donors when compared to Severe COVID-19 and deceased patients (P < 0.001). VWF levels were also significantly different between severe patients (153.5 ± 24.3 UI/dl) and non-severe ones (133.9 ± 20.2 UI/dl) (P < 0.0001). WBC and glucose levels were also significantly elevated in patients with Severe COVID-19. Plasma concentrations of all prothrombotic biomarkers were significantly higher in patients with a fatal outcome.

Resumen Introducción: La reducción de las hormonas tiroideas, triyodotironina total (T3) y triyodotironina libre (T3L) en pacientes en hemodiálisis, es un marcador de malnutrición e inflamación y son predictores de mortalidad. El objetivo del estudio fue determinar la prevalencia del síndrome complejo de malnutrición e inflamación en hemodiálisis y su asociación con las hormonas tiroideas: tirotropina, T3, T3L y tiroxina libre (T4L); además de evaluar la incidencia del síndrome de T3L y su correlación con marcadores nutricionales e inflamatorios. Materiales y métodos: Estudio transversal, analítico y comparativo, incluyó 128 pacientes en HD, 50,8% mujeres, edad 45,05 ± 17,01 años, 45,4 ± 38,8 meses en hemodiálisis, 29,7% diabéticos y 79,7% hipertensos. Se determinó en suero la concentración de tirotropina, T3, T3L y T4L, se aplicó la encuesta Malnutrition-Inflammation Score para diagnosticar malnutrición e inflamación. Resultados: La media de valores de las hormonas tiroideas fueron: tirotropina 2,48 ± 1,8 mUI/mL (rango 0,015-9,5), T3 1,18 ± 0,39 ng/mL (0,67-2,64), T3L 5,21 ± 0,96 pmol/l (3,47-9,75), T4L 1,35 ± 0,4 ng/mL (0,52-2,57). La prevalencia de síndrome complejo de malnutrición e inflamación es 53,9%; un 11,7% mostró T3L baja. Las concentraciones séricas de T3 y T3L correlacionan negativamente con Malnutrition-Inflammation Score y T4L correlaciona positivamente con Malnutrition-Inflammation Score. El análisis de regresión lineal de T3L baja fue asociado con IL-6 (β=0,265 p = 0,031), proteína C reactiva (β=-0,313 p = 0,018) y albúmina (β=0,276 p = 0,002). Conclusiones: Bajos niveles de T3 y T3L correlacionan con parámetros de inflamación y nutrición. El síndrome complejo de malnutrición e inflamación puede afectar la concentración sérica de hormonas tiroideas. Abstract Introduction: Low levels of thyroid hormones, total triiodothyronine (T3) and free triiodothyronine (FT3) in haemodialysis patients is a marker of malnutrition and inflammation and are predictors of mortality. The aim of this study was to determine the prevalence of malnutrition-inflammation complex syndrome in haemodialysis and its relationship with the thyroid hormones thyrotropin, T3, FT3 and free thyroxine (FT4), as well as to evaluate the prevalence of low FT3 syndrome and its correlation with nutritional and inflammatory markers. Materials and methods: Cross-sectional, analytical and comparative study that enrolled 128 haemodialysis patients: 50.8% females; mean age 45.05 ± 17.01 years; mean time on haemodialysis 45.4 ± 38.8 months; 29.7% diabetics; 79.7% with hypertension. Serum thyroid hormones thyrotropin, T3, FT3 and FT4 concentrations were measured and Malnutritition-Inflammation Score (MIS) was applie to diagnostic. Results: Mean thyroid hormone values were: thyroid hormones thyrotropin 2.48 ± 1.8 mIU/ml (range: 0.015-9.5), T3 1.18 ± 0.39 ng/ml (range 0.67-2.64), FT3 5.21 ± 0.96 pmol/l (range: 3.47-9.75); FT4 1.35 ± 0.4 ng/ml (range: 0.52-2.57). Malnutrition-inflammation complex syndrome prevalence was 53.9%; 11.7% presented low FT3 levels. Serum T3 and FT3 concentrations inversely correlated with Malnutritition-Inflammation Score (MIS), while FT4 correlated positively with Malnutrition-Inflammation Score. In the linear regression analysis, low FT3 was associated with IL-6 (β= 0.265, p = .031), C-reactive protein (CRP) (β= -0.313, p = .018) and albumin (β= 0.276, p = .002). Conclusion: Low T3 and FT3 levels are correlated with malnutrition and inflammation parameters. Malnutrition-inflammation complex syndrome can affect serum concentrations of thyroid hormones.

Resumen Introducción: La reducción de las hormonas tiroideas, triyodotironina total (T3) y triyodotironina libre (T3L) en pacientes en hemodiálisis, es un marcador de malnutrición e inflamación y son predictores de mortalidad. El objetivo del estudio fue determinar la prevalencia del síndrome complejo de malnutrición e inflamación en hemodiálisis y su asociación con las hormonas tiroideas: tirotropina, T3, T3L y tiroxina libre (T4L); además de evaluar la incidencia del síndrome de T3L y su correlación con marcadores nutricionales e inflamatorios. Materiales y métodos: Estudio transversal, analítico y comparativo, incluyó 128 pacientes en HD, 50,8% mujeres, edad 45,05 ± 17,01 años, 45,4 ± 38,8 meses en hemodiálisis, 29,7% diabéticos y 79,7% hipertensos. Se determinó en suero la concentración de tirotropina, T3, T3L y T4L, se aplicó la encuesta Malnutrition-Inflammation Score para diagnosticar malnutrición e inflamación. Resultados: La media de valores de las hormonas tiroideas fueron: tirotropina 2,48 ± 1,8 mUI/mL (rango 0,015-9,5), T3 1,18 ± 0,39 ng/mL (0,67-2,64), T3L 5,21 ± 0,96 pmol/l (3,47-9,75), T4L 1,35 ± 0,4 ng/mL (0,52-2,57). La prevalencia de síndrome complejo de malnutrición e inflamación es 53,9%; un 11,7% mostró T3L baja. Las concentraciones séricas de T3 y T3L correlacionan negativamente con Malnutrition-Inflammation Score y T4L correlaciona positivamente con Malnutrition-Inflammation Score. El análisis de regresión lineal de T3L baja fue asociado con IL-6 (β=0,265 p = 0,031), proteína C reactiva (β=-0,313 p = 0,018) y albúmina (β=0,276 p = 0,002). Conclusiones: Bajos niveles de T3 y T3L correlacionan con parámetros de inflamación y nutrición. El síndrome complejo de malnutrición e inflamación puede afectar la concentración sérica de hormonas tiroideas.

Emergency Medical Technicians are health professionals commonly exposed to dangerous traumatic scenarios which can lead to an altered hypothalamus-pituitary-adrenal (HPA) axis and stress related symptoms; however, the stress intensity may be mediated by personality traits such as self-esteem. We tested, in a population of 96 EMTs from the National Red Cross, the hypothesis of reduced cortisol secretion after the exposition to traumatic content images that could represent a stressor in non-paramedic healthy volunteers (59 non-EMTs). We took three saliva samples to measure cortisol (basal, 15 and 30 min after the images) and quantified the levels of self-esteem, perceived and physiological stress. Results showed a peak of cortisol 15 min after the images in the non-EMTs population, whereas a decreased cortisol profile was observed in EMTs, suggesting a higher sensitivity for a negative feedback regulation of cortisol. EMTs had lower levels of perceived stress but higher physiological stress symptoms than non-EMTs. The most important predictors of cortisol levels in EMTs were the number of working days per week and self-esteem which also had a negative correlation with perceived and physiological stress. We suggested that, in general, this paramedic population is habituated and predisposed to accidental scenarios.

Zika virus (ZIKV) is an emerging mosquito-borne flavivirus associated with congenital Zika syndrome and Guillain-Barré syndrome in adults. The recombinant ZIKV envelope (E) antigen can be useful for serodiagnosis of ZIKV infection and for monitoring immune responses during preclinical and clinical ZIKV vaccine development. In this study, we describe production of ZIKV E using the modified polyethyleneimine (PEI) transfection in HEK293 cells to improve cost-effective large-scale production. We show that the secretion of ZIKV E in HEK293 cells is dependent on cell culture incubation temperatures where incubation at a low temperature of 28 °C improved protein secretion of both, E-CD4 and E, whereas a substantial decrease in secretion was observed at 37 °C. The resulting E-CD4 produced at low temperature yielded similar binding profiles in ELISAs in comparison with a commercially available E protein using human seropositive sera to ZIKV. We also show that ZIKV NS1 and NS1 β-ladder antigens produced in HEK293 cells, have similar binding profiles in ELISA which suggests that both NS1 or NS1 β-ladder can be used for serodiagnosis of ZIKV. In conclusion, we propose a cost-effective production of the ZIKV E and NS1, suitable for both, clinical and research applications in endemic countries.

von Willebrand factor (vWF) is a multimeric glycoprotein present in blood plasma. It is synthesized in megakaryocytes and endothelial cells, secreted into circulation in the form of high-molecular-weight multimers (HMWMs), and cleaved into shorter, less active multimers by ADAMTS13. It is essential for platelet adhesion and aggregation. Previous studies have investigated the relationship between vWF levels and thromboembolic events with little regard to vWF multimeric structure. Patients with atrial fibrillation (AF) exhibit higher plasma vWF and lower ADAMTS13 levels. One hundred seven patients with AF, 51 anticoagulated and 56 nonanticoagulated, were eligible for the study. Plasma samples were analyzed for vWF antigen, vWF activity, and ADAMTS13; vWF multimers were analyzed by Western blot in 1% to 1.3% sodium dodecyl sulfate agarose gel electrophoresis. Patients with AF without oral anticoagulation (OAC) had significantly higher vWF plasma levels (154.00 [75-201] UI/dL) and vWF activity (60.00% [20%-210%]) compared to patients with OAC (133.50 [90-192] UI/dL, P =

In this chapter, we describe production and purification of the Zika virus NS1 glycoprotein in human embryonic kidney (HEK293T) cells at small, research laboratory scale. The expression of secreted NS1 (sNS1) and the C-terminal β-ladder domain in HEK293T cells were tested in a small-scale transfection before scaling up to a larger-scale transfection using roller bottles. Two different purification approaches have been applied to obtain purified NS1 (sNS1) and the C-terminal β-ladder domain ready for clinical applications.

Chikungunya fever is a debilitating disease caused by Chikungunya virus (CHIKV) that can result in long-lasting arthralgias. The early diagnosis of CHIKV relies on PCR during the acute infection phase to allow differential diagnosis with other co-circulating arboviruses such as dengue and Zika. Alternatively, serology can support diagnosis and provide epidemiological information on current and past outbreaks. Many commercial serological ELISA assays are based on the inactivated whole CHIKV, but their sensitivity and specificity show great variability. We produced recombinant CHIKV E2 that is suitable for ELISA assays, which was used for the serodiagnosis of CHIKV infections occurring in an arbovirus endemic Mexican region within Michoac&aacute, n state. A cross-sectional study was conducted in 2016&ndash, 2017, sera was obtained from 15 healthy donors and 68 patients presenting undifferentiated febrile illness. Serum samples were screened by RT-PCR and by our in-house ELISA assay. Our results indicate that IgM and IgG anti-CHIKV E2 antibodies were detected with our ELISA assay with higher sensitivity than a commercially available CHIKV ELISA kit. Our simple and sensitive ELISA assay for the serodiagnosis of CHIKV infections can be applied to population-based seroprevalence surveys and has potential for monitoring vaccine immunogenicity in CHIKV vaccine clinical trials.

Objective Drug inhibition of platelet P2Y12 adenosine diphosphate receptor has reduced the incidence of adverse cardiovascular events after percutaneous coronary interventions. The analysis of the phosphorylation status of vasodilator-stimulated phosphoprotein by flow cytometry has shown a predictive value for adverse events and stent thrombosis. Polymorphisms of CYP2C19 in high risk patients may also relate to adverse cardiovascular events. Methods Ninety patients were enrolled. Patients received a 600 mg clopidogrel loading dose. Blood samples were obtained at the time of the procedure and 24 h later, platelet reactivity was assessed by vasodilator-stimulated phosphoprotein phosphorylation measurement using flow cytometry. Low response to clopidogrel was defined as a platelet reactivity index ≥ 50%. The presence of CYP2C19*2 was identified with the restriction enzyme Smal. Results Mean platelet reactivity index: 53.45 ± 22.48% in the baseline sample and 57.14 ± 23.08% at 24 h (p = 0.183); 40% of patients behaved as good responders, the rest behaved as non-responders with 38% of patients showing platelet reactivity indexes between 50-70% and 22% showing indexes above 70%. The CYP2C19*2 polymorphism was found in 17% of patients, with a 3.9% AA homozygous genotype carriers. Conclusion Response to the clopidogrel loading dose showed a wide variability among patients with 40% responding to the drug according to previously established cut-off values. Our results showed that 3.9% of patients show the AA genotype. To our knowledge, this is the first study involving clopidogrel response by flow citometry and genotype typification in Mexican Mestizo population

The thyroid gland and the kidney are closely related. Thyroid hormones (TH) contribute to the homeostasis of the human being through complex interactions of fluids and electrolytes, protein synthesis, etc. The effects on the kidney of TH may be pre renal or direct actions. Decreasing glomerular filtration (GF) this balance especially in advanced stages of chronic kidney disease (CKD) is altered. CKD is linked to alterations in TH levels and/or metabolism, resulting in a high prevalence of subclinical hypothyroidism and low free triiodothyronine (FT3) syndrome. These alterations are linked in micro inflammation, endothelial damage, cardiac abnormalities, and high mortality. In this study, we describe the most common thyroid abnormalities reported in CKD with dialytic stage approach.La glándula tiroides y el riñón están estrechamente relacionados. Las hormonas tiroideas (HT) contribuyen en la homeostasis del ser humano a través de complejas interacciones de líquidos y electrolitos, síntesis de proteínas, etc. Los efectos de las HT sobre el riñón pueden ser pre-renales o directos. Está demostrado que al disminuir la filtración glomerular (FG) se altera este equilibrio, sobre todo en estadios avanzados de la enfermedad renal crónica (ERC). La ERC está vinculada con alteraciones en los niveles de HT y/o su metabolismo, lo que resulta en una alta prevalencia de hipotiroidismo subclínico y el síndrome de T3 libre baja. Estas alteraciones están relacionadas con micro inflamation, daño endotelial, alteraciones cardiacas y alta mortalidad. El presente estudio, describe las alteraciones tiroideas más frecuentes reportadas en ERC con enfoque en la etapa dialítica.

Background Zika virus (ZIKV) has become a global threat with immediate need for accurate diagnostics, efficacious vaccines and therapeutics. Several ZIKV envelope (Env)-based vaccines have been developed recently. However, many commercially available ZIKV Env are based on the African lineage and produced in insect cells. Here, we sought to produce Asian-lineage ZIKV Env in mammalian cells for research and clinical applications. Methods We designed various gene expression constructs to optimize the production of ZIKV using prM-Env and full or C-terminal truncations of Env; with or without a rat CD4 fusion partner to allow large-scale production of soluble protein in mammalian HEK293 cells. Protein expression was verified by mass spectrometry and western-blot with a pan-flavivirus antibody, a ZIKV Env monoclonal antibody and with immune sera from adenoviral (ChAdOx1) ZIKV Env-vaccinated mice. The resulting Env-CD4 was used as a coating reagent for immunoassay (ELISA) using both mouse and human seropositive sera. Results Replacement of the C-terminus transmembrane Env domain by a rat CD4 and addition of prM supported optimal expression and secretion of Env. Binding between the antigens and the antibodies was similar to binding when using commercially available ZIKV Env reagents. Furthermore, antibodies from ZIKV patients bound ZIKV Env-CD4 in ELISA assays, whereas sera from healthy blood donors yielded minimal OD background. The serological outcomes of this assay correlated also with ZIKV neutralisation capacity in vitro. Conclusions Results obtained from this study indicate the potential of the Asian-lineage Zika Env-CD4 and Env proteins in ELISA assays to monitor humoral immune responses in upcoming clinical trials as well as a sero-diagnostic tool in ZIKV infection. Electronic supplementary material The online version of this article (10.1186/s12985-018-1104-6) contains supplementary material, which is available to authorized users.

ResumenAntecedentesLas citoquinas proinflamatorias desempeñan un papel importante en la retinopatía diabética; existe evidencia contradictoria sobre la elevación sérica de las mismas en esta condición y el posible rol que tienen como su marcador sérico.ObjetivoEvaluar la presencia de citoquinas séricas proinflamatorias en pacientes con y sin retinopatía diabética.Material y métodosSeries de casos comparativas. Se incluyeron 36 pacientes divididos en 3 grupos: 12 pacientes con diabetes mellitus con retinopatía diabética (grupo 1), 12 pacientes con diabetes mellitus sin retinopatía diabética (grupo 2) y 12 pacientes sin diabetes mellitus como grupo control. Se midieron los niveles séricos de las siguientes citoquinas proinflamatorias en todos los pacientes: TNF-α, IL-1β e IL-6. También se realizó medición de biomarcadores proinflamatorios o reactantes de fase aguda, como velocidad de sedimentación globular y proteína C reactiva.ResultadosLos niveles de TNF-α e IL-6 resultaron mayores en el grupo 1 (TNF-α: 19.4±10.9pg/ml, IL-6: 5.75±7pg/ml) comparados con los de los otros 2 grupos, aunque la diferencia resultó estadísticamente significativa solo en el caso del TNF-α (grupo 1: 19.4±10.9pg/ml, grupo 2: 14±4.3pg/ml y grupo control: 8.49±3.69pg/ml, p=0.001). No hubo diferencias entre biomarcadores proinflamatorios como velocidad de sedimentación globular y proteína C reactiva entre los 3 grupos (p>0.05).ConclusionesLos niveles séricos de citoquinas proinflamatorias fueron mayores en el grupo 1. Se requieren mayores estudios para establecer mejor el impacto de este hallazgo.AbstractBackgroundPro-inflammatory cytokines play an important role in diabetic retinopathy. There is conflicting evidence about their serum elevation in this condition and that they also may be possible serum inflammatory biomarkers of diabetic retinopathy.ObjectiveTo evaluate the presence of serum pro-inflammatory cytokines and acute phase reactants in the serum of patients with and without diabetic retinopathy.Material and methodsComparative case series with 36 patients divided into three groups were included: 12 patients with diabetes mellitus and diabetic retinopathy (group 1), 12 diabetic patients without diabetic retinopathy (group 2), and 12 healthy patients as a control group. Serum levels of the following pro-inflammatory cytokines were measured in all patients: TNF-α, IL-1β and IL-6. Pro-inflammatory biomarkers measurements were also performed, such as erythrocyte sedimentation rate and C-reactive protein.ResultsThe levels of TNF-α and IL-6 were higher in group 1 (TNF-α: 19.4 ± 10.9 pg/ml, IL-6: 5.75 ± 7 pg/ml) compared to the other two groups, although the difference was statistically significant only in the case of TNF-α (group 1: 19.4 ± 10.9 pg/ml, group 2: 14 ± 4.3 pg/ml and control: 8.49 ± 3.69 pg/ml, p = 0.001). There were no differences among pro-inflammatory biomarkers such as erythrocyte sedimentation rate and C reactive protein. among the three groups (p > 0.05).ConclusionsPro-- inflammatory serum cytokine levels were higher in the diabetes mellitus with diabetic retinopathy group. Larger studies are warranted to establish the real impact of this finding.

Childhood obesity is a public health problem in Mexico and worldwide because of the later clinical consequences including diabetes mellitus, hypertension, and cardiovascular disease (CVD).1 The rapid increase in the prevalence and severity of obesity in children likely lowers the age of onset and increases the incidence of CVD. Childhood obesity is associated with endothelial dysfunction, one of the earliest changes in the development of atherosclerosis,2,3 and evidence supports atherosclerotic cardiovascular disease beginning in childhood.4 Additionally, the high serum levels of low density lipoprotein (LDL)-cholesterol and total cholesterol in childhood were associated in adults with carotid intima-media thickness5 and subclinical atherosclerosis.6 In adults, inflammation, endothelial dysfunction, and hyperuricemia are factors that contribute to a link between obesity and CVD. Leptin has been shown to represent an important candidate linking these disorders,7,8 because of the potential role in the regulated functioning of the immune system,9 on platelet activation and segregation,10 and on relation with uric acid (UA).11 In the presence of obesity, inflammation leads to platelet activation and increased plasma levels of prothrombotic proteins stored in platelet α-granules including soluble P-selectin (sP-selectin), von Willebrand Factor (vWF), and plasminogen activator inhibitor-1 (PAI-1). High levels of these proteins are believed to play a central role in accelerating the risk of atherothrombosis.12 sP-selectin is not only expressed on activated endothelial cells, but also on activated platelets, and it mediates rosetting of the platelets with monocytes and neutrophils that contribute to atherosclerotic lesion.13 It is, therefore, considered a plasma marker of platelet activation and endothelial dysfunction in the atherogenic process that has been related to adverse cardiovascular events in adults.14,15 vWF levels have been significantly associated with insulin resistance (IR).16 Increased PAI-1 levels have been associated with the risk of thrombosis and fibrosis, and it has been shown to have a direct effect in the development of IR and type 2 diabetes.17 Disorders of UA metabolism are often seen in conjunction with factors associated with lifestyles such as an unbalanced diet abundant in purine, obesity, and alcohol consumption.18 In adolescents, UA levels are significantly increased with obesity,19 and some studies suggests that UA stimulates vascular inflammation and endothelial dysfunction, and it predicts adult blood pressure.20–22 The aim of our study was to investigate the factors associated with platelet activation in obese children.

Four different vaccine platforms, each targeting the human malaria parasite Plasmodium vivax cell-traversal protein for ookinetes and sporozoites (PvCelTOS), were generated and assessed for protective efficacy. These platforms consisted of a recombinant chimpanzee adenoviral vector 63 (ChAd63) expressing PvCelTOS (Ad), a recombinant modified vaccinia virus Ankara expressing PvCelTOS (MVA), PvCelTOS conjugated to bacteriophage Qβ virus-like particles (VLPs), and a recombinant PvCelTOS protein expressed in eukaryotic HEK293T cells (protein). Inbred BALB/c mice and outbred CD-1 mice were immunized using the following prime-boost regimens: Ad-MVA, Ad-VLPs, and Ad-protein. Protective efficacy against sporozoite challenge was assessed after immunization using a novel chimeric rodent Plasmodium berghei parasite (Pb-PvCelTOS). This chimeric parasite expresses P. vivax CelTOS in place of the endogenous P. berghei CelTOS and produces fully infectious sporozoites. A single Ad immunization in BALB/c and CD-1 mice induced anti-PvCelTOS antibodies which were boosted efficiently using MVA, VLP, or protein immunization. PvCelTOS-specific gamma interferon- and tumor necrosis factor alpha-producing CD8+ T cells were induced at high frequencies by all prime-boost regimens in BALB/c mice but not in CD-1 mice; in CD-1 mice, they were only marginally increased after boosting with MVA. Despite the induction of anti-PvCelTOS antibodies and PvCelTOS-specific CD8+ T-cell responses, only low levels of protective efficacy against challenge with Pb-PvCelTOS sporozoites were obtained using any immunization strategy. In BALB/c mice, no immunization regimens provided significant protection against a Pb-PvCelTOS chimeric sporozoite challenge. In CD-1 mice, modest protective efficacy against challenge with chimeric P. berghei sporozoites expressing either PvCelTOS or P. falciparum CelTOS was observed using the Ad-protein vaccination regimen.

The above article, published online on 29 July 2016 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the Journal Editor in Chief, Journal Production Manager, and John WileySons, Ltd. The retraction has been agreed due to 48% similar significant between this article and an article published in Nature Reviews Cancer journal.

Antinuclear antibodies (ANA) are frequently found in healthy populations. To define the prevalence, pattern, and titer of ANA in different groups of the healthy Mexican population, we studied 304 individuals, classified into 3 groups: 104 blood donors, 100 hospital personnel working at The State General Hospital, which included doctors, laboratory technicians, and nurses; and 100 relatives of patient diagnosed either with systemic lupus erythematosus or rheumatoid arthritis, all of them apparently healthy at the time of study. We determined ANA using immunofluorescence microscopy performed on HEp-2 cells. Fluorescence was detected in 165 serum samples (54.3%). The most frequent pattern was the speckled (50.3%). The most frequent dilution was 1:40 (35.4%), followed by 1:80 (13.4%), 1:160 (3.2%), and 1:320 (1.3%).Regarding the results by study group, we found a trend toward higher ANA levels in group 2 (hospital personnel), compared with group 1 (blood donors) and group 3 (relatives of patients), a trend also reflected by the increasing frequency of serum titers of 1:80 and higher (P = 0.074). According to occupation, medical doctors showed a higher incidence of speckled pattern when compared with other occupations (P = 0.022). Medical doctors (n = 75) showed also higher titers of this particular pattern (P = 0.03). In group 3, relatives of patients with systemic lupus erythematosus showed the speckled pattern more frequently than relatives of patients with rheumatoid arthritis, in low titers (P = 0.017). We suggest that ANA tests showing speckled pattern should be at a 1:160 titer or higher to be considered positive; other patterns such as homogeneous, peripheral, or centromeric might be considered positive even at low titers (

The above article, published online on 29 July 2016 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the Journal Editor in Chief, Journal Production Manager, and John Wiley & Sons, Ltd. The retraction has been agreed due to 48% similar significant between this article and an article published in Nature Reviews Cancer journal.

Introduccion: El laboratorio en reumatologia tiene un papel importante en la evaluacion, el diagnostico y el seguimiento de diversos padecimientos. Los anticuerpos antinucleares (ANA), los anticuerpos anti-ADN de cadena sencilla o doble (ss o dsAnti-ADN) y anticuerpos anticardiolipinicos (AcACL) se usan frecuentemente y su utilidad diagnostica es bien conocida en centros de atencion de tercer nivel. Nuestro hospital es un centro de atencion de segundo nivel que implemento hace 2 anos estas pruebas. Despues de 1 ano de su introduccion, decidimos evaluar la frecuencia en su uso, quien solicita estas pruebas, su utilidad diagnostica en lupus eritematoso generalizado (LEG) y sindrome antifosfolipidico (SAF). Pacientes y metodo: Se evaluo a todos los pacientes con cuadro clinico de estas enfermedades y solicitud de estas pruebas del 1 de septiembre de 2005 al 30 de junio de 2006. De manera prospectiva, los analizo un evaluador con un formato estandarizado que contenia informacion clinica, diagnostico inicial, datos del medico solicitante, servicio, diagnostico tras resultado y los cambios en la terapeutica. Analisis estadistico: se utilizo estadistica descriptiva y tablas de 2 * 2 para evaluar la utilidad diagnostica en las indicaciones mas comunes. Resultados: De un total de 286 solicitudes recibidas, se analizaron 157. Reumatologia y medicina interna enviaron 63 y 31 solicitudes respectivamente. Con respecto a los ANA en LEG, se calculo la sensibilidad (70%); la especificidad (92%); el valor predictivo positivo (VPP) (81%); el valor predictivo negativo (VPN) (86%); la razon de verosimilitud positiva (RVsP) (8,73) y la razon de verosimilitud negativa (RVsN) (0,33). En relacion con los anti-ADN en LEG, la sensibilidad (78%); la especificidad (50%); el VPP (80%); el VPN (46%); la RVsP (1,56) y la RVsN (0,44). Respecto a los AcACL en SAF, la sensibilidad (78%); la especificidad (92%), el VPP (78%), el VPN (92%), la RVsP (10) y la RVsN (0,24). Conclusiones: En nuestro hospital hay poca frecuencia en la solicitud de estos estudios. La sensibilidad y la especificidad parecen no estar acordes con lo publicado. Es necesaria la elaboracion de lineamientos que en nuestro medio regulen la solicitud de estudios especializados en reumatologia y aumenten su utilidad clinica.

Von Willebrand disease is an inherited disease in which the structure, function, and concentration of von Willebrand factor are altered, as well as the platelet von Willebrand factor endothelium interaction. In Mexico there are no epidemiological records of the disease. Only a few isolated studies have been reported from the clinical and hematological standpoint.We studied 155 Mexican Mestizos: 75 with presumptive diagnosis of von Willebrand disease, 15 with suspected diagnosis ofhemophilia A and 65 healthy donors (controls). Basic coagulation tests, special tests and classification test (analysis of multimeric composition) were carried out.There were 15 patients with clinical diagnosis of hemophilia A, 75 patients with suspected von Willebrand disease of which 50 were diagnosed as the following types and subtypes: Type 1 (62%), Type 2 (22%) [subtypes: 2A (14%), 2B (2%), and 2N (6%)] and Type 3 (16%).It has been reported that analysis of von Willebrand factor is a method that meets the characteristics for the diagnosis of von Willebrand disease. It is necessary to implement this methodology to study and improve the specific diagnoses.Antecedentes: la enfermedad de von Willebrand es un padecimiento hereditario en el que la estructura, función y concentración del factor de von Willebrand están alteradas y, en consecuencia, también la interacción plaqueta-factor de von Willebrand-endotelio. En México no hay registros epidemiológicos de la enfermedad, sólo se han efectuado algunos estudios aislados desde el punto de vista clínico y hematológico. Material y métodos: estudio retrospectivo efectuado en 155 mexicanos mestizos, 75 de ellos con diagnóstico presuntivo de enfermedad de von Willebrand, 15 con sospecha de hemofilia A y 65 donadores sanos (testigos). Se realizaron pruebas: básicas de coagulación, especiales y de clasificación: análisis de la composición multimérica. Resultados: 15 pacientes se diagnosticaron con hemofilia A; de los 75 sujetos con sospecha de enfermedad de von Willebrand se diagnosticaron 50 de la manera siguiente: tipo 1 (62%), tipo 2 (22%) [subtipos: 2A (14%), 2B (2%) y 2N (6%)] y tipo 3 (16%). Conclusión: el análisis de los multímeros del factor de von Willebrand es un método que cumple con las características adecuadas para el diagnóstico de la enfermedad de von Willebrand, por lo que es necesario implementar esta metodología para su estudio y mejorar su diagnóstico específico.

Background Pro-inflammatory cytokines play an important role in diabetic retinopathy. There is conflicting evidence about their serum elevation in this condition and that they also may be possible serum inflammatory biomarkers of diabetic retinopathy. Objective To evaluate the presence of serum pro-inflammatory cytokines and acute phase reactants in the serum of patients with and without diabetic retinopathy. Material and methods Comparative case series with 36 patients divided into three groups were included: 12 patients with diabetes mellitus and diabetic retinopathy (group 1), 12 diabetic patients without diabetic retinopathy (group 2), and 12 healthy patients as a control group. Serum levels of the following pro-inflammatory cytokines were measured in all patients: TNF-α, IL-1β and IL-6. Pro-inflammatory biomarkers measurements were also performed, such as erythrocyte sedimentation rate and C-reactive protein. Results The levels of TNF-α and IL-6 were higher in group 1 (TNF-α: 19.4 ± 10.9 pg/ml, IL-6: 5.75 ± 7 pg/ml) compared to the other two groups, although the difference was statistically significant only in the case of TNF-α (group 1: 19.4 ± 10.9 pg/ml, group 2: 14 ± 4.3 pg/ml and control: 8.49 ± 3.69 pg/ml, p = 0.001). There were no differences among pro-inflammatory biomarkers such as erythrocyte sedimentation rate and C reactive protein. Among the three groups ( p > 0.05). Conclusions Pro-inflammatory serum cytokine levels were higher in the diabetes mellitus with diabetic retinopathy group. Larger studies are warranted to establish the real impact of this finding.

Atrial Fibrillation (AF) is the most common cardiac arrhythmia of clinical significance; it increases the risk of mortality due to stroke. The mechanisms behind cerebral thromboembolism in AF are associated with a prothrombotic state, demonstrated by higher levels of von Willebrand Factor (vWF), a multimeric glycoprotein that plays a crucial role in platelet adhesion and aggregation and it has been proposed as a biomarker of endothelial dysfunction. Plasma vWF levels are elevated in patients with nonvalvular Atrial Fibrillation (NVAF) associated to the presence of cardiovascular risk factors. The variability in vWF plasma levels in healthy subjects has a wide distribution, but there is no description available of the variability in AF patients and among types of AF. The aim of this study was to determine the variability of vWF plasma concentrations in patients with NVAF, associated to cardiovascular risk factors. Search strategy included PubMed and Ovid. Keywords used were “Atrial Fibrillation” and “von Willebrand Factor”. It includes original articles, with analysis of plasma vWF levels by ELISA, without acute stroke. Review articles and meta-analysis were excluded. Reviewed studies include 22 trials and 6542 patients with nonvalvular AF associated to cardiovascular disease risk factors: age, sex, hypertension, heart failure, diabetes mellitus, prior stroke, coronary artery disease. Variability in vWF plasma levels was wide, with minimum values of 77 IU/dl and maximum values of 245 IU/dl and a mean of 146 IU/dl. Age of patients ranged between 54 and 78 years, and the percentage of males ranged between 23% and 80%. According to type of AF vWF levels were as follows, in paroxysmal AF: 92-264 IU/dl; persistent AF: 76-234 IU/dl; permanent AF: 91-247 IU/dl. The variability in vWF plasma levels is affected by risk factors and the AF type, however vWF levels in AF patients are higher when compared with healthy subjects.

"Antecedentes: la enfermedad de von Willebrand es un padecimiento hereditario en el que la estructura, función y concentración del factor de von Willebrand están alteradas y, en consecuencia, también la interacción plaqueta-factor de von Willebrand-endotelio. En México no hay registros epidemiológicos de la enfermedad, sólo se han efectuado algunos estudios aislados desde el punto de vista clínico y hematológico. Material y métodos: estudio retrospectivo efectuado en 155 mexicanos mestizos, 75 de ellos con diagnóstico presuntivo de enfermedad de von Willebrand, 15 con sospecha de hemofilia A y 65 donadores sanos (testigos). Se realizaron pruebas: básicas de coagulación, especiales y de clasificación: análisis de la composición multimérica. Resultados: 15 pacientes se diagnosticaron con hemofilia A; de los 75 sujetos con sospecha de enfermedad de von Willebrand se diagnosticaron 50 de la manera siguiente: tipo 1 (62%), tipo 2 (22%) [subtipos: 2A (14%), 2B (2%) y 2N (6%)] y tipo 3 (16%). Conclusión: el análisis de los multímeros del factor de von Willebrand es un método que cumple con las características adecuadas para el diagnóstico de la enfermedad de von Willebrand, por lo que es necesario implementar esta metodología para su estudio y mejorar su diagnóstico específico."

Introduction Laboratory tests have an important role in rheumatology for evaluation, diagnosis, and follow up in several diseases. Specialized tests such as antinuclear antibodies (ANA), anti single, or double stranded DNA antibodies (anti-DNA), and anticardiolipin antibodies (ACL) are frequently used and its diagnostic performance is well known in tertiary care centers. Our setting is a secondary care center that implemented these tests 2 years ago. After 1 year of implementation, we decided to evaluate the frequency of use, who orders these tests, and their diagnostic properties for the diagnosis of systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APLS). Patienst and method All patients with clinical charts and a request for these tests were evaluated from September 1, 2005 to June 30, 2006. These evaluations were done prospectively by a single, trained evaluator following a standardized format looking at pretest clinical information such as pretest diagnosis, physician's level of training, service, and posttest results as well as therapeutic changes after results. Statistical analysis: descriptive statistics and 2 by 2 tables to estimate diagnostic performance of most common indications. Results Two hundred and eighty-six requests were reviewed and only 157 were evaluated. Rheumatology and Internal Medicine services sent 63 and 31 requests for these tests respectively. Diagnostic properties of ANA for SLE were sensitivity (sen) 70%, specificity (spec): 92%, positive predictive value (PPV): 81%, negative predictive value (NPP): 86%, positive likelihood ratio (PLR): 8.73, and negative likelihood ratio (NLR): 0.33. Anti-double stranded DNA, Sen: 78%, spec: 50%, PPV: 80%, NPP: 46%, PLR: 1.56, NLR: 0.44; ACACL y Sen: 78%, spec: 92%, PPV: 78%, NPV 92%, PLR: 10, NLR 0,24. Conclusions These specialized tests are not frequently used in our setting. Their diagnostic properties are not as accurate as those published in medical literature. Guidelines are needed in our hospital to improve their diagnostic performance.

Background Rheumatoid Arthritis (RA) is characterized by increased cardiovascular (CV) morbimortality due to higher prevalence of traditional CV risk factors such as diabetes, dyslipidemia, hypertension, sedentarism, increased body mass index (BMI) that is often undertreated 1 . RA is considered a state of accelerated atherosclerosis, which has been recognized as a dynamic inflammatory process that begins with endothelial activation/dysfunction. Diabetes Mellitus Type 2 (DM2) is the prototype of disease with a high degree of endothelial dysfunction, atherosclerosis and CV risk. CV risk could be similar in RA and DM2. 2,3 Objectives To determine the degree of endothelial dysfunction in RA patients compared to DM2 patients with and without glycemic control, obese patients and healthy subjects. Methods Descriptive, prolective, transversal, observational clinical study, with 5 groups: 1) AR, 2) UDM: Uncontrolled DM2, 3) CDM: Controlled DM2, 4) Obesity, 5) Control Group. All matched with group 1 for age (±2 years), gender, period of clinical diagnosis. Complete medical history, Carotid Intima-Media Thickness, blood cell count, erythrosedimentation rate, C-reactive Protein, fasting & 2 hours postprandial glucose, fasting insulin, urea, creatinine, ureic nitrogen, uric acid, glycosylated haemoglobin (HbA1c%), Triglycerides (TG), total cholesterol (TC), C-HDL, C-LDL, C-VLDL, urianalysis, micro-albuminuria. Endothelial dysfunction biomarkers (ELISA): TNF-α, IL-6, vWF, PAI-1. Descriptive statistical analyses:mean & standard deviation. Kolmogorov-Smirnov test to determine distribution type. Comparison between groups (parametric distribution): ANOVA; non-parametric distribution: Kruskal-Wallis. Spearman´s correlation coefficient to associate biomarkers with cardiovascular risk variables. Results 190 participants in five groups, main results showed. Conclusions Patients with RA share traditional factors of CV risk with diabetic patients (obesity, arterial hypertension, low levels of C-HDL), present non-traditional risk factors that are characteristic of their inflammatory status (SGS, CRP, TNF-α, IL-6) and also appear to present a prothrombotic status (PAI-1, vWF) that is greater than that of DM2 patients. This confers an increased risk of accelerated atherosclerosis upon the RA patients. It is therefore imperative that these patients are managed appropriately in a comprehensive manner. References Liao KP, Solomon DH. Rheumatology 2012; 2: 123-129. Van Halm VP et al. Ann Rheum Dis 2009; 68: 1395-1400. Stamatelopoulos SK et al. Arterioscler Thromb Vasc Biol 2009: 5:1-7. Acknowledgements The research team is grateful to Bristol Myers Squibb for the provision of funding with which to conduct this study. Disclosure of Interest None Declared

The Zika virus (ZIKV) was first isolated in 1947 in Uganda. While it took 60 years for this virus to cause major outbreaks, an important shift in its ability to cause epidemics took place in the first and second decades of the this century: in 2007 in Yap Island, Micronesia, followed by French Polynesia in 2013 and, finally in 2015 and 2016, when ZIKV infections occurred throughout South America, Central America and the Caribbean, spreading rapidly to reach North America in just a single year. No licensed prophylactic vaccine is yet available but recent efforts towards the development of a vaccine have been remarkable from both the private and public sectors and include new candidate vaccines ranging from the classical live-attenuated or inactivated vaccines to more sophisticated approaches such as mRNA or genetically engineered viral platforms. Previous successes with licensed flavivirus vaccines indicate that a protective ZIKV vaccine should be an achievable goal. Nevertheless, numerous pre- and post-licensure challenges need to be taken into account, such as the interaction of vaccine-induced immune responses with other flaviviruses, in particular with dengue, where antibody-dependent enhancement could become an issue, and the importance of a rapid induction of protective responses during pregnancy.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Background: Thrombotic risk increases in patients with respiratory diseases such as lung cancer and SARS-Cov-2 infection, due to endothelial dysfunction that develops due to the activation of thrombotic and inflammatory processes. The von Willebrand Factor maintains hemostatic interactions, in addition; it is a biomarker of endothelial dysfunction, vascular damage causes the release of VWF from the Weibel-Palade bodies, the increase in its plasma concentration is related to thrombotic risk. The development of inflammatory processes accompanied by the mechanisms of evasion to the immune response of these respiratory pathologies, directly to the constitutive release of high molecular weight structures of VWF; as well as the increase in plasma concentrations and the decrease of its regulatory enzyme ADAMTS-13 favor a procoagulant state that maintains feedback cascades causing hemodynamic alterations until they become pathological. Objective: To establish the usefulness of plasma concentrations of VWF protein and ADAMTS 13 enzyme as biomarkers of thrombotic risk in lung cancer and SARS-CoV-2 infection. Materials and methods: Patients with lung cancer, patients with SARS-CoV-2 (COVID-19) infection and healthy subjects. Determination of plasma concentration of VWF and ADAMTS-13 by means of enzyme-linked immunoassay (ELISA) and analysis of multimeric structure by Western blot technique. Results: A total of 49 patients were included, distributed as follows; lung cancer patients (n = 8), COVID-19 patients (n = 25), healthy subjects (n = 16). Obtaining an average age of 52 years and where 61.2% belong to the male gender and 38.8% to the female gender. An increase in plasma VWF concentrations was observed in the group with lung cancer (p = 0.002) and the group with COVID-19 (p = 0.000) compared to the control group, as well as an increase in the densitometric mean of (p = 0.006) and (p = 0.001) respectively. Antecedentes: El riesgo trombótico aumenta en los pacientes con enfermedades respiratorias como cáncer pulmonar e infección por SARS-Cov-2, debido a la disfunción endotelial que se desarrolla por la activación de procesos trombóticos e inflamatorios. El Factor von Willebrand mantiene numerosas interacciones hemostáticas; además, es un biomarcador de disfunción endotelial, el daño vascular causa la liberación de FvW de los cuerpos de Weibel- Palade, el incremento en su concentración plasmática está relacionada con el riesgo trombótico. El desarrollo de procesos inflamatorios acompañados de los mecanismos de evasión a la respuesta inmune de estas patologías respiratorias, contribuyen directamente a la liberación constitutiva de estructuras de elevado peso molecular de FvW; así como el incremento en las concentraciones plasmáticas y la disminución de su enzima reguladora ADAMTS-13 favorecen un estado procoagulante que mantiene cascadas de retroalimentación provocando las alteraciones hemodinámicas hasta convertirlas en patológicas. Objetivo: Establecer la variación de las concentraciones plasmáticas de la proteína FvW y enzima ADAMTS 13 como biomarcadores de riesgo trombótico en Cáncer pulmonar e infección por SARS-CoV-2. Materiales y métodos: Pacientes con cáncer pulmonar, pacientes con infección por SARS-CoV-2 (COVID-19) y sujetos sanos. Determinación de la concentración plasmática de FvW y ADAMTS-13 por medio de inmunoensayo ligado a enzima (ELISA) y análisis de la estructura multimérica mediante técnica de Western blot. Resultados: Se incluyeron un total de 49 pacientes distribuidos de la siguiente manera; pacientes con cáncer de pulmón (n=8), pacientes con COVID-19 (n=25), sujetos sanos (n=16). Obteniendo una edad promedio de 52 años y donde el 61.2% pertenece al género masculino y un 38.8% al género femenino. Se observó un incremento en las concentraciones plasmáticas del FvW en el grupo con cáncer pulmonar (p=0.002) y grupo con COVID-19 (p=0.000) en comparación al grupo control, así como un incremento en la media densitométrica de (p=0.006) y (p=0.001) respectivamente.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Diseases caused by the emerging arboviruses chikungunya (CHIKV), dengue (DENV) and Zika (ZIKV) are a public health problem with broad distribution in tropical and sub-tropical urban areas. These viruses share a common vector, female mosquitoes of the genus Aedes (Aedes aegypti and Aedes albopictus). Human mobility, urbanization and the inconsistent use of prevention strategies have favoured an increase in their distribution. Clinical manifestations vary from mild or asymptomatic, to chronic sequels and death in some cases. Furthermore, clinical manifestations can be similar, in particular joint pain and fever, making diagnosis difficult by clinical methods. Epidemiological data suggests that at least half of the world’s population (3.6 billion people) is at risk of acquiring the infection by DENV, which is the most widely distributed arbovirosis. CHIKV distribution has increased due to air traffic and mobility to reach the Americas, causing outbreaks since 2013. In the case of ZIKV, which was confined to the equatorial zone of Africa and Asia; outbreaks have emerged in the Americas between 2015 and 2016. There is no specific treatment for any of these arboviral infections. The development of vaccines is considered now as the strategy with highest potential for the control of these diseases. Vaccine development requires multiple experimental and epidemiological approaches to evaluate the real incidence and prevalence of these infections in endemic areas. General objective. To determine seroprevalence and seroconversion of antibodies against CHIKV, DENV and ZIKV in healthy individuals from the General Hospital “Dr. Miguel Silva”, the Faculty of Medical and Biological Sciences “Dr. Ignacio Chávez” of the Universidad Michoacana de San Nicolas de Hidalgo and volunteers from general population in the city of Morelia, Michoacan, during a one-year period. Las enfermedades causadas por arbovirus emergentes chikungunya (CHIKV), dengue (DENV) y Zika (ZIKV), son un problema de salud pública a nivel mundial, con amplia distribución en zonas urbanas de regiones tropicales y subtropicales. Estos virus comparten un vector común, las hembras del mosquito del género Aedes (Aedes aegypti y Aedes albopictus). La movilidad humana, la urbanización y el uso inconsistente de estrategias de prevención han favorecido el incremento en su distribución. Las manifestaciones clínicas varían desde un curso asintomático o leve, hasta secuelas crónicas y muerte, en algunos casos. Además, estas manifestaciones pueden ser similares entre sí, particularmente en cuanto a síntomas como dolor articular y la fiebre, dificultando la distinción por métodos clínicos. Los datos epidemiológicos sugieren que al menos la mitad de la población mundial (3.6 mil millones de personas) está en riesgo para adquirir infección por DENV, siendo la arbovirosis más ampliamente distribuida. CHIKV, ha aumentado su distribución debido al tráfico aéreo y la movilidad hasta alcanzar las Américas, causando brotes en 2013. En el caso de ZIKV, originalmente presente en la zona ecuatorial de África y Asia; se presentaron brotes en América entre 2015 y 2016. No existe tratamiento específico para ninguna de éstas infecciones por arbovirus. El desarrollo de vacunas es considerado actualmente como la estrategia con mayor potencial para el control de estas enfermedades. Este último requiere múltiples enfoques experimentales y epidemiológicos que evalúen la incidencia y prevalencia reales en áreas endémicas. Objetivo general. Determinar seroprevalencia y seroconversión de anticuerpos frente a virus CHIKV, DENV y ZIKV en individuos sanos del Hospital General “Dr. Miguel Silva”, la Facultad de Ciencias Médicas y Biológicas “Dr. Ignacio Chávez” de la UMSNH y voluntarios de la población general en la ciudad de Morelia, Michoacán, en el período de un año.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud The protein STAT3 has been involved in the development of breast cancer because it modulates genes involved in carcinogenesis. Bcl-xL is an anti-apoptotic protein that promotes greater survival to neoplastic cells. Both proteins are associated with poor prognosis. The aim of the present study was to associate the histological, molecular and clinical stage type with the expression of nuclear STAT3 and Bcl-xL proteins in breast cancer. In tumor microarrays (TMA) of 310 cases of breast cancer, the nuclear STAT3 protein phosphorylated (pSTAT3) and cytoplasmic and nuclear Bcl-xL, were analyzed by inmunohistochemistry. Nuclear expression was found in 78.2% and 72% of pSTAT3 and Bcl-xL cases, respectively. In analyzing the expression of nuclear pSTAT3 in relation to the histological type, statistically significant differences (P = 0.001) and with the clinical stage (P = 0.03) were found, but not with the molecular subtype (P = 0.52). A protective factor of nuclear pSTAT3 was found between IIA and IIB stages (RR, 0.68; IC 95%, 0.517-0.893; P = 0.014), and relative risk between stages IIA and IIIA (RR, 2,696; IC 95%, 1,169-6.218; P = 0.006); as well as between stages N0 and N1 (RR 1,845; IC 95%, 1.123-3.029; P = 0.006). The expression of total Bcl-xL (nuclear and cytoplasmic) showed no statistically significant differences with the histological type (P = 0.06 and P = 0.20), with the molecular subtype (P = 0.14 and P = 0.70) or with the clinical stage (P = 0.56 and P = 0.15). The results of this study suggest that STAT3 has a dual function in breast cancer, as it presents both as a protective factor in early clinical stages and as a risk factor in locally advanced clinical stages. La proteína STAT3 se ha visto implicada en el desarrollo del cáncer de mama porque modula genes implicados en la carcinogénesis. Bcl-xL es una proteína antiapoptótica que favorece mayor supervivencia a las células neoplásicas. Ambas proteínas se asocian con mal pronóstico. El objetivo de este estudio fue asociar el tipo histológico, molecular y estadio clínico con la expresión de las proteínas STAT3 nuclear y Bcl-xL en cáncer de mama. Mediante inmunohistoquímica en microarreglos de tumores (TMA) de 310 casos de cáncer de mama, se analizaron la proteína STAT3 nuclear fosforilada (pSTAT3) y Bcl-xL citoplasmática y nuclear. Se encontró expresión nuclear en 78.2% y 72% de los casos de pSTAT3 y Bcl-xL, respectivamente. Al analizar la expresión de pSTAT3 nuclear en relación al tipo histológico, se encontraron diferencias estadísticamente significativas (P = 0.001) y con el estadio clínico (P = 0.03), pero no con el subtipo molecular (P = 0.52). Se encontraron un factor protector de pSTAT3 nuclear entre los estadios IIA y IIB (RR, 0.68; IC 95%, 0.517-0.893; P = 0.014) y riesgo relativo entre los estadios IIA y IIIA (RR, 2.696; IC 95%, 1.169-6.218; P = 0.006); así como entre los estadios N0 y N1 (RR 1.845; IC 95%, 1.123-3.029; P = 0.006). La expresión de Bcl-xL total (nuclear y citoplasmática) no mostró diferencias estadísticamente significativas con el tipo histológico (P=0.06 y P=0.20), con el subtipo molecular (P=0.14 y P=0.70) ni con el estadio clínico (P=0.56 y P=0.15). Los resultados de este estudio sugieren que STAT3 tiene una función dual en el cáncer de mama, ya que se presenta como un factor de protección en estadios clínicos tempranos y como factor de riesgo en estadios clínicos localmente avanzados.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Discoid and anucleate cells, platelets have traditionally been associated with the processes of haemostasis and thrombosis; however, its involvement in other biological processes has recently been described including the immune response against viruses. Dengue (DENV) and Zika (ZIKV) are arboviruses that have a genome that encodes 3 structural and 7 non-structural proteins. Non-structural protein 1 (NS1) is mainly involved in the replication of viral genetic material. NS1 can also be secreted by infected cells and their circulating presence has been associated with the evasion of the immune response against these viruses. Objective: Study platelet reactivity in response to stimulus with the NS1 protein of dengue and Zika virus. Materials and methods: Blood sample was obtained by venepuncture with Vacutainer system® volunteers who agree to participate in the protocol and meet the criteria of inclusion. Determination of the serological status of NS1 samples of dengue and Zika using ELISA method. Obtention of platelet-rich plasma (PRP) 1000, and platelet-poor-plasma (PPP). Determination of platelet stimulus conditions by flow cytometry (CytoFLEX, Beckman Coulter®). Standardization of atomic force microscopy (AFM) and quartz crystal microbalance (QCM) for further evaluation of post-stimulus cell reactivity with NS1 protein. NT-MDT microscope® NTEGRA and a QCMopen, Novaetech® microbalance was used. Results: The seronegativeness of the 3 working samples was confirmed. The increased expression of markers P-Selectin and GP aIIßI by flow cytometry were observed at 60 min with a protein concentration of 2.5 μg/mL and this was higher when compared to known agonists of activation (ADP, collagen and epinephrine), and the basal expression in inactive platelets. We determined PRP/Buffered Tyrodes onto glass surface for the assessment of platelet by AFM. Células discoides y anucleadas, las plaquetas han estado asociadas tradicionalmente a los procesos de hemostasia y trombosis; sin embargo, recientemente se ha descrito su participación en otros procesos biológicos incluida la respuesta inmunológica contra virus. El dengue (DENV) y el Zika (ZIKV) son arbovirus que poseen un genoma que codifica 3 proteínas estructurales y 7 no estructurales. La proteína no estructural 1 (NS1) participa principalmente en la replicación del material genético viral. NS1 puede también ser secretada por células infectadas y su presencia en circulación se ha asociado a la evasión de la respuesta inmune contra estos virus. Objetivo: Estudiar la reactividad plaquetaria en respuesta a estímulo con la proteína NS1 del virus de dengue y Zika. Materiales y métodos: Obtención de muestra sanguínea por venopunción con sistema Vacutainer® de voluntarios que acepten participar en el protocolo y cumplan con los criterios de inclusión. Determinación del estado serológico de las muestras contra NS1 de dengue y Zika mediante ELISA. Obtención de plasma rico en plaquetas (PRP) y de plasma pobre en plaquetas (PPP). Determinación de las condiciones de estímulo plaquetario mediante citometría de flujo (CytoFLEX, Beckman Coulter®). Estandarización de microscopia de fuerza atómica (AFM) y microbalanza de cristal de cuarzo (QCM) para posterior evaluación de la reactividad celular posterior a estímulo con proteína NS1. Se utilizó un microscopio NT-MDT® NTEGRA y una microbalanza QCMopen, Novaetech®. Resultados: Se confirmó la seronegatividad de las 3 muestras de trabajo. La mayor expresión de los marcadores P-Selectina y GP αIIβI por citometría de flujo se observaron a los 60 minutos con una concentración de proteína de 2.5 μg/mL y que esta fue mayor al compararse con ADP, colágeno y epinefrina y la expresión basal en plaquetas inactivas. Se determinó PRP/Buffer Tyrodes sobre superficie de vidrio para la evaluación de plaquetas mediante AFM.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Zika is a viremia that has a nonspecific clinical picture and that has been associated with other complications such as Guillain-Barré Syndrome and Congenital Syndrome. The immune response displayed before infections. includes an inflammatory state that is characterized by an elevation of cytokines. On the other hand, fever is a related but not exclusive sign of the infections that is included in the Zika infection. Fever is caused by cytokines such as IL-1, IL-6, TNF and IFN γ. On the other hand, platelets are cells that participate in the immune response in bacterial and viral infections, and as they are also mediators of inflammation. Platelets are activated in infections and vascular lesions, releasing activation biomarkers such as PF-4, B-TG, CD40Ls and P-selectin, FvW. Objective: To determine if there is platelet activation in Zika virus infection through the quantification of platelet biomarkers (MPV, PF-4, B-thromboglobulin, CD40Ls, P-selectin) and endothelial damage (VWF) Material and Methods: Blood samples were collected from serum and citrated plasma from patients with probable Zika infection, with acute onset fever and apparently healthy subjects as control. El Zika, es una viremia que tiene un cuadro clínico inespecífico y que se ha asociado con otras complicaciones como Síndrome de Guillain-Barré y Síndrome Congénito. La respuesta inmune desplegada ante infecciones. incluye un estado inflamatorio que se caracteriza por una elevación de citocinas Por otra parte la fiebre, es un signo relacionado, pero no exclusivo de las infecciones que está incluido en la infección Zika. La fiebre es originada por citocinas como IL-1, IL-6, TNF  e IFN γ. Por otra parte, las plaquetas son células que participen en la respuesta inmune en infecciones bacterianas y virales, y como también son mediadoras de la inflamación. Las plaquetas se activan en infecciones y las lesiones vasculares, liberando biomarcadores de activación como el PF-4, B- TG, CD40Ls y P- selectina, FvW. Objetivo: Determinar si existe activación plaquetaria en la infección por virus Zika a través de la cuantificación de biomarcadores plaquetarios (VPM, PF-4, B- tromboglobulina, CD40Ls, P- selectina) y daño endotelial (FvW) Material y Métodos: Se colectaron muestras sanguíneas de suero y plasma citratado de pacientes con probable infección por Zika, con fiebre de inicio agudo y de sujetos aparentemente sanos como control.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Arbovirus infections are a problem of public health by complications presenting. Immune response is still not well defined; it is thought that platelets are involved in this response. However, there are no studies that determine its activation in these infections. Our goal is to investigate the presence of Platelet Activation in the immune response to infections by DENV, CHIKV and ZIKV. Methodology. A study of cases and controls was conducted, qRT-PCR was performed for diagnosis and by ELISA biomarkers were evaluated. Differences levels of biomarkers were assessed using Kruskal-Wallis test Dunn and determined the correlation between inflammatory biomarkers and Platelet Activation by spearman test. Results. We confirmed 20 cases which was performed Cytometry hematic and measurement of platelet and inflammatory biomarkers. We found modest elevation of platelet and inflammatory biomarkers measured, enough to depurate the virus, also indicates a platelet activation. We found significantly decreased of Hb, HCT and lymphocyte, as well as increase the VPM, leukocytes, FvW and PF-4 in the ZIKV group. In the Group of CHIKV we obtain a decrease of Hb and HCT also an increase of VPM, leukocytes and IL-6. Las infecciones por arbovirus son un problema de salud pública por las complicaciones que presentan. La respuesta inmune aún no está bien definida, se piensa que las plaquetas participan en esta respuesta. Sin embargo, no hay estudios que determinen su activación en estas infecciones. Nuestro objetivo es investigar la presencia de activación plaquetaria en la respuesta inmune ante infecciones por DENV, CHIKV Y ZIKV. Metodología. Se realizó un estudio de casos y controles, se realizó diagnóstico definitivo por qRT-PCR y se evaluaron los biomarcadores mediante ELISA. Se realizó medianas con rangos. Se analizaron las diferencias mediante Kruskal-Wallis con prueba Dunn y se determinó la correlación entre los biomarcadores inflamatorios y activación plaquetaria mediante una prueba de spearman. Resultados. Se confirmaron 20 casos a los cuales se les realizo citometría hemática y medición de biomarcadores inflamatorios y plaquetarios. Se encontró elevación modesta de los biomarcadores inflamatorio y plaquetarios medidos, suficiente para eliminar al virus, también indica una activación plaquetaria. De forma significativa encontramos en el grupo de ZIKV disminución de Hb, Hto y linfocitos incremento del VPM, leucocitos, FvW y PF-4.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Obesity(OB) is a systemic disease, characterized by fat tissue remodeling that triggers a low-grade chronic inflammatory state, which is manifested by the expression of pro-inflammatory biomarkers that correlate with endothelial dysfunction(DE) and insulin resistance(IR). BAFF has been linked with IR development, suggesting its role in the chronic inflammatory state related to OB. Objective: To evaluate the association between BAFF and inflammation and ED biomarkers in OB patients. Methods: Cross-sectional comparative study. Three groups of patients (Normal-weight, Overweight and Obesity) underwent physical examination and venous blood collection: blood count, blood chemistry (glucose, triglycerides, total cholesterol, HDL-c and LDL-c) TH1/TH2/TH17 cytokines, ED markers (vWF, sCD40L), insulin and BAFF. Results: Serum BAFF increased significantly with BMI(p

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Malnutrition is a common complication in chronic kidney disease. In dialysis this prevalence are estimated between 18-75%; multifactorial causes in the inflammatory status of patients on hemodialysis (HD), named "complex syndrome of malnutrition and inflammation," also known as MIA syndrome: Malnutrition, inflammation, atherosclerosis syndrome. In chronic inflammation the most studied biomarkers for monitoring are: C reactive protein (CRP), interleukin-6 (IL-6), ferritin, transferrin and albumin; free triiodothyronine (FT3) and mean platelet volume (MPV) are proposed as inflammatory biomarkers. Aim: Determine the correlation between VPM and FT3 with MIA syndrome. Material and Methods: Cross-sectional, descriptive and analytical study of patients in chronic HD in Hospital General Regional No. 1 (HGR No.1) IMSS, after signing the informed consent, MIS survey, nutrition assessment, doppler ultrasound carotid to measure thickness of the carotid intima was taken blood for determination of FT3, complete blood count, glucose, urea, creatinine, albumin, parathormone, CRP and IL-6. Results: The prevalence of MIA was 53.9%; We didn't found association of the MPV with the MIA Syndrome. La desnutrición es una complicación comunmente crónica; en diálisis, su prevalencia se ha estimado entre 18 a 75%; las causas son multifactoriales e influyen en el estado inflamatorio de los pacientes en hemodiálisis (HD), lo que en conjunto se ha denominado “síndrome complejo de malnutrición e inflamación”, también denominado como síndrome MIA: Malnutrition inflamation, atherosclerosis syndrome. Dentro de esta inflamación crónica las biomarcadores más estudiadas para su vigilancia son la proteína C reactiva (PCR) e interleucina 6 (IL-6), ferritina, transferrina y albúmina; se propone triyodotironina libre (T3L) y volumen plaquetario medio (VPM) como biomarcadores inflamatorios. Objetivo: Conocer la prevalencia de Síndrome MIA y la correlación entre el VPM- T3L con el Síndrome MIA. Material y métodos: Estudio transversal, descriptivo y analítico de pacientes en HD crónica en el Hospital General Regional No. 1 (HGR No.1) del IMSS, previa firma del consentimiento informado se realizó aplicación de encuesta MIS, valoración por nutrición, ultrasonido doppler carotideo para medir grosor de la íntima carotídea (GMIc), y se tomó de sangre para determinación de T3L, biometría hemática completa, glucosa, urea, creatinina, albúmina, parathormona, PCR e IL-6. Resultados: La prevalencia de MIA es 53.9%; no encontramos asociación del VPM con el Síndrome MIA.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud Ischemic cardiovascular disease is the leading cause of death worldwide and the second in Mexico, atherosclerosis and coronary atherosclerosis are the main pathophysiological substrate of ischemic heart disease. Secondary ischemia triggers release of growth factors that stimulate the bone marrow to release endothelial progenitor cells. These cells are involved in neoangiogenesis and rejuvenation of the endothelium, plus there is a close interaction between these cells and cardiovascular risk factors, so it is very important study as a new diagnostic and prognostic marker in this disease, relating quantifying with other biomarkers, as well as the clinical features of patients. OBJECTIVE: To correlate the number of circulating endothelial progenitor cells with plasma levels of prothrombotic biomarkers of endothelial damage in patients with ischemic cardiovascular disease. METHODOLOGY: endothelial progenitor cells were quantified by flow cytometry, are determined prothrombotic biomarkers (vWF, P-selectin, LCD40, PAI-1, adiponectin) by ELISA, which are correlated with each other and clinical parameters of the patients. RESULTS: A total of 88 patients, 69 were included in the study with a diagnosis of ischemic heart disease: 22 patients in the stable angina group, 24 in the NSTEMI and STEMI 23 in addition to 19 healthy subjects. Finding elevated levels of PAI-1 in patients with ischemic heart disease unlike the control group, the number of EPCs is reduced in patients with stable angina, in uncontrolled diabetes, in women and in elderly patients, in addition to adiponectin is elevated in patients with better cardiovascular Framingham risk and patients with higher numbers of HDL. CONCLUSIONS: The amount of CPE is decreased in patients with stable angina and uncontrolled diabetes, which is a reflection of chronic endothelial dysfunction in these patients. Adiponectin is a useful biomarker that correlates inversely with cardiovascular risk patients La enfermedad cardiovascular isquémica es la primera causa de muerte en el mundo y la segunda en México, la ateroesclerosis y la aterotrombosis coronaria son el principal sustrato fisiopatológico de la cardiopatía isquémica. La isquemia secundaria desencadena liberación de factores de crecimiento que estimulan a la médula ósea para la liberación de células progenitoras endoteliales. Estas células participan en la neoangiogénesis y el rejuvenecimiento del endotelio, además de que hay una estrecha interacción entre dichas células y los factores de riesgo cardiovascular, por lo que es de suma importancia su estudio como un nuevo marcador diagnóstico y pronóstico en esta patología, relacionándolo con la cuantificación de otros biomarcadores, así como con las características clínicas de los pacientes. OBJETIVO: Correlacionar la cantidad de células progenitoras endoteliales circulantes con los niveles plasmáticos de biomarcadores protrombóticos de daño endotelial en pacientes con enfermedad cardiovascular isquémica. METODOLOGÍA: Se cuantificaron células progenitoras endoteliales mediante citometría de flujo, se determinarán biomarcadores protrombóticos (FvW, P-Selectina, LCD40, PAI-1, Adiponectina) por técnica de ELISA, los cuales se correlacionarán entre sí y con parámetros clínicos de los pacientes. RESULTADOS: Se incluyeron en el estudio un total de 88 pacientes, 69 con diagnóstico de cardiopatía isquémica: 22 pacientes incluidos en el grupo de angina estable, 24 en el de IAMSEST y 23 en el de IAMCEST además de 19 sujetos sanos. Encontrando niveles elevados de PAI-1 en los pacientes con cardiopatía isquémica a diferencia del grupo control, que la cantidad de CPE está disminuida en pacientes con Angina estable, en diabéticos descontrolados, en mujeres y en pacientes de edad avanzada, además de que la adiponectina se encuentra elevada en pacientes con mejor riesgo cardiovascular de Framingham y en pacientes con cifras mayores de HDL.

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud The rheumatoid arthritis ( RA) is an autoimmune disease that involves chronic widespread inflammatory process that affects 1.6% -2% of the Mexican population ( Sang- Cheol et al, 2009 ; . Brennan and McInnes 2008; Calder 2008; Peláez - Ballestas et al. , 2011). The pathophysiology of RA involves an imbalance between the production of inflammatory cytokines such as tumor necrosis factor alpha ( TNF - α ) , interleukin 1 beta ( IL - 1β ) , interleukin 6 (IL -6) , interleukin 8 (IL -8 ) and the colony-stimulating factor granulocyte (GM -CSF ) , and inflammatory and transforming growth factor - β ( TGF- β ) , interleukin 10 (IL- 10) , Interleukin 1 alpha ( IL - 1α ) compared and TNF receptors ( sTNFR ) , which puts damage at the level of articulation as other organ systems ( Calder 2008,. Sang- Cheol et al, 2009) such as the vascular endothelium favoring accelerated atherogenesis ( Bisoendial et al. 2011: Hannawi et al, 2007 ) . so the CV risk is double the general population ( Cortes et al, 2007 ; . González and González 2009,. James et al, 2010 ) . with a mortality rate of 35-50 % ( Karakoc et al, 2012 , Gonzalez and Gonzalez 2009; Pattison et al, 2004) La Artritis reumatoide (AR) es una enfermedad autoinmune que involucra un proceso inflamatorio crónico y generalizado que afecta al del 1.6%-2% de la población Mexicana (Sang-Cheol et al., 2009; Brennan y Mclnnes 2008; Calder 2008; Peláez-Ballestas et al., 2011). La fisiopatología de la AR implica un desequilibrio entre la producción de citocinas inflamatorias como el factor de necrosis tumoral alfa (TNF-α), Interleucina 1 beta (IL-1β), Interleucina 6 (IL-6), Interleucina 8 (IL-8) y el factor de estimulación de colonias de granulocitos (GM-CSF) y respecto las antiinflamatorias como el factor de crecimiento transformante-β (TGF-β), Interleucina 10 (IL- 10), Interleucina 1 alfa (IL-1α) y receptores para TNF (sTNFR), lo que ejerce daño tanto a nivel de articulación como a otros órganos y sistemas (Calder 2008; Sang-Cheol et al., 2009) como lo es el endotelio vascular favoreciéndose una aterogénesis acelerada (Bisoendial et al., 2011: Hannawi et al., 2007); por lo que el riesgo CV es del doble que la población general (Cortés et al., 2007; González y González 2009; James et al., 2010) con una tasa de mortalidad del 35-50% (Karakoc et al., 2012; González y González 2009; Pattison et al., 2004).

Facultad de Ciencias Médicas y Biológicas. Maestría en Ciencias de la Salud The percutaneous coronary intervention (PCI) is the therapy of choice for revascularization in coronary artery disease (CAD), usually caused by atherosclerosis. Attached to the PCI, antiplatelet therapy by inhibition of P2Y12 ADP receptor by clopidogrel reduce the incidence of mayor cardiovascular adverse events (MACE). However, there is a group of patients with low response to clopidogrel, identified as "nonresponders." This variability may be due to cellular, clinical and genetic factors. Genetic polymorphisms in hepatic enzymes that are responsible for the biotransformation of the drug, particularly in the cytochrome P450 family have been study. Polymorphisms in CYP2C19 isoenzyme (particularly the loss of function cytochorme CYP2C19*2 variant) have accounted for Clopidogrel nonresponse in patients with ischaemic heart disease. There are several methods to assess platelet function. The analysis of the phosphorylation of vasodilator-stimulated phosphoprotein (VASP) by flow cytometry, evaluates the degree of inhibition of the P2Y12 receptor and thereby determine the Platelet Reactivity Index (PRI) as a messure of effectiveness of Clopidogrel. La Intervención Coronaria Percutánea (ICP) es la terapia de elección para la revascularización en la Enfermedad Arterial Coronaria, ocasionada generalmente por la ateroesclerosis. Unida a la ICP, la terapia antiplaquetaria por inhibición del receptor de ADP P2Y12 mediante Clopidogrel ha reducido la incidencia de eventos adversos cardio-cerebro-vasculares-mayores (ECCVM). Sin embargo existe un grupo de pacientes que presentan baja respuesta a Clopidogrel, identificados como ?no respondedores?. Esta variabilidad puede deberse a factores celulares, clínicos y genéticos; dentro de éstos últimos se ha estudiado los polimorfismos en la familia del citocromo P450, enzimas que se encargan de la biotransformación de fármacos. Destaca el estudio de la isoforma CYP2C19, enzima que participa en los dos pasos de la transformación del Clopidogrel en su metabolito activo; y su defecto CYP2C19*2, que su presencia está relacionada con un fenotipo de una pobre metabolización del fármaco. Existen varios métodos para evaluar la función plaquetaria, dentro de los cuales el análisis de la fosforilación de la fosfoproteína estimulada por vasodilatadores (VASP) mediante citometría de flujo permite evaluar el grado de inhibición del receptor P2Y12 y por tanto la eficacia del Clopidogrel determinando del índice de Reactividad Plaquetaria (IRP).