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1.
Eur J Immunol ; 52(1): 123-137, 2022 01.
Article in English | MEDLINE | ID: mdl-34599760

ABSTRACT

The immunopathogenesis of multisystem inflammatory syndrome (MIS-C) in children that may follow exposure to SARS-CoV-2 is incompletely understood. Here, we studied SARS-CoV-2-specific T cells in MIS-C, Kawasaki disease (KD), and SARS-CoV-2 convalescent controls using peptide pools derived from SARS-CoV-2 spike or nonspike proteins, and common cold coronaviruses (CCC). Coordinated CD4+ and CD8+ SARS-CoV-2-specific T cells were detected in five MIS-C subjects with cross-reactivity to CCC. CD4+ and CD8+ T-cell responses alone were documented in three and one subjects, respectively. T-cell specificities in MIS-C did not correlate with disease severity and were similar to SARS-CoV-2 convalescent controls. T-cell memory and cross-reactivity to CCC in MIS-C and SARS-CoV-2 convalescent controls were also similar. The chemokine receptor CCR6, but not CCR9, was highly expressed on SARS-CoV-2-specific CD4+ but not on CD8+ T cells. Only two of 10 KD subjects showed a T-cell response to CCC. Enumeration of myeloid APCs revealed low cell precursors in MIS-C subjects compared to KD. In summary, children with MIS-C mount a normal T-cell response to SARS-CoV-2 with no apparent relationship to antecedent CCC exposure. Low numbers of tolerogenic myeloid DCs may impair their anti-inflammatory response.


Subject(s)
CD4-Positive T-Lymphocytes/immunology , CD8-Positive T-Lymphocytes/immunology , COVID-19/complications , Immunity, Cellular , Immunologic Memory , Mucocutaneous Lymph Node Syndrome , SARS-CoV-2/immunology , Systemic Inflammatory Response Syndrome/immunology , Adolescent , COVID-19/immunology , Child , Child, Preschool , Female , Humans , Infant , Male , Mucocutaneous Lymph Node Syndrome/complications , Mucocutaneous Lymph Node Syndrome/immunology
2.
BMC Pediatr ; 23(1): 465, 2023 09 15.
Article in English | MEDLINE | ID: mdl-37715152

ABSTRACT

BACKGROUND: Children with neurodevelopmental disorders have a very wide clinical variability. A common prevalent factor is problems with stool and sleep quality. Currently, there are multiple studies related to their evaluation, but not so much related to a specific intervention. The aim was to evaluate the effectiveness and safety of the application of non-invasive neuromodulation as a treatment in children with neurodevelopmental disorders to improve constipation and quality of sleep. METHODS: A total of 23 minors aged between 2 and 16 were included in this cross-sectional study. All participants were applied the microcurrent device for 60 min, 3 times per week for a total of 4 weeks. The technique was based on non-invasive neuromodulation using a surface-applied microcurrent electrostimulation device that administers an external, imperceptible, pulsed electrical stimulation. It is applied to the extremities, in a coordinated manner, using gloves and anklets connected with electrodes to a control console. Sleep latency and microarousals were evaluated through a sleep diary. To assess the evolution and type of defecation, the adapted and validated version in Spanish of the Bristol Stool Form Scale was used. RESULTS: No adverse events occurred during the study and no incidences were registered. Clinically relevant improvements were registered in defecation frequency and type as well as in sleep related parameters. An increase in the hours of sleep was registered, from 7,35 (0,83) to 9,09 (1,35), and sleep interruptions decreased from 3,83 (1,95) to 1,17 (1,11), (p < .001). CONCLUSION: Microcurrents can be used as an effective and safe treatment to improve quality of sleep and constipation in children with neurodevelopmental disorders. More studies are needed in order to obtain statistically significant results. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT05265702. FIRST REGISTRATION: 03/03/2022 PROTOCOL: https://clinicaltrials.gov/ct2/show/NCT05265702?term=baez+suarez&draw=2&rank=4.


Subject(s)
Neurodevelopmental Disorders , Sleep Quality , Child , Humans , Child, Preschool , Adolescent , Cross-Sectional Studies , Sleep , Constipation/therapy , Neurodevelopmental Disorders/therapy
3.
J Pediatr ; 243: 173-180.e8, 2022 04.
Article in English | MEDLINE | ID: mdl-34953816

ABSTRACT

OBJECTIVES: To determine the safety, pharmacokinetics, and immunomodulatory effects of 2-6 weeks of anakinra therapy in patients with acute Kawasaki disease with a coronary artery aneurysm (CAA). STUDY DESIGN: We performed a Phase I/IIa dose-escalation study of anakinra (2-11 mg/kg/day) in 22 patients with acute Kawasaki disease with CAA. We measured interleukin (IL)-1RA concentrations after the first dose and trough levels up to study week 6. Markers of inflammation and coronary artery z-scores were assessed pretreatment and at 48 hours, 2 weeks, and 6 weeks after initiation of therapy. RESULTS: Up to 6 weeks of anakinra (up to 11 mg/kg/day) was safe and well tolerated by the 22 participants (median age, 1.1 years), with no serious adverse events attributable to the study drug. All participants were treated with intravenous immunoglobulin (IVIG), and 20 also received infliximab (10 mg/kg) before initiation of anakinra. Serum levels of IL-6, IL-8, and tumor necrosis factor α decreased similarly in patients with Kawasaki disease treated with IVIG, infliximab, and anakinra compared with age- and sex-matched patients with Kawasaki disease treated only with IVIG and infliximab. Anakinra clearance increased with illness day at diagnosis. Simulations demonstrated that more frequent intravenous (IV) dosing may result in more sustained concentrations without significantly increasing the peak concentration compared with subcutaneous (SC) dosing. CONCLUSIONS: Both IV and SC anakinra are safe in infants and children with acute Kawasaki disease and CAA. IV dosing every 8-12 hours during the acute hospitalization of patients with Kawasaki disease may result in a sustained concentration while avoiding frequent SC injections. The efficacy of a short course of IV therapy during hospitalization should be studied. TRIAL REGISTRATION CLINICALTRIALS.GOV: NCT02179853.


Subject(s)
Coronary Aneurysm , Interleukin 1 Receptor Antagonist Protein , Mucocutaneous Lymph Node Syndrome , Acute Disease , Coronary Aneurysm/complications , Coronary Aneurysm/drug therapy , Female , Humans , Immunoglobulins, Intravenous/therapeutic use , Infant , Infliximab/therapeutic use , Interleukin 1 Receptor Antagonist Protein/adverse effects , Male , Mucocutaneous Lymph Node Syndrome/complications , Mucocutaneous Lymph Node Syndrome/drug therapy
4.
Parasitology ; 145(9): 1161-1169, 2018 08.
Article in English | MEDLINE | ID: mdl-29526166

ABSTRACT

American tegumentary leishmaniasis (ATL) samples obtained from the lesions of patients with typical (n = 25, 29%), atypical (n = 60, 69%) or both (n = 2%) clinical manifestations were analysed by multilocus enzyme electrophoresis, hsp70 restriction-fragment length polymorphism (PCR-RFLP), hsp70 sequencing and phylogenetics methods. The hsp70 PCR-RFLP analysis revealed two different profiles whose the most samples differed from those expected for Leishmania braziliensis and the other Leishmania species tested: of 39 samples evaluated, two (5%) had a restriction profile corresponding to L. braziliensis, and 37 (95%) had a restriction profile corresponding to a variant pattern. A 1300-bp hsp70 gene fragment was sequenced to aid in parasite identification and a phylogenetic analysis was performed including 26 consensus sequences from the ATL patient's samples and comparing to other Leishmania and trypanosomatids species. The dendrogram allowed to observe a potential population structure of L. braziliensis complex in the studied region, emphasizing that the majority of clinical samples presented a variant genetic profile. Of interest, the L. braziliensis diversity was associated with different clinical manifestations whose parasites with hsp70 variant profile were associated with atypical lesions. The results may be helpful to improve the diagnosis, treatment and control measures of the ATL in endemic areas.


Subject(s)
Genetic Variation , Leishmania braziliensis/genetics , Leishmaniasis, Cutaneous/epidemiology , Skin/parasitology , Brazil/epidemiology , DNA, Protozoan/genetics , Endemic Diseases , HSP70 Heat-Shock Proteins/genetics , Humans , Phylogeny , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Skin/pathology
5.
Transfusion ; 56(11): 2857-2867, 2016 11.
Article in English | MEDLINE | ID: mdl-27716930

ABSTRACT

BACKGROUND: There has been increased worldwide emphasis on the many benefits of human immunodeficiency virus (HIV) serostatus awareness for both infection prevention and improved treatment outcomes. Previous studies indicate that donors may use blood donation to be tested; the objectives of this analysis were to assess, among donors with previously undisclosed risk behavior in the 12 months before donation, the frequency of those who have previously been tested for HIV and the demographic and behavioral factors associated with such testing. STUDY DESIGN AND METHODS: In this secondary analysis from an HIV case-control study of blood donors in Brazil, we analyzed the response to the question, "Other than blood donation, have you ever been tested for HIV?" Demographic and disclosed risk behaviors associated with previous testing were determined. RESULTS: The study included 341 HIV-positive cases and 791 HIV-negative controls (1:2 case/control ratio). Overall, 31% of blood donors (40% of cases and 26% of controls) reported having been tested for HIV outside of blood donation. History of HIV testing varied according to sex, HIV status, and reported sexual risk behavior. CONCLUSION: Although it is encouraging that previous testing was more frequent in donors with acknowledged sexual risk behavior in Brazil, 60% still had not been tested for HIV outside of the blood donation setting. Educating donors on the importance of not using blood centers as a means to get tested for HIV in Brazil, especially if they engage in higher risk behaviors, and seeking alternate testing venues instead could improve the safety of donated blood.


Subject(s)
Blood Donors , HIV Infections/diagnosis , Self Report , Adolescent , Adult , Aged , Brazil , Case-Control Studies , Female , HIV Infections/transmission , Humans , Male , Middle Aged , Risk-Taking , Sexual Behavior , Young Adult
6.
Virol J ; 13: 121, 2016 07 04.
Article in English | MEDLINE | ID: mdl-27377374

ABSTRACT

BACKGROUND: Bovine vaccinia (BV) is a zoonosis caused by Vaccinia virus, a virus from Orthopoxvirus genus (OPV) that affects mainly cattle herds and humans in rural areas in Brazil. Because most studies have focused on outbreaks situations, data on BV epidemiology is limited. A cross sectional study in Brazilian rural areas during 2012-2013 was conducted to determine the neutralizing antibodies seroprevalence and risk factors for BV. METHODS: A structured questionnaire was applied to elicit demographics data and farming practices considered risk factors for BV exposure. Neutralizing anti-OPV antibodies were investigated using plaque reduction neutralization test. The neutralizing antibodies prevalence rates were calculated and the risk factor analysis was performed using multivariate logistic regression. RESULTS: Two hundred and forty participants were enrolled in this study with a prevalence of neutralizing antibodies of 30.8 % (95 % confidence interval [CI], 25.3-36.9). In multivariate analysis, age > 35 years (Odds Ratio [OR] = 18.2; CI 95 % = 7.7 - 43.2) and previous outbreak in property (OR = 3.9; C I95 % = 1.2 - 12.6) were independently associated with anti-OPV neutralizing antibodies. CONCLUSIONS: In this study, anti-OPV protective immunity (neutralizing antibody titers) was assessed in an endemic BV Brazilian rural area. Our findings indicate that epidemiological surveillance is required and should be applied by public health authorities to create interventions and/or prevention strategies to avoid viral spread causing future outbreaks among individuals who are under risk of infection.


Subject(s)
Antibodies, Viral/blood , Orthopoxvirus/immunology , Poxviridae Infections/blood , Zoonoses/blood , Adolescent , Adult , Aged , Aged, 80 and over , Agricultural Workers' Diseases/blood , Agricultural Workers' Diseases/immunology , Agricultural Workers' Diseases/virology , Animals , Antibodies, Neutralizing/blood , Brazil/epidemiology , Child , Child, Preschool , Cross-Sectional Studies , Female , Humans , Livestock/virology , Male , Middle Aged , Orthopoxvirus/genetics , Orthopoxvirus/isolation & purification , Poxviridae Infections/immunology , Poxviridae Infections/virology , Rural Population , Seroepidemiologic Studies , Young Adult , Zoonoses/epidemiology , Zoonoses/immunology , Zoonoses/virology
7.
Transfus Apher Sci ; 53(2): 238-41, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26138911

ABSTRACT

This study aimed to evaluate the trends of HCV seropositivity rates in first time blood donors from a reference blood centre in Southeast Brazil. Data from the period of 2007-2010 were analysed according to anti-HCV antibodies, donor demographic characteristics and type of donation. There was a marked and continuous decline in prevalence in the analysed period, and in 93,534 first time donors, the prevalence of anti-HCV was 0.09%. Anti-HCV were associated with less education and older age (≥ 35 years). The rates of anti-HCV observed in the present study were lower than in Brazil, but considerably higher than developed countries.


Subject(s)
Blood Donors , Hepatitis C Antibodies/blood , Hepatitis C/blood , Hepatitis C/epidemiology , Adult , Biomarkers/blood , Brazil/epidemiology , Female , Humans , Male , Middle Aged , Prevalence , Risk Factors
8.
Transfusion ; 53(6): 1291-301, 2013 Jun.
Article in English | MEDLINE | ID: mdl-22998740

ABSTRACT

BACKGROUND: Studies analyzing motivation factors that lead to blood donation have found altruism to be the primary motivation factor; however, social capital has not been analyzed in this context. Our study examines the association between motivation factors (altruism, self-interest, and response to direct appeal) and social capital (cognitive and structural) across three large blood centers in Brazil. STUDY DESIGN AND METHODS: We conducted a cross-sectional survey of 7635 donor candidates from October 15 through November 20, 2009. Participants completed self-administered questionnaires on demographics, previous blood donation, human immunodeficiency virus testing and knowledge, social capital, and donor motivations. Enrollment was determined before the donor screening process. RESULTS: Among participants, 43.5 and 41.7% expressed high levels of altruism and response to direct appeal, respectively, while only 26.9% expressed high levels of self-interest. More high self-interest was observed at Hemope-Recife (41.7%). Of participants, 37.4% expressed high levels of cognitive social capital while 19.2% expressed high levels of structural social capital. More high cognitive and structural social capital was observed at Hemope-Recife (47.3 and 21.3%, respectively). High cognitive social capital was associated with high levels of altruism, self-interest, and response to direct appeal. Philanthropic and high social altruism were associated with high levels of altruism and response to direct appeal. CONCLUSION: Cognitive and structural social capital and social altruism are associated with altruism and response to direct appeal, while only cognitive social capital is associated with self-interest. Designing marketing campaigns with these aspects in mind may help blood banks attract potential blood donors more efficiently.


Subject(s)
Altruism , Blood Banks/statistics & numerical data , Blood Donors/psychology , Blood Donors/supply & distribution , Motivation , Social Values , Adolescent , Adult , Attitude to Health , Blood Donors/statistics & numerical data , Brazil/epidemiology , Cross-Sectional Studies , Educational Status , Female , Humans , Male , Multivariate Analysis , Self Efficacy , Surveys and Questionnaires , Young Adult
9.
Rev Med Inst Mex Seguro Soc ; 61(Suppl 1): S59-S64, 2023 01 01.
Article in Spanish | MEDLINE | ID: mdl-36378144

ABSTRACT

For the proper functioning of the Immunohematology Area, an External Quality Control was established since 1973 through a program that evaluates the performance of the laboratories of the Blood Bank and transfusion services that carry out immunohematology tests. This program consists of sending panel cells to participating blood banks or services, which are phenotyped erythrocyte samples previously studied as problem cases but whose results are unknown by the participating laboratories. The processes in which the program is of most importance are determination of the ABO group, determination of Rh, performance of the direct and indirect Coombs test, and pre-transfusion compatibility tests. It was carried out an observational and retrospective study of the results obtained in the 2020 period from 104 units participating in the Immunohematology Quality Control Program of the National Medical Center's Blood Bank. A panel of cells was sent for external quality control of immunohematology every 45 days, resulting in 9 panels for each unit in the studied period. Compliance with the program was observed in the general result (79.6%), i.e., there was a decrease in the participation of the registered units. Of a maximum score of 100% to be obtained, it was observed a general result of 95.3% compliance of the participating units. The results obtained confirm the good general training of the immunohematology laboratories of the participating units. Yet, as in any external control program, it becomes clear that obtaining an erroneous result is a risk that can occur in any laboratory.


Para el buen funcionamiento del Área de Inmunohematología, desde 1973 se instauró el Control de Calidad Externo con un programa que evalúa los laboratorios del Banco de Sangre y los servicios de transfusión que hacen pruebas de inmunohematología. El programa consiste en enviar a los bancos de sangre, o servicios participantes, células panel, que son muestras de eritrocitos fenotipados y previamente estudiados como casos problema, pero cuyos resultados son desconocidos por los laboratorios participantes. Los procesos en los que el programa es de suma importancia son determinación del grupo ABO, determinación del Rh, realización de la prueba de Coombs directa e indirecta y las pruebas de compatibilidad pretransfusionales. Se hizo un estudio observacional y retrospectivo de los resultados de 2020 de 104 unidades participantes en el Programa de Control de Calidad de Inmunohematología del Banco de Sangre del Centro Médico Nacional Siglo XXI. Se envió un panel de células para el control de calidad externo de inmunohematología cada 45 días y dio como resultado nueve paneles para cada unidad. El cumplimiento del programa se observó en el resultado general (79.6%), es decir, disminuyó la participación de las unidades inscritas. De una calificación máxima de 100%, hubo un resultado general de las unidades participantes del 95.3% de cumplimiento. Los resultados confirman la buena capacitación general de los laboratorios de inmunohematología de las unidades participantes. Aun así, como en cualquier programa de control externo, obtener un resultado erróneo es un riesgo que puede presentarse en cualquier laboratorio.


Subject(s)
Blood Banks , Laboratories , Humans , Retrospective Studies , Quality Control
10.
Public Health Nutr ; 15(2): 285-90, 2012 Feb.
Article in English | MEDLINE | ID: mdl-21806859

ABSTRACT

OBJECTIVE: California Assembly Bill 1535 awarded $US 15 million to California public schools to promote, develop and sustain instructional school gardens through the California Instructional School Garden Program (CISGP). The present study was designed to assess the effectiveness of the CISGP at assisting schools in implementing, maintaining and sustaining an academic school garden programme, determine how schools utilized the funding they received and assess the impact of the California state budget crisis on the CISGP. DESIGN: A mid-term evaluation was used to assess the degree to which schools achieved their instructional garden-related goals. SETTING: California. SUBJECTS: Only schools that applied for the CIGSP grant as part of a school district and also provided a contact email and had a unique contact person were included in the study (n 3103, 80·6 %). RESULTS: In general, many schools reported not achieving their predicted goals with regard to the CISGP grant. Only 39·4 % of schools reported accomplishing all of their garden-related goals. Over one-third (37·8 %) of schools reported that their school gardens were negatively affected by the California budget deficit. CONCLUSIONS: The difference between predicted and actual utilization of the CISGP grants may be due to a combination of the effects of budget shortfall and insufficiency of the grant award amount.


Subject(s)
Child Nutrition Sciences/education , Gardening/education , Schools , Budgets , California , Child , Child Nutritional Physiological Phenomena , Economic Recession , Female , Gardening/economics , Humans , Male , Program Evaluation , Schools/economics
11.
Mem Inst Oswaldo Cruz ; 107(3): 348-55, 2012 May.
Article in English | MEDLINE | ID: mdl-22510830

ABSTRACT

We investigated the cytokine profile of peripheral mononuclear cells from chronic osteomyelitis (OST) patients following in vitro stimulation with staphylococcal enterotoxin A (SEA). We demonstrate that stimulation with SEA induced prominent lymphocyte proliferation and high levels of tumour necrosis factor (TNF)-α, interleukin (IL)-4 and IL-10 secretion in both OST and non-infected individuals (NI). Even though stimulation with SEA had no impact on IL-6 production in either patient group, the baseline level of IL-6 production by cells from OST patients was always significantly less than that produced by cells from NI. After classifying the osteomyelitic episodes based on the time after the last reactivation event as "early" (1-4 months) or "late" osteomyelitis (5-12 months), we found that increased levels of TNF-α and IL-4 in combination with decreased levels of IL-6 were observed in the early episodes. By contrast, increased levels of IL-10, IL-2 and IL-6 were hallmarks of late episodes. Our data demonstrate that early osteomyelitic episodes are accompanied by an increased frequency of "high producers" of TNF-α and IL-4, whereas late events are characterised by increased frequencies of "high producers" of IL-10, IL-6 and IL-2. These findings demonstrate the distinct cytokine profiles in chronic osteomyelitis, with a distinct regulation of IL-6 production during early and late episodes.


Subject(s)
Cytokines/biosynthesis , Enterotoxins/immunology , Leukocytes, Mononuclear/immunology , Nitric Oxide/biosynthesis , Osteomyelitis/immunology , Staphylococcal Infections/immunology , Staphylococcus aureus/immunology , Adolescent , Adult , Case-Control Studies , Child , Chronic Disease , Female , Humans , Interleukins/biosynthesis , Lymphocyte Activation , Male , Middle Aged , Osteomyelitis/microbiology , Tumor Necrosis Factor-alpha/biosynthesis , Young Adult
12.
Int J Cardiol Heart Vasc ; 40: 101039, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35573651

ABSTRACT

Background: We compared the long-term health of adults with prior Kawasaki disease (KD) to controls and determined whether outcomes varied by coronary artery (CA) status. Methods: We conducted a prospective cohort study of 258 KD subjects (mean 19 ± 9 years since KD) and 148 age-similar controls who completed extensive health questionnaires. KD subjects were divided into 2 groups, Cohort 1: 109 subjects followed since KD diagnosis at our institution; Cohort 2: 149 KD subjects diagnosed elsewhere. Results: KD subjects and controls were of similar age at the time of questionnaire completion (p = 0.50). Overall, 128 subjects (including 60 in Cohort 1) reported normal CAs during and after KD. Compared to controls, KD subjects with normal CAs reported several medical conditions with increased prevalence including migraine headaches, shortness of breath, and leg pain with walking, among others. When limited to Cohort 1, KD subjects were significantly more likely to report chest pain (47% vs 16%, p < 0.001) or palpitations (23% vs 10%, p = 0.01) compared to controls. Prevalence of depression was similar (7% vs 5%, p = 0.73). Conclusions: Despite always having normal CAs in the acute and subacute phases of KD, young adults with a history of KD with normal coronaries were more likely than controls to experience cardiovascular symptoms. These differences could be influenced by anxiety or depression, but report of depression was similar between groups. Whether these health differences reflect a heightened awareness of symptoms among KD subjects, or underlying vascular pathology (i.e. vasospasm, microvascular dysfunction, other) merits further study.

13.
Hematol Transfus Cell Ther ; 44(2): 169-176, 2022.
Article in English | MEDLINE | ID: mdl-33371972

ABSTRACT

INTRODUCTION: Leg ulcers (LUs) are relatively common in patients with sickle cell anemia (SCA). The role of inflammation and nitric oxide (NO) pathways in the pathophysiology of the LU is not understood. OBJECTIVE: The aim of this study was to verify the association between inflammatory molecules and nitric oxide metabolites (NOx) and the occurrence of the LU in patients with SCA. METHOD: It was a cross-sectional study on adult participants with SCA followed at Fundação Hemominas, a public blood center in Brazil. Eligible participants were recruited and included in one of two groups: Group 1, comprised of cases with SCA (Hb SS) and at least one LU at the time of inclusion in the study and Group 2, comprised of controls with SCA without a history of LU, matched by sex and age to cases. Participants were interviewed to obtain sociodemographic data and blood samples were collected. Clinical and laboratory data were abstracted from medical records. Nitric oxide metabolites (NOx) and inflammatory molecules were quantified using an immunoassay and Multiplex xMAP® technology, respectively. Eighty-seven individuals were included, ranging in age from 17 to 61 years (mean 40 ±â€¯10.7 years); 30 had LU and 57 were controls without LU. RESULTS: Participants with LU had significantly higher levels of interleukin 8 (IL-8), IL-10, IL-15, NOx and platelet and white blood cell (WBC) counts, when compared to those without LU. Participants with LU had a significantly higher risk of having a history of osteomyelitis and a higher use of antiseptic soap in bathing, when compared to those without LU. CONCLUSION: In conclusion, our results showed that NOx, inflammatory molecules and hematological features were associated with LU in Brazilian adults with SCA.

14.
PLoS One ; 16(6): e0252377, 2021.
Article in English | MEDLINE | ID: mdl-34077475

ABSTRACT

Genomic sequence analysis of Acinetobacter baumannii revealed the presence of a putative Acid Phosphatase (AcpA; EC 3.1.3.2). A plasmid construct was made, and recombinant protein (rAcpA) was expressed in E. coli. PAGE analysis (carried out under denaturing/reducing conditions) of nickel-affinity purified protein revealed the presence of a near-homogeneous band of approximately 37 kDa. The identity of the 37 kDa species was verified as rAcpA by proteomic analysis with a molecular mass of 34.6 kDa from the deduced sequence. The dependence of substrate hydrolysis on pH was broad with an optimum observed at 6.0. Kinetic analysis revealed relatively high affinity for PNPP (Km = 90 µM) with Vmax, kcat, and Kcat/Km values of 19.2 pmoles s-1, 4.80 s-1(calculated on the basis of 37 kDa), and 5.30 x 104 M-1s-1, respectively. Sensitivity to a variety of reagents, i.e., detergents, reducing, and chelating agents as well as classic acid phosphatase inhibitors was examined in addition to assessment of hydrolysis of a number of phosphorylated compounds. Removal of phosphate from different phosphorylated compounds is supportive of broad, i.e., 'nonspecific' substrate specificity; although, the enzyme appears to prefer phosphotyrosine and/or peptides containing phosphotyrosine in comparison to serine and threonine. Examination of the primary sequence indicated the absence of signature sequences characteristic of Type A, B, and C nonspecific bacterial acid phosphatases.


Subject(s)
Acid Phosphatase/metabolism , Acinetobacter baumannii/enzymology , Proteome/analysis , Recombinant Proteins/metabolism , Acid Phosphatase/genetics , Amino Acid Sequence , Hydrogen-Ion Concentration , Hydrolysis , Kinetics , Molecular Weight , Recombinant Proteins/genetics , Sequence Homology , Substrate Specificity
15.
J Clin Invest ; 131(20)2021 10 15.
Article in English | MEDLINE | ID: mdl-34464357

ABSTRACT

BACKGROUNDMultisystem inflammatory syndrome in children (MIS-C) is a rare but potentially severe illness that follows exposure to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Kawasaki disease (KD) shares several clinical features with MIS-C, which prompted the use of intravenous immunoglobulin (IVIG), a mainstay therapy for KD. Both diseases share a robust activation of the innate immune system, including the IL-1 signaling pathway, and IL-1 blockade has been used for the treatment of both MIS-C and KD. The mechanism of action of IVIG in these 2 diseases and the cellular source of IL-1ß have not been defined.METHODSThe effects of IVIG on peripheral blood leukocyte populations from patients with MIS-C and KD were examined using flow cytometry and mass cytometry (CyTOF) and live-cell imaging.RESULTSCirculating neutrophils were highly activated in patients with KD and MIS-C and were a major source of IL-1ß. Following IVIG treatment, activated IL-1ß+ neutrophils were reduced in the circulation. In vitro, IVIG was a potent activator of neutrophil cell death via PI3K and NADPH oxidase, but independently of caspase activation.CONCLUSIONSActivated neutrophils expressing IL-1ß can be targeted by IVIG, supporting its use in both KD and MIS-C to ameliorate inflammation.FUNDINGPatient Centered Outcomes Research Institute; NIH; American Asthma Foundation; American Heart Association; Novo Nordisk Foundation; NIGMS; American Academy of Allergy, Asthma and Immunology Foundation.


Subject(s)
COVID-19/complications , Immunoglobulins, Intravenous/therapeutic use , Mucocutaneous Lymph Node Syndrome/immunology , Mucocutaneous Lymph Node Syndrome/therapy , Systemic Inflammatory Response Syndrome/immunology , Systemic Inflammatory Response Syndrome/therapy , COVID-19/blood , COVID-19/immunology , COVID-19/therapy , Case-Control Studies , Cell Death/immunology , Cell Lineage/immunology , Child , Child, Preschool , Fas Ligand Protein/immunology , Female , Humans , Infant , Interleukin-1beta/antagonists & inhibitors , Interleukin-1beta/blood , Leukocyte Count , Male , Mucocutaneous Lymph Node Syndrome/blood , Neutrophil Activation , Neutrophils/classification , Neutrophils/immunology , Neutrophils/pathology , Systemic Inflammatory Response Syndrome/blood
17.
BMC Immunol ; 11: 56, 2010 Nov 15.
Article in English | MEDLINE | ID: mdl-21078176

ABSTRACT

BACKGROUND: During the pre-patent stage of infection, juvenile Schistosoma blood flukes co-opt signals from the adaptive immune system to facilitate parasite development, but the types of responses that are induced at this early stage of infection, and the parasite antigens they target, have not been characterized. RESULTS: Through analysis of experimental pre-patent infections, we show that the S. mansoni cysteine protease SmCB1 is rapidly targeted by an antigen-specific IgE response. The induction of this response is independent of schistosome eggs as infection with male or female worms alone also induced SmCB1-specific IgE. We also show that the SmCB1-specific IgE response is dependent on cognate CD4+ T cell help and IL-4, suggesting that pre-patent Th2 responses provide T cell help for the SmCB1-specific IgE response. Finally, exposed human subjects also produced IgE against SmCB1. CONCLUSIONS: Our data demonstrate that, like eggs, schistosome worms also induce functional type 2 responses and that a parasite cysteine protease is an inducer of type 2 responses during the early stages of schistosome infection.


Subject(s)
Cathepsin B/metabolism , Helminth Proteins/metabolism , Interleukin-4/metabolism , Schistosoma mansoni/immunology , Schistosomiasis/immunology , Th2 Cells/immunology , Adaptive Immunity , Animals , Cathepsin B/immunology , Female , Helminth Proteins/immunology , Humans , Immunoglobulin E/blood , Interleukin-4/genetics , Interleukin-4/immunology , Life Cycle Stages , Male , Mice , Schistosoma mansoni/pathogenicity , Schistosomiasis/blood , Th1-Th2 Balance
19.
Front Pediatr ; 8: 384, 2020.
Article in English | MEDLINE | ID: mdl-32766186

ABSTRACT

Objective: To characterize the clinical presentation and outcomes of Kawasaki disease (KD) in infants <6 months of age as compared to those ≥6 months in Latin America. Methods: We evaluated 36 infants <6 months old and 940 infants ≥6 months old diagnosed with KD in Latin America. We compared differences in laboratory data, clinical presentation, treatment response, and coronary artery outcomes between the two cohorts. Results: The majority (78.1%) of infants and children ≥6 months of age were initially diagnosed with KD, as compared to only 38.2% of infants <6 months. Clinical features of KD were more commonly observed in the older cohort: oral changes (92 vs. 75%, P = 0.0023), extremity changes (74.6 vs. 57.1%, P = 0.029), and cervical lymphadenopathy (67.6 vs. 37.1%, P = 0.0004). Whether treated in the first 10 days of illness or after the 10th day, infants <6 months were at greater risk of developing a coronary artery aneurysm compared to KD patients ≥6 months treated at the same point in the course of illness [ ≤ 10 days (53.8 vs. 9.4%, P = 0.00012); >10 days (50 vs. 7.4%, P = 0.043)]. Conclusion: Our data show that despite treatment in the first 10 days of illness, infants <6 months of age in Latin America have a higher risk of developing a coronary artery aneurysm. Delay in the diagnosis leads to larger coronary artery aneurysms disproportionately in these infants. Thus, suspicion for KD should be high in this vulnerable population.

20.
Hematol Transfus Cell Ther ; 41(3): 229-235, 2019.
Article in English | MEDLINE | ID: mdl-31085151

ABSTRACT

BACKGROUND: Test-seeking is associated with HIV in Brazilian blood donors. This study sought to investigate the frequency with which three different donor groups: deferred donors, accepted donors who tested HIV positive [HIV (+)], and accepted donors who tested infectious disease markers negative [IDM (-)], came to the blood bank at the suggestion of a health care professional. STUDY DESIGN AND METHODS: Donors deferred for reporting high-risk behaviors and participants in an HIV risk factor case-control study completed a confidential audio computer-assisted self-interview (ACASI) that included two questions related to health care professionals and test-seeking. RESULTS: Of 4013 enrolled deferred donors, 468 (11.8%) reported a health care professional suggested donation as a way to be tested for infection. Of 341 HIV (+) and 791 IDM (-) participants, 43 (12.6%) and 11 (1.4%), respectively, reported a health care professional suggested donation as a way to be tested for infection. Physicians were the most frequently reported source of referral: [(61.5% of deferred, 69.1% of HIV (+), and 9.1% of IDM (-) donors)]. CONCLUSION: HIV (+) donors and deferred donors were 10 times more likely to report test-seeking behavior by suggestion of health care professional than IDM (-) donors. If true, efforts should be made to educate health care professionals and blood donors on how to safeguard the blood supply, emphasizing that HIV testing should be done at volunteer testing centers rather than at the blood centers.

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