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1.
Anal Methods ; 16(22): 3539-3550, 2024 Jun 06.
Article in English | MEDLINE | ID: mdl-38780022

ABSTRACT

Dengue virus (DENV) is the most prevalent global arbovirus, exhibiting a high worldwide incidence with intensified severity of symptoms and alarming mortality rates. Faced with the limitations of diagnostic methods, an optical and electrochemical biosystem was developed for the detection of DENV genotypes 1 and 2, using cysteine (Cys), cadmium telluride (CdTe) quantum dots, and anti-DENV antibodies. Cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), surface plasmon resonance (SPR), atomic force microscopy (AFM), and Fourier transform infrared spectroscopy (FTIR) were employed to characterize the immunosensor. The AFM and SPR results demonstrated discernible topographic and angular changes confirming the biomolecular recognition. Different concentrations of DENV-1 and DENV-2 were evaluated (0.05 × 106 to 2.0 × 106 PFU mL-1), resulting in a maximum anodic shift (ΔI%) of 263.67% ± 12.54 for DENV-1 and 63.36% ± 3.68 for DENV-2. The detection strategies exhibited a linear response to the increase in viral concentration. Excellent linear correlations, with R2 values of 0.95391 for DENV-1 and 0.97773 for DENV-2, were obtained across a broad concentration range. Data analysis demonstrated high reproducibility, displaying relative standard deviation values of 3.42% and 3.62% for Cys-CdTe-antibodyDENV-1-BSA and Cys-CdTe-antibodyDENV-2-BSA systems. The detection limits were 0.34 × 106 PFU mL-1 and 0.02 × 106 PFU mL-1, while the quantification limits were set at 1.49 × 106 PFU mL-1 and 0.06 × 106 PFU mL-1 for DENV-1 and DENV-2, respectively. Therefore, the biosensing apparatus demonstrates analytical effectiveness in viral screening and can be considered an innovative solution for early dengue diagnosis, contributing to global public health.


Subject(s)
Biosensing Techniques , Dengue Virus , Dengue , Tellurium , Dengue Virus/isolation & purification , Dengue Virus/immunology , Biosensing Techniques/methods , Tellurium/chemistry , Humans , Dengue/diagnosis , Electrochemical Techniques/methods , Electrochemical Techniques/instrumentation , Quantum Dots/chemistry , Surface Plasmon Resonance/methods , Cysteine/chemistry , Cadmium Compounds/chemistry , Antibodies, Viral/immunology , Antibodies, Viral/analysis , Immunoassay/methods , Immunoassay/instrumentation , Limit of Detection , Microscopy, Atomic Force
2.
ACS Appl Mater Interfaces ; 14(1): 41-48, 2022 Jan 12.
Article in English | MEDLINE | ID: mdl-34932313

ABSTRACT

Zika virus (ZIKV) infection is associated with the Guillain-Barré syndrome, and when non-vector congenital transmission occurs, fetal brain abnormalities are expected. After ZIKV infection, the blood, breast milk, and other body fluids contain low viral loads. Their detection is challenging as it requires the processing of larger input volumes of the clinical samples. Pre-enrichment is a valuable strategy to increase the analyte concentration. Therefore, the authors propose the use of a hierarchal composite polyaniline-(electrospun nanofiber) hydrogel mat (ENM) for the simultaneous enrichment and impedimetric sensing of ZIKV viral particles. The electrospinning conditions of polyvinyl alcohol and alginate, including blend formulation, were optimized through a factorial design. Disintegration and gelatinization were controlled via cross-linking to improve the hydrogel properties. Hierarchization was achieved by in situ chemical deposition of conductive polyaniline. The carboxyl groups of the ENM were used for the covalent immobilization of anti-ZIKV polyclonal antibodies used in the specific recognition of ZIKV within the medium of Vero cell culture. The specific capture and desorption of virions were studied at different pHs. ENMs were characterized by scanning electron microscopy and FTIR. Atomic force microscopy along with UV-vis and electrochemical impedance spectroscopies was used to monitor the antibody immobilization, ZIKV capture, and elution processes. Our results show that 14.2 mg (0.25 cm3) of ENM can capture 38.7 ± 2.5 µg of ZIKV with a desorption rate of 99.97% (38.29 ± 2.7 µg ZIKV), which is reusable for at least three times. Therefore, the capture capacity (micrograms of ZIKV captured per milligram of ENM) of polyaniline-hierarchized mats was 2.72 µg ZIKV/mg. The impedance LOD value was determined to be 2.76 µg of ZIKV particles (approximately 6.6 × 103 PFU/mL). As a result, we present a fast small-scale purification system that can simultaneously monitor ZIKV electrochemically and optically.


Subject(s)
Alginates/chemistry , Aniline Compounds/chemistry , Biosensing Techniques/methods , Nanofibers/chemistry , Viral Load/methods , Zika Virus/isolation & purification , Animals , Antibodies, Immobilized/immunology , Antibodies, Viral/immunology , Blood/virology , Chlorocebus aethiops , Electrochemical Techniques , Hydrogels/chemistry , Immunoassay/methods , Limit of Detection , Vero Cells , Zika Virus/immunology
3.
Rev. medica electron ; 41(4): 959-978, jul.-ago. 2019.
Article in Spanish | CUMED | ID: cum-76345

ABSTRACT

RESUMEN La ontogenia humana está basada en fundamentos genéticos y epigenéticos. Con el objetivo de estructurar los referentes teóricos sobre el papel relevante de la epigenética en la ontogenianormal y defectuosa que contribuyan a la promoción de salud y prevención de enfermedad, se realizó la revisión de 37 referencias bibliográficas. La epigenética es el conjunto de procesos químicos dependientes del ambiente que modifican la expresión del ácido desoxirribonucleico, sin alterar su secuencia. Su acción está presente durante toda la vida, especialmente en la prenatal cuando, por modificaciones ambientales intraútero ocurre la programación epigenética que hace al humano susceptible a defectos en la ontogenia, incluso a padecer ulteriormente de enfermedades crónicas no transmisibles. Se han reportado factores ambientales inductores de marcas epigenéticas, entre ellos: alimentación, hábitos tóxicos, estrés, consumo inadecuado de ácido fólico y técnicas de reproducción asistida, todos modificables; su conocimiento constituye un baluarte inestimable en la promoción de salud y prevención de enfermedad (AU).


ABSTRACT Human ontogeny is based in genetic and epigenetic fundaments. 37 bibliographic references were reviewed with the objective of structuring the theoretical referents on the relevant role of epigenetics in normal and defective ontogeny to contribute to health promotion and disease prevention. Epigenetics is the whole of chemical processes depending from the environment that modify the deoxyribonucleic acid expression without modifying its sequence. Its action is present during all lifetime, especially at pre-natal times; when due to intrauterine environmental modifications the epigenetic programming takes place, making humans susceptible to defects in ontogeny, even to subsequently suffer non-communicable chronic diseases. Environmental factors inducing epigenetic marks have been reported: food, toxic habits, stress, folic acid inadequate intake and assisted reproduction techniques, all modifiable. Its knowledge is an invaluable bulkward in health promotion and disease prevention (AU).


Subject(s)
Humans , Preventive Health Services , Fetal Development/genetics , Disease Prevention , Epigenomics , Human Genetics , Genetics, Medical , Health Promotion , Impacts of Polution on Health , Environmental Hazards , Genetic Code
4.
Rev. medica electron ; 41(4): 959-978, jul.-ago. 2019.
Article in Spanish | LILACS, CUMED | ID: biblio-1094101

ABSTRACT

RESUMEN La ontogenia humana está basada en fundamentos genéticos y epigenéticos. Con el objetivo de estructurar los referentes teóricos sobre el papel relevante de la epigenética en la ontogenianormal y defectuosa que contribuyan a la promoción de salud y prevención de enfermedad, se realizó la revisión de 37 referencias bibliográficas. La epigenética es el conjunto de procesos químicos dependientes del ambiente que modifican la expresión del ácido desoxirribonucleico, sin alterar su secuencia. Su acción está presente durante toda la vida, especialmente en la prenatal cuando, por modificaciones ambientales intraútero ocurre la programación epigenética que hace al humano susceptible a defectos en la ontogenia, incluso a padecer ulteriormente de enfermedades crónicas no transmisibles. Se han reportado factores ambientales inductores de marcas epigenéticas, entre ellos: alimentación, hábitos tóxicos, estrés, consumo inadecuado de ácido fólico y técnicas de reproducción asistida, todos modificables; su conocimiento constituye un baluarte inestimable en la promoción de salud y prevención de enfermedad.


ABSTRACT Human ontogeny is based in genetic and epigenetic fundaments. 37 bibliographic references were reviewed with the objective of structuring the theoretical referents on the relevant role of epigenetics in normal and defective ontogeny to contribute to health promotion and disease prevention. Epigenetics is the whole of chemical processes depending from the environment that modify the deoxyribonucleic acid expression without modifying its sequence. Its action is present during all lifetime, especially at pre-natal times; when due to intrauterine environmental modifications the epigenetic programming takes place, making humans susceptible to defects in ontogeny, even to subsequently suffer non-communicable chronic diseases. Environmental factors inducing epigenetic marks have been reported: food, toxic habits, stress, folic acid inadequate intake and assisted reproduction techniques, all modifiable. Its knowledge is an invaluable bulkward in health promotion and disease prevention.


Subject(s)
Humans , Preventive Health Services , Fetal Development/genetics , Disease Prevention , Epigenomics , Human Genetics , Genetics, Medical , Health Promotion , Impacts of Polution on Health , Environmental Hazards , Genetic Code
5.
PLoS Negl Trop Dis ; 11(2): e0005363, 2017 02.
Article in English | MEDLINE | ID: mdl-28231241

ABSTRACT

The teratogenic mechanisms triggered by ZIKV are still obscure due to the lack of a suitable animal model. Here we present a mouse model of developmental disruption induced by ZIKV hematogenic infection. The model utilizes immunocompetent animals from wild-type FVB/NJ and C57BL/6J strains, providing a better analogy to the human condition than approaches involving immunodeficient, genetically modified animals, or direct ZIKV injection into the brain. When injected via the jugular vein into the blood of pregnant females harboring conceptuses from early gastrulation to organogenesis stages, akin to the human second and fifth week of pregnancy, ZIKV infects maternal tissues, placentas and embryos/fetuses. Early exposure to ZIKV at developmental day 5 (second week in humans) produced complex manifestations of anterior and posterior dysraphia and hydrocephalus, as well as severe malformations and delayed development in 10.5 days post-coitum (dpc) embryos. Exposure to the virus at 7.5-9.5 dpc induces intra-amniotic hemorrhage, widespread edema, and vascular rarefaction, often prominent in the cephalic region. At these stages, most affected embryos/fetuses displayed gross malformations and/or intrauterine growth restriction (IUGR), rather than isolated microcephaly. Disrupted conceptuses failed to achieve normal developmental landmarks and died in utero. Importantly, this is the only model so far to display dysraphia and hydrocephalus, the harbinger of microcephaly in humans, as well as arthrogryposis, a set of abnormal joint postures observed in the human setting. Late exposure to ZIKV at 12.5 dpc failed to produce noticeable malformations. We have thus characterized a developmental window of opportunity for ZIKV-induced teratogenesis encompassing early gastrulation, neurulation and early organogenesis stages. This should not, however, be interpreted as evidence for any safe developmental windows for ZIKV exposure. Late developmental abnormalities correlated with damage to the placenta, particularly to the labyrinthine layer, suggesting that circulatory changes are integral to the altered phenotypes.


Subject(s)
Arthrogryposis/virology , Disease Models, Animal , Hydrocephalus/virology , Pregnancy Complications, Infectious/virology , Zika Virus Infection/virology , Zika Virus/physiology , Animals , Arthrogryposis/embryology , Arthrogryposis/immunology , Arthrogryposis/pathology , Female , Humans , Hydrocephalus/embryology , Hydrocephalus/immunology , Hydrocephalus/pathology , Male , Mice , Mice, Inbred C57BL , Placenta/abnormalities , Placenta/immunology , Placenta/virology , Pregnancy , Pregnancy Complications, Infectious/immunology , Pregnancy Complications, Infectious/pathology , Teratogens/analysis , Zika Virus Infection/embryology , Zika Virus Infection/immunology , Zika Virus Infection/pathology
6.
Am J Trop Med Hyg ; 85(4): 739-47, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21976581

ABSTRACT

From September 2005 to March 2007, 238 individuals being vaccinated for the first time with the yellow fever (YF) -17DD vaccine were enrolled in a cohort established in Recife, Brazil. A prospective study indicated that, after immunization, anti-YF immunoglobulin M (IgM) and anti-YF IgG were present in 70.6% (IgM) and 98.3% (IgG) of the vaccinated subjects. All vaccinees developed protective immunity, which was detected by the plaque reduction neutralization test (PRNT) with a geometric mean titer of 892. Of the 238 individuals, 86.6% had IgG antibodies to dengue virus; however, the presence of anti-dengue IgG did not interfere significantly with the development of anti-YF neutralizing antibodies. In a separate retrospective study of individuals immunized with the 17DD vaccine, the PRNT values at 5 and 10 years post-vaccination remained positive but showed a significant decrease in neutralization titer (25% with PRNT titers < 100 after 5 years and 35% after 10 years).


Subject(s)
Yellow Fever Vaccine/administration & dosage , Antibodies, Viral/blood , Brazil , Enzyme-Linked Immunosorbent Assay , Humans , Neutralization Tests , Prospective Studies , Viral Plaque Assay , Yellow Fever Vaccine/immunology
7.
J Toxicol Environ Health A ; 68(11-12): 951-65, 2005.
Article in English | MEDLINE | ID: mdl-16020186

ABSTRACT

Embryos (stage 8-47, Nieuwkoop and Faber) of the African clawed frog (Xenopus laevis) were subjected to water-borne depleted uranium (DU) concentrations that ranged from 4.8 to 77.7 mg/L using an acute 96-h frog embryo teratogenesis assay-Xenopus (FETAX). In a chronic 64-d assay, X. laevis (from embryo through metamorphosis; stages 8-66) were subjected to concentrations of DU that ranged from 6.2 to 54.3 mg/L. Our results indicate DU is a non teratogenic metal. No effects on mortality, malformations, or growth were observed in the 96-h FETAX with concentrations of DU that ranged from 4.8 to 77.7 mg/L. From stage 8 to stage 47, X. laevis tadpoles do not actively feed and the gills are not well developed. Thus, uptake of DU was reduced despite exposure to elevated concentrations. The 64-d assay resulted in no concentration response for either mortality or malformations; however, a delay in metamorphosis was observed in tadpoles subjected to elevated DU concentrations (from 13.1 to 54.3 mg/L) compared to tadpoles in both the well-water control and reference. The delay in metamorphosis was likely due to increasing body burden of DU that ranged from 0.98 to 2.82 mg/kg.


Subject(s)
Embryo, Nonmammalian/drug effects , Metamorphosis, Biological/drug effects , Uranium/toxicity , Water Pollutants, Radioactive/toxicity , Animals , Dose-Response Relationship, Drug , Embryonic Development/drug effects , Teratogens/toxicity , Xenopus laevis/embryology , Xenopus laevis/growth & development
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