RESUMO
Immunity that controls parasitemia and inflammation during Plasmodium falciparum (Pf) malaria can be acquired with repeated infections. A limited understanding of this complex immune response impedes the development of vaccines and adjunctive therapies. We conducted a prospective systems biology study of children who differed in their ability to control parasitemia and fever following Pf infection. By integrating whole-blood transcriptomics, flow-cytometric analysis, and plasma cytokine and antibody profiles, we demonstrate that a pre-infection signature of B cell enrichment, upregulation of T helper type 1 (Th1) and Th2 cell-associated pathways, including interferon responses, and p53 activation associated with control of malarial fever and coordinated with Pf-specific immunoglobulin G (IgG) and Fc receptor activation to control parasitemia. Our hypothesis-generating approach identified host molecules that may contribute to differential clinical outcomes during Pf infection. As a proof of concept, we have shown that enhanced p53 expression in monocytes attenuated Plasmodium-induced inflammation and predicted protection from fever.
Assuntos
Linfócitos B/imunologia , Proteínas Sanguíneas/metabolismo , Inflamação/metabolismo , Malária Falciparum/metabolismo , Plasmodium falciparum/fisiologia , Células Th1/imunologia , Células Th2/imunologia , Proteína Supressora de Tumor p53/metabolismo , Adolescente , Adulto , Animais , Anticorpos Antiprotozoários/metabolismo , Criança , Pré-Escolar , Resistência à Doença , Feminino , Perfilação da Expressão Gênica , Humanos , Lactente , Interferons/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Estudos Prospectivos , Receptores Fc/metabolismo , Transdução de Sinais , Proteína Supressora de Tumor p53/genética , Adulto JovemRESUMO
Despite having the highest risk of progressing to severe disease due to lack of acquired immunity, the youngest children living in areas of highly intense malaria transmission have long been observed to be infected at lower rates than older children. Whether this observation is due to reduced exposure to infectious mosquito bites from behavioral and biological factors, maternally transferred immunity, genetic factors, or enhanced innate immunity in the young child has intrigued malaria researchers for over half a century. Recent evidence suggests that maternally transferred immunity may be limited to early infancy and that the young child's own immune system may contribute to control of malarial symptoms early in life and prior to the development of more effective adaptive immunity. Prospective studies of active and passive detection of Plasmodium falciparum blood-stage infections have identified young children (<5 years old) who remain uninfected through a defined surveillance period despite living in settings of highly intense malaria transmission. Yet, little is known about the potential immunological basis for this 'aparasitemic' phenotype. In this review, we summarize the observational evidence for this phenotype in field studies and examine potential reasons why these children escape detection of parasitemia, covering factors that are either extrinsic or intrinsic to their developing immune system. We discuss the challenges of distinguishing malaria protection from lack of malaria exposure in field studies. We also identify gaps in our knowledge regarding cellular immunity in the youngest age group and propose directions that researchers may take to address these gaps.
Assuntos
Malária Falciparum , Parasitemia , Plasmodium falciparum , Humanos , Pré-Escolar , Malária Falciparum/transmissão , Plasmodium falciparum/imunologia , Lactente , Malária/transmissão , Imunidade Inata , AnimaisRESUMO
BACKGROUND: TP53 has been shown to play a role in inflammatory processes, including malaria. We previously found that p53 attenuates parasite-induced inflammation and predicts clinical protection to Plasmodium falciparum infection in Malian children. Here, we investigated whether p53 codon 47 and 72 polymorphisms are associated with differential risk of P. falciparum infection and uncomplicated malaria in a prospective cohort study of malaria immunity. METHODS: p53 codon 47 and 72 polymorphisms were determined by sequencing TP53 exon 4 in 631 Malian children and adults enrolled in the Kalifabougou cohort study. The effects of these polymorphisms on the prospective risk of febrile malaria, incident parasitemia, and time to fever after incident parasitemia over 6 months of intense malaria transmission were assessed using Cox proportional hazards models. RESULTS: Confounders of malaria risk, including age and hemoglobin S or C, were similar between individuals with or without p53 S47 and R72 polymorphisms. Relative to their respective common variants, neither S47 nor R72 was associated with differences in prospective risk of febrile malaria, incident parasitemia, or febrile malaria after parasitemia. CONCLUSIONS: These findings indicate that p53 codon 47 and 72 polymorphisms are not associated with protection against incident P. falciparum parasitemia or uncomplicated febrile malaria.
Assuntos
Malária Falciparum , Malária , Criança , Adulto , Humanos , Estudos de Coortes , Estudos Prospectivos , Parasitemia/genética , Proteína Supressora de Tumor p53/genética , Plasmodium falciparum/genética , Malária/complicações , Malária Falciparum/epidemiologia , Malária Falciparum/genética , Malária Falciparum/complicações , Febre/etiologiaRESUMO
Airborne influenza viruses are responsible for serious respiratory diseases, and most detection methods for airborne viruses are based on extraction of nucleic acids. Herein, vertical-flow-assay-based electrochemical paper immunosensors were fabricated to rapidly quantify the influenza H1N1 viruses in air after sampling with a portable electrostatic particle concentrator (EPC). The effects of antibodies, anti-influenza nucleoprotein antibodies (NP-Abs) and anti-influenza hemagglutinin antibodies (HA-Abs), on the paper sensors as well as nonpulsed high electrostatic fields with and without corona charging on the virus measurement were investigated. The antigenicity losses of the surface (HA) proteins were caused by H2O2 via lipid oxidation-derived radicals and 1O2 via direct protein peroxidation upon exposure of a high electrostatic field. However, minimal losses in antigenicity of NP of the influenza viruses were observed, and the concentration of the H1N1 viruses was more than 160 times higher in the EPC than the BioSampler upon using NP-Ab based paper sensors after 60 min collection. This NP-Ab-based paper sensors with the EPC provided measurements comparable to quantitative polymerase chain reaction (qPCR) but much quicker, specific to the influenza H1N1 viruses in the presence of other airborne microorganisms and beads, and more cost-effective than enzyme-linked immunosorbent assay and qPCR.
Assuntos
Vírus da Influenza A Subtipo H1N1 , Influenza Humana , Ensaio de Imunoadsorção Enzimática , Humanos , Peróxido de Hidrogênio , Eletricidade EstáticaRESUMO
Although many studies concerning the detection of influenza virus have been published, a paper-based, label-free electrochemical immunosensor has never been reported. Here, we present a cost-effective, handmade paper-based immunosensor for label-free electrochemical detection of influenza virus H1N1. This immunosensor was prepared by modifying paper with a spray of hydrophobic silica nanoparticles, and using stencil-printed electrodes. We used a glass vaporizer to spray the hydrophobic silica nanoparticles onto the paper, rendering it super-hydrophobic. The super-hydrophobicity, which is essential for this paper-based biosensor, was achieved via 30-40 spray coatings, corresponding to a 0.39-0.41 mg cm-2 coating of nanoparticles on the paper and yielding a water contact angle of 150° ± 1°. Stencil-printed carbon electrodes modified with single-walled carbon nanotubes and chitosan were employed to increase the sensitivity of the sensor, and the antibodies were immobilized via glutaraldehyde cross-linking. Differential pulse voltammetry was used to assess the sensitivity of the sensors at various virus concentrations, ranging from 10 to 104 PFU mL-1, and the selectivity was assessed against MS2 bacteriophages and the influenza B viruses. These immunosensors showed good linear behaviors, improved detection times (30 min), and selectivity for the H1N1 virus with a limit of detection of 113 PFU mL-1, which is sufficiently sensitive for rapid on-site diagnosis. The simple and inexpensive methodologies developed in this study have great potential to be used for the development of a low-cost and disposable immunosensor for detection of pathogenic microorganisms, especially in developing countries.
Assuntos
Imunoensaio , Anticorpos Imobilizados , Técnicas Biossensoriais , Técnicas Eletroquímicas , Eletrodos , Ouro , Vírus da Influenza A Subtipo H1N1 , Limite de Detecção , Nanotubos de CarbonoRESUMO
Inspired by the availability of de novo transcriptome of horse gram (Macrotyloma uniflorum) and recent developments in systems biology studies, the first ever global protein-protein interactome (PPI) map was constructed for this highly drought-tolerant legume. Large-scale studies of PPIs and the constructed database would provide rationale behind the interplay at cascading translational levels for drought stress-adaptive mechanisms in horse gram. Using a bidirectional approach (interolog and domain-based), a high-confidence interactome map and database for horse gram was constructed. Available transcriptomic information for shoot and root tissues of a sensitive (M-191; genotype 1) and a drought-tolerant (M-249; genotype 2) genotype of horse gram was utilized to draw comparative PPI subnetworks under drought stress. High-confidence 6804 interactions were predicted among 1812 proteins covering about one-fourth of the horse gram proteome. The highest number of interactions (33.86%) in horse gram interactome matched with Arabidopsis PPI data. The top five hub nodes mostly included ubiquitin and heat-shock-related proteins. Higher numbers of PPIs were found to be responsive in shoot tissue (416) and root tissue (2228) of genotype 2 compared with shoot tissue (136) and root tissue (579) of genotype 1. Characterization of PPIs using gene ontology analysis revealed that kinase and transferase activities involved in signal transduction, cellular processes, nucleocytoplasmic transport, protein ubiquitination, and localization of molecules were most responsive to drought stress. Hence, these could be framed in stress adaptive mechanisms of horse gram. Being the first legume global PPI map, it would provide new insights into gene and protein regulatory networks for drought stress tolerance mechanisms in horse gram. Information compiled in the form of database (MauPIR) will provide the much needed high-confidence systems biology information for horse gram genes, proteins, and involved processes. This information would ease the effort and increase the efficacy for similar studies on other legumes. Public access is available at http://14.139.59.221/MauPIR/ .
Assuntos
Arabidopsis/fisiologia , Secas , Mapeamento de Interação de Proteínas , Estresse Fisiológico , Arabidopsis/química , Bases de Dados de Proteínas , Redes Reguladoras de Genes , Genótipo , Proteínas de Plantas , Proteoma , Estresse Fisiológico/genética , Biologia de SistemasRESUMO
Measurements of airborne viruses via sampling have been critical issues. Most electrostatic samplers have been assessed for bacterial aerosols or micrometer-sized viral particles; however, sampling of submicrometer-sized airborne viruses is necessary, especially because of the high probability of their staying airborne and their deposition in the lower respiratory tract. Here, we present a novel personal electrostatic particle concentrator (EPC) for gentle sampling of submicrometer airborne virus particles. Owing to the enhanced electric field designed in this EPC, the collection efficiencies reached values as high as 99.3-99.8% for 0.05-2 µm diameter polystyrene particles at a flow rate of 1.2 L/min. Submicrometer-sized MS2 and T3 virus particles were also collected in the EPC, and the concentrations relative to their respective initial suspensions were more than 10 times higher than those in the SKC BioSampler. Moreover, the recovery rate of T3 was 982 times higher in the EPC (-2 kV) than in the BioSampler at 12.5 L/min because of the gentle sampling of the EPC. Gentle sampling is desirable because many bioaerosols suffer from significant viability losses during sampling. The influence of ozone generated, applied electrostatic field, and the flow rate on the viability of the viruses will also be discussed.
Assuntos
Microbiologia do Ar , Vírion , Aerossóis , Monitoramento Ambiental , Tamanho da Partícula , Manejo de Espécimes , Eletricidade EstáticaRESUMO
In plants, epigenetic changes have been identified as regulators of developmental events during normal growth as well as environmental stress exposures. Flavonoid biosynthetic and antioxidant pathways play a significant role in plant defence during their exposure to environmental cues. The aim of this study was to unravel whether genes encoding enzymes of flavonoid biosynthetic and antioxidant pathways are under epigenetic regulation, particularly DNA methylation, during salt stress. For this, a repressor of silencing from Arabidopsis, AtROS1, was overexpressed in transgenic tobacco. Generated transgenics were evaluated to examine the influence of AtROS1 on methylation status of promoters as well as on coding regions of genes encoding enzymes of flavonoids biosynthesis and antioxidant pathways. Overexpression of AtROS1 increases the demethylation levels of both promoters as well as coding regions of genes encoding chalcone synthase, chalcone isomerase, flavanone 3-hydroxylase, flavonol synthase, dihydroflavonol 4-reductase, and anthocyanidin synthase of the flavonoid biosynthetic pathway, and glutathione S-transferase, ascorbate peroxidase, glutathione peroxidase, and glutathione reductase of the antioxidant pathway during control conditions. The level of demethylation was further increased at promoters as well as coding regions of these genes during salt-stress conditions. Transgenic tobacco overexpressing AtROS1 showed tolerance to salt stress that could have been due to the higher expression levels of the genes encoding enzymes of the flavonoid biosynthetic and antioxidant pathways. This is the first comprehensive study documenting the epigenetic regulation of flavonoid biosynthetic and antioxidant pathways during salt-stress exposure of plants.
Assuntos
Antioxidantes/metabolismo , Proteínas de Arabidopsis/genética , Epigênese Genética , Flavonoides/biossíntese , Regulação da Expressão Gênica de Plantas , Nicotiana/genética , Proteínas Nucleares/genética , Cloreto de Sódio/farmacologia , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas Nucleares/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Estresse Fisiológico , Nicotiana/enzimologia , Nicotiana/metabolismoRESUMO
Use of highly specific, sensitive and quantitative Real-Time PCR (qRT-PCR) based methods greatly facilitate the monitoring of experimental drug intervention and vaccination efficacy targeting liver stage malaria parasite. Here, in this study we have used qRT-PCR to detect the growing liver stage parasites following inoculation of Plasmodium yoelii sporozoite. Route of sporozoite administration and size of the sporozoite inoculums are two major determinants that affect the liver stage parasite load and therefore its detection and quantification. Thus, these factors need to be addressed to determine the accuracy of detection and quantification of Real-Time PCR method. Furthermore, applicability of quantitative RT-PCR system needs to be confirmed by analyzing the effect of different antimalarials on liver stage parasite burden. We have observed that parasite burden in mice infected via intravenous route was higher compared to that in subcutaneous, intradermal and intraperitoneal route infected mice. Moreover, this method detected liver stage parasite load with as low as 50 sporozoites. The inhibition studies with primaquine and atovaquone revealed inhibition of liver stage parasite and well correlated with patency and course of blood stage infection. This study characterized the simplicity, accuracy, and quantitative analysis of liver stage parasite development by real time PCR under different experimental conditions. Use of real time PCR method greatly improves the reproducibility and applicability to estimate the efficacy and potency of vaccine or drug candidates targeting liver stage parasite.
Assuntos
Fígado/parasitologia , Malária/parasitologia , Carga Parasitária/métodos , Plasmodium yoelii/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Antimaláricos/administração & dosagem , Atovaquona/administração & dosagem , Malária/tratamento farmacológico , Camundongos , Plasmodium yoelii/crescimento & desenvolvimento , Primaquina/administração & dosagem , Esporozoítos/crescimento & desenvolvimento , Resultado do TratamentoRESUMO
Cytokines and immune effector cells play an important role in determining the outcome of infection with various intracellular pathogens, including protozoan parasites. However, their role during lethal and nonlethal malaria needs further validation. In the present study, we examined the role of cytokines and various immune effector cells during lethal and nonlethal malaria caused by Plasmodium vinckei in AKR mice. We show that lethal P. vinckei infection (PvAS) in AKR mice is characterized by increased parasite growth, decreased production of pro-inflammatory cytokines, and attenuated cell proliferation and nitric oxide (NO) synthesis resulting in increased parasitemia which ultimately leads to death of all animals by day 5 post infection. In contrast, AKR mice infected with lethal parasite (PvAR) showed elevated levels of pro-inflammatory cytokines, heightened cell proliferation, and NO synthesis leading to complete parasite clearance by day 22 post infection. Flow cytometric analysis performed on splenocytes from PvAS- and PvAR-infected mice shows that host immunity is severely compromised in PvAS-infected mice as was evident by decreased percentages of CD4(+) and CD8(+) T cells, B cells, plasma cells, dendritic cells (DCs), and macrophages (MΦs) which was in complete contrast to PvAR-infected animals which exhibited elevated numbers of all the cell types analyzed. Taken together, findings of the present study show that coordinated actions of pro-inflammatory cytokines and other immune effector cells are essential to control lethal malarial infection and their attenuation leads to increased parasite growth and, ultimately, death of animals.
Assuntos
Malária/imunologia , Malária/parasitologia , Plasmodium/classificação , Animais , Linfócitos B , Linfócitos T CD8-Positivos , Citocinas/metabolismo , Células Dendríticas , Macrófagos/imunologia , Malária/mortalidade , Camundongos , Camundongos Endogâmicos AKR , Parasitemia/imunologia , Plasmodium/imunologiaRESUMO
The anti-cancer activity of saponins and phenolic compounds present in green tea was previously reported. However, the immunomodulatory and adjuvanticity activity of tea saponin has never been studied. In this study, we investigated the immunomodulatory effect of tea saponin in T-lymphocytes and EL4 cells via regulation of cytokine response and mitogen-activated protein kinases (MAPK) signaling pathway. Quantitative analysis of mRNA expression level of cytokines were performed by reverse transcription polymerase chain reaction following stimulation with tea saponin, ovalbumin (OVA) alone or tea saponin in combination with OVA. Tea saponin inhibited the proliferation of EL4 cells measured in a dose-dependent manner. No cytotoxicity effect of tea saponin was detected in T-lymphocytes; rather, tea saponin enhanced the proliferation of T-lymphocytes. Tea saponin with OVA increased the expression of interleukin (IL)-1, IL-2, IL-12, interferon-γ and tumor necrosis factor (TNF)-α and decreased the expression level of IL-10 and IL-8 in T-lymphocytes. Furthermore, tea saponin, in the presence of OVA, downregulated the MAPK signaling pathway via inhibition of IL-4, IL-8 and nuclear factor kappaB (NF-κB) in EL4 cells. Th1 cytokines enhancer and Th2 cytokines and NF-κB inhibitor, tea saponin can markedly inhibit the proliferation and invasiveness of T-lymphoma (EL4) cells, possibly due to TNF-α- and NF-κB-mediated regulation of MAPK signaling pathway.
Assuntos
Antineoplásicos/farmacologia , Fatores Imunológicos/farmacologia , Linfoma de Células T/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Saponinas/farmacologia , Linfócitos T/efeitos dos fármacos , Chá/química , Animais , Hemólise/efeitos dos fármacos , L-Lactato Desidrogenase/metabolismo , Sistema de Sinalização das MAP Quinases/fisiologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteína Quinase 1 Ativada por Mitógeno/fisiologia , Linfócitos T/imunologia , Células Th1/imunologia , Células Th2/imunologia , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologiaRESUMO
A systems analysis was conducted to determine the potential molecular mechanisms underlying differential immunogenicity and protective efficacy results of a clinical trial of the radiation-attenuated whole-sporozoite PfSPZ vaccine in African infants. Innate immune activation and myeloid signatures at prevaccination baseline correlated with protection from P. falciparum parasitemia in placebo controls. These same signatures were associated with susceptibility to parasitemia among infants who received the highest and most protective PfSPZ vaccine dose. Machine learning identified spliceosome, proteosome, and resting DC signatures as prevaccination features predictive of protection after highest-dose PfSPZ vaccination, whereas baseline circumsporozoite protein-specific (CSP-specific) IgG predicted nonprotection. Prevaccination innate inflammatory and myeloid signatures were associated with higher sporozoite-specific IgG Ab response but undetectable PfSPZ-specific CD8+ T cell responses after vaccination. Consistent with these human data, innate stimulation in vivo conferred protection against infection by sporozoite injection in malaria-naive mice while diminishing the CD8+ T cell response to radiation-attenuated sporozoites. These data suggest a dichotomous role of innate stimulation for malaria protection and induction of protective immunity by whole-sporozoite malaria vaccines. The uncoupling of vaccine-induced protective immunity achieved by Abs from more protective CD8+ T cell responses suggests that PfSPZ vaccine efficacy in malaria-endemic settings may be constrained by opposing antigen presentation pathways.
Assuntos
Imunidade Inata , Vacinas Antimaláricas , Malária Falciparum , Plasmodium falciparum , Esporozoítos , Vacinas Atenuadas , Vacinas Antimaláricas/imunologia , Vacinas Antimaláricas/administração & dosagem , Imunidade Inata/imunologia , Humanos , Animais , Malária Falciparum/prevenção & controle , Malária Falciparum/imunologia , Plasmodium falciparum/imunologia , Camundongos , Vacinas Atenuadas/imunologia , Vacinas Atenuadas/administração & dosagem , Esporozoítos/imunologia , Esporozoítos/efeitos da radiação , Linfócitos T CD8-Positivos/imunologia , Lactente , Proteínas de Protozoários/imunologia , Anticorpos Antiprotozoários/imunologia , Feminino , Parasitemia/imunologia , Parasitemia/prevenção & controle , Imunoglobulina G/imunologia , Imunoglobulina G/sangue , Eficácia de VacinasRESUMO
BACKGROUND: Drought tolerance is an attribute maintained in plants by cross-talk between multiple and cascading metabolic pathways. Without a sequenced genome available for horse gram, it is difficult to comprehend such complex networks and intercalated genes associated with drought tolerance of horse gram (Macrotyloma uniflorum). Therefore, de novo transcriptome discovery and associated analyses was done for this highly drought tolerant yet under exploited legume to decipher its genetic makeup. RESULTS: Eight samples comprising of shoot and root tissues of two horse gram genotypes (drought-sensitive; M-191 and drought-tolerant; M-249) were used for comparison under control and polyethylene glycol-induced drought stress conditions. Using Illumina sequencing technology, a total of 229,297,896 paired end read pairs were generated and utilized for de novo assembly of horse gram. Significant BLAST hits were obtained for 26,045 transcripts while, 3,558 transcripts had no hits but contained important conserved domains. A total of 21,887 unigenes were identified. SSRs containing sequences covered 16.25% of the transcriptome with predominant tri- and mono-nucleotides (43%). The total GC content of the transcriptome was found to be 43.44%. Under Gene Ontology response to stimulus, DNA binding and catalytic activity was highly expressed during drought stress conditions. Serine/threonine protein kinase was found to dominate in Enzyme Classification while pathways belonging to ribosome metabolism followed by plant pathogen interaction and plant hormone signal transduction were predominant in Kyoto Encyclopedia of Genes and Genomes analysis. Independent search on plant metabolic network pathways suggested valine degradation, gluconeogenesis and purine nucleotide degradation to be highly influenced under drought stress in horse gram. Transcription factors belonging to NAC, MYB-related, and WRKY families were found highly represented under drought stress. qRT-PCR validated the expression profile for 9 out of 10 genes analyzed in response to drought stress. CONCLUSIONS: De novo transcriptome discovery and analysis has generated enormous information over horse gram genomics. The genes and pathways identified suggest efficient regulation leading to active adaptation as a basal defense response against drought stress by horse gram. The knowledge generated can be further utilized for exploring other underexploited plants for stress responsive genes and improving plant tolerance.
Assuntos
Secas , Fabaceae/genética , Fabaceae/fisiologia , Estresse Fisiológico/genética , Transcriptoma/genética , Adaptação Fisiológica/genética , Composição de Bases/genética , Sequência de Bases , Análise por Conglomerados , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ontologia Genética , Genes de Plantas , Redes e Vias Metabólicas/genética , Anotação de Sequência Molecular , Sequências Repetitivas de Ácido Nucleico , Reprodutibilidade dos Testes , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/metabolismoRESUMO
DNA methylation is known as an epigenetic modification that affects gene expression in plants. Variation in CpG methylation behavior was studied in two natural horse gram (Macrotyloma uniflorum [Lam.] Verdc.) genotypes, HPKC2 (drought-sensitive) and HPK4 (drought-tolerant). The methylation pattern in both genotypes was studied through methylation-sensitive amplified polymorphism. The results revealed that methylation was higher in HPKC2 (10.1%) than in HPK4 (8.6%). Sequencing demonstrated sequence homology with the DRE binding factor (cbf1), the POZ/BTB protein, and the Ty1-copia retrotransposon among some of the polymorphic fragments showing alteration in methylation behavior. Differences in DNA methylation patterns could explain the differential drought tolerance and the epigenetic signature of these two horse gram genotypes.
Assuntos
Metilação de DNA , Epigênese Genética , Fabaceae/genética , Regulação da Expressão Gênica de Plantas , Polimorfismo Genético , Clonagem Molecular , Primers do DNA/genética , Elementos de DNA Transponíveis , DNA de Plantas/genética , Secas , Genes de Plantas , Genótipo , Fenótipo , Folhas de Planta/genética , Ligação Proteica , Análise de Sequência de DNARESUMO
Conventional airborne virus measurement usually requires appreciable sampling and detection times. Viral aerosols should also be collected or prepared in a liquid medium whose volume typically ranges from milliliters to tens of milliliters; hence, many sampling and detection steps need to be taken with the unit horizontal or immobile. Moreover, viral aerosols need to be sufficiently enriched, which makes real-time monitoring difficult. Herein, we present a near real-time enrichment and quantification system of airborne viruses that consists of a wet-paper-based electrochemical immunosensor with a gel electrolyte and a modified electrostatic particle concentrator. A small amount of phosphate-buffered saline flowed on the electrode, which resulted in sensor electrodes that are barely wet (covered in a thin buffer film measuring several micrometers) to ensure antigen-antibody interaction and the removal of non-target particles on the electrode surface. This system ensures that airborne viruses are highly enriched on the working electrode of the immunosensor, and it is possible to measure the MS2 virus particle concentrations every 10 min for 60 min stably and selectively against non-target airborne viruses and bacteria at horizontal and tilted measurement configurations. This system thus has the potential to be used in the real-time mobile monitoring of airborne microorganisms.
Assuntos
Técnicas Biossensoriais , Vírus , Eletricidade Estática , Imunoensaio , Aerossóis , FosfatosRESUMO
Malaria and schistosomiasis are two major parasitic diseases that remain leading causes of morbidity and mortality worldwide. Co-infections of these two parasites are common in the tropics, where both diseases are endemic. The clinical consequences of schistosomiasis and malaria are determined by a variety of host, parasitic, and environmental variables. Chronic schistosomiasis causes malnutrition and cognitive impairments in children, while malaria can cause fatal acute infections. There are effective drugs available to treat malaria and schistosomiasis. However, the occurrence of allelic polymorphisms and the rapid selection of parasites with genetic mutations can confer reduced susceptibility and lead to the emergence of drug resistance. Moreover, the successful elimination and complete management of these parasites are difficult due to the lack of effective vaccines against Plasmodium and Schistosoma infections. Therefore, it is important to highlight all current vaccine candidates undergoing clinical trials, such as pre-erythrocytic and erythrocytic stage malaria, as well as a next-generation RTS,S-like vaccine, the R21/Matrix-M vaccine, that conferred 77% protection against clinical malaria in a Phase 2b trial. Moreover, this review also discusses the progress and development of schistosomiasis vaccines. Furthermore, significant information is provided through this review on the effectiveness and progress of schistosomiasis vaccines currently under clinical trials, such as Sh28GST, Sm-14, and Sm-p80. Overall, this review provides insights into recent progress in malarial and schistosomiasis vaccines and their developmental approaches.
RESUMO
Cytokines play an important role in the defense against malaria and some have long been documented to influence the course of malaria infection in rodents and humans. The present study was conducted to determine the mRNA expression pattern of a few prominent cytokines at different time points during the course of infection with a nonlethal and lethal Plasmodium vinckei rodent malaria parasite, using highly sensitive real-time PCR. Analysis of mRNA expression of cytokines in spleen from infected mice revealed that the principal difference was an early depletion in pro-inflammatory cytokine's mRNA expression in mice infected with lethal P. vinckei (PvAS) parasites. In addition, an increase in anti-inflammatory cytokines particularly IL-10 mRNA expression levels was found in the same group of mice. In contrast, the significant rise in pro-inflammatory cytokine's mRNA expression levels was recorded at day 1 onwards after infection with nonlethal P. vinckei (PvAR). The maximum fold change was recorded for IFN-γ and IL-10, when compared to baseline value. TGF-ß did not seem to play any major role in P. vinckei infection.
Assuntos
Interleucina-10/metabolismo , Malária/imunologia , Plasmodium yoelii/patogenicidade , RNA Mensageiro/genética , Fator de Crescimento Transformador beta/metabolismo , Animais , Interferon gama/metabolismo , Malária/parasitologia , Camundongos , Camundongos Endogâmicos AKR , Plasmodium yoelii/isolamento & purificação , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de RNA , Baço/imunologia , Baço/metabolismo , Baço/parasitologiaRESUMO
Many studies have been conducted on measuring avian influenza viruses and their hemagglutinin (HA) antigens via electrochemical principles; most of these studies have used gold electrodes on ceramic, glass, or silicon substrates, and/or labeling for signal enhancement. Herein, we present a paper-based immunosensor for label-free measurement of multiple avian influenza virus (H5N1, H7N9, and H9N2) antigens using flexible screen-printed carbon nanotube-polydimethylsiloxane electrodes. These flexible electrodes on a paper substrate can complement the physical weakness of the paper-based sensors when wetted, without affecting flexibility. The relative standard deviation of the peak currents was 1.88% when the electrodes were repeatedly bent and unfolded twenty times with deionized water provided each cycle, showing the stability of the electrodes. For the detection of HA antigens, approximately 10-µl samples (concentration: 100 pg/ml-100 ng/ml) were needed to form the antigen-antibody complexes during 20-30 min incubation, and the immune responses were measured via differential pulse voltammetry. The limits of detections were 55.7 pg/ml (0.95 pM) for H5N1 HA, 99.6 pg/ml (1.69 pM) for H7N9 HA, and 54.0 pg/ml (0.72 pM) for H9N2 HA antigens in phosphate buffered saline, and the sensors showed good selectivity and reproducibility. Such paper-based sensors are economical, flexible, robust, and easy-to-manufacture, with the ability to detect several avian influenza viruses.
Assuntos
Antígenos Virais/análise , Técnicas Biossensoriais/métodos , Dimetilpolisiloxanos , Técnicas Eletroquímicas/métodos , Eletrodos , Imunoensaio/métodos , Virus da Influenza A Subtipo H5N1/imunologia , Subtipo H7N9 do Vírus da Influenza A/imunologia , Vírus da Influenza A Subtipo H9N2/imunologia , Nanotubos de Carbono , Papel , Virologia/métodos , Animais , Aves , Humanos , Influenza Aviária/diagnóstico , Influenza Aviária/virologia , Influenza Humana/diagnóstico , Influenza Humana/virologia , Limite de Detecção , Reprodutibilidade dos TestesRESUMO
Growth tube collectors (GTCs) are used to sample virus aerosols because of their superior viable virus recovery among air samplers. However, a major limitation of such samplers is that they operate at low flow rates compared to many inertia-based air samplers. Herein, we demonstrated efficient measurements of airborne MS2 and T3 viruses using a GTC that can implement high flow velocities for higher flow rates per tube, which we refer to as the growth-based virus aerosol concentrator (GVC), via qPCR and the plaque assay technique. The GVC exhibited a flow rate of up to 6 L/min, where the average sampling flow velocity was 5.09 m/s, 22 times higher than those used in the GTCs, for a single tube with a diameter of 5 mm. The count median diameter of the size-increased particles at the exit of the initiator was measured to be 1.44 µm at 6 L/min, considerably smaller than those observed in conventional GTCs. Nevertheless, the measurement of airborne MS2 and T3 viruses using the GVC showed a high concentration (high enrichment ratio of 109,458 at 10-min sampling) of viruses in a sampling medium, with a high viable virus percentage (> 90%) and physical collection efficiency (> 90%) at 6 L/min, which shows the potential for rapid on-site detection of airborne viruses.
Assuntos
Microbiologia do Ar , Vírus , Aerossóis/análise , Monitoramento Ambiental/métodos , Tamanho da PartículaRESUMO
In the present study, we have investigated the role of antimalarial drug halofantrine (HF) in inducing the sterile protection against challenges with sporozoites of the live infectious Plasmodium yoelii (Killick-Kendrick, 1967) in Swiss mice malaria model. We observed that during the first to third sequential sporozoite inoculation cycles, blood-stage patency remains the same in the control and chemoprophylaxis under HF drug cover (CPS-HF) groups. However, a delayed blood-stage infection was observed during the fourth and fifth sporozoite challenges and complete sterile protection was produced following the sixth sporozoite challenge in CPS-HF mice. We also noticed a steady decline in liver stage parasite load after 3th to 6th sporozoite challenge cycle in CPS-HF mice. CPS-HF immunisation results in a significant up-regulation of pro-inflammatory cytokines (IFN-γ, TNF-α, IL-12 and iNOS) and down-regulation of anti-inflammatory cytokines (IL-10 and TGF-ß) mRNA expression in hepatic mononuclear cells (HMNC) and spleen cells in the immunised CPS-HF mice (after 6th sporozoite challenge) compared to control. Overall, our study suggests that the repetitive sporozoite inoculation under HF drug treatment develops a strong immune response that confers protection against subsequent challenges with sporozoites of P. yoelii.