RESUMO
BACKGROUND: RNA editing in chloroplast and mitochondrion transcripts of plants is an important type of post-transcriptional RNA modification in which members of the multiple organellar RNA editing factor gene family (MORF) play a crucial role. However, a systematic identification and characterization of MORF members in Brassica napus is still lacking. RESULTS: In this study, a total of 43 MORF genes were identified from the genome of the Brassica napus cultivar "Zhongshuang 11". The Brassica napus MORF (BnMORF) family members were divided into three groups through phylogenetic analysis. BnMORF genes distributed on 14 chromosomes and expanded due to segmental duplication and whole genome duplication repetitions. The majority of BnMORF proteins were predicted to be localized to mitochondria and chloroplasts. The promoter cis-regulatory element analysis, spatial-temporal expression profiling, and co-expression network of BnMORF genes indicated the involvement of BnMORF genes in stress and phytohormone responses, as well as growth and development. CONCLUSION: This study provides a comprehensive analysis of BnMORF genes and lays a foundation for further exploring their physiological functions in Brassica napus.
Assuntos
Brassica napus , Família Multigênica , Filogenia , Proteínas de Plantas , Brassica napus/genética , Brassica napus/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Edição de RNA , Perfilação da Expressão Gênica , Cloroplastos/genética , Cloroplastos/metabolismoRESUMO
The chloroplast and mitochondrion are semi-autonomous organelles that play essential roles in cell function. These two organelles are embellished with prokaryotic remnants and contain many new features emerging from the co-evolution of organelles and the nucleus. A typical plant chloroplast or mitochondrion genome encodes less than 100 genes, and the regulation of these genes' expression is remarkably complex. The regulation of chloroplast and mitochondrion gene expression can be achieved at multiple levels during development and in response to environmental cues, in which, RNA metabolism, including: RNA transcription, processing, translation, and degradation, plays an important role. RNA metabolism in plant chloroplasts and mitochondria combines bacterial-like traits with novel features evolved in the host cell and is regulated by a large number of nucleus-encoded proteins. Among these, pentatricopeptide repeat (PPR) proteins are deeply involved in multiple aspects of the RNA metabolism of organellar genes. Research over the past decades has revealed new insights into different RNA metabolic events in plant organelles, such as the composition of chloroplast and mitochondrion RNA editosomes. We summarize and discuss the most recent knowledge and biotechnological implications of various RNA metabolism processes in plant chloroplasts and mitochondria, with a focus on the nucleus-encoded factors supporting them, to gain a deeper understanding of the function and evolution of these two organelles in plant cells. Furthermore, a better understanding of the role of nucleus-encoded factors in chloroplast and mitochondrion RNA metabolism will motivate future studies on manipulating the plant gene expression machinery with engineered nucleus-encoded factors.
RESUMO
RNA editing is the process of modifying RNA molecules by inserting, deleting, or substituting nucleotides. In flowering plants, RNA editing occurs predominantly in RNAs encoded by the organellar genomes of mitochondria and chloroplasts, and the main type of editing involves the substitution of cytidine with uridine at specific sites. Abnormal RNA editing in plants can affect gene expression, organelle function, plant growth, and reproduction. In this study, we report that ATPC1, the gamma subunit of ATP synthase in Arabidopsis chloroplasts, has an unexpected role in the regulation of editing at multiple sites of plastid RNAs. The loss of function of ATPC1 severely arrests chloroplast development, causing a pale-green phenotype and early seedling lethality. Disruption of ATPC1 increases the editing of matK-640, rps12-i-58, atpH-3'UTR-13210, and ycf2-as-91535 sites while decreasing the editing of rpl23-89, rpoA-200, rpoC1-488, and ndhD-2 sites. We further show that ATPC1 participates in RNA editing by interacting with known multiple-site chloroplast RNA editing factors, including MORFs, ORRM1, and OZ1. The transcriptome in the atpc1 mutant is profoundly affected, with a pattern of defective expression of chloroplast development-related genes. These results reveal that the ATP synthase γ subunit ATPC1 is involved in multiple-site RNA editing in Arabidopsis chloroplasts.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , ATPases de Cloroplastos Translocadoras de Prótons , Trifosfato de Adenosina/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Cloroplastos/genética , Cloroplastos/metabolismo , Regulação da Expressão Gênica de Plantas , Óxido Nítrico Sintase/metabolismo , Edição de RNA , RNA de Plantas/genética , ATPases de Cloroplastos Translocadoras de Prótons/metabolismoRESUMO
Arsenic (As) a non-essential element is of particular concern with respect to harmful effects on plant metabolism. While extensive studies have been conducted on the physiological responses of plants to increase As concentrations, however, molecular differences elucidating species-specific changes remain largely unknown. In the present experiment, two oilseed Brassica napus (B. napus) cultivars, ZS758 and ZD622, were treated by elevated As concentration. Their responses to the As stress have been investigated through pulse amplitude modulated fluorometer and isobaric tags based proteomic (iTRAQ) analysis. The chlorophyll fluorescence attributes showed that As stress significantly decrease the photochemical efficiency of photosystem II (PSII) and photosystem I (PSI) as well as the comparatively closed stomata observed under scanning electron microscopy (SEM). In this study, 65 proteins displayed increased abundance and 52 down-regulated were found in the control vs As comparison in cultivar ZS758, while 44 up and 67 down-regulated proteins were found in the control vs As comparison in ZD622. Metabolic pathways, followed by ribosome and biosynthesis of secondary metabolites were the dominant functional annotation categories among the differentially expressed protein (DEPs). Many genes involved in primary metabolism, stress and defense were found to be As-responsive DEPs and/or DEPs between these two cultivars. Based on these results, a schematic description of key processes involved in As tolerance in ZS758 and ZD622 is proposed, which suggests that higher tolerance in ZS758 depends on a multilevel coordination of efficient defense and energy metabolism. Real-time quantitative PCR supported the expression patterns of several genes encoding a protein similar to their corresponding DEPs. In addition, these findings could shed light in unraveling the molecular mechanisms of B. napus exposed to As stress and provide or improve essential understandings in the development of advanced B. napus cultivars against As resistance.
Assuntos
Arsênio/toxicidade , Brassica napus/efeitos dos fármacos , Proteoma/metabolismo , Arsênio/metabolismo , Metabolismo Energético , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Proteômica/métodosRESUMO
Orobanche cumana is an obligate root parasite causing severe damage to many economically important crops, including sunflowers worldwide. For efficient control measures, it is necessary to understand the resistant mechanism during interaction at molecular level. The present study emphasizes on comparative proteomics to investigate the mechanistic basis of compatible and incompatible interaction of O. cumana with resistant (JY207) and susceptible (TK0409) sunflowers. More than 3500 proteins were identified from two cultivars by iTRAQ analysis. Identified proteins associated with general functions, posttranslational modification, energy production and conversion, carbohydrate transport and metabolism, and signal transduction mechanisms were the most represented category of induced proteins in both cultivars. The resistant interaction was characterized by alteration of defense-related proteins involved in recognition of parasites, accumulation of pathogenesis-related proteins, biosynthesis of lignin, and detoxification of toxic metabolites in JY207 after inoculation. The susceptible interaction was characterized by decreased abundance of proteins involved in biosynthesis and signaling of plant growth regulators including auxin, gibberellin, brassinosteroid, and ethylene in TK0409 after inoculation. The present study provides comprehensive details of proteins and differential modulation of pathways regulated under compatible and incompatible interaction, allowing the identification of important molecular components for development of sustainable resistance against this parasite.
Assuntos
Helianthus/imunologia , Orobanche/crescimento & desenvolvimento , Orobanche/imunologia , Doenças das Plantas/parasitologia , Proteínas de Plantas/metabolismo , Proteômica/métodos , Resistência à Doença , Helianthus/crescimento & desenvolvimento , Helianthus/parasitologia , Interações Hospedeiro-Parasita , Marcação por Isótopo , Orobanche/fisiologia , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Sementes/parasitologia , Espectrometria de Massas em Tandem/métodosRESUMO
Blockage of pathogen transmission through water decontamination is considered an important strategy for the prevention of schistosome infection. Many believe that this strategy is feasible, but it has yet to be achieved. Silver has a long history of use as a disinfectant. With the emergence of nanotechnology, silver can be shaped into nanoparticles which have been found to possess superb antimicrobial activities. In this light, we investigated the effects of silver nanoparticles (AgNPs) on Schistosoma japonicum cercariae. AgNPs rapidly induced cercarial tail-shedding, agitated behaviour and a decrease in cercarial secretion in a dose-dependent manner. Prolonged treatment was found to be cercariocidal, which nevertheless might be attributable to AgNP-induced cercarial tail loss rather than to toxicity. Higher concentrations of AgNPs (125 µg mL-1 and above) completely blocked cercarial infectivity. Despite decreased infectivity, cercariae exposed to lower concentrations of AgNPs for 30 min were still found capable of infecting hosts even without their tails, suggesting that tail loss does not necessarily signify a total loss of infective ability. We also found that silver ions (Ag+) were heavily involved in the observed cercarial responses of AgNPs. Our observations provide insight into the interactions between the larvae of helminth parasites and nanoparticles.
Assuntos
Nanopartículas Metálicas/química , Schistosoma japonicum/efeitos dos fármacos , Esquistossomicidas/farmacologia , Prata/farmacologia , Animais , Relação Dose-Resposta a Droga , Schistosoma japonicum/ultraestrutura , Esquistossomicidas/administração & dosagem , Esquistossomicidas/química , Prata/administração & dosagem , Prata/químicaRESUMO
Concomitant infections of different species of parasites are common in the field. Infection with one parasite species likely triggers host responses that may influence the subsequent infection of another species and alter disease outcomes. So far, the majority of studies have focused on single species parasite infection, and the mechanisms of protection induced by the first parasite infection against the secondary infection remain poorly defined. In this study, we assess the impact of trematode Clonorchis sinensis infection on the course of another tissue nematode Trichinella spiralis challenge. We observed that mice with preexisting C. sinensis infection had lower worm burden of intestinal T. spiralis than those infected with T. spiralis alone; mice with preexisting C. sinensis also had severe enteric histopathological changes and higher counts of intestinal Paneth cells in responses to T. spiralis challenge. The mRNA levels of interleukin (IL)-4, IL-10, IL-13, and tumor necrosis factor (TNF)-α from the small intestine and spleen of the different groups were analyzed using quantitative real-time polymerase chain reaction. Compared with that in mice infected with T. spiralis alone, the mRNA expression of IL-13 was significantly increased in the small intestine tissues and IL-4, IL-13, and TNF-α were significantly increased in the spleen tissues in the dually infected mice. Our findings suggest that a "preexisting" trematode infection of C. sinensis is a factor which contributes to reducing the establishment of T. spiralis adult worms in the small intestine.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Clonorquíase/imunologia , Trichinella spiralis/imunologia , Triquinelose/imunologia , Animais , Clonorquíase/parasitologia , Clonorquíase/patologia , Clonorchis sinensis/imunologia , Clonorchis sinensis/fisiologia , Coinfecção , Duodeno/imunologia , Duodeno/parasitologia , Duodeno/patologia , Eosinófilos/citologia , Eosinófilos/imunologia , Feminino , Células Caliciformes/citologia , Células Caliciformes/imunologia , Interleucinas/genética , Mucosa Intestinal/imunologia , Mucosa Intestinal/parasitologia , Mucosa Intestinal/patologia , Masculino , Mastócitos/citologia , Mastócitos/imunologia , Camundongos , Celulas de Paneth/citologia , Celulas de Paneth/imunologia , RNA Mensageiro/genética , Organismos Livres de Patógenos Específicos , Baço/imunologia , Baço/parasitologia , Triquinelose/parasitologia , Triquinelose/patologia , Fator de Necrose Tumoral alfa/genéticaRESUMO
Chromium (Cr) is a serious environmental contaminant that drastically limited the crop yields. Nitric oxide (NO) and spermine (Spm) portrayal significance in improving the plant tolerance against abiotic stresses. Therefore, we investigate the protective efficacy of seed priming with NO (100µM) and/or Spm (0.01mM) in minimising the Cr-induced toxic effects in rice (Oryza sativa L.) plants. Our outcomes revealed that Cr alone treatments (100µM) notably reduced the seed germination rate, plant growth, photosynthetic apparatus, nutrients uptake and antioxidant defence system, but extra generation of reactive oxygen species (ROS). Interestingly, the combine applications of NO and Spm significantly reversed the Cr-induced toxic effects by reducing the Cr-accumulation, maintaining the nutrient balance, improving the germination indices, levels of photosynthetic pigments (chl a by 24.6%, chl b by 36.3%, chl (a+b ) by 57.2% and carotenoids by 79.4%), PSII, photosynthesis gas exchange parameters and total soluble sugar (74.9%) by improving antioxidative enzyme activities. As a result, NO+Spm lowered the accumulation of oxidative markers (H2 O2 by 93.9/70.4%, O2 Ë- by 86.3/69.9% and MDA by 97.2/73.7% in leaves/roots), electrolyte leakage (71.4% in leaves) and improved the plant growth traits. Based on these findings, it can be concluded that NO triggers Spm to minimise the Cr-accumulation and its adverse effects on rice plants. Additionally, combined treatments (NO+Spm) were more effective in minimising the Cr-induced toxic effects in comparison to NO and Spm alone treatments. Thus, co-exposure of NO and Spm may be utilised to boost rice tolerance under Cr stress conditions.
Assuntos
Oryza , Plântula , Óxido Nítrico/farmacologia , Espermina/farmacologia , Cromo/toxicidade , Estresse Oxidativo , Antioxidantes/farmacologia , Antioxidantes/metabolismo , SementesRESUMO
CD4(+) T-helper (Th) cell is widely recognized to be capable of influencing worm development and egg granuloma formation after schistosome infection. Interleukin (IL)-12 and IL-4 play key roles in regulation of Th cell differentiation. In the present study, we subcutaneously inoculated mice with hybridoma cells secreting monoclonal antibodies to neutralize IL-12 and IL-4 and explored the effects of IL-12 and IL-4 deficiency on the worm development and granuloma formation in mice infected with cercariae of Schistosoma japonicum. It was found that deficiency of host IL-12 and IL-4 supported normal parasite survival and fecundity. However, worm development (length and female fecundity) was significantly enhanced in anti-IL-12-treated mice. Mean length of worms in anti-IL-12-treated group was significantly greater than that of intact controls on day 28 after infection (females, 11.84 ± 1.20 mm vs. 9.45 ± 1.34; males, 9.35 ± 1.21 mm vs. 8.10 ± 0.85 mm, p < 0.05). Liver egg load per pair of worms (1,770.12 ± 470.67 vs. 806.08 ± 232.37, p < 0.05) and uterine egg load of ovigerous females (93.08 ± 27.85 vs. 46.05 ± 34.24, p < 0.05) in anti-IL-12-treated mice were significantly higher than those in intact control 28 days postinfection. But these effects diminished 42 days postinfection (p > 0.05). Granuloma size in anti-IL-12-treated mice was significantly larger than that in intact mice 42 days postinfection (398.3 ± 80.7 µm vs. 294.4 ± 72.2 µm, p < 0.05). Granuloma fibrosis dramatically intensified in anti-IL-12-treated mice but diminished in anti-IL-4-treated mice. The results suggest that IL-12 may play an impeditive role in the development of S. japonicum and in granuloma formation as well as fibrosis. IL-4 may promote granuloma formation but have no effect on worm development.
Assuntos
Granuloma/patologia , Interleucina-12/deficiência , Interleucina-4/deficiência , Schistosoma japonicum/imunologia , Schistosoma japonicum/patogenicidade , Esquistossomose Japônica/imunologia , Esquistossomose Japônica/patologia , Animais , Modelos Animais de Doenças , Feminino , Granuloma/imunologia , Histocitoquímica , Interleucina-12/imunologia , Interleucina-4/imunologia , Fígado/parasitologia , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia , Contagem de Ovos de Parasitas , Útero/parasitologiaRESUMO
Toxoplasma gondii infections are prevalent in a wide range of mammalian hosts including humans. Infection in pregnant women may cause the transmission of parasite to the fetus that makes serious problems. IgM antibodies against Toxoplasma (Toxo-IgM) have been believed to be significant indicators for both recently acquired and congenital toxoplasmosis. So far, however, there has not been any recognized protein of T. gondii that specifically reacts to IgM antibodies. Here, an antigen exclusively for detection of IgM antibodies screened by two-dimensional electrophoresis and mass spectrometry has been reported. The study identified 13 Toxoplasma proteins probed by IgG antibodies and one (rhpotry protein 2 [ROP2]) by IgM antibodies with human sera of Toxo-IgM(-)-IgG(+) and -IgM(+)-IgG(-), respectively, which had been prescreened by Toxo-IgM and -IgG commercial kits from the suspected cases. Following cloning, expression, and purification of the fragment of ROP2(186-533), an enzyme-linked immunosorbent assay with rROP2(186-533) to measure IgM and IgG antibodies was developed. As a result, 100%(48/48) of sera with Toxo-IgM(+)-IgG(-)showed positive Toxo-IgM but none of them (0%) showed positive Toxo-IgG when rROP2(186-533) was used as antigen. Neither Toxo-IgG nor Toxo-IgM antibodies were found when tested with 59 sera of Toxo-IgM(-)-IgG(+). These results indicate that rROP2(186-533) could be used as an antigen that specifically capture Toxo-IgM antibodies and may have a high potential in the serological diagnosis of both acute acquired and congenital toxoplasmosis.
Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários , Complicações Parasitárias na Gravidez/diagnóstico , Toxoplasma/imunologia , Toxoplasmose Congênita/diagnóstico , Toxoplasmose/diagnóstico , Animais , Especificidade de Anticorpos , Antígenos de Protozoários/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina M/sangue , Proteínas de Membrana , Peptídeos , Gravidez , Complicações Parasitárias na Gravidez/imunologia , Proteínas de Protozoários , Proteínas Recombinantes , Sensibilidade e Especificidade , Toxoplasma/isolamento & purificação , Toxoplasmose/imunologia , Toxoplasmose/parasitologia , Toxoplasmose Congênita/imunologia , Toxoplasmose Congênita/parasitologiaRESUMO
INTRODUCTION: Mature seeds deteriorate gradually and die eventually during long-term storage. Controlled deterioration is often used to accelerate the seed deterioration rate to assess the seed vigor and physiological quality of seed lots. OBJECTIVES: Although it is well known that the process of seed deterioration produced by controlled deterioration is distinct from that caused by long-term storage, the differences in transcriptional levels have not been reported. Clarifying the mechanism of seed deterioration is critical for identifying, conserving and utilizing germplasm resources. METHODS: Tobacco (Nicotiana tabacum L.) seeds were studied thoroughly using transcriptome, small RNA, and degradome sequencing after long-term storage (LS) and controlled deterioration (CD). Co-expression trend analysis identified transcripts involved in tobacco seed deterioration, while phylogenetic analysis helped to uncover comparable targets in rice (Oryza sativa L.) for further verification and utilization. RESULTS: In LS and CD, a total of 2,112 genes and 164 miRNAs were differentially expressed, including 20 interaction miRNA-mRNA pairs with contrasting expression. Transcriptional multiomics found that the main causes of LS were plant hormone signal transduction and protein processing in the endoplasmic reticulum, whereas the primary cause of CD was nucleotide excision repair dysfunction. The homeostatic balance of RNA degradation and the spliceosome occurred in both modes of seed deterioration. Additionally, co-expression trend analysis identified two coherent pairs, nta-miR160b-NtARF18 and nta-miR396c-NtMBD10, as being significant in LS and CD, respectively. For utilization, rice homologous targets OsARF18 and OsMBD707 were verified to play similar roles in LS and CD, respectively. CONCLUSION: This study demonstrated the transcriptional mechanism of tobacco and key genes in seed deterioration. And the application of key genes in rice also verified the feasibility of the multiomics method, guiding the identification of candidate genes to precisely delay seed deterioration in other species of seed research.
Assuntos
MicroRNAs , Oryza , Oryza/genética , Oryza/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Regulação da Expressão Gênica de Plantas , Filogenia , Multiômica , Sementes/genética , MicroRNAs/metabolismoRESUMO
BACKGROUND: The digestibility of starch is important for the nutritive value of staple food. Although several genes are responsible for resistant starch (RS) and slowly digestible starch (SDS), gaps persist concerning the molecular basis of RS and SDS formation due to the complex genetic mechanisms of starch digestibility. OBJECTIVES: The objective of this study was to identify new genes for starch digestibility in rice and interprete the genetic mechanisms of RS and SDS by GWAS. METHODS: Genome-wide association studies were conducted by associating the RS and SDS phenotypes of 104 re-sequenced rice lines to an SNP dataset of 2,288,867 sites using a compressed mixed linear model. Candidate genes were identified according to the position of the SNPs based on data from the MSU Rice Genome Annotation Project. RESULTS: Seven quantitative trait loci (QTLs) were detected to be associated with the RS content, among which the SNP 6 m1765761 was located on Waxy. Starch branching enzymes IIa (BEIIa) close to QTL qRS-I4 was detected and further identified as a specific candidate gene for RS in INDICA. Two QTLs were associated with SDS, and the LOC_Os09g09360 encoding lipase was identified as a causal gene for SDS. CONCLUSIONS: GWAS is a valid strategy to genetically dissect the formation of starch digestion properties in rice. RS formation in grains is dependent on the rice type; lipid might also contribute to starch digestibility and should be an alternative factor to improve rice starch digestibility.
Assuntos
Enzima Ramificadora de 1,4-alfa-Glucana/genética , Lipase/genética , Oryza/genética , Amido Resistente , Amido/genética , Digestão/genética , Genoma de Planta/genética , Estudo de Associação Genômica Ampla , Humanos , Oryza/química , Oryza/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Locos de Características Quantitativas/genética , Amido/metabolismoRESUMO
A series of novel phenacylimidazolium derivatives, bearing an aryl or alkyl substituent at position-1 and a phenacyl substituent at position-3 of the imidazole ring, has been prepared and evaluated in vitro against a panel of human tumor cell lines. Phenacylimidazolium bromides bearing a highly sterically hindered aryl group at position-1 and an electron-rich phenacyl or naphthylacyl substituent at position-3 of imidazole ring proved to be more active than imidazolium bromides with other substituted groups. In particular, compound 5j was found to be the most potent compounds with IC(50) values lower than 5.0 microM against 8 strains human tumor cell lines and more active than cisplatin (DDP).
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Antineoplásicos/síntese química , Antineoplásicos/toxicidade , Imidazóis/síntese química , Imidazóis/toxicidade , Antineoplásicos/química , Linhagem Celular Tumoral , Cisplatino/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Imidazóis/química , Relação Estrutura-AtividadeRESUMO
Liver biopsy still remains the gold standard for diagnosing nonalcoholic steatohepatitis (NASH), but with limitations. There is an urgent need to develop noninvasive tests that accurately distinguish NASH from simple steatosis. The purpose of this meta-analysis was to evaluate the diagnostic value of serum biomarkers including cytokeratin 18 (CK-18), fibroblast growth factor 21 (FGF-21), and combined biomarker panel (CBP) in the diagnosis of NAFLD, especially NASH. A total of 25 studies met the inclusion criteria. Pooled sensitivity and specificity values for chosen serum markers for diagnosing NASH are as follows: CK-18 (M30), 0.75 and 0.77; CK-18 (M65), 0.71 and 0.77; FGF-21, 0.62 and 0.78; and CBP, 0.92 and 0.85. CBP demonstrated better accuracy with higher sensitivity and specificity than those tested individually. Furthermore, the AUROC of CBP was 0.94 (95% CI, 0.92-0.96), compared to CK-18 or FGF-21 assay, which showed the most significant ability to distinguish NASH from simple steatosis. The results suggest that increased circulating CK-18 and FGF-21 are associated with NASH and may be used for initial assessment, but not enough. Importantly, CBP is potentially used as accurate diagnostic tools for NASH. Further prospective designed studies are warranted to confirm our findings.
Assuntos
Biomarcadores/sangue , Fatores de Crescimento de Fibroblastos/sangue , Queratina-18/sangue , Hepatopatia Gordurosa não Alcoólica/sangue , Biópsia , Fígado Gorduroso/patologia , Humanos , Fígado/patologia , Hepatopatia Gordurosa não Alcoólica/patologiaRESUMO
The performance of indirect haemagglutination assays (IHA), enzyme-linked immunosorbent assays (ELISA) and indirect immunofluorescent antibody tests (IFAT) were compared with 450 sera from a Schistosoma mansoni-endemic area in Burkina Faso. All participants in this survey provided at least one sample each of stool, urine and serum. From those with an egg-negative Kato-Katz thick smear, a second stool sample was examined. IHA was based on either extracts of adult S. mansoni worms (SmIHA) or S. japonicum egg antigen (SjIHA). For ELISA, three antigen preparations were used, namely: (i) soluble S. mansoni adult worm antigens (SWAP); (ii) soluble S. mansoni egg antigens (SEA); and (iii) a cationic exchange fraction of S. mansoni eggs (CEF6). IFAT was performed with S. mansoni male worm sections. Among the egg-excretors, the sensitivity of ELISA was high and egg antigens performed slightly better (SEA, 96%; CEF6, 97%) than worm antigen (94%). Sensitivity of IHA was satisfactory with homologous (Sm, >85%), but not heterologous (Sj, 56%) parasite antigen. In IFAT, the parenchyma-associated fluorescence showed high sensitivity (95%), but gut-associated fluorescence, which is known to be a sensitive diagnostic marker for schistosome-infected European travelers, was observed only in 76% of a sub-sample of 100 of the endemic sera. Among sera from egg-negative individuals, many gave positive reactions in several or all of the tests employed. These reactions (formally "false positive") are considered to represent true infections, since chemotherapy had not yet been delivered to this population. For the purpose of further surveys in Burkina Faso or other resource-poor settings, we suggest IHA as an accurate diagnostic test and propose to further improve its performance by including egg rather than worm antigens.
Assuntos
Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/diagnóstico , Adolescente , Adulto , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/química , Burkina Faso/epidemiologia , Criança , Estudos Transversais , Doenças Endêmicas , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Testes de Hemaglutinação , Humanos , Masculino , Contagem de Ovos de Parasitas , Esquistossomose mansoni/epidemiologia , Esquistossomose mansoni/imunologiaRESUMO
OBJECTIVE: To explore possible associations between host polymorphism of HLA class II genotypes and advanced hepatosplenic schistosomiasis japonica. METHODS: 45 advanced schistosomiasis patients (experimental group) and 44 age- and sex-matched patients with chronic schistosomiasis (control group) from the same area were investigated for their HLA class II gene DRB genotypes by genotyping the alleles using microarray DNA chip. The correlation of allele frequencies to advanced hepatosplenic schistosomiasis was compared for the two groups. RESULTS: HLA-DRB1*04x exhibited markedly higher frequency in advanced patients than that in control group (P < 0.01, RR = 3.928). In contrast, the frequency of HLA-DRB1*15x in advanced patients was much lower when compared with that in control group (P < 0.01, RR = 0.050). Besides, the significant allele HLA-DRB1*15x displayed concurrence with allele DRB5*010x/020x. The linked gene haplotype DRB1*15x-DRB5*010x/020x showed significantly higher incidence in control group than in experimental group (P < 0.01). CONCLUSION: Allele HLA-DRB1*04x is positively, while HLA-DRB1*15x is negatively, correlated with advanced hepatosplenic schistosomiasis.
Assuntos
Antígenos HLA-DR/genética , Análise de Sequência com Séries de Oligonucleotídeos , Esquistossomose Japônica/genética , Adulto , Idoso , Alelos , Feminino , Frequência do Gene , Genótipo , Cadeias HLA-DRB1 , Humanos , Fígado/parasitologia , Masculino , Pessoa de Meia-Idade , Polimorfismo Genético , Baço/parasitologiaRESUMO
OBJECTIVE: To study the impact of chronic schistosomiasis on the protective immunity induced by vaccine against hepatitis B virus. METHODS: 24 patients with chronic or advanced schistosomiasis (experimental group) and 26 healthy volunteers (control group) all without hepatitis B virus infection were selected for the study. Sera of the subjects in the two groups were collected before inoculation and on the 35th day after inoculation with yeast-derived recombinant hepatitis B vaccine. The level of anti-Hbs, IL-2 and TNF-alpha in sera was examined by ELISA respectively. RESULTS: Anti-Hbs in both groups were negative before inoculation, with an average absorbance (A value) of 0.134 and 0.150 respectively. After inoculation, positive rate of anti-Hbs was 17% (4/24, average A value 0.145 ) in experimental group and 92% (24/26, average A value was 1.210) in control group. The vaccine against hepatitis B induced significantly higher level of anti-Hbs in healthy volunteers compared with that in schistosomiasis patients (P < 0.01). The level of IL-2 and TNF-alpha increased in both groups after inoculation without significant difference compared with the level before inoculation. CONCLUSION: The results suggest that the protective immunity of patients with chronic schistosomiasis is deficient to the stimulation of hepatitis B virus and it may involve in a higher incidence of hepatitis B among schistosomiasis patients.
Assuntos
Vacinas contra Hepatite B/imunologia , Vírus da Hepatite B/imunologia , Esquistossomose Japônica/imunologia , Adulto , Doença Crônica , Ensaio de Imunoadsorção Enzimática , Feminino , Hepatite B/imunologia , Anticorpos Anti-Hepatite B/sangue , Humanos , Interleucina-2/sangue , Masculino , Pessoa de Meia-Idade , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Chilling stress is an important constraint for maize seed establishment in the field. In this study, a type of "on-off" thermoresponsive coating agent containing poly (N-isopropylacrylamide-co-butylmethacrylate) (Abbr. P(NIPAm-co-BMA)) hydrogel was developed to improve the chilling tolerance of coated maize seed. The P(NIPAm-co-BMA) hydrogel was synthesized by free-radical polymerization of N-isopropylacrylamide (NIPAm) and butylmethacrylate (BMA). Salicylic acid (SA) was loaded in the hydrogel as the chilling resistance agent. SA-loaded P(NIPAm-co-BMA) was used for seed film-coating of two maize varieties, Huang C (HC, chilling-tolerant) and Mo17 (chilling-sensitive), to investigate the coated seed germination and seedling growth status under chilling stress. The results showed that the hydrogel obtained a phase transition temperature near 12°C with a NIPAM to MBA weight ratio of 1: 0.1988 (w/w). The temperature of 12°C was considered the "on-off" temperature for chilling-resistant agent release; the SA was released from the hydrogel more rapidly at external temperatures below 12°C than above 12°C. In addition, when seedlings of both maize varieties suffered a short chilling stress (5°C), higher concentrations of SA-loaded hydrogel resulted in increased germination energy, germination percentage, germination index, root length, shoot height, dry weight of roots and shoots and protective enzyme activities and a decreased malondialdehyde content in coated maize seeds compared to single SA treatments. The majority of these physiological and biochemical parameters achieved significant levels compared with the control. Therefore, SA-loaded P(NIPAm-co-BMA), a nontoxic thermoresponsive hydrogel, can be used as an effective material for chilling tolerance in film-coated maize seeds.
Assuntos
Germinação/efeitos dos fármacos , Ácido Salicílico/farmacologia , Plântula/efeitos dos fármacos , Sementes/efeitos dos fármacos , Estresse Fisiológico/efeitos dos fármacos , Zea mays/efeitos dos fármacos , Zea mays/fisiologia , Acrilamidas/farmacologia , Fenômenos Bioquímicos/efeitos dos fármacos , Temperatura Baixa , Germinação/fisiologia , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Malondialdeído/farmacologia , Fenômenos Fisiológicos/efeitos dos fármacos , Fenômenos Fisiológicos/fisiologia , Plântula/fisiologia , Sementes/fisiologiaRESUMO
BACKGROUND: Hepatitis B and schistosomiasis are most prevalent in Africa and Asia, and co-infections of both are frequent in these areas. The immunomodulation reported to be induced by schistosome infections might restrict immune control of hepatitis B virus (HBV) leading to more severe viral infection. Vaccination is the most effective measure to control and prevent HBV infection, but there is evidence for a reduced immune response to the vaccine in patients with chronic schistosomiasis japonica. METHODOLOGY/PRINCIPAL FINDINGS: In this paper, we demonstrate in a mouse model that a chronic Schistosoma japonicum infection can inhibit the immune response to hepatitis B vaccine (HBV vaccine) and lead to lower production of anti-HBs antibodies, interferon-γ (IFN-γ) and interleukin-2 (IL-2). After deworming with Praziquantel (PZQ), the level of anti-HBs antibodies gradually increased and the Th2-biased profile slowly tapered. At 16 weeks after deworming, the levels of anti-HBs antibodies and Th1/Th2 cytokines returned to the normal levels. CONCLUSIONS/SIGNIFICANCE: The results suggest that the preexisting Th2-dominated immune profile in the host infected with the parasite may down-regulate levels of anti-HBs antibodies and Th1 cytokines. To improve the efficacy of HBV vaccination in schistosome infected humans it may be valuable to treat them with praziquantel (PZQ) some time prior to HBV vaccination.
Assuntos
Vacinas contra Hepatite B/metabolismo , Schistosoma japonicum/imunologia , Esquistossomose Japônica/imunologia , Esquistossomose Japônica/parasitologia , Animais , Anti-Helmínticos/farmacologia , Doença Crônica , Citocinas/metabolismo , Sistema Imunitário , Interferon gama/metabolismo , Interleucina-2/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Praziquantel/farmacologia , RNA Mensageiro/metabolismo , Baço/imunologia , Células Th2/imunologiaRESUMO
T cell immunoglobulin domain and mucin domain (Tim) family, a new gene that expresses on the surface of T cells, plays a critical role in regulation of T cells response. Previous data have shown that Tim-3 expressed on Th1 cells promotes itself apoptosis. Tim-2 is preferentially up-regulated during Th2 differentiation and functions as a potent costimulatory molecule for T-cell immunity. The present study aims to learn whether Tims are responsible for Th2-biased response evoked by Schistosoma japonicum infection. The expressions of Tim-2 and Tim-3 in spleen lymphocytes from S. japonicum-infected mice were examined, and the possible role of galectin-9-Tim-3 pathway in Th2-biased response triggered by schistosome infection was discussed. Our results showed that Tim-2 mRNAs were up-regulated in the spleen of schistosome-infected mice, which coincided with elevated IL-4 gene expression. Administration of galectin-9 significantly induced apoptosis of naïve spleen lymphocytes with down-regulation IFN-γexpression in vitro. Additionally, Tim-3-Fc fusion protein notably enhanced Th1 cells and decreased Th2 cells in vitro. Thus, we concluded that pro-apoptotic effects on Th1 population through galectin-9-Tim-3 pathway and the up-regulation of Tim-2 on Th2 cells might be critical to Th2-biased response of host with schistosomiasis japonica.