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1.
J Insect Sci ; 19(5)2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31606748

RESUMO

Dengue, yellow fever, and Zika are viruses transmitted by yellow fever mosquito, Aedes aegypti [Linnaeus (Diptera: Culicidae)], to thousands of people each year. Mosquitoes transmit these viruses while consuming a blood meal that is required for oogenesis. Iron, an essential nutrient from the blood meal, is required for egg development. Mosquitoes receive a high iron load in the meal; although iron can be toxic, these animals have developed mechanisms for dealing with this load. Our previous research has shown iron from the blood meal is absorbed in the gut and transported by ferritin, the main iron transport and storage protein, to the ovaries. We now report the distribution of iron and ferritin in ovarian tissues before blood feeding and 24 and 72 h post-blood meal. Ovarian iron is observed in specific locations. Timing post-blood feeding influences the location and distribution of the ferritin heavy-chain homolog, light-chain homolog 1, and light-chain homolog 2 in ovaries. Understanding iron deposition in ovarian tissues is important to the potential use of interference in iron metabolism as a vector control strategy for reducing mosquito fecundity, decreasing mosquito populations, and thereby reducing transmission rates of vector-borne diseases.


Assuntos
Aedes/metabolismo , Ferritinas/metabolismo , Ferro/metabolismo , Ovário/metabolismo , Animais , Sangue/metabolismo , Feminino , Ferritinas/química , Suínos
2.
Zhongguo Zhong Yao Za Zhi ; 44(16): 3569-3575, 2019 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-31602924

RESUMO

To further investigate the metabolism of Tripterygium wilfordii and Paeonia lactiflora micro-emulsion gel in vivo,an LCMS/MS method was established for the determination of triptolide and paeoniflorin in T. wilfordii and P. lactiflora micro-emulsion gel.The extracorporeal recovery rate of blood probe was measured by concentration difference methods( incremental method and decremental method). Meanwhile,the skin and blood micro-dialysis methods of tripterine and paeoniflorin were established,and the pharmacokinetics of T. wilfordii microemulsion gel in skin and blood was studied by micro-dialysis combined with LC-MS/MS quantitative analysis. The results showed that the established method for the determination of triptolide and paeoniflorin in T. wilfordii microemulsion gel was well linear within the required range,and the specificity,recovery rate and degree of precision of the chromatography all conformed to the research requirements of micro-dialysis samples. The stability of freeze-thawing and the residual effect all conformed to the criteria of biological sample methodology. The probe recovery rates measured by incremental method and decremental method were almost consistent with the extracorporeal recovery rate test. The recovery rates of paeoniflorin in skin and blood micro-dialysis were( 30. 60±1. 09) % and( 28. 01± 1. 75) %,respectively. And the recovery rates of skin and blood micro-dialysis were( 26. 79 ± 2. 78) % and( 25. 39±1. 86) %,respectively. The intraday recovery rate of probes was stable within 11 h. The results of pharmacokinetic study showed that the Cmaxvalues of triptolide in skin and blood were( 148. 03±41. 51) and( 76. 77±15. 27) µg·L-1,respectively. And the Tmaxvalues were( 2. 33±0. 29) and( 3. 00± 0) h,respectively. The AUC0-11 hvalues were( 2 814. 05± 1 070. 37) and( 1 580. 63±208. 27) µg·h·L-1,respectively. The MRT0-11 hvalues were( 4. 20± 0. 33) and( 4. 54± 0. 34) h,respectively. The T1/2 values were( 4. 61±4. 11) and( 1. 07± 0. 13) h,respectively. The Cmaxvalues of paeoniflorin in skin and blood were( 991. 88 ± 152. 22) and( 407. 02±120. 06) µg·L-1,respectively. The Tmaxvalues were( 2. 00±0) h and( 2. 83±0. 29) h,respectively. The AUC0-11 hvalues were( 18 430. 27±3 289. 35) and( 6 338. 59 ± 1 659. 32) µg·h·L-1,respectively. The MRT0-11 hvalues were( 4. 29 ± 0. 16) and( 4. 00±0. 05) h,respectively. The T1/2 values were( 2. 16±0. 43) and( 1. 78±0. 48) h,respectively. The results suggested that micro-emulsion gel played a role in forming skin reservoir through percutaneous penetration. It not only could improve drug transdermal efficiency,but also control the sustained release of drug and form a long-term effect.


Assuntos
Sangue/metabolismo , Medicamentos de Ervas Chinesas/farmacocinética , Paeonia/química , Pele/metabolismo , Tripterygium/química , Cromatografia Líquida , Emulsões , Géis , Humanos , Espectrometria de Massas em Tandem
3.
Nature ; 574(7779): 543-548, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31645720

RESUMO

Multicellular organisms have co-evolved with complex consortia of viruses, bacteria, fungi and parasites, collectively referred to as the microbiota1. In mammals, changes in the composition of the microbiota can influence many physiologic processes (including development, metabolism and immune cell function) and are associated with susceptibility to multiple diseases2. Alterations in the microbiota can also modulate host behaviours-such as social activity, stress, and anxiety-related responses-that are linked to diverse neuropsychiatric disorders3. However, the mechanisms by which the microbiota influence neuronal activity and host behaviour remain poorly defined. Here we show that manipulation of the microbiota in antibiotic-treated or germ-free adult mice results in significant deficits in fear extinction learning. Single-nucleus RNA sequencing of the medial prefrontal cortex of the brain revealed significant alterations in gene expression in excitatory neurons, glia and other cell types. Transcranial two-photon imaging showed that deficits in extinction learning after manipulation of the microbiota in adult mice were associated with defective learning-related remodelling of postsynaptic dendritic spines and reduced activity in cue-encoding neurons in the medial prefrontal cortex. In addition, selective re-establishment of the microbiota revealed a limited neonatal developmental window in which microbiota-derived signals can restore normal extinction learning in adulthood. Finally, unbiased metabolomic analysis identified four metabolites that were significantly downregulated in germ-free mice and have been reported to be related to neuropsychiatric disorders in humans and mouse models, suggesting that microbiota-derived compounds may directly affect brain function and behaviour. Together, these data indicate that fear extinction learning requires microbiota-derived signals both during early postnatal neurodevelopment and in adult mice, with implications for our understanding of how diet, infection, and lifestyle influence brain health and subsequent susceptibility to neuropsychiatric disorders.


Assuntos
Extinção Psicológica/fisiologia , Medo/fisiologia , Metabolômica , Microbiota/fisiologia , Neurônios/fisiologia , Animais , Antibacterianos/farmacologia , Transtorno Autístico/metabolismo , Sangue/metabolismo , Cálcio/metabolismo , Líquido Cefalorraquidiano/química , Líquido Cefalorraquidiano/metabolismo , Sinais (Psicologia) , Espinhas Dendríticas/efeitos dos fármacos , Espinhas Dendríticas/patologia , Espinhas Dendríticas/fisiologia , Extinção Psicológica/efeitos dos fármacos , Medo/efeitos dos fármacos , Fezes/química , Vida Livre de Germes , Indicã/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Microbiota/efeitos dos fármacos , Microbiota/imunologia , Inibição Neural , Neuroglia/patologia , Neuroglia/fisiologia , Neurônios/efeitos dos fármacos , Neurônios/imunologia , Neurônios/patologia , Fenilpropionatos/metabolismo , Córtex Pré-Frontal/citologia , Córtex Pré-Frontal/efeitos dos fármacos , Córtex Pré-Frontal/imunologia , Córtex Pré-Frontal/fisiologia , Esquizofrenia/metabolismo , Transcriptoma , Nervo Vago/fisiologia
4.
BMC Genomics ; 20(1): 704, 2019 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-31506065

RESUMO

BACKGROUND: It is well-known that methylation changes occur as humans age, however, understanding how age-related changes in DNA methylation vary by sex is lacking. In this study, we characterize the effect of age on DNA methylation in a sex-specific manner and determine if these effects vary by genomic context. We used the Illumina HumanMethylation 450 K array and DNA derived from whole blood for 400 adult participants (189 males and 211 females) from Bangladesh to identify age-associated CpG sites and regions and characterize the location of these age-associated sites with respect to CpG islands (vs. shore, shelf, or open sea) and gene regions (vs. intergenic). We conducted a genome-wide search for age-associated CpG sites (among 423,604 sites) using a reference-free approach to adjust for cell type composition (the R package RefFreeEWAS) and performed an independent replication analysis of age-associated CpGs. RESULTS: The number of age-associated CpGs (p < 5 x 10- 8) were 986 among men and 3479 among women of which 2027(63.8%) and 572 (64.1%) replicated (using Bonferroni adjusted p < 1.2 × 10- 5). For both sexes, age-associated CpG sites were more likely to be hyper-methylated with increasing age (compared to hypo-methylated) and were enriched in CpG islands and promoter regions compared with other locations and all CpGs on the array. Although we observed strong correlation between chronological age and previously-developed epigenetic age models (r ≈ 0.8), among our top (based on lowest p-value) age-associated CpG sites only 12 for males and 44 for females are included in these prediction models, and the median chronological age compared to predicted age was 44 vs. 51.7 in males and 45 vs. 52.1 in females. CONCLUSIONS: Our results describe genome-wide features of age-related changes in DNA methylation. The observed associations between age and methylation were generally consistent for both sexes, although the associations tended to be stronger among women. Our population may have unique age-related methylation changes that are not captured in the established methylation-based age prediction model we used, which was developed to be non-tissue-specific.


Assuntos
Envelhecimento/genética , Sangue/metabolismo , Metilação de DNA , Adulto , Idoso , Bangladesh , Ilhas de CpG/genética , Epigênese Genética , Feminino , Predisposição Genética para Doença/genética , Genoma Humano/genética , Humanos , Masculino , Pessoa de Meia-Idade , Caracteres Sexuais
5.
Analyst ; 144(20): 6011-6018, 2019 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-31538156

RESUMO

Serotonin and kynurenine are formed following metabolism of the essential amino acid tryptophan. Both molecules play important biological roles and the balance of how tryptophan metabolism varies to either the serotonin or kynurenine pathway may provide key insight into the inflammatory status of the biological region. At present complex chromatographic methods are utilised which predominately focus on either monitoring analytes in the serotonin or kynurenine pathway rather than both. Our study develops a simple yet robust methodology for the monitoring of tryptophan metabolism. We utilised isocratic reverse phase high-performance liquid chromatography with simultaneously dual electrochemical detection. This approach allowed for separation of co-eluted analytes and identification of analytes from both pathways within 14 minutes. For all analytes, limits of detection were <35 nM. No crosstalk was observed when dual simultaneous detection was conducted in a radial flow cell. Responses from the hippocampus, blood and ileum mucosa highlighted that each region had a varying ratio of serotonin to kynurenine pathway, indicating varied approaches to tryptophan metabolism. The developed method can monitor how the metabolism of tryptophan varies between the two pathways which can provide insight into the inflammatory state of reach region with age and disease.


Assuntos
Sangue/metabolismo , Hipocampo/metabolismo , Íleo/metabolismo , Triptofano/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Técnicas Eletroquímicas , Cinurenina/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Serotonina/metabolismo
6.
Radiat Res ; 192(4): 399-409, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31373872

RESUMO

In a large-scale radiological incident, rapid and high-throughput biodosimetry would be needed. Gene expression-based biodosimetry is a promising approach to determine the dose received after radiation exposure. We previously identified 35 candidate genes as biodosimetry markers based on a systematic review. The goal of the current study was to establish and validate a specific gene expression-based radiological biodosimetry using a panel of highly radioresponsive genes in human peripheral blood for improving the accuracy of dose estimation. Human peripheral blood samples from 30 adult donors were irradiated to 0, 0.5, 1, 2, 3, 4, 6 and 8 Gy with 60Co γ rays at a dose rate of 1 Gy/min. We examined the expression patterns of candidate genes using real-time polymerase chain reaction (qRT-PCR) at 6, 12, 24 and 48 h postirradiation. Stepwise regression analysis was employed to develop the gene expression-based dosimetry models at each time point. Samples from another 10 healthy donors (blind samples) and four total-body irradiated (TBI) patients were used to validate the radiation dosimetry models. We observed significant linear dose-response relationships of CDKN1A, BAX, MDM2, XPC, PCNA, FDXR, GDF-15, DDB2, TNFRSF10B, PHPT1, ASTN2, RPS27L, BBC3, TNFSF4, POLH, CCNG1, PPM1D and GADD45A in human peripheral blood at the various time points. However, the expression levels of these genes were affected by inter-individual variations and gender. We found that the gender-dependent regression models could explain 0.85 of variance at 24 h postirradiation and could also accurately estimate the absorbed radiation doses with dose range of 0-5 Gy, in human peripheral blood samples irradiated ex vivo and from TBI patients, respectively. This study demonstrates that developing gender-specific biodosimetry based on a panel of highly radioresponsive genes may help advance the application of gene expression signature for dose estimation in the event of a radiological accident or in clinical treatment.


Assuntos
Sangue/metabolismo , Sangue/efeitos da radiação , Radiometria/métodos , Caracteres Sexuais , Transcriptoma/efeitos da radiação , Relação Dose-Resposta à Radiação , Feminino , Humanos , Masculino , Modelos Biológicos , Fatores de Tempo
7.
Nucl Med Commun ; 40(10): 1029-1035, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31469806

RESUMO

BACKGROUND: Measurement of glomerular filtration rate from a single blood sample using filtration markers may be based on empirical or model-based formulae. The former do not require a whole body metric but are sample time-inflexible. The latter include time as a continuous variable but require a whole body metric to estimate extracellular fluid volume. This study describes a new empirical method that is time-flexible and requires no whole body metric. METHODS: Data comprise 982 slope-intercept glomerular filtration rate measurements from a single institution in which the correlation coefficient of the three-sample fit was ≥0.999. In 582 measurements, samples were taken at 118-122, 144-148, 178-182, 202-206 or 238-242 minutes. Within each of these five sample time groups, slope, k(t), of the regression of apparent volume of distribution (Vt) against slope-intercept glomerular filtration rate was recorded following exclusion of outliers. The five sampling times correlated closely and exponentially with k(t) (=5.4.e), allowing calculation of k(t) for any sampling time. In a further 341 measurements, glomerular filtration rate from a single blood sample was calculated as [k(t).V(t)] and compared with slope-intercept glomerular filtration rate. Sample time for glomerular filtration rate from a single blood sample is critical, so the most appropriately timed sample was chosen according to slope-intercept glomerular filtration rate/1.73 m. RESULTS: The current method compared favourably with three previously published model-based methods (Fleming et al., Jacobsson and Gref and Karp), showing better precision, and unlike the other three, no correlation between difference and average on Bland-Altman analysis. CONCLUSION: This completely time-flexible, empirically-derived approach to glomerular filtration rate from a single blood sample requires no whole body metric, is simple, equally applicable to children and adults and superior to three model-based methods.


Assuntos
Sangue/metabolismo , Taxa de Filtração Glomerular , Testes de Função Renal/métodos , Humanos , Fatores de Tempo
8.
Radiat Res ; 192(2): 189-199, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31237816

RESUMO

In the possible event of a detonation of an improvised nuclear device (IND), the immediate radiation would consist of both photons (gamma rays) and neutrons. Since neutrons generally have a high relative biological effectiveness (RBE) for most physiological end points, it is important to understand the effect that neutrons would have on the biodosimetry methods that are being developed for medical triage purposes. We previously compared the transcriptomic response in human blood after neutron and photon irradiation. In this study, we analyzed the effect of mixed-field-neutron-photon radiation on gene expression responses in human peripheral blood, to elucidate the neutron contribution in the setting of a radiation exposure from an IND detonation. We used four combinations of mixed neutron-photon exposures, with increasing percentages of neutrons, to a cumulative dose of 3 Gy. The mixed-field exposures consisted of 0%, 5%, 15% and 25% of neutrons, where 0% corresponds to 3 Gy of pure X rays. A maximum neutron exposure, corresponding to 83% neutrons (0.75 Gy) was also used in the study. Increases were observed in both the number and expression level of genes, with increasing percentages of neutrons from 0% to 25% in the mixed-field exposures. Gene ontology analysis showed an overall predominance of TP53 signaling among upregulated genes across all exposures. Some TP53 regulated genes, such as EDA2R, GDF15 and VWCE, demonstrated increased expression with increasing neutron percentages in mixed-field exposures. Immune response, specifically natural-killer-cell mediated signaling, was the most significant biological process associated with downregulated genes. We observed significant suppression of T-cell-mediated signaling in mixed-field exposures, which was absent in the response to pure photons. In this first study investigating gene expression in human blood cells exposed to mixed neutron-photon fields similar to an actual IND explosion, we have identified a number of genes responding to the 3 Gy dose that showed increasing expression as the neutron percentage increased. Such genes may serve as better indicators of the expected biological damage than a report of total physical dose, and thus provide more relevant information for treating physicians.


Assuntos
Nêutrons/efeitos adversos , Fótons/efeitos adversos , Exposição à Radiação/efeitos adversos , Transcriptoma/efeitos da radiação , Sangue/metabolismo , Sangue/efeitos da radiação , Ontologia Genética , Voluntários Saudáveis , Humanos , Eficiência Biológica Relativa
9.
J Pharmacol Exp Ther ; 370(2): 288-298, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31152006

RESUMO

In mouse pharmacokinetic (PK) studies, current standard methods often require large numbers of animals to support collection of blood samples serially over a defined time range. We have developed and validated a noninvasive fluorescence molecular tomography (FMT) heart imaging approach for blood PK quantification that uses small numbers of mice and has the advantage of repeated, longitudinal live imaging. This method was validated using a variety of near infrared (NIR) fluorescent-labeled molecules, ranging in size from 1.3 to 150 kDa, that were assessed by microplate blood assays as well as by noninvasive FMT 4000 imaging. Excellent agreement in kinetic profiles and calculated PK metrics was seen for the two methods, establishing the robustness of this noninvasive optical imaging approach. FMT heart imaging was further assessed in the challenging application of inulin-based glomerular filtration rate (GFR) measurement. After a single bolus injection of an NIR fluorescent-labeled inulin probe in small cohorts of mice (n = 5 per group), 2-minute heart scans (at 2, 6, 15, 30, and 45 minutes) were performed by FMT imaging. GFR was calculated using two-compartment PK modeling, determining an average rate of 240 ± 21 µl/min in normal mice, in agreement with published mouse GFR ranges. Validation of GFR assessment in unilaterally nephrectomized mice and cyclosporin A-treated mice both measured ∼50% decreases in GFR. Imaging results correlated well with ex vivo plasma microplate assays for inulin blood kinetics, and the decreases in GFR were accompanied by increases in plasma creatinine and blood urea nitrogen.


Assuntos
Sangue/metabolismo , Taxa de Filtração Glomerular/efeitos dos fármacos , Coração/diagnóstico por imagem , Imagem Óptica , Tomografia , Animais , Sangue/efeitos dos fármacos , Creatinina/sangue , Feminino , Camundongos , Nitrogênio/urina , Distribuição Tecidual
10.
Forensic Sci Int ; 300: e44-e49, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31126709

RESUMO

MicroRNAs (miRNAs) have been of interest in forensic science for body fluid identification with recent years. However, there is no study investigating the species specificity of miRNA markers by the SYBR Green method. Due to the conservation of miRNAs across species, miRNA markers maybe less species-specific than mRNA markers, and in forensic cases, animal buccal swabs are more likely to appear. Therefore, in this study we addressed the influence of 8 kinds of animal buccal swabs on human saliva, semen, vaginal secretion swabs and blood identification with 10 candidate specific miRNA markers by the SYBR Green quantitative PCR. Our data showed that the expression levels of the candidate specific miRNA markers miR-124a and 372 in the cat, dog, mouse and rabbit buccal swabs were in the same range as the human vaginal secretion swabs; buccal swabs from these animals also showed similar expression levels to human saliva for the candidate specific miRNA markers miR-200c, 205 and 658. These results indicated that biomaterials of buccal swabs from cats, dogs, mice and rabbits may be mistaken for human saliva or human vaginal secretion swabs, both of which could result in false positives for human body fluids. Thus, the interpretation of these miRNA profiles for human body fluid identification can be inaccurate in the presence of these animal buccal swabs. Therefore, we suggested performing species tests before human body identification with miRNA markers.


Assuntos
Marcadores Genéticos , MicroRNAs/genética , Mucosa Bucal/metabolismo , Animais , Sangue/metabolismo , Gatos , Muco do Colo Uterino/metabolismo , Cães , Feminino , Genética Forense/métodos , Humanos , Masculino , Camundongos , MicroRNAs/metabolismo , Reação em Cadeia da Polimerase , Coelhos , Saliva/metabolismo , Sêmen/metabolismo , Especificidade da Espécie , Manejo de Espécimes
11.
Anim Genet ; 50(3): 217-227, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30957254

RESUMO

Indigenous cattle breeds in northern Eurasia have adapted to harsh climate conditions. The local breeds are important genetic resources with cultural and historical heritages, and therefore, their preservation and genetic characterization are important. In this study, we profiled the whole-blood transcriptome of two native breeds (Northern Finncattle and Yakutian cattle) and one commercial breed (Holstein) using high-throughput RNA sequencing. More than 15 000 genes were identified, of which two, 89 and 162 genes were significantly upregulated exclusively in Northern Finncattle, Yakutian cattle and Holstein cattle respectively. The functional classification of these significantly differentially expressed genes identified several biological processes and pathways related to signalling mechanisms, cell differentiation and host-pathogen interactions that, in general, point towards immunity and disease resistance mechanisms. The gene expression pattern observed in Northern Finncattle was more similar to that of Yakutian cattle, despite sharing similar living conditions with the Holstein cattle included in our study. In conclusion, our study identified unique biological processes in these breeds that may have helped them to adapt and survive in northern and sub-arctic environments.


Assuntos
Sangue/metabolismo , Bovinos/genética , Perfilação da Expressão Gênica , Animais , Bovinos/classificação , Bovinos/metabolismo , Regulação da Expressão Gênica , Redes e Vias Metabólicas
12.
Environ Sci Pollut Res Int ; 26(16): 16274-16278, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30977003

RESUMO

This study evaluated the effects of Bacillus subtilis, Saccharomyces boulardii, oregano, and calcium montmorillonite on the physical growth, intestinal histomorphology, and blood metabolites in Salmonella-challenged birds during the finisher phase. In this study, a total of 600 chicks (Ross 308) were randomly distributed into the following dietary treatments: basal diet with no treatment; infected with Salmonella; T1, infected + avilamycin; T2, infected + Bacillus subtilis; T3, Saccharomyces boulardii; T4, infected + oregano; T5, infected + calcium montmorillonite. Our results indicated that feed consumption, body weight gain, total body weight, and feed conversion ratio increased significantly (P < 0.01) in T1 and T2. Villus width increased significantly (P < 0.01) in T1 while the total area was significantly (P < 0.01) higher in T1 and T2 among the treatment groups. Blood protein was significantly (P < 0.01) high in T3 and T4; however, the glucose concentration was significantly (P < 0.01) high in T2, T3, and T4. The treatments increased significantly (P< 0.01) in the treatment groups compared to the negative control. Aspirate aminotransferase (AST) was significantly (P < 0.05) low in T3 compared to the positive control. In conclusion, the results indicated that supplementation of Bacillus subtilis and calcium montmorillonite improved the production performance compared to other feed additives in broiler chicks infected with Salmonella during the finisher phase.


Assuntos
Bentonita/farmacologia , Galinhas/crescimento & desenvolvimento , Intestinos/efeitos dos fármacos , Origanum , Salmonelose Animal/dietoterapia , Ração Animal/análise , Animais , Bacillus subtilis , Sangue/metabolismo , Sangue/microbiologia , Peso Corporal/efeitos dos fármacos , Cálcio/farmacologia , Galinhas/metabolismo , Galinhas/microbiologia , Intestinos/microbiologia , Intestinos/patologia , Oligossacarídeos/farmacologia , Doenças das Aves Domésticas/microbiologia , Probióticos/farmacologia , Saccharomyces boulardii , Salmonelose Animal/metabolismo , Salmonella enterica/patogenicidade , Hormônios Tireóideos/sangue
13.
Mol Nutr Food Res ; 63(9): e1801095, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30725537

RESUMO

SCOPE: The Mediterranean (MED) diet has been associated with a decreased risk of cardiovascular diseases. It is unclear whether this health effect can be mainly contributed to high intakes of monounsaturated fatty acids (MUFA), characteristic for the MED diet, or whether other components of a MED diet also play an important role. METHODS AND RESULTS: A randomized fully controlled parallel trial is performed to examine the effects of the consumption of a saturated fatty acid rich diet, a MUFA-rich diet, or a MED diet for 8 weeks on metabolite profiles, in 47 subjects at risk of the metabolic syndrome. A total of 162 serum metabolites are assessed before and after the intervention by using a targeted NMR platform. Fifty-two metabolites are changed during the intervention (false discovery rate [FDR] p < 0.05). Both the MUFA and MED diet decrease exactly the same fractions of LDL, including particle number, lipid, phospholipid, and free cholesterol fraction (FDR p < 0.05). The MED diet additionally decreases the larger subclasses of very-low-density lipoprotein (VLDL), related VLDL fractions, VLDL-triglycerides, and serum-triglycerides (FDR p < 0.05). CONCLUSION: The findings clearly demonstrate that the MUFA component is responsible for reducing LDL subclasses and fractions, and therefore causes an antiatherogenic lipid profile. Interestingly, consumption of the other components in the MED diet show additional health effects.


Assuntos
Dieta Mediterrânea , Ácidos Graxos Monoinsaturados/farmacologia , Lipídeos/sangue , Lipoproteínas/sangue , Síndrome Metabólica/prevenção & controle , Biomarcadores/sangue , Sangue/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
14.
PLoS One ; 14(2): e0212679, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30802258

RESUMO

OBJECTIVES: To evaluate the accuracy of Spectral Photon-Counting Computed Tomography (SPCCT) in the quantification of iodine concentrations and its potential for the differentiation between blood and iodine. METHODS: Tubes with blood and a concentration series of iodine were scanned with a preclinical SPCCT system (both in vitro and in an ex vivo bovine brain tissue sample). Iodine density maps (IDM) and virtual non-contrast (VNC) images were generated using the multi-bin spectral information to perform material decomposition. Region-of-interest (ROI) analysis was performed within the tubes to quantitatively determine the absolute content of iodine (mg/ml). RESULTS: In conventional CT images, ROI analysis showed similar Hounsfield Unit (HU) values for the tubes with blood and iodine (59.9 ± 1.8 versus 59.2 ± 1.5). Iodine density maps enabled clear differentiation between blood and iodine in vitro, as well as in the bovine brain model. Quantitative measurements of the different iodine concentrations matched well with those of actual known concentrations even for very small iodine concentrations with values below 1mg/ml (RMSE = 0.19). CONCLUSIONS: SPCCT providing iodine maps and virtual non-contrast images allows material decomposition, differentiation between blood and iodine in vitro and ex vivo in a bovine brain model and reliably quantifies the iodine concentration.


Assuntos
Sangue/metabolismo , Encéfalo/diagnóstico por imagem , Meios de Contraste/farmacocinética , Iodo/farmacocinética , Fótons , Tomografia Computadorizada de Emissão , Animais , Encéfalo/metabolismo , Bovinos , Meios de Contraste/farmacologia , Iodo/farmacologia
15.
Am J Trop Med Hyg ; 100(4): 798-807, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30793681

RESUMO

American cutaneous leishmaniasis (ACL) is a common and important vector-borne parasitic zoonosis in Panamá. Here, we study Leishmania spp. infection rates and blood-feeding patterns among common sand flies in Trinidad de Las Minas, a rural community with hyperendemic ACL transmission, and where a deltamethrin fogging trial was performed. Sand flies were collected from April 2010 to June 2011 with light traps installed inside and in the peridomicile of 24 houses. We restricted our analysis to the most abundant species at the study site: Lutzomyia trapidoi, Lutzomyia gomezi, Lutzomyia panamensis, Lutzomyia triramula, and Lutzomyia dysponeta. We detected Leishmania spp. infection in sand flies by polymerase chain reaction (PCR) amplification of the internal transcribed spacer region 1 (ITS-1) in pooled females (1-10 females per pool). Host species of engorged sand flies were identified using a cytochrome b PCR. From 455 sand fly pools analyzed, 255 pools were positive for Leishmania spp., with an estimated infection rate (confidence interval) of 0.096 [0.080-0.115] before the deltamethrin fogging which slightly, but not significantly (P > 0.05), increased to 0.116 [0.098-0.136] after the deltamethrin fogging. Blood meal analysis suggested that pigs, goats, and birds were the most common sand fly blood sources, followed by humans and domestic dogs. DNA sequencing from a subsample of ITS-1 positive pools suggests that Leishmania panamensis, Leishmania naiffi, and other Leishmania spp. were the parasite species infecting the most common vectors at the study site. Our data confirm an association between sand fly species, humans, domestic dogs, and pigs and Leishmania spp. parasites in rural Panamá.


Assuntos
Doenças Endêmicas , Comportamento Alimentar , Leishmaniose Cutânea/epidemiologia , Psychodidae/fisiologia , Psychodidae/parasitologia , Animais , Sangue/metabolismo , DNA Intergênico/genética , DNA de Protozoário/genética , Cães , Feminino , Humanos , Insetos Vetores/parasitologia , Insetos Vetores/fisiologia , Leishmania/genética , Panamá/epidemiologia , População Rural , Análise de Sequência de DNA , Suínos
16.
Methods Mol Biol ; 1929: 739-754, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30710308

RESUMO

S100A8/A9 represents a novel biomarker and therapeutic target in sterile inflammatory diseases. Among the various S100 proteins, S100A8 and S100A9 have been shown to be the most important of all the damage-associated molecular pattern (DAMP) proteins in sterile inflammatory conditions such as diabetes, cardiovascular disease, autoimmune disorders, etc. We present here methods to quantify S100A8/A9 expression in various tissues in mouse models of myocardial infarction (MI) using flow cytometry (FC), immunofluorescence, quantitative real-time polymerase chain reaction (q-RT-PCR), and enzyme-linked immunosorbent assays (ELISA).


Assuntos
Calgranulina A/genética , Calgranulina A/metabolismo , Calgranulina B/genética , Calgranulina B/metabolismo , Infarto do Miocárdio/metabolismo , Animais , Biomarcadores/sangue , Biomarcadores/metabolismo , Sangue/metabolismo , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Imunofluorescência , Humanos , Camundongos , Monócitos/metabolismo , Infarto do Miocárdio/genética , Miocárdio/metabolismo , Neutrófilos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
17.
PLoS One ; 14(1): e0211743, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30703133

RESUMO

Camels are livestock with unique adaptations to hot-arid regions. To effectively study camel traits, a biobank of camel DNA specimens with associated biological information is needed. We examined whole-blood, saliva (buccal swabs), and tail-hair follicle samples to determine which is the best source for establishing a DNA biobank. We inspected five amounts of each of whole-blood, buccal swabs, and tail-hair follicles in nine camels, both qualitatively via gel electrophoresis and quantitatively using a NanoDrop spectrophotometer. We also tested the effects of long term-storage on the quality and quantity of DNA, and measured the rate of degradation, by analyzing three buccal swab samples and 30 tail-hair follicles over a period of nine months. Good quality DNA, in the form of visible large size DNA bands, was extracted from all three sources, for all five amounts. The five volumes of whole-blood samples (20-100µl) provided ~0.4-3.6 µg, the five quantities of buccal swabs (1-5) produced ~0.1-12 µg, while the five amounts of tail-hair follicles (10-50) resulted in ~0.7-25 µg. No differences in the rate of degradation of buccal swab and tail-hair follicle DNA were detected, but there was clearly greater deterioration in the quality of DNA extracted from buccal swabs when compared to tail-hair follicles. We recommend using tail-hair samples for camel DNA biobanking, because it resulted in both an adequate quality and quantity of DNA, along with its ease of collection, transportation, and storage. Compared to its success in studies of other domesticated animals, we anticipate that using ~50 tail-hair follicles will provide sufficient DNA for sequencing or SNP genotyping.


Assuntos
Bancos de Espécimes Biológicos/normas , Sangue/metabolismo , Camelus/genética , DNA/análise , Cabelo/metabolismo , Saliva/metabolismo , Manejo de Espécimes/métodos , Animais , DNA/genética , Cabelo/química , Saliva/química
18.
Oxid Med Cell Longev ; 2019: 7198484, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30733852

RESUMO

The purpose of an experimental design is to improve the productivity of experimentation. It is an efficient procedure for planning experiments, so the data obtained can be analyzed to yield a valid and objective conclusion. This approach has been used as an important tool in the optimization of different analytical approaches. A D-optimal experimental design was used here, for the first time, to optimize the experimental conditions for the detection of reactive oxygen species (ROS) produced by human blood from healthy donors, a biological matrix that better resembles the physiologic environment, following stimulation by a potent inflammatory mediator, phorbol-12-myristate-13-acetate (PMA). For that purpose, different fluorescent probes were used, as 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA), 2-[6-(4'-amino)-phenoxy-3H-xanthen-3-on-9-yl] benzoic acid (APF), and 10-acetyl-3,7-dihydroxyphenoxazine (amplex red). The variables tested were the human blood dilution, and the fluorescent probe and PMA concentrations. The experiments were evaluated using the Response Surface Methodology and the method was validated using specific compounds. This model allowed the search for optimal conditions for a set of responses simultaneously, enabling, from a small number of experiments, the evaluation of the interaction between the variables under study. Moreover, a cellular model was implemented and optimized to detect the production of ROS using a yet nonexplored matrix, which is human blood.


Assuntos
Sangue/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Humanos
19.
Bioorg Med Chem ; 27(5): 748-759, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30683552

RESUMO

To explore the application potential of dual prodrug strategies in the development of anti-HCV agents, a variety of sofosbuvir derivatives with modifications at the C4 or N3 position of the uracil moiety were designed and synthesized. Some compounds exhibited potent anti-HCV activities, such as 4e and 8a-8c with similar EC50 values (0.20-0.22 µM) comparative to that of sofosbuvir (EC50 = 0.18 µM) in a genotype 1b based replicon Huh-7 cell line. Moreover, 8b displayed a good human plasma stability profile, and was easily metabolized in human liver microsomes expectantly. On the other hand, aiming to discover novel anti-HCV nucleosides, pyrazin-2(1H)-one nucleosides and their phosphoramidate prodrugs were investigated. Several active compounds were discovered, such as 25e (EC50 = 7.3 µM) and S-29b (EC50 = 19.5 µM). This kind of nucleosides were interesting and would open a new avenue for the development of antiviral agents.


Assuntos
Antivirais/farmacologia , Hepacivirus/efeitos dos fármacos , Pró-Fármacos/farmacologia , Pirazinas/farmacologia , Sofosbuvir/análogos & derivados , Sofosbuvir/farmacologia , Antivirais/síntese química , Sangue/metabolismo , Linhagem Celular Tumoral , Descoberta de Drogas , Estabilidade de Medicamentos , Humanos , Microssomos Hepáticos/metabolismo , Pró-Fármacos/síntese química , Pirazinas/síntese química , Sofosbuvir/síntese química
20.
Rapid Commun Mass Spectrom ; 33(6): 561-568, 2019 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-30614103

RESUMO

RATIONALE: Metabonomics investigating perturbation to endogenous metabolism in response to external stimuli is emerging as a powerful tool for clinical diagnosis as well as in many other areas. The ability to retrieve reliable and reproducible information from complex biological fluids such as serum is crucial for its further applications. METHODS: In this study, the performance of the commonly used sample preparation methods for ultra-performance liquid chromatography/mass spectrometry (UPLC/MS)-based metabonomics was investigated. Specifically, we compared the extraction efficiencies, the method reproducibility, and the ability to identify potential biomarkers using solvent-based protein precipitation and solid-phase extraction (SPE) for serum metabonomic studies. Differences between extraction methods were explored using principal component analysis (PCA) and orthogonal partial least squares-discrimination analysis (OPLS-DA). RESULTS: Among the sample preparation methods tested, solvent-based protein precipitation using methanol has demonstrated the best analytical precision and extraction efficiency. Furthermore, this study revealed, for the first time, gender-specific differences in levels of two lysophosphatidylcholines (lysoPC 18:0 and lysoPC 18:1) in rat serum samples. CONCLUSIONS: The performance of sample preparation methods for UPLC/MS-based serum metabonomics was evaluated systematically. Results showed sample preparation by solvent precipitation using methanol provided the best analytical precision and extraction efficiency for UPLC/MS-based serum metabonomics.


Assuntos
Sangue/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Metabolômica/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Biomarcadores/sangue , Precipitação Química , Feminino , Masculino , Metaboloma , Análise Multivariada , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Extração em Fase Sólida/métodos , Solventes/química
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