Colchicine exerts anti-atherosclerotic and -plaque-stabilizing effects targeting foam cell formation.

Colchicine is a broad-acting anti-inflammatory agent that has attracted interest for repurposing in atherosclerotic cardiovascular disease. Here, we studied its ability at a human equivalent dose of 0.5 mg/day to modify plaque formation and composition in murine atherosclerosis and investigated its actions on macrophage responses to atherogenic stimuli in vitro. In atherosclerosis induced by high-cholesterol diet, Apoe-/- mice treated with colchicine had 50% reduction in aortic oil Red O+ plaque area compared to saline control (p = .001) and lower oil Red O+ staining of aortic sinus lesions (p = .03). In vitro, addition of 10 nM colchicine inhibited foam cell formation from murine and human macrophages after treatment with oxidized LDL (ox-LDL). Mechanistically, colchicine downregulated glycosylation and surface expression of the ox-LDL uptake receptor, CD36, and reduced CD36+ staining in aortic sinus plaques. It also decreased macrophage uptake of cholesterol crystals, resulting in lower intracellular lysosomal activity, inhibition of the NLRP3 inflammasome, and reduced secretion of IL-1ß and IL-18. Colchicine's anti-atherosclerotic actions were accentuated in a mouse model of unstable plaque induced by carotid artery tandem stenosis surgery, where it decreased lesion size by 48% (p = .01), reduced lipid (p = .006) and necrotic core area (p = .007), increased collagen content and cap-to-necrotic core ratio (p = .05), and attenuated plaque neutrophil extracellular traps (p < .001). At low dose, colchicine's effects were not accompanied by the evidence of microtubule depolymerization. Together, these results show that colchicine exerts anti-atherosclerotic and plaque-stabilizing effects at low dose by inhibiting foam cell formation and cholesterol crystal-induced inflammation. This provides a new framework to support its repurposing for atherosclerotic cardiovascular disease.

Eukaryotic elongation factor 2 kinase regulates foam cell formation via translation of CD36.

Eukaryotic elongation factor 2 kinase (eEF2K) is an atypical protein kinase that controls protein synthesis in cells under stress. Although well studied in cancer, less is known about its roles in chronic inflammatory diseases. Here, we examined its regulation of macrophage cholesterol handling in the context of atherosclerosis. eEF2K mRNA expression and protein activity were upregulated in murine bone marrow-derived macrophages (BMDMs) exposed to oxidized low-density lipoprotein cholesterol (oxLDL). When incubated with oxLDL, BMDMs from eEF2K knockout (Eef2k-/- ) mice formed fewer Oil Red O+ foam cells than Eef2k+/+ BMDMs (12.5% ± 2.3% vs. 32.3% ± 2.0%, p < .01). Treatment with a selective eEF2K inhibitor, JAN-384, also decreased foam cell formation for C57BL/6J BMDMs and human monocyte-derived macrophages. Disabling eEF2K selectively decreased protein expression of the CD36 cholesterol uptake receptor, mediated by a reduction in the proportion of translationally active Cd36 mRNA. Eef2k-/- mice bred onto the Ldlr-/- background developed aortic sinus atherosclerotic plaques that were 30% smaller than Eef2k+/+ -Ldlr-/- mice after 16 weeks of high cholesterol diet (p < .05). Although accompanied by a reduction in plaque CD36+ staining (p < .05) and lower CD36 expression in circulating monocytes (p < .01), this was not associated with reduced lipid content in plaques as measured by oil red O staining. Finally, EEF2K and CD36 mRNA levels were higher in blood mononuclear cells from patients with coronary artery disease and recent myocardial infarction compared to healthy controls without coronary artery disease. These results reveal a new role for eEF2K in translationally regulating CD36 expression and foam cell formation in macrophages. Further studies are required to explore therapeutic targeting of eEF2K in atherosclerosis.

Origins and functional evolution of Y chromosomes across mammals.

Y chromosomes underlie sex determination in mammals, but their repeat-rich nature has hampered sequencing and associated evolutionary studies. Here we trace Y evolution across 15 representative mammals on the basis of high-throughput genome and transcriptome sequencing. We uncover three independent sex chromosome originations in mammals and birds (the outgroup). The original placental and marsupial (therian) Y, containing the sex-determining gene SRY, emerged in the therian ancestor approximately 180 million years ago, in parallel with the first of five monotreme Y chromosomes, carrying the probable sex-determining gene AMH. The avian W chromosome arose approximately 140 million years ago in the bird ancestor. The small Y/W gene repertoires, enriched in regulatory functions, were rapidly defined following stratification (recombination arrest) and erosion events and have remained considerably stable. Despite expression decreases in therians, Y/W genes show notable conservation of proto-sex chromosome expression patterns, although various Y genes evolved testis-specificities through differential regulatory decay. Thus, although some genes evolved novel functions through spatial/temporal expression shifts, most Y genes probably endured, at least initially, because of dosage constraints.

Non-invasive genetic sexing technique for analysis of short-beaked echidna (Tachyglossus aculeatus) populations.

Identifying male and female echidnas is challenging due to the lack of external genitalia or any other differing morphological features. This limits studies of wild populations and is a major problem for echidna captive management and breeding. Non-invasive genetic approaches to determine sex minimise the need for handling animals and are used extensively in other mammals. However, currently available approaches cannot be applied to monotremes because their sex chromosomes share no homology with sex chromosomes in other mammals. In this study we used recently identified X and Y chromosome-specific sequences to establish a non-invasive polymerase chain reaction-based technique to determine the sex of echidnas. Genomic DNA was extracted from echidna hair follicles followed by amplification of two Y chromosome (male-specific) genes (mediator complex subunit 26 Y-gametolog (CRSPY) and anti-Müllerian hormone Y-gametolog (AMHY)) and the X chromosome gene (anti-Müllerian hormone X-gametolog (AMHX)). Using this technique, we identified the sex of 10 juvenile echidnas born at Perth Zoo, revealing that eight of the 10 echidnas were female. Future use of the genetic sexing technique in echidnas will inform captive management, continue breeding success and can be used to investigate sex ratios and population dynamics in wild populations.

Fine-mapping scan of bipolar disorder susceptibility loci in Latino pedigrees.

We previously identified bipolar disorder (BD) susceptibility loci on 8q24, 14q32, and 2q12-14 in a genome-wide nonparametric linkage screen in a Latino cohort. We now perform a fine mapping analysis using a dense map of additional SNPs to identify BD susceptibility genes within these regions. One thousand nine hundred and thirty-eight individuals with Latino ancestry (880 individuals with BD Type I or Schizoaffective, Bipolar Type) from 416 Latino pedigrees from the United States, Mexico, Costa Rica, and Guatemala were genotyped with 3,074 SNPs to provide dense coverage of the 8q24 (11.5 cM), 14q32 (7.5 cM), and 2q12-14 (6.5 cM) chromosomal loci. Single-marker association tests in the presence of linkage were performed using the LAMP software. The top linkage peak (rs7834818; LOD = 5.08, p = 3.30E - 5) and associated single marker (rs2280915, p = 2.70E - 12) were located within FBXO32 on 8q24. On chromosome 2, the top linkage peak (rs6750326; LOD = 5.06, p = 3.50E - 5) and associated single marker (rs11887088, p = 2.90E - 6) were located in intragenic regions near ACTR3 and DPP10. None of the additional markers in the region around chromosome 14q32 met significance levels for linkage or association. We identified six SNPs on 2q12-q14 and one SNP in FBXO32 on 8q24 that were significantly associated with BD in this Latino cohort.

A genome-wide quantitative trait locus (QTL) linkage scan of NEO personality factors in Latino families segregating bipolar disorder.

Personality traits have been suggested as potential endophenotypes for Bipolar Disorder (BP), as they can be quantitatively measured and show correlations with BP. The present study utilized data from 2,745 individuals from 686 extended pedigrees originally ascertained for having multiplex cases of BP (963 cases of BPI or schizoaffective BP). Subjects were assessed with the NEO Personality Inventory, Revised (NEO PI-R) and genotyped using the Illumina HumanLinkage-24 Bead Chip, with an average genetic coverage of 0.67 cM. Two point linkage scores were calculated for each trait as a quantitative variable using SOLAR (Sequential Oligogenic Linkage Analysis Routines). Suggestive evidence for linkage was found for neuroticism at 1q32.1 (LOD = 2.52), 6q23.3 (2.32), 16p12 (2.79), extraversion at 4p15.3 (2.33), agreeableness at 4q31.1 (2.37), 5q34 (2.80), 7q31.1 (2.56), 16q22 (2.52), and conscientiousness at 4q31.1 (2.50). Each of the above traits have been shown to be correlated with the broad BP phenotype in this same sample. In addition, for the trait of openness, we found significant evidence of linkage to chromosome 3p24.3 (rs336610, LOD = 4.75) and suggestive evidence at 1q43 (2.74), 5q35.1 (3.03), 11q14.3 (2.61), 11q21 (2.30), and 19q13.1 (2.52). These findings support previous linkage findings of the openness trait to chromosome 19q13 and the agreeableness trait to 4q31 and identify a number of new loci for personality endophenotypes related to bipolar disorder.

Replication of genome-wide association study (GWAS) susceptibility loci in a Latino bipolar disorder cohort.

OBJECTIVES: Recent genome-wide association studies (GWASs) have identified numerous putative genetic polymorphisms associated with bipolar disorder (BD) and/or schizophrenia (SC). We hypothesized that a portion of these polymorphisms would also be associated with BD in the Latino American population. To identify such regions, we tested previously identified genetic variants associated with BD and/or SC and ancestral haploblocks containing these single nucleotide polymorphisms (SNPs) in a sample of Latino subjects with BD. METHODS: A total of 2254 Latino individuals were genotyped for 91 SNPs identified in previous BD and/or SC GWASs, along with selected SNPs in strong linkage disequilibrium with these markers. Family-based single marker and haplotype association testing was performed using the PBAT software package. Empirical P-values were derived from 10 000 permutations. RESULTS: Associations of eight a priori GWAS SNPs with BD were replicated with nominal (P≤.05) levels of significance. These included SNPs within nuclear factor I A (NFIA), serologically defined colon cancer antigen 8 (SDCCAG8), lysosomal associated membrane protein 3 (LAMP3), nuclear factor kappa B subunit 1 (NFKB1), major histocompatibility complex, class I, B (HLA-B) and 5'-nucleotidase, cytosolic II (NT5C2) and SNPs within intragenic regions microRNA 6828 (MIR6828)-solute carrier family 7 member 14 (SLC7A14) and sonic hedgehog (SHH)-long intergenic non-protein coding RNA 1006 (LINC01006). Of the 76 ancestral haploblocks that were tested for associations with BD, our top associated haploblock was located in LAMP3; however, the association did not meet statistical thresholds of significance following Bonferroni correction. CONCLUSIONS: These results indicate that some of the gene variants found to be associated with BD or SC in other populations are also associated with BD risk in Latinos. Variants in six genes and two intragenic regions were associated with BD in our Latino sample and provide additional evidence for overlap in genetic risk between SC and BD.

A genome-wide linkage scan of bipolar disorder in Latino families identifies susceptibility loci at 8q24 and 14q32.

A genome-wide nonparametric linkage screen was performed to localize Bipolar Disorder (BP) susceptibility loci in a sample of 3757 individuals of Latino ancestry. The sample included 963 individuals with BP phenotype (704 relative pairs) from 686 families recruited from the US, Mexico, Costa Rica, and Guatemala. Non-parametric analyses were performed over a 5 cM grid with an average genetic coverage of 0.67 cM. Multipoint analyses were conducted across the genome using non-parametric Kong & Cox LOD scores along with Sall statistics for all relative pairs. Suggestive and significant genome-wide thresholds were calculated based on 1000 simulations. Single-marker association tests in the presence of linkage were performed assuming a multiplicative model with a population prevalence of 2%. We identified two genome-wide significant susceptibly loci for BP at 8q24 and 14q32, and a third suggestive locus at 2q13-q14. Within these three linkage regions, the top associated single marker (rs1847694, P = 2.40 × 10(-5)) is located 195 Kb upstream of DPP10 in Chromosome 2. DPP10 is prominently expressed in brain neuronal populations, where it has been shown to bind and regulate Kv4-mediated A-type potassium channels. Taken together, these results provide additional evidence that 8q24, 14q32, and 2q13-q14 are susceptibly loci for BP and these regions may be involved in the pathogenesis of BP in the Latino population.

Discovery of an embryonically derived bipotent population of endothelial-macrophage progenitor cells in postnatal aorta.

Converging evidence indicates that extra-embryonic yolk sac is the source of both macrophages and endothelial cells in adult mouse tissues. Prevailing views are that these embryonically derived cells are maintained after birth by proliferative self-renewal in their differentiated states. Here we identify clonogenic endothelial-macrophage (EndoMac) progenitor cells in the adventitia of embryonic and postnatal mouse aorta, that are independent of Flt3-mediated bone marrow hematopoiesis and derive from an early embryonic CX3CR1+ and CSF1R+ source. These bipotent progenitors are proliferative and vasculogenic, contributing to adventitial neovascularization and formation of perfused blood vessels after transfer into ischemic tissue. We establish a regulatory role for angiotensin II, which enhances their clonogenic and differentiation properties and rapidly stimulates their proliferative expansion in vivo. Our findings demonstrate that embryonically derived EndoMac progenitors participate in local vasculogenic responses in the aortic wall by contributing to the expansion of endothelial cells and macrophages postnatally.

Suggestive evidence for association between L-type voltage-gated calcium channel (CACNA1C) gene haplotypes and bipolar disorder in Latinos: a family-based association study.

OBJECTIVES: Through recent genome-wide association studies (GWASs), several groups have reported significant association between variants in the calcium channel, voltage-dependent, L-type, alpha 1C subunit (CACNA1C) and bipolar disorder (BP) in European and European-American cohorts. We performed a family-based association study to determine whether CACNA1C is associated with BP in the Latino population. METHODS: This study included 913 individuals from 215 Latino pedigrees recruited from the USA, Mexico, Guatemala, and Costa Rica. The Illumina GoldenGate Genotyping Assay was used to genotype 58 single-nucleotide polymorphisms (SNPs) that spanned a 602.9-kb region encompassing the CACNA1C gene including two SNPs (rs7297582 and rs1006737) previously shown to associate with BP. Individual SNP and haplotype association analyses were performed using Family-Based Association Test (version 2.0.3) and Haploview (version 4.2) software. RESULTS: An eight-locus haplotype block that included these two markers showed significant association with BP (global marker permuted p = 0.0018) in the Latino population. For individual SNPs, this sample had insufficient power (10%) to detect associations with SNPs with minor effect (odds ratio = 1.15). CONCLUSIONS: Although we were not able to replicate findings of association between individual CACNA1C SNPs rs7297582 and rs1006737 and BP, we were able to replicate the GWAS signal reported for CACNA1C through a haplotype analysis that encompassed these previously reported significant SNPs. These results provide additional evidence that CACNA1C is associated with BP and provides the first evidence that variations in this gene might play a role in the pathogenesis of this disorder in the Latino population.

An Academic Practice Partnership: Helping New Registered Nurses to Advance Quality and Patient Safety.

Significant changes in the healthcare environment have occurred that offer challenges for quality improvement and nursing education programs, and thus impact both nursing practice and education. We formed an academic-practice partnership to actively engage students enrolled in an undergraduate nursing research course in quality processes with participation in a medical center's performance improvement program. This article describes the development of the partnership; and projects, results, and implications for practice. Students worked collaboratively in groups with hospital staff performance improvement preceptors and a course faculty member. Using the Plan, Do, Check, Act (PDCA) model, students collected, analyzed, and disseminated data from existing projects, or those for which the organization had identified a need. Leaders involved in the inception of this partnership agreed that it achieved its goals of enabling the college to effectively teach recently mandated quality improvement methodologies to achieve competency and enhancing the medical center's capabilities to obtain data for quality improvement purposes. The academic-practice partnership continues to evolve, and we offer discussion about lessons learned and partnership growth.

Effect of corn stigma extract on physical and antioxidant properties of biodegradable and edible gelatin and corn starch films.

The development of bio-based food packaging with antioxidant properties is an important research topic and has gained prominence these days. In this study, bioactive films were developed based gelatin-corn starch (GCS) incorporated with corn stigma extract (CSE) at different concentrations (15% and 25%; w/v). In preliminary tests, the extract maintained cell viability above 90% indicating that it is safe for application as an active ingredient. Insertion of the extract did not influence the thickness of the films but caused a slight change in optical properties. Scanning electron microscopy (SEM) analysis revealed interactions between the extract's bioactive compounds with gelatin and corn starch compounds, which may have improved the mechanical properties (elongation at break, Young's modulus). The addition of 25% corn stigma extract increased the contact angle, giving the film a hydrophobic character. Furthermore, at this concentration, a 15% reduction in water vapor permeability was observed. The elaborated films showed complete biodegradability before the tenth day of the study. It can be inferred that the films with corn stigma extract have good antioxidant properties, indicating that they can be used as an ingredient for food packaging.

The use of profanity during letter fluency tasks in frontotemporal dementia and Alzheimer disease.

OBJECTIVE: To assess whether the production of profanity during letter fluency testing distinguishes frontotemporal dementia (FTD) and Alzheimer disease (AD) patients. BACKGROUND: Alterations in language and social behavior typify FTD spectrum disorders. Nonetheless, in can be difficult to distinguish pathologically defined frontotemporal lobar degeneration (FTLD) from AD clinically. Assessing verbal fluency by having patients generate words beginning with specific letters in a given period of time can yield diverse information of diagnostic use. METHOD: Words produced during FAS letter fluency testing were reviewed, and instances of the use of "f*ck," "*ss," and "sh*t" and other words felt to be inappropriate were sought. The frequency of these words was compared between clinically diagnosed FTD and AD patients using χ(2) tests. RESULTS: We found that 6/32 (18.8%) patients with FTD generated the word "f*ck" during the "F" trial as opposed to none of 38 patients with AD (P=0.007). Patients who said "f*ck" had diagnoses of either behavioral variant FTD (3/15), progressive nonfluent aphasia (2/8), or semantic dementia (1/3). CONCLUSIONS: Though the specific neuropathology in these cases is uncertain, generation of "f*ck" during letter fluency testing seems to have use in differentiating FTD from AD.

Phytochemical characterization and antimicrobial activity of Cymbopogon citratus extract for application as natural antioxidant in fresh sausage.

The development of natural additives is considered an important research topic. In this work, the use of Cymbopogon citratus (CC) extract as a natural additive for chicken sausage refrigerated was investigated. The CC extract was characterized by electrospray ionization with high-resolution time-of-flight mass spectrometry (ESI-ToF-MS) and the identified compounds were directly related to the antioxidant activity demonstrated by CC in the fresh sausage. In total, 31 phytochemical compounds were identified, and 27 of these still were not described in the literature for CC. The antimicrobial activity showed that CC extract is a potential antibacterial agent. Besides, the results showed that CC extract reduced lipid oxidation compared to synthetic additive. The sensorial characteristics were maintained, demonstrating good acceptability by the consumer. The results confirmed that CC can keep the quality of chicken sausage refrigerated for up to 42 days of storage.

Vasculogenic properties of adventitial Sca-1+CD45+ progenitor cells in mice: a potential source of vasa vasorum in atherosclerosis.

The cellular origins of vasa vasorum are ill-defined and may involve circulating or local progenitor cells. We previously discovered that murine aortic adventitia contains Sca-1+CD45+ progenitors that produce macrophages. Here we investigated whether they are also vasculogenic. In aortas of C57BL/6 mice, Sca-1+CD45+ cells were localised to adventitia and lacked surface expression of endothelial markers (<1% for CD31, CD144, TIE-2). In contrast, they did show expression of CD31, CD144, TIE-2 and VEGFR2 in atherosclerotic ApoE-/- aortas. Although Sca-1+CD45+ cells from C57BL/6 aorta did not express CD31, they formed CD31+ colonies in endothelial differentiation media and produced interconnecting vascular-like cords in Matrigel that contained both endothelial cells and a small population of macrophages, which were located at branch points. Transfer of aortic Sca-1+CD45+ cells generated endothelial cells and neovessels de novo in a hindlimb model of ischaemia and resulted in a 50% increase in perfusion compared to cell-free control. Similarly, their injection into the carotid adventitia of ApoE-/- mice produced donor-derived adventitial and peri-adventitial microvessels after atherogenic diet, suggestive of newly formed vasa vasorum. These findings show that beyond its content of macrophage progenitors, adventitial Sca-1+CD45+ cells are also vasculogenic and may be a source of vasa vasorum during atherogenesis.

Cellular Therapy for Heart Failure.

The pathogenesis of cardiomyopathy and heart failure (HF) is underpinned by complex changes at subcellular, cellular and extracellular levels in the ventricular myocardium. For all of the gains that conventional treatments for HF have brought to mortality and morbidity, they do not adequately address the loss of cardiomyocyte numbers in the remodeling ventricle. Originally conceived to address this problem, cellular transplantation for HF has already gone through several stages of evolution over the past two decades. Various cell types and delivery routes have been implemented to positive effect in preclinical models of ischemic and nonischemic cardiomyopathy, with pleiotropic benefits observed in terms of myocardial remodeling, systolic and diastolic performance, perfusion, fibrosis, inflammation, metabolism and electrophysiology. To a large extent, these salubrious effects are now attributed to the indirect, paracrine capacity of transplanted stem cells to facilitate endogenous cardiac repair processes. Promising results have also followed in early phase human studies, although these have been relatively modest and somewhat inconsistent. This review details the preclinical and clinical evidence currently available regarding the use of pluripotent stem cells and adult-derived progenitor cells for cardiomyopathy and HF. It outlines the important lessons that have been learned to this point in time, and balances the promise of this exciting field against the key challenges and questions that still need to be addressed at all levels of research, to ensure that cell therapy realizes its full potential by adding to the armamentarium of HF management.

Identification of circadian gene variants in bipolar disorder in Latino populations.

BACKGROUND: Variations in circadian genes can impact biological rhythms. Given the rhythm disturbances that characterize bipolar disorder (BD), genes encoding components of molecular clocks are good candidate genes for the illness. METHODS: A family based association analysis of circadian gene single nucleotide polymorphisms (SNPs) and BD was conducted in Latino pedigrees. 884 individuals from 207 pedigrees (473BP phenotype and 411 unaffected family members) were genotyped. Family based single marker association testing was performed. Ancestral haplotypes (SNPs found to be in strong LD defined using confidence intervals) were also tested for association with BD. RESULTS: Multiple suggestive associations between circadian gene SNPs and BD were noted. These included CSNK1E (rs1534891, p=0.00689), ARNTL (rs3789327, p=0.021172), CSNK1D (rs4510078, p=0.022801), CLOCK (rs17777927, p=0.031664). Individually, none of the SNPs were significantly associated with BD after correction for multiple testing. However, a 4-locus CSNK1E haplotype encompassing the rs1534891 SNP (Z-score=2.685, permuted p=0.0076) and a 3-locus haplotype in ARNTL (Z-score=3.269, permuted p=0.0011) showed a significant association with BD. LIMITATIONS: Larger samples are required to confirm these findings and assess the relationship between circadian gene SNPs and BD in Latinos. CONCLUSIONS: The results suggest that ARNTL and CSKN1E variants may be associated with BD. Further studies are warranted to assess the relationships between these genes and BD in Latino populations.

Clinical outcomes and genome-wide association for a brain methylation site in an antidepressant pharmacogenetics study in Mexican Americans.

OBJECTIVE: The authors compared the effectiveness of fluoxetine and desipramine treatment in a prospective double-blind pharmacogenetics study in first-generation Mexican Americans and examined the role of whole-exome functional gene variations in the patients' antidepressant response. METHOD: A total of 232 Mexican Americans who met DSM-IV criteria for major depressive disorder were randomly assigned to receive 8 weeks of double-blind treatment with desipramine (50-200 mg/day) or fluoxetine (10-40 mg/day) after a 1-week placebo lead-in period. Outcome measures included the Hamilton Depression Rating Scale (HAM-D), the Hamilton Anxiety Rating Scale, and the Beck Depression Inventory. At week 8, whole-exome genotyping data were obtained for 36 participants who remitted and 29 who did not respond to treatment. RESULTS: Compared with desipramine treatment, fluoxetine treatment was associated with a greater reduction in HAM-D score, higher response and remission rates, shorter time to response and remission, and lower incidences of anticholinergic and cardiovascular side effects. Pharmacogenetics analysis showed that exm-rs1321744 achieved exome-wide significance for treatment remission. This variant is located in a brain methylated DNA immunoprecipitation sequencing site, which suggests that it may be involved in epigenetic regulation of neuronal gene expression. This and two other common gene variants provided a highly accurate cross-validated predictive model for treatment remission of major depression (receiver operating characteristic integral=0.95). CONCLUSIONS: Compared with desipramine, fluoxetine treatment showed a more rapid reduction of HAM-D score and a lower incidence of side effects in a population comprising primarily first-generation Mexican Americans with major depression. This study's pharmacogenetics approach strongly implicates the role of functional variants in antidepressant treatment response.

Extraction of bioactive compounds of lemongrass, antioxidant activity and evaluation of antimicrobial activity in fresh chicken sausage / Extração de compostos bioativos de capim-limão, atividade antioxidante e avaliação da atividade antimicrobiana em linguiça frescal de frango

ABSTRACT: The aim of this work was to determine the best extraction condition of bioactive compounds from lemongrass (Cymbopogon citratus), using the conventional method and ultrasonic assisted extraction, varying the temperature, in order to evaluate the antioxidant activity and the antimicrobial activity of the extract with higher antioxidant power in fresh chicken sausages during the storage period. The extracts were obtained by the conventional method (solvent extraction) and by ultrasound assisted extraction, varying the temperature (20°C, 40°C and 60°C). Phenolic compounds, total flavonoids and antioxidant activity were measured by the DPPH, FRAP, ORAC methods. Conventional extraction and ultrasound methods influenced the phenolic and total flavonoid content at all tested temperatures. Conventional and ultrasonic methods did not influence the IC50 at temperatures of 40°C and 60°C. The antioxidant activity by the DPPH method and by the FRAP method was superior in the conventional method at the temperature of 60°C, however by the ORAC method the best results were in the extraction by ultrasound. The results demonstrate that the conventional extraction at 60ºC was better to obtain extracts of lemongrass with greater amount of bioactive compounds. The antimicrobial capacity evaluated in sausage of fresh chicken showed that in the concentration of 1.0% of the extract protected the product as the growth of mesophilic aerobes and against the growth of psychrotrophic bacteria. Lemongrass can be considered as a natural alternative to obtain extracts rich in bioactive compounds, with antioxidant activity and high antimicrobial capacity.

RESUMO: O objetivo deste trabalho foi determinar a melhor condição de extração de compostos bioativos do capim-limão (Cymbopogon citratus), usando o método convencional e extração assistida por ultrassom, em diferentes temperaturas, a fim de avaliar a atividade antioxidante e a atividade antimicrobiana do extrato com maior poder antioxidante em linguiças de frango frescal durante o período de armazenamento. Os extratos foram obtidos pelo método convencional (extração com solvente) e por extração assistida por ultrassom, variando a temperatura (20°C, 40°C e 60°C). Foram medidos os compostos fenólicos, flavonoides totais e atividade antioxidante pelos métodos DPPH, FRAP, ORAC. Os métodos de extração convencional e ultrassom influenciaram no teor de fenólicos e flavonoides totais em todas as temperaturas testadas. Os métodos convencional e ultrassom não influenciaram no IC50 nas temperaturas de 40oC e 60oC. A atividade antioxidante pelo método DPPH e pelo método FRAP foi superior no método convencional na temperatura de 60°C, entretanto pelo método ORAC os melhores resultados foram na extração por ultrassom. Os resultados demonstram que a extração convencional a 60ºC foi melhor para obter extratos de capim-limão com maior quantidade de compostos bioativos. A capacidade antimicrobiana avaliada em linguiça frescal mostrou que na concentração de 1,0% do extrato protegeu o produto quanto o crescimento de aeróbios mesófilos e contra o crescimento de bactérias psicrotróficas. O capim-limão pode ser considerado uma alternativa natural para obtenção de extratos ricos em compostos bioativos, com atividade antioxidante e elevada capacidade antimicrobiana.

Ultrasonic assisted extraction to obtain bioactive, antioxidant and antimicrobial compounds from marcela / Extração assistida por ultrassom para obtenção de compostos bioativos, antioxidantes e antimicrobianos de marcela

ABSTRACT: The aim of this study was to evaluate the effect of extraction conditions on bioactive compounds, as well as on antioxidant activity, and the antimicrobial activity of the extracts with the highest antioxidant characteristics. The extracts were obtained by conventional method and ultrasound-assisted extraction at various temperatures (20°C, 40°C and 60°C). Total phenolics, total flavonoids, antimicrobial activity and antioxidant activity were quantified by the methods of DPPH, FRAP, and ORAC, respectively. The conventional extraction method and ultrasound method influenced the phenolic content at all the tested temperatures. Flavonoids were not influenced by extraction methods. The antioxidant activity (DPPH) was highest in the ultrasonic method at temperatures of 40°C and 60°C; however, in the case of the FRAP method the best results were for the conventional extraction method. The conventional and ultrasonic methods did not influence the IC50 at temperatures of 20°C and 40°C, but using ORAC the antioxidant activity was influenced by the methods at all temperatures. The extract obtained at 60°C by the ultrasound method had high antimicrobial action in relation to the strains of Salmonella sp., Escherichia coli, and Staphylococcus aureus. Extraction ultrasonic-assisted can be considered adequate to obtain extracts of marcela, which are rich in bioactive compounds with high antioxidant activity.

RESUMO: O objetivo deste trabalho foi avaliar o efeito das condições de extração sobre os compostos bioativos, bem como sobre a atividade antioxidante e atividade antimicrobiana do extrato com maior característica antioxidante. Os extratos foram obtidos pelo método convencional e extração assistida por ultrassom, variando a temperatura (20oC, 40oC e 60oC). Foram quantificados os fenólicos totais, flavonóides totais, a atividade antimicrobiana, e atividade antioxidante pelos métodos DPPH, FRAP, e ORAC. Os métodos de extração convencional e ultrassom influenciaram no teor de fenólicos em todas as temperaturas testadas. Os flavonóides não sofreram influência dos métodos de extração. A atividade antioxidante (DPPH) foi superior no método ultrassom nas temperaturas de 40°C e 60°C, entretanto pelo método FRAP os melhores resultados foram na extração convencional. Os métodos convencional e ultrassom não influenciaram no IC50 nas temperaturas de 20oC e 40oC, mas a atividade antioxidante pelo método ORAC sofreu influencia dos métodos em todas as temperaturas. O extrato obtido a 60°C pelo método ultrassom possui elevada ação antimicrobiana frente a cepas de Salmonella sp., Escherichia coli, e Staphylococcus aureus. A extração assistida por ultrassom pode ser considerada adequada para obtenção de extratos de marcela, que são ricos em compostos bioativos com alta atividade antioxidante.