Impacto da dieta cetogênica rica em MUFAS e PUFAS e suplementada com DHA sobre parâmetros oxidativos, inflamatórios e comportamentais / Impact of a ketogenic diet rich in MUFAS and PUFAS and supplemented with DHA on oxidative, inflammatory and behavioral parameters

Introdução - A dieta cetogênica (DC) tem sido utilizada como tratamento adjuvante da epilepsia fármaco resistente e tem se popularizado no manejo de outras doenças neurológicas e metabólicas (obesidade, diabetes e dislipidemias). Entretanto, também está associada a efeitos adversos que precisam ser melhor compreendidos. Objetivo - Avaliar o potencial efeito protetor de uma dieta cetogênica modificada (DCM), rica em ácidos graxos monoinsaturados (MUFAs) e ácidos graxos poli-insaturados (PUFAs), e suplementada com ácido docosahexaenóico (DHA), sobre a neuroinflamação e o estresse oxidativo e suas repercussões no comportamento de ratos. Materiais e Métodos - Estudo experimental com 18 ratos Wistar, alocados em 3 grupos de 6 animais. No grupo da dieta padrão (DP), os animais consumiram dieta comercial. No grupo da dieta cetogênica clássica (DCC), os animais consumiram uma dieta rica em gorduras saturadas. No grupo DCM, os animais consumiram dieta rica em MUFAs e PUFAs e enriquecida com DHA. Após 100 dias de intervenção foram realizados testes comportamentais e, após a eutanásia, foram feitos extratos lipídico e aquoso do lobo frontal e do hipocampo para análise de ácidos graxos e parâmetros oxidativos e inflamatórios. Resultados - Os animais DCC apresentaram um comportamento mais ansioso que ao animas DP, segundo os testes de Labirinto em Cruz Elevado e Medo ao Contexto. O grupo DCC teve mais incorporação de ácidos graxos saturados (AGs) no lobo frontal e no hipocampo em relação ao DCM. O grupo DCM teve maior incorporação de PUFAs, com destaque para o aumento de DHA no hipocampo. As citocinas inflamatórias IL1-ß e IL-6 tiveram menor concentração nos grupos DCM e DCC em relação ao grupo DP no lobo frontal, perfil semelhante ao observado com a IL-6 no hipocampo, e diferente da IL1-ß que foi reduzida significativamente apenas no DCM. Não houve diferença da concentração da Neuroprotectina D (NPD) e da enzima antioxidante Superóxido Dismutase (SOD) entre os grupos. A LDLox teve menor concentração nos grupos DCC e DCM em relação ao DP. A enzima catalase (CAT) foi menor no grupo DCM em relação ao grupo DP. Conclusão - Embora a modulação da composição de ácidos graxos da DC possa ser promissora, uma vez que a DCC induziu a um estado mais ansioso, não foi possível explicar essa diferença pelas vias inflamatória e antioxidante avaliadas.

Introduction - The ketogenic diet (KD) has been applied as an adjuvant treatment for drug-resistant epilepsy and has become popular in the management of other neurological and metabolic diseases (obesity, diabetes and dyslipidemia). However, it is also associated with adverse effects that need to be better understood. Objective - To evaluate the potential protective effect of a modified ketogenic diet (MKD), rich in monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs), and supplemented with docosahexaenoic acid (DHA), on neuroinflammation and oxidative stress and their repercussions on the behavior of rats. Materials and Methods - Experimental study with 18 Wistar rats, allocated into 3 groups of 6 animals. In the standard diet (SD) group, the animals consumed a commercial diet. In the classical ketogenic diet (CKD) group, the animals consumed a diet rich in saturated fats. In the MKD group, the animals consumed a diet rich in MUFAs and PUFAs and enriched with DHA. After 100 days of intervention, behavioral tests were carried out and, after euthanasia, lipid and aqueous extracts were made from the frontal lobe and hippocampus for analysis of fatty acids and oxidative and inflammatory parameters. Results - The CKD animals showed more anxious behavior than the DP animals, according to the Elevated Cross Maze and Context Fear tests. The CKD group had more incorporation of saturated fatty acids (SFAs) in the frontal lobe and hippocampus compared to MKD. The MKD group had greater incorporation of PUFAs, with emphasis on the increase in DHA in the hippocampus. The inflammatory cytokines IL1-ß and IL-6 had lower concentrations in the MKD and CKD groups compared to the PD group in the frontal lobe, a profile similar to that observed with IL-6 in the hippocampus, and different from IL1-ß, which was significantly reduced in the MKD. There was no difference in the concentration of Neuroprotectin D (NPD) and the antioxidant enzyme Superoxide Dismutase (SOD) between the groups. LDLox had a lower concentration in the CKD and MKD groups compared to the DP. The catalase enzyme was lower in the MKD group compared to the DP group. Conclusion - Although the modulation of the fatty acid composition of KD may be promising, since CKD induced a more anxious state, it was not possible to explain this difference through the inflammatory and antioxidant pathways evaluated.

Docosahexaenoic fatty acid nanoencapsulated with Anti-PECAM-1 as strategy to increase atherosclerotic plaque stability / Ácido graxo docosahexaenoico nanoencapsulado com anti-PECAM-1 como uma estratégia para aumentar a estabilidade de placas ateroscleróticas

Cardiovascular diseases (CVDs) are the main cause of mortality worldwide, being the ischemic heart disease responsible for 85% of deaths. Atherosclerosis is a chronic inflammation of the arteries that underlies ischemic forms of CVD and involves the innate and adaptive immune systems, from initial fatty streak formation to atherosclerotic plaque ruptures, which defines the beginning and end stages of disease, respectively. Recent research on the reduction of systemic inflammation in order to treat CVD is controversial, since results show that this reduced inflammation can also increase patient susceptibility to general infection. Therefore, new tissue-targeting strategies are necessary. Docosahexaenoic fatty acid (DHA) is a natural bioactive precursor of pro-resolving oxylipins that can reduce inflammation. Based on these factors, the objective of this study was to develop a nanocapsule containing algae oil as a DHA source and apply anti-PECAM-1 on its surface to drive it to the inflamed endothelium. Initially, a surface-functionalized metal-complex multi-wall nanocapsule containing algae oil in its nucleus (MLNC-DHA-a1) was developed. This nanocapsules presented a mean diameter of 163 ± 5 nm, was spherical in shape, showed 94.80% conjugation efficiency using 200 µg/mL of anti-PECAM-1 on the surface, and did not show significant toxicity toward HUVECs at concentrations from 0.14 to 2.90x1011 nanocapsules/mL. The nanocapsules were also stable for 2 h, sufficient time to allow for clinical applications. In cell viability assays, concentrations of 0.14 to 1.40x1011 nanocapsules/mL did not significantly affect the viability of immortalized murine macrophages (RAW 264.7) and U-937 cells after 24, 48, and 72 h of treatment. Finally, macrophages were incubated with 0.75x1011 MLNC-DHA-a1 nanocapsules/mL for 4 h and showed a significant uptake, observed using dark-field hyperspectral microscopy (CytoViva®). Once inside murine macrophages (RAW 264.7), MLNC-DHA-a1 nanocapsules promoted a strong increase in M2 phenotype polarization compared to non-treated control cells. Our results suggest that DHA-enriched algae oil, as part of a lipid core nanocapsules, does not reduce cell viability and improves macrophage phenotype, making it a promising potential therapy for controlling chronic inflammation and healing or stabilizing atherosclerotic plaques

As doenças cardiovasculares (DCVs) são a principal causa de mortalidade no mundo, sendo os eventos isquêmicos responsáveis por 85% das mortes. A aterosclerose é uma inflamação crônica das artérias associada aos eventos isquêmicos das DCVs, na qual o sistema imunológico inato e adaptativo estão envolvidos desde a formação inicial das estrias gordurosas até a ruptura das placas ateroscleróticas. Pesquisas recentes direcionadas à redução da inflamação sistêmica têm mostrado resultados controversos, pois essa abordagem pode aumentar a susceptibilidade do paciente a infecções. Nesse sentido, novas estratégias direcionadas ao tecido lesionado são necessárias. No que se refere a medicamentos anti-inflamatórios ou suplementos alimentares, o ácido docosaexaenóico (DHA) tem sido relatado como um precursor natural de oxilipinas pró- resolutivas. Baseado nesse contexto, o objetivo deste estudo foi desenvolver nanocápsulas contendo óleo de alga como fonte de DHA e vetorizar essas nanopartículas com o anticorpo antiPECAM-1 em sua superfície, visando direcioná-las ao endotélio inflamado. Inicialmente, a nanocápsula multiparede metal-complexa funcionalizada contendo óleo de alga em seu núcleo (MLNC-DHA-a1) foi desenvolvida, apresentando um diâmetro médio de 163 ± 5 nm, formato esférico, onde a eficiência de conjugação do anti-PECAM-1 (200 µg/mL) foi de 94,80% sem toxicidade significativa em HUVECs nas concentrações de 1.14 a 2.9 x 1011 nanocápsulas/mL. As nanocápsulas apresentaram uma estabilidade de 2h, o que representa tempo suficiente para a sua aplicação clínica. A seguir, ensaios de viabilidade celular foram realizados em outras linhagens de células para avaliar a toxicidade das nanocápsulas. As concentrações de 0.14 a 1.40 x 1011 de nanocápsulas/mL não afetaram significativamente a viabilidade celular de macrófagos murinos imortalizados (RAW 264.7) e U-937 após 24, 48 e 72 h de tratamento. Por fim, os macrófagos (RAW 264.7) foram incubados com 0.75 x 1011 MLNC-DHA-a1/mL durante 4 h e apresentam uma captação significativa das nanocápsulas, observada por microscopia hiperespectral de campo escuro (CytoViva®). Uma vez captadas pelos macrófagos murinos imortalizados (RAW 264.7), as nanoformulações MLNC-DHA-a1 promoveram um forte aumento da polarização do fenótipo M2 em comparação com as células controle não tratadas. Nossos resultados sugerem que o óleo de alga rico em DHA presente no núcleo lipídico das nanocápsulas, não reduziu a viabilidade celular e estimulou uma maior polarização de macrófagos para o tipo M2, sendo assim uma terapia potencial para controlar a inflamação crônica e cicatrizar ou estabilizar placas ateroscleróticas

Efeito do ácido docosa-hexaenoico e do Trolox® no diluidor de refrigeração de sêmen de garanhões da raça Mangalarga Marchador / Effect of docosahexaenoic acid and Trolox® in extenders for cooling semen from Mangalarga Marchador stallion

Objetivou-se avaliar o efeito do ácido docosa-hexaenoico (DHA), associado ou não ao Trolox®, na refrigeração de sêmen de garanhões da raça Mangalarga Marchador. Foram refrigerados 10 ejaculados nos diluidores: D1) BotuSêmen® (BS; controle); D2) BS + 30ngmL-1 de DHA (BS30DHA); D3) BS30DHA + 40µM de Trolox® (BS30DHA40T); D4) BS + 50ngmL-1 de DHA (BS50DHA); D5) BS50DHA + 40µM de Trolox® (BS50DHA40T). Após 48 horas de refrigeração, foram avaliados os parâmetros de movimento espermático, a integridade estrutural e funcional da membrana plasmática e a longevidade espermática. Todos os diluidores testados preservaram, de forma semelhante, a motilidade, a linearidade, a retilinearidade, a amplitude do deslocamento lateral da cabeça, a frequência do batimento flagelar cruzado, o percentual de hiperativos e a integridade estrutural e funcional da membrana espermática (P>0,05). O diluidor BS50DHA foi superior ao BS30DHA40T em preservar a VCL e a VSL e foi superior ao BS30DHA40T e ao BS50DHA40T em preservar a VAP e o índice de oscilação (P<0,05). Conclui-se que o uso do Trolox® em diluidores utilizados para refrigeração de sêmen de garanhões contendo ácido docosa-hexaenoico, nas concentrações propostas, não é indicado por alterar parâmetros de movimento espermático considerados importantes para a fertilidade.(AU)

The objective of this study was to evaluate the effect of docosahexaenoic acid (DHA), associated or not with Trolox®, in extenders for cooling semen from Mangalarga Marchador stallions. Ten ejaculates were cooled in the following diluents: D1) BotuSemen® (BS; control); D2) BS + 30ngmL-1 DHA (BS30DHA); D3) BS30DHA + 40µM Trolox® (BS30DHA40T); D4) BS + 50ngmL-1 DHA (BS50DHA); D5) BS50DHA + 40µM Trolox® (BS50DHA40T). After 48 hours of refrigeration, the sperm movement, structural and functional integrity of the plasma membrane and sperm longevity were evaluated. All extenders tested similarly preserved motility, linearity, straightness of trajectory, amplitude of lateral head displacement, beat cross frequency, hyperactive, the structural and functional integrity of the sperm membrane (P> 0.05). The BS50DHA extender was superior to BS30DHA40T in preserving VCL and VSL and was superior to BS30DHA40T and BS50DHA40T in preserving VAP and oscillation index (P< 0.05). It is concluded that the use of Trolox® in extenders for cooling stallion sperm containing docosahexaenoic acid at the proposed concentrations is not indicated because it alters sperm movement parameters considered important for fertility.(AU)

High levels of docosahexaenoic acid are present in eight New World silversides (Pisces: Atherinopsidae)

Docosahexaenoic acid (DHA) is the most critical and least available omega-3 fatty acid in the Western human diet. Currently, the source of omega-3 long chain polyunsaturated fatty acids (LC-PUFA) is mainly dependent on wild fisheries, making this resource unsustainable in the foreseeable future. In recent years, a high rate of biosynthesis and accumulation of DHA has been discovered in a freshwater species (Chirostoma estor) belonging to the Atherinopsidae family. Interest in evaluating fatty acid composition in other members of the family has emerged, so this study compiles original data of flesh composition of eight atherinopsid species from freshwater and brackish environments, either wild or cultured. High levels of DHA (16 to 31%) were found in all analyzed members of the family, except in C. grandocule, independently of their habitat or origin. The analyzed species of the Jordani group (C. estor, C. promelas and C. humboldtianum) showed high DHA and low EPA levels (<0.5%) as previously reported for cultured C. estor. The low trophic niche of these atherinopsids and their fatty acid accumulation capabilities are factors that make these species noteworthy candidates for sustainable aquaculture.(AU)

O ácido docosahexaenóico (DHA) é o ácido graxo ômega-3 mais importante e menos disponível na dieta humana ocidental. Atualmente, a fonte de ácidos graxos poliinsaturados de cadeia longa ômega-3 (LC-PUFA) depende principalmente da pesca extrativista, tornando esse recurso insustentável em um futuro próximo. Nos últimos anos, uma alta taxa de biossíntese e acúmulo de DHA foi descoberta em uma espécie de água doce (Chirostoma estor) pertencente à família Atherinopsidae. Deste modo, surgiu o interesse em avaliar a composição de ácidos graxos em outros membros da família. Portanto, este estudo compila dados originais da composição de carne de oito espécies de aterinopsídeos de ambientes de água doce e salobra, selvagens ou cultivadas. Altos níveis de DHA (16 a 31%) foram encontrados em todos os membros da família analisados, exceto em C. grandocule, independentemente de seu habitat ou origem. As espécies analisadas do grupo Jordani (C. estor, C. promelas e C. humboldtianum) apresentaram altos níveis de DHA e EPA baixos (<0,5%), como relatado anteriormente para C. estor cultivado. O baixo nicho trófico desses aterinopsídeos e sua capacidade de acumulação de ácidos graxos são fatores que tornam essas espécies notáveis candidatas à aquicultura sustentável.(AU)

Deficiency or activation of peroxisome proliferator-activated receptor α reduces the tissue concentrations of endogenously synthesized docosahexaenoic acid in C57BL/6J mice

BACKGROUND/OBJECTIVES: Docosahexaenoic acid (DHA), an n-3 long chain polyunsaturated fatty acid (LCPUFA), is acquired by dietary intake or the in vivo conversion of α-linolenic acid. Many enzymes participating in LCPUFA synthesis are regulated by peroxisome proliferator-activated receptor alpha (PPARα). Therefore, it was hypothesized that the tissue accretion of endogenously synthesized DHA could be modified by PPARα. MATERIALS/METHODS: The tissue DHA concentrations and mRNA levels of genes participating in DHA biosynthesis were compared among PPARα homozygous (KO), heterozygous (HZ), and wild type (WT) mice (Exp I), and between WT mice treated with clofibrate (PPARα agonist) or those not treated (Exp II). In ExpII, the expression levels of the proteins associated with DHA function in the brain cortex and retina were also measured. An n3-PUFA depleted/replenished regimen was applied to mitigate the confounding effects of maternal DHA. RESULTS: PPARα ablation reduced the hepatic Acox, Fads1, and Fads2 mRNA levels, as well as the DHA concentration in the liver, but not in the brain cortex. In contrast, PPARα activation increased hepatic Acox, Fads1, Fads2 and Elovl5 mRNA levels, but reduced the DHA concentrations in the liver, retina, and phospholipid of brain cortex, and decreased mRNA and protein levels of the brain-derived neurotrophic factor in brain cortex. CONCLUSIONS: LCPUFA enzyme expression was altered by PPARα. Either PPARα deficiency or activation-decreased tissue DHA concentration is a stimulus for further studies to determine the functional significance.

Screening for hyper-accumulating lipid mutants in Aurantiochytrium limacinum using high-throughput fluorescence-based method / 生物工程学报

Docosahexaenoic acid (DHA) has many unique physiological functions such as promoting the development of brain and retina in infants. Therefore, it is widely applied to food, pharmacy, breeding and other industries. To obtain engineered strains of Aurantiochytrium limacinum SR21 suitable for industrial application with increased lipid and DHA production, we designed a simple, fast, accurate and high-throughput screening method based on Nile red staining of oil droplets. First, ultraviolet C (UVC) mutagenesis was used to generate a random mutant library of A. limacinum. Second, screening conditions were optimized including staining conditions of Nile red and the sorting criterion. Thereby, three putative high-lipid mutants (D03432, D05106 and D01521) were selected from the mutant library containing 3 648 mutated clones. The three mutants grew faster and produced higher amounts of lipids and DHA compared to wild type (WT). In 100 mL cultures, the lipid content of D03432 and D05106 mutants reached 64.74% and 63.13% of dry cell weight respectively, whereas the wild strain exhibited only 43.19%. DHA yield in these two mutants were even 2.26-fold and 2.37-fold higher than that of the wild strain. Experiment with 5 L fermentor further confirmed that D03432 and D05106 mutants had better performance than the wild strain on DHA yield (45.51% and 66.46% more than that of the wild strain, respectively), and demonstrated their high potential for industrial application. This work not only generated several high-DHA content mutants with high potential for industrial use, but also provided vital guidance for high-throughput screening of lipid hyper-accumulating mutants in other oil-producing microorganisms.

Efeito da suplementação de ômega 3 isolado ou associado em pacientes com doença de Alzheimer: uma revisão sistemática da literatura científica / Effect of omega 3 supplementation alone or associated in patients with Alzheimer's disease: a systematic literature review

Introdução: O envelhecimento da população está associado ao aumento da incidência da doença de Alzheimer (DA), a qual causa graves complicações ao paciente. Os nutrientes imunomoduladores, como os ácidos graxos poli-insaturados (PUFAs) da série ômega 3 (w-3) podem auxiliar na melhora do quadro clínico da DA. Objetivo: Analisar o efeito da suplementação de PUFAs w-3 isolado ou associado na população idosa com DA. Material e Métodos: Trata-se de uma revisão sistemática de literatura científica nas bases de dados Pubmed e Science Direct, que englobou ensaios clínicos em idosos com provável e/ou comprovado diagnóstico de DA e suplementados com PUFAS w-3 isolado ou associado, no idioma inglês e com os seguintes descritores em saúde (DECs): brain, Alzheimer's disease, fatty docosahexaenoic acid(DHA), polyunsaturated fatty acids (PUFA), older and elderly people e termo de pesquisa: eicosapentaenoic acid (EPA). O recorte temporal das publicações foi delimitado de 2006 a 2017. Resultados: foram selecionados 10 ensaios clínicos, cuja suplementação de w-3 favoreceu menor declínio no score de miniexame de estado mental (MEEM), retardo da disfunção, melhora no domínio de agitação do inventário neuropsiquiátrico (NPI) e melhora dos sintomas depressivos pela Escala de Depressão de Montgomery-Asberg (MADRS). Foram observadas mudanças significativas como aumento de apetite, peso, índice de massa corporal (IMC), EPA e DHA, além de reduções nos níveis séricos de albumina, ácido araquidônico (AA), ácido mirístico, interleucina-6 (IL- 6), interleucina 1 ß (IL-1ß) e fator estimulante de colônias de granulócitos (G-CSF) e redução na liberação de prostaglandina F2α (PGF2α). Foram relatadas alterações positivas em alguns genes e em outros, redução de sua expressão, além de hipometilação de importantes genes. Conclusão: A suplementação de PUFAs w-3 exerceu efeito positivo em pacientes com DA grau leve a moderado.

Introduction: Aging of the population is associated with an increased incidence of Alzheimer's disease (AD), which causes severe complications to the patient. Immunomodulatory nutrients such as polyunsaturated fatty acids (PUFAs) of the omega-3 (w-3) series may help improve the clinical picture of AD. Objective: To analyze the effect of supplementation of isolated or associated w-3 PUFAs in patients with AD. Material and Methods: This is a systematic review of the scientific literature in the Pubmed and Science Direct databases, which included clinical trials in elderly people with probable and / or proven diagnosis of AD supplemented with isolated or associated PUFAS w-3, in English and with the following health descriptors (DECs): brain, Alzheimer's disease, fatty docosahexaenoic acid (DHA), polyunsaturated fatty acids (PUFA), older and elderly people and search term: eicosapentaenoic acid (EPA).The time cut of publications was delimited from 2006 to 2017. Results: a total of 10 clinical trials were selected, whose w-3 supplementation favored a smaller decline in the Mini Mental Status Examination (MMSE) score, delayed dysfunction, improved neuropsychiatric inventory (NPI) agitation, and improved depressive symptoms by the Montgomery-Asberg Depression Scale (MADRS). Significant changes were observed as increased appetite, weight, body mass index (BMI), EPA and DHA, as well as reductions in serum albumin levels, arachidonic acid (AA), myristic acid, interleukin-6 (IL-6), interleukin 1ß (IL-1ß), and granulocyte colony stimulating factor (G-CSF) and reduced clearance of prostaglandin F2α (PGF2α). Positive alterations have been reported in some genes and in others, reduction of their expression, besides hypomethylation of important genes. Conclusion: Supplementation of PUFAs w-3 had a positive effect in patients with mild to moderate AD.

Oral omega 3 in different proportions of EPA, DHA, and antioxidants as adjuvant in treatment of keratoconjunctivitis sicca in dogs / Ômega 3 oral em diferentes proporções de EPA, DHA e antioxidantes como adjuvante no tratamento de ceratoconjuntivite seca em cães

ABSTRACT Purpose: To compare the efficacy of 0.03% to­pical tacrolimus in combination with oral omega (ω) 3 with different ratios of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and antioxidants as adjuvant in the treatment of keratoconjunctivitis sicca (KCS) in dogs. Methods: Forty-five dogs with KCS were evaluated monthly for 6 months. Evaluations included performance of the Schirmer tear, fluorescein, and lissamine green tests. Tear film break-up time (TBUT) was assessed. Conjunctival cytology was evaluated at the beginning, middle, and end of the study. Conjunctiva was biopsied at the beginning and end of the study. Dogs were randomly assigned to one of the three groups (n=15): Group T (topical tacrolimus 0.03%), Group TO (topical tacrolimus + ω-1.5 EPA: 1 DHA), or Group TOA (topical tacrolimus + ω-1 EPA:4.5 DHA + antioxidants). Results: There was a significant improvement in clinical signs in all groups. TBUT increased throughout treatment in all groups; this effect was most pronounced in Group TO. Cytological analysis performed at the end of the study period, showed decreased levels of lymphocytes, neutrophils, and metaplastic and squamous cells in Groups T, TO, and TOA. Histological analysis performed at the end of the study period showed decreased levels of lymphocytes and neutrophils and increased levels of goblet cells. These effects were most pronounced in Group TO. Conclusion: Oral treatment with ω-3 containing a higher proportion of EPA than DHA increased the effectiveness of topical tacrolimus 0.03% in the treatment of keratoconjunctivitis sicca in dogs.

RESUMO Objetivo: Comparar a eficácia do tacrolimus 0,03% tópico associado ao ômega 3 oral, com diferentes proporções de ácido eicosapentaenoico (EPA), ácidos docosa-hexaenoicos (DHA) e antioxidantes, como adjuvante no tratamento de cães acometidos por ceratoconjuntivite seca. Métodos: Quarenta e cinco cães atendidos no Hospital Veterinário da UNOESTE portadores de ceratoconjuntivite seca foram avaliados mensalmente por 6 meses pelo Teste Lacrimal de Schirmer, Teste de Fluoresceína, Tempo de Ruptura do Filme Lacrimal, Teste de Rosa Bengala, citologia da conjuntiva no início, meio e fim do projeto e biopsia da conjuntiva no início e final do projeto. Os cães foram distribuídos aleatoriamente em 3 grupos (n=15): grupo T (tacrolimus 0,03% tópico), grupo TO (tacrolimus + ômegas 1.5 EPA/1 DHA oral) e grupo TOA (tacrolimus + ômegas 1 EPA/4,5 DHA + antioxidantes oral). Resultados: Houve uma melhora significativa nos sinais clínicos em ambos os grupos. No tempo de ruptura do filme lacrimal todos os grupos apresentaram aumento no decorrer do tratamento, sendo que o grupo TO foi o que apresentou melhor resultado em todos momentos quando comparado aos demais grupos. Ao final do experimento, os grupos T, TO e TOA apresentaram na análise citológica, diminuição de linfócitos, neutrófilos, células metaplásicas e escamosas, e na análise histopatológica, diminuição de linfócitos e neutrófilos e aumento das células caliciformes, com o grupo TO com melhor desempenho. Conclusão: O tratamento oral com ω-3 contendo uma maior proporção de EPA do que o DHA aumentou a eficácia do tacrolimus tópico 0,03% no tratamento de ceratoconjuntivite sicca em cães.

Alanine mother liquor as a nitrogen source for docosahexaenoic acid production by Schizochytrium sp. B4D1

Alanine mother liquor, a type of industrial waste from alanine fermentation, was used as a nitrogen source to produce docosahexaenoic acid (DHA) by Schizochytrium sp. B4D1. The results indicated that yeast extract could trigger the utilization of the alanine mother liquor. Additionally, the alanine can be quenched during the culture, which aids in DHA accumulation. The medium components were optimized via response surface methodology as follows: 99.98-g/L glucose, 0.05-g/L yeast extract and a 183.17 dilution factor of the alanine mother liquid (v/v, with an alanine content of 0.72 g/L) and 17.98% inoculum concentration (v/v). Finally, in a 50-mL shake-flask fermentation, the DHA yield was 2.29 g/L.

Ômega 3 na gestação e seus benefícios / Omega 3 during pregnancy and its benefits

O objetivo desta pesquisa é levantar a literatura científica sobre os benefícios do Ômega-3 na gestação, que se deu pelas bases de dados Scientifc Electronic Library Online (Scielo), Literatura Latino-Americana e do Caribe em Ciências da Saúde (Lilacs), Literatura Internacional em Ciências de Saúde (Medline) e National Library of Medicine (Pubmed) e Cochrane em literaturas nacionais e internacionais. Nesta pesquisa pode-se verificar que o ácido docosahexaenoico (DHA) é considerado como o principal tipo de Ômega-3 pelo fato de proporcionar benefícios para a saúde, que vão desde o desenvolvimento do cérebro e da retina do bebê, os quais têm início a partir da suplementação da mãe já na gestação. O acúmulo do Ômega-3 ocorre no último trimestre da gestação e o transporte se dá através da placenta, sendo depositado no cérebro e na retina do concepto. Ocorre também um acúmulo simultâneo nas glândulas mamárias durante esta fase. O recomendado pelo consenso é de 200mg/dia, independentemente da fonte utilizada para suplementação. O adequado aporte de Ômega-3 na gestação e no pós-natal tem influência positiva no desenvolvimento visual e do sistema nervoso do recém-nascido, influenciando também na inteligência e na intelectualidade do indivíduo na vida adulta. O Ômega-3 é importante também na prevenção e tratamento de diversas doenças como obesidade, doenças cardiovasculares, imunológicas, câncer de cólon, entre outras.(AU)

The objective of this research is to raise the scientific literature on the benefits of omega-3 during pregnancy, which occurred in research in databases Scientifc Electronic Library Online (Scielo), Latin American and Caribbean Health Sciences (Lilacs) , International Literature in Health Sciences (MEDLINE) and national Library of Medicine (PubMed) and the Cochrane in national and international literature, which can be seen that docosahexaenoic acid (DHA) is considered as the main type of Omega-3, the fact provide health benefits ranging from brain development and the baby's retina, which begins from the mother during pregnancy supplementation already. The Omega-3 accumulation occurs in the last trimester of pregnancy and the transport is through the placenta being deposited in the brain and retina of the fetus. It is also a simultaneous accumulation in the mammary glands during this phase. The recommended by consensus is 200 mg / day, regardless of the source used for supplementation. Adequate intake of omega-3 during pregnancy and in the postnatal has positive influences on visual development and the nervous system of the newborn, influencing also the intelligence and the individual intellect in adulthood. The Omega-3 is also important in the prevention and treatment of various diseases as obesity, cardiovascular, immune disorders, colon cancer, among others.(AU)

Protectin DX Exhibits Protective Effects in Mouse Model of Lipopolysaccharide-Induced Acute Lung Injury / 中华医学杂志(英文版)

<p><b>Background</b>Acute lung injury (ALI) is a severe disease with high mortality and poor prognosis. Protectin DX (PDX), a pro-resolving lipid mediator, exhibits protective effects in ALI. Our experiment aimed to explore the effects and related mechanisms of PDX in mice with ALI induced by lipopolysaccharide (LPS).</p><p><b>Methods</b>BALB/c mice were randomly divided into five groups: sham, LPS, LPS plus 1 ng of PDX (LPS + PDX-1 ng), LPS plus 10 ng of PDX (LPS + PDX-10 ng), and LPS plus 100 ng of PDX (LPS + PDX-100 ng). Bronchoalveolar lavage fluids (BALFs) were collected after 24 h, and total cells, polymorphonuclear leukocytes, monocyte-macrophages, and lymphocytes in BALF were enumerated. The concentration of interleukin (IL)-1β, IL-6, IL-10, tumor necrosis factor-alpha (TNF-α), macrophage inflammatory protein (MIP)-1α, and MIP-2 in BALF was determined, and histopathological changes of the lung were observed. The concentration of protein in BALF and lung wet/dry weight ratios were detected to evaluate pulmonary edema. After determining the optimal dose of PDX, neutrophil-platelet interactions in whole blood were evaluated by flow cytometry.</p><p><b>Results</b>The highest dose of PDX (100 ng/mouse) failed to provide pulmonary protective effects, whereas lower doses of PDX (1 ng/mouse and 10 ng/mouse), especially 1 ng PDX, alleviated pulmonary histopathological changes, mitigated LPS-induced ALI and pulmonary edema, inhibited neutrophil infiltration, and reduced pro-inflammatory mediator (IL-1β, IL-6, TNF-α, and MIP-1α) levels. Meanwhile, 1 ng PDX exhibited pro-resolving functions in ALI including upregulation of monocyte-macrophage numbers and anti-inflammatory mediator IL-10 levels. The flow cytometry results showed that PDX could inhibit neutrophil-platelet interactions in ALI.</p><p><b>Conclusion</b>PDX exerts protective effects in LPS-induced ALI by mitigating pulmonary inflammation and abrogating neutrophil-platelet interactions.</p>

Xuebijing Injection () and Resolvin D1 Synergize Regulate Leukocyte Adhesion and Improve Survival Rate in Mice with Sepsis-Induced Lung Injury / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effect of combined application of Xuebijing Injection ( , XBJ) and resolvin D1 (RvD1) on survival rate and the underlying mechanisms in mice with sepsisinduced lung injury.</p><p><b>METHODS</b>The cecal ligation and puncture (CLP) method was used to develop a mouse sepsis model. Specific pathogen free male C57BL/6 mice were randomly divided into 5 groups (n=20 each): sham, CLP, CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1. After surgery, mice in the CLP+XBJ, CLP+RvD1 and CLP+XBJ+RvD1 groups were given XBJ (25 μL/g body weight), RvD1 (10 ng/g body weight), and their combination (the same dose of XBJ and RvD1), respectively. In each group, 12 mice were used to observe 1-week survival rate, while the rest were executed at 12 h. Whole blood was collected for flow cytometric analysis of leukocyte adhesion molecules CD18, lung tissues were harvested for observing pathological changes, and testing the activity of myeloperoxidase (MPO) and the expression of intercellular cell adhesion molecule 1 (ICAM-1).</p><p><b>RESULTS</b>Compared with the CLP group, the histopathological damage of the lung tissues was mitigated, MPO activity was decreased in the CLP+XBJ and CLP+RvD1 groups (P<0.05). In addition, the 1-week survival rate was improved, proportion of CD18-expressing cells in whole blood and ICAM-1 protein expression in lung tissue were decreased in the CLP+XBJ+RvD1 group (P<0.05 or P<0.01).</p><p><b>CONCLUSIONS</b>XBJ together with RvD1 could effectively inhibit leukocyte adhesion, reduce lung injury, and improve the survival rate of mice with sepsis.</p>

Effect of docosahexenoic acid supplementation on infant's growth and body mass index during maternal pregnancy / 中华流行病学杂志

Objective: To investigate the effects of docosahexenoic acid (DHA) supplementation on infant's growth and BMI during pregnancy. Methods: A total of 1 516 healthy pregnant women delivered their babies in two maternal and child health care hospitals in Beijing and were chosen as the subjects in this cohort study from May to October 2015. Self-developed questionnaires were used to gather general information of the subjects, including age, height, weight, weight gain during pregnancy, delivery mode, DHA supplementation etc., before giving birth. Information on body length, weight, head circumference and BMI at birth and 6 months postnatal, of the infants were recorded. Breast milk was collected to test the fatty acid profiles by using the gas chromatography (GC) method at one to three months postnatally. Results: The overall rate of DHA supplementation was 47.76% among the pregnant women, in which introduction of DHA from the early and second stage of the pregnancy accounted for 49.31% and 39.64% respectively. When DHA supplementation began from the early pregnant stage, the DHA concentration showed an increase in the milk (P<0.05), whereas the supplementation began from the second and third stages did not affect the milk DHA concentration (P>0.05). Higher height and lower BMI were seen in the infants at birth and 6 months in the supplementation group when comparing to the non-supplementary group (P<0.05), with the greatest effects noticed in the earliest supplementation group. Specifically, the head circumference appeared larger from the early pregnant stage in the DHA supplementary group, than that in the non-supplement group (P=0.001). The increment of head circumference was larger than that in the other groups when the infants were 6-month old (P<0.01). Results from the partial regression analysis showed that during pregnancy, there were positive correlations between DHA supplementation and height (r=0.324, r=0.216), head circumference (r=0.221, r=0.302) as well as the increment of head circumference (r=0.276) at birth and 6 months (P<0.05). Whereas, a negative correlation was shown between DHA and the infants' BMI (r=-0.310, r=-0.371) (P<0.05) when supplementation was given during maternal pregnancy. Conclusions: When DHA supplementation program was carried out during maternal pregnancy, it could increase the height and head circumference and inhibit the rapid increase of BMI in the infants BMI. Our findings seemed helpful in promoting brain development and preventing the childhood obesity.

Effectiveness of omega-3 polyunsaturated fatty acids against microbial pathogens / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Microorganisms provide both beneficial and harmful effects to human beings. Beneficial effects come from the symbiotic relationship that exists between humans and microbiota, but then several human illnesses have turned some friendly microbes into opportunistic pathogens, causing several microbial-related diseases. Various efforts have been made to create and utilize antimicrobial agents in the treatment and prevention of these infections, but such efforts have been hampered by the emergence of antimicrobial resistance. Despite extensive studies on drug discovery to alleviate this problem, issues with the toxicity and tolerance of certain compounds and continuous microbial evolution have forced researchers to focus on screening various phytochemical dietary compounds for antimicrobial activity. Linolenic acid and its derivatives (eicosapentaenoic acid and docosahexaenoic acid) are omega-3 fatty acids that have been studied due to their role in human health, being important for the brain, the eye, the cardiovascular system, and general human growth. However, their utilization as antimicrobial agents has not been widely appreciated, perhaps due to a lack of understanding of antimicrobial mechanisms, toxicity, and route of administration. Therefore, this review focuses on the efficacy, mechanism, and toxicity of omega-3 fatty acids as alternative therapeutic agents for treating and preventing diseases associated with pathogenic microorganisms.

Association of serum inflammatory cytokines and Resolvin D1 concentration with pathological stage of colon cancer / 中华胃肠外科杂志

OBJECTIVE@#To investigate the levels of serum inflammatory cytokines and Resolvin D1 (RvD1) and their association with pathological staging of colon cancer.@*METHODS@#Clinical data of 50 colon cancer patients (colon cancer group) admitted to the General Surgery Department of Zhongshan Hospital of Fudan University from January to December 2016 and 5 ml of whole blood specimen were collected at admission. During the same period, 50 healthy volunteers were enrolled (healthy volunteer group). Inclusion criteria for the colon cancer group: colon cancer diagnosed by preoperative colonoscopy and pathology; no recent enteral or parenteral nutrition support treatment or use of oral nutrition preparation; age ≤85 years; no surgical contraindications by preoperative evaluation; no history of taking fish oil-related preparations; no radiotherapy or chemotherapy before surgery. Healthy volunteer group enrollment criteria: no history of malignant tumors; no organ with organic lesions detected by the healthy examination center of our hospital; detection indicators in normal reference range; no administration of fish oil-related preparations; age ≤ 85 years. Serum inflammatory factors(IL-1β, IL-6, IL-10 and TNF-α) concentrations were detected by chemiluminescence immunoassay; serum RvD1 concentration was measured by enzyme-linked immunosorbent assay. The levels of inflammatory factors and RvD1 were compared between the two groups, and their associations with TNM staging of colon cancer patients were analyzed.@*RESULTS@#There were no significant differences in age, gender and nutrition-related indicators between the two groups (all P>0.05). There were 31 males and 19 females in the healthy volunteer group with age of (61.8±11.6) years. There were 23 males and 27 females in the colon cancer group with age of (65.4±12.4) years. According to the 7th edition of the American Cancer Society TNM staging criteria, 10 cases were stage I, 13 cases stage II, 17 cases stage III, and 10 cases stage IV. Compared with healthy volunteer group, colon cancer group had higher serum IL-1β [(3.89±0.24)×10 μg/L vs.(1.55±0.37)×10 μg/L, t=37.52, P<0.01], higher IL-6 [(129.14±3.07)×10 μg/L vs.(51.46±3.14)×10 μg/L, t=125.08, P<0.01], higher IL-10 [(100.59±8.69)×103 μg/L vs.(27.57±4.77)×10 μg/L, t=52.09, P<0.01] and higher TNF-α [(114.31±4.43)×10 μg/L vs.(41.04±5.27)×10 μg/L, t=75.25, P<0.01], while lower RvD1 [(34.19±1.93)×10 μg/L vs.(77.76±1.02)×10 μg/L, t=140.56, P<0.01], all the differences were statistically significant. Subgroup analysis revealed that concentrations of IL-6, IL-1β, IL-10 and TNF-α gradually increased with the advancement of TNM staging (P<0.01). In stage III, concentrations of IL-6, IL-1β, and IL-10 were the highest, TNF-α concentration was the highest in stage IV. RvD1 concentration gradually decreased with the advancement of TNM staging(P<0.01).@*CONCLUSIONS@#Compared with healthy volunteers, the levels of serum inflammatory cytokines in colon cancer patients increase significantly while the level of RvD1 decreases significantly. Both are associated with higher TNM stage of colon cancer.

Efectos neuroprotectores del ácido araquidónico y del ácido docosahexaenoico en las etapas extremas de la vida: Una visión integradora / Neuroprotective effects of arachidonic and docosahexaenoic acid in the extreme stages of life: An integrative view

RESUMEN El estudio focalizado en dilucidar el rol neuroprotector del ARA y del DHA a lo largo del ciclo vital ha cobrado cada vez más interés puesto que se continúan descubriendo mecanismos mediante los cuales estos ácidos grasos poliinsaturados de cadena larga (AGPICL) modulan el metabolismo. Tanto el ARA como el DHA se encuentran depositados en los lípidos de las membranas de las células que forman la materia gris y representan aproximadamente el 25% del contenido total de ácidos grasos cerebrales. El ARA y el DHA tienen efectos sobre el crecimiento y la diferenciación neuronal a través de la modulación de las propiedades físicas de la membrana, de la transducción de señales asociada a proteínas G y la modulación de la expresión génica, adquiriendo un rol relevante en la neuro-génesis y el desarrollo cerebral. Además, se les atribuye un rol neuroprotector en patologías neurodegenerativas como la enfermedad de Alzheimer y la enfermedad de Parkinson, pudiendo disminuir la disfunción mitocondrial, la neuro-inflamación y el estrés oxidativo, expresiones características de estas patologías. La presente revisión analiza y discute acerca del rol del ARA y del DHA en la neuro-protección y en la neurodegeneración a través de una visión integradora.

ABSTRACT The study focused on elucidating the neuro-protective effects of ARA and DHA throughout the life cycle has become of increasingly interest since the continue discovering of mechanisms by which these long-chain polyunsaturated fatty acids (LCPUFA) modulate the metabolism. Both ARA and DHA are deposited into the membrane lipids of the cells that form the gray matter of the brain and represent approximately 25% of the total content of cerebral fatty acids. ARA and DHA have effects on the growth and neuronal differentiation through the modulation of the physical properties of the membrane, the signal transduction associated to G proteins and by the modulation of gene expression, acquiring a relevant role in neurogenesis and brain development. In addition, it is attributed to these fatty acids a neuro-protective role in neurodegenerative pathologies such as Alzheimer's disease and Parkinson's disease by decreasing the mitochondrial dysfunction, neuroinflammation and oxidative stress, characteristic of these pathologies. This review analyzes and discusses the role of ARA and DHA in neuro-protection and neuro-degeneration through an integrative vision.

Selection of Schizochytrium limacinum mutants based on butanol tolerance

Background: Mutation breeding is one of the most important routes to achieving high docosahexaenoic acid (DHA) productivity using Schizochytrium. However, few selection strategies have been reported that aim to generate a high DHA content in Schizochytrium lipids. Results: First, culture temperature altered the butanol tolerance of Schizochytrium limacinum B4D1. Second, S. limacinum E8 was obtained by selecting mutants with high butanol tolerance. This mutant exhibited a 17.97% lower proportion of DHA than the parent strain S. limacinum B4D1. Third, a negative selection strategy was designed in which S. limacinum F6, a mutant with poor butanol tolerance, was obtained. The proportion of DHA in S. limacinum F6 was 11.22% higher than that of parent strain S. limacinum B4D1. Finally, the performances of S. limacinum B4D1, E8 and F6 were compared. These three strains had different fatty acid profiles, but there was no statistical difference in their biomasses and lipid yields. Conclusion: It was feasible to identified the relative DHA content of S. limacinum mutants based on their butanol tolerance.

Influência dos ácidos graxos ômega-3 e vitamina D na depressão: uma breve revisão / Influence of omega 3 fatty acids and vitamin D in depression: a short review

Objetivo: descrever uma revisão da literatura sobre a importância dos ácidos graxos ômega-3 e da vitamina D na prevenção e tratamento da depressão. Metodologia: essa revisão foi feita no período de agosto, setembro de 2016, incluindo as publicações de 2000 a 2016. Procedeu-se a busca nas bases de dados Pubmed, Science Direct, Bireme e periódicos CAPES com a utilização dos descritores: "ômega-3", "docosahexaenoic acid", "DHA", "eicosapentaenoic acid", "EPA", "vitamin D" e "depression". Resultados: a literatura reporta vários estudos sobre a eficácia dos ácidos graxos ômega-3 no tratamento da depressão. No entanto, existe um número considerável de trabalhos que não demonstram eficácia dos ácidos graxos ômega-3 na depressão. Os fatores responsáveis pelos resultados variados são: modelos experimentais, diferenças de tamanho da amostra, diferenças biológicas e genéticas entre os doentes, a variabilidade ambiental, e variabilidade na resposta à ácidos graxos ômega-3. Em relação a vitamina D, os resultados também são contraditórios, com evidências sugerindo que baixos níveis dessa vitamina podem estar associados a um maior risco de depressão. Conclusão: não é possível afirmar que consumir alimentos ricos em ômega-3 e vitamina D, juntamente com o uso de fármacos possam aliviar os sintomas depressivos.

Objective: to describe a literature review on the importance of omega-3 fatty acids and vitamin D in the depression prevention and treatment. Methodology: this review was done in August / September, 2016, including publications from 2000 to 2016. Pubmed, Science Direct, Bireme and Journals CAPES databases were searched using the descriptors: "omega-3", " Docosahexaenoic acid "," DHA "," eicosapentaenoic acid "," EPA "," vitamin D" and "depression ". Results: the literature report several studies on the efficacy of omega-3 fatty acids in the depression treatment. However, there are a number of studies that don't demonstrate efficacy of omega-3 fatty acids in depression. The factors responsible for the varied results are: experimental models, differences in sample size, biological and genetics differences among patients, environmental variability, and variability in response to omega-3 fatty acids. There are many researches with contradictory results regarding vitamin D, with evidence suggesting that low levels of this vitamin may be associated with an increased risk of depression. Conclusion: it can't be affirm that consume foods rich in omega-3 and vitamin D along with the use of pharmaceuticals products may alleviate depressive symptoms.

Production of docosahexaenoic acid by Aurantiochytrium sp. ATCC PRA-276

Abstract The high costs and environmental concerns associated with using marine resources as sources of oils rich in polyunsaturated fatty acids have prompted searches for alternative sources of such oils. Some microorganisms, among them members of the genus Aurantiochytrium, can synthesize large amounts of these biocompounds. However, various parameters that affect the polyunsaturated fatty acids production of these organisms, such as the carbon and nitrogen sources supplied during their cultivation, require further elucidation. The objective of this investigation was to study the effect of different concentrations of carbon and total nitrogen on the production of polyunsaturated fatty acids, particularly docosahexaenoic acid, by Aurantiochytrium sp. ATCC PRA-276. We performed batch system experiments using an initial glucose concentration of 30 g/L and three different concentrations of total nitrogen, including 3.0, 0.44, and 0.22 g/L, and fed-batch system experiments in which 0.14 g/L of glucose and 0.0014 g/L of total nitrogen were supplied hourly. To assess the effects of these different treatments, we determined the biomass, glucose, total nitrogen and polyunsaturated fatty acids concentration. The maximum cell concentration (23.9 g/L) was obtained after 96 h of cultivation in the batch system using initial concentrations of 0.22 g/L total nitrogen and 30 g/L glucose. Under these conditions, we observed the highest level of polyunsaturated fatty acids production (3.6 g/L), with docosahexaenoic acid and docosapentaenoic acid ω6 concentrations reaching 2.54 and 0.80 g/L, respectively.

Combination therapy with butyrate and docosahexaenoic acid for keloid fibrogenesis: an in vitro study

Abstract: Background: A single, effective therapeutic regimen for keloids has not been established yet, and the development of novel therapeutic approaches is expected. Butyrate, a short-chain fatty acid, and docosahexaenoic acid (DHA), a ω-3 polyunsaturated fatty acid, play multiple anti-inflammatory and anticancer roles via their respective mechanisms of action. Objective: In this study, we evaluated the antifibrogenic effects of their single and combined use on keloid fibroblasts. Methods: Keloid fibroblasts were treated with butyrate (0-16 mM) and/or DHA (0-100 µM) for 48 or 96 h. Results: Butyrate inhibited cell proliferation, and α-smooth muscle actin (α-SMA) and type III collagen expressions, with inhibition of the transforming growth factor (TGF)-β1 and TGF-β type I receptor expressions and increased prostaglandin E2 with upregulation of cyclooxygenase-1 expression with induction of histone acetylation. DHA inhibited α-SMA, type III collagen, and TGF-β type I receptor expressions. Then, the butyrate/DHA combination augmented the antifibrogenic effects, resulting in additional inhibition of α-SMA, type I and III collagen expressions, with strong disruption of stress fiber and apoptosis induction. Moreover, the butyrate/DHA combination inhibited the cyclooxygenase-2 expression, suggesting stronger anti-inflammatory effect than each monotherapy. Study limitations: Activation in keloid tissue is affected not only by fibroblasts but also by epithelial cells and immune cells. Evaluation of the effects by butyrate and DHA in these cells or in an in vivo study is required. Conclusion: This study demonstrated that butyrate and docosahexaenoic acid have antifibrogenic effects on keloid fibroblasts and that these may exert therapeutic effects for keloid.