Breast cancer molecular subtypes, based on expression of estrogen, progesterone and human epidermal growth factor 2 receptors, alter prognosis and treatment options. α-linolenic acid (ALA) is a complementary therapy, however its effectiveness across breast cancer types and estrogen environments is unclear. This research determined the effect of ALA on growth, apoptosis, fatty acid profile, and gene changes in four breast cancer cell lines with varying receptor expression with or without (±) estradiol (E2). ALA (50-200uM) ± E2 reduced growth in all cell lines. 75μM ALA +E2 increased phospholipid % ALA in all cell lines and induced apoptosis in cell lines lacking the three receptors. Cellular % ALA was positively associated with apoptosis and inversely associated with cell growth. ALA altered expression of cell cycle, apoptosis and signal transduction genes. In conclusion, ALA incorporates into breast cancer cells, reduces growth and induces apoptosis regardless of receptor status or E2 level.

Breast cancer molecular subtypes, based on expression of estrogen, progesterone and human epidermal growth factor 2 receptors, alter prognosis and treatment options. α-linolenic acid (ALA) is a complementary therapy, however its effectiveness across breast cancer types and estrogen environments is unclear. This research determined the effect of ALA on growth, apoptosis, fatty acid profile, and gene changes in four breast cancer cell lines with varying receptor expression with or without (±) estradiol (E2). ALA (50-200uM) ± E2 reduced growth in all cell lines. 75μM ALA +E2 increased phospholipid % ALA in all cell lines and induced apoptosis in cell lines lacking the three receptors. Cellular % ALA was positively associated with apoptosis and inversely associated with cell growth. ALA altered expression of cell cycle, apoptosis and signal transduction genes. In conclusion, ALA incorporates into breast cancer cells, reduces growth and induces apoptosis regardless of receptor status or E2 level.

Orientador: Vera Lucia Garcia Rehder / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T17:07:35Z (GMT). No. of bitstreams: 1 Santos_PauloEuzebioRubbodos_M.pdf: 8287880 bytes, checksum: 81821aafe849e03ea5711f1686603c72 (MD5) Previous issue date: 2013 / Resumo: O resumo poderá ser visualizado no texto completo da tese digital / Abstract: The abstract is available with the full electronic document / Mestrado / Fármacos, Medicamentos e Insumos para Saúde / Mestre em Biociências e Tecnologia de Produtos Bioativos

Pre-treatment with polyunsaturated fatty acids or bioactive lipid mediators has been shown to reduce neuronal injury in rodent models of focal ischaemia, but the molecular mechanisms underlying this neuroprotection are unclear. In this study, we aimed to investigate whether systemic administration of alpha linolenic acid (ALA) leads to changes in the profile of mouse brain phospholipid and bioactive lipid mediators in both mouse brain and plasma within the previously determined neuroprotection time window. Mass spectrometry (MS) and tandem mass spectrometry (MS/MS) allowed us to detect and identify 47 phospholipids in mouse cerebral cortex, including several phospholipid species not previously reported in brain lipidomic studies. These included a phosphatidylethanolamine species with m/z 720 that has been associated with retinal stem cells. No widespread changes in cerebral cortex phospholipid composition were observed following intravenous ALA. Several significant changes in lipid mediators (P<0.05 with two-way ANOVA and post hoc Dunnett's t test) were detected in ALA-treated animals compared to untreated and vehicle-injected animals. Many of the affected lipid mediators are ligands for prostanoid receptors which have been demonstrated to play a role in the development of brain injury following cerebral ischaemia, implying that changes in bioactive lipid mediators or modulation of prostanoid receptors may occur following ALA pre-treatment in mice. This study illustrates the potential of advanced lipidomic analysis as a novel tool for neurochemists.

The objectives were to 1) conduct a meta-analysis to assess the association between alpha-linolenic acid (ALA) and prostate cancer; 2) analyze a trial of ALA on coronary heart disease with PSA as a post hoc outcome; 3) assess the effect of trial serum and also ALA directly on LNCaP cell growth. 1) The ALA meta-analysis of prospective and case-control studies showed no overall effect on prostate cancer. However, removal of one study from the analysis of prospective studies changed the result to a significant protective effect (RR=0.91; 95%CI:0.83,0.99). 2) No significant treatment difference was seen in the change in PSA in the randomized controlled trial. 3) The ALA treatment serum from the clinical trial did not affect LNCaP cell growth. However, ALA decreased LNCaP cell growth in a dose dependent manner when added to cell culture. The results provide no positive evidence for an effect of ALA on prostate cancer.

The objectives were to 1) conduct a meta-analysis to assess the association between alpha-linolenic acid (ALA) and prostate cancer; 2) analyze a trial of ALA on coronary heart disease with PSA as a post hoc outcome; 3) assess the effect of trial serum and also ALA directly on LNCaP cell growth. 1) The ALA meta-analysis of prospective and case-control studies showed no overall effect on prostate cancer. However, removal of one study from the analysis of prospective studies changed the result to a significant protective effect (RR=0.91; 95%CI:0.83,0.99). 2) No significant treatment difference was seen in the change in PSA in the randomized controlled trial. 3) The ALA treatment serum from the clinical trial did not affect LNCaP cell growth. However, ALA decreased LNCaP cell growth in a dose dependent manner when added to cell culture. The results provide no positive evidence for an effect of ALA on prostate cancer.

Orientador: Fernando Antonio Cabral / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-08T22:12:06Z (GMT). No. of bitstreams: 1 FollegattiRomero_LuisAlberto_M.pdf: 666792 bytes, checksum: 9ae97012c5bd9df738d34f847f6ef5b7 (MD5) Previous issue date: 2007 / Resumo: A extração supercrítica do óleo das sementes de sacha inchi (plukenetia volubilis L.) usando CO2 como solvente é apresentada neste estudo. O objetivo foi investigar os principais efeitos das variáveis de processo como a pressão, a temperatura e o diâmetro médio de partícula (0,387, 0,595 e 0,842 mm) no rendimento da extração. O rendimento de extração aumentou com a diminuição do diâmetro de partícula, indicando a importância do efeito da difusão intra-partícula nas partículas maiores (0,842 mm). A extração supercrítica foi realizada a 30 e 40 MPa, temperaturas de 40, 50 e 60°C, utilizando 0,842 mm de diâmetro de partícula a uma vazão de 1,5L de CO2/min. O rendimento máximo de extração para as sementes trituradas foi de 92%, obtido a 40MPa, 60°C e 0,842 mm de diâmetro médio de partícula. Para extração por prensagem a frio, seguida da extração supercrítica a 40Mpa, 60ºC e 0,716 mm de diâmetro médio geométrico da partícula o rendimento total de extração foi de 99%. A solubilidade do óleo da semente de Sacha inchi em dióxido de carbono supercrítico (SC-CO2) foi medida nas temperaturas de 40 e 60 °C e pressões entre 10 e 40 MPa. Os valores de solubilidade mostraram um comportamento retrógrado, abaixo de 35 MPa a solubilidade diminuiu com o aumento da temperatura.A tendência observada acima desta pressão foi um aumento da solubilidade com o aumento da temperatura a pressão constante. O modelo matemático de Chrastil foi usado para correlacionar os valores experimentais de solubilidade. O conteúdo de óleo determinado com hexano em equipamentos soxhlet foi de 54,32% e o rendimento de óleo extraído por prensagem a frio foi de 46,8%. As características físicas e químicas do óleo foram obtidas. As composições em ácidos graxos do óleo obtido por SC-CO2 e do óleo obtido com hexano não apresentaram diferenças significativas. O principal ácido graxo foi o ácido alfa-linolênico (Ômega-3) aproximadamente 50,5%, seguido peloácido linoléico (34,12%) e ácido aléico (14%). O conteúdo de ácidos graxos livres (AGL) e de tocoferóis (tocoferol e tocotrienol) dos óleos obtidos por SC-CO2, solvente e prensagem a frio foram determinados. O conteúdo de tocoferóis so óleo extraído com SC-CO2 variou de 222,8 a 306,8 mg/100g, do óleo extraído por prensagem a frio foi 278,4 mg/100g e do óleo extraído com hexano foi 238,7 mg/100g. O teor de AGL no óleo extraído com hexano foi de 2,98% (medido como % de ác. Linolênico), ligeiramente menor do que aquele extraído por prensagem a frio (3,75%) e SC-CO2 (3,97-4,26%). O óleo de sementes de Sacha inchi pode ter grande potencial debtro do mercado crescente de óleos especiais / Abstract: The supercritical fluid extraction of oil from sacha inchi (plukenetia volubilis L.) seeds, using Co2 as a solvent, it is presented in this study. The objective was to investigate the main process parameters such as extraction pressure and temperature, mean particle size of (0,387, 0,595 and 0,842 mm) on the extraction yield. The extraction yield increased as the particle size decreased, indicating the importance of the effect of the intraparticle diffusion in larger particles (0,84 mm). The extraction was carried out at operating pressures of 30 and 40 MPa, operating temperatures of 40, 50 and 60°C, particle 0,84 mm and flow rate of CO2 1,5L/min. Maximum extraction yield was 92% obtained at 40 MPa, 60°C, 0.84 mm of particle sizes and 1.5 L of CO2/min. For the cold-pressed extraction, followed of supercritical extraction 40 MPa, 60ºC and 0,716 mm of geometric average diameter of the particle, the maximum extraction yield was 99%. The solubility of Sacha inchi seed oil in supercritical carbon dioxide (SC-CO2) were measured in the temperatures from 40 to 60 °C and pressures between 10 and 40 MPa. Under working conditions, the values of solubility showed retrograde behavior. Solubility, increase with pressure and temperature above the crossover pressure, which found between 30 and 40 MPa, and decreased with temperature below the crossover pressure. Mathematical modeling (Chrastil) of the solubility. The oil content was 54,32% and 46.80% determined bu solvent extraction and cold-pressed, respectively. Physical and chemical characteristics of the oil were obtained. The results in terms of fatty acid compositions of oil obtained with SC-CO2 were compared with those obtained when hexano was used as the solvent, and no significant differences between the oils extracted by both methods were found. The main fatty acid was alpha-linolênico acid (Ômega-3) making up 50.5%, followed by linoleic acid (34,12%) and oleic acid (14%). The level of FFA and tocol composition (tocoferol and tocotrienol) contents of the oils obtained by SC-CO2, solvent and cold-pressed extraction was determined. Tocol content of the SC-CO2 - extracted oil varied from 222.85 to 306.82 mg/100g, which was higher than that of hexane-extraced oil (238.72 mg/100g) and cold-pressed oil (278.4 mg/100g). The level of FFA in the solvent-extraced oils (2.98%) (% as ác. Linolenic) was lower than cold-pressed extraced oil (3.75%) and SC-CO2)-extraced oils (3.97-4.26%). Sacha inchi seed oil may have potential in the growing specialty oil market / Mestrado / Mestre em Engenharia de Alimentos

Pre-treatment with polyunsaturated fatty acids or bioactive lipid mediators has been shown to reduce neuronal injury in rodent models of focal ischaemia, but the molecular mechanisms underlying this neuroprotection are unclear. In this study, we aimed to investigate whether systemic administration of alpha linolenic acid (ALA) leads to changes in the profile of mouse brain phospholipid and bioactive lipid mediators in both mouse brain and plasma within the previously determined neuroprotection time window. Mass spectrometry (MS) and tandem mass spectrometry (MS/MS) allowed us to detect and identify 47 phospholipids in mouse cerebral cortex, including several phospholipid species not previously reported in brain lipidomic studies. These included a phosphatidylethanolamine species with m/z 720 that has been associated with retinal stem cells. No widespread changes in cerebral cortex phospholipid composition were observed following intravenous ALA. Several significant changes in lipid mediators (P<0.05 with two-way ANOVA and post hoc Dunnett¿s t test) were detected in ALA-treated animals compared to untreated and vehicle-injected animals. Many of the affected lipid mediators are ligands for prostanoid receptors which have been demonstrated to play a role in the development of brain injury following cerebral ischaemia, implying that changes in bioactive lipid mediators or modulation of prostanoid receptors may occur following ALA pre-treatment in mice. This study illustrates the potential of advanced lipidomic analysis as a novel tool for neurochemists.

Atualmente, a obesidade é considerada uma epidemia mundial. Está associada a um estado de inflamação crônica e ativação do estresse do retículo endoplasmático (ERE), relacionados à patogênese de diversas doenças como diabetes mellitus tipo 2, doenças cardiovasculares, câncer, hipertensão, dentre outras. Nesse contexto, são necessários estudos para encontrar alternativas que melhorem o processo inflamatório. Vários estudos em humanos e animais já demostraram as propriedades anti-inflamatórias do ácido graxo ômega-3. O objetivo do nosso trabalho foi avaliar os efeitos da suplementação do ácido alfa-linolênico (ALA) no metabolismo e no estresse do retículo endoplasmático em obesos grau III. Foi conduzido um estudo prospectivo, randomizado, placebo-controlado, duplo-cego. No total, 52 pacientes foram randomizados para a suplementação com 3g/dia de ALA ou placebo, sendo 27 indivíduos do grupo ômega-3/ALA e 25 do grupo controle. Foi avaliado perfil lipídico, glicídico e inflamatório antes e após suplementação. O tecido adiposo visceral (TAV) foi coletado durante cirurgia bariátrica após suplementação. O perfil de ácidos graxos incorporados no TAV foi avaliado por cromatografia gasosa. Os genes foram avaliados através de PCR em tempo real. Não houve alteração nos níveis séricos de IL-6 (p=0,2201), TNF-? (0.7703) e PCR (p=0,57) após suplementação com ômega-3/ALA, porém observamos diminuição nos níveis séricos de Leptina (p=0.0154) e IP-10 (p=0.0410), citocinas inflamatórias, e aumento na IL-4 (p=0,0211), citocina anti-inflamatória. Foi observado incorporação significativa do ALA (p=0,0002), EPA (p<0,0001) e DHA (p=0,0005) no TAV. Avaliação molecular evidenciou um aumento da expressão gênica do XBP1 (p=0,0013), sXBP1 (p<0,0001), EIF2-? (p=0,0063) e da chaperona CCT4 (p=0,0001) e diminuição na expressão gênica da leptina (p=0,0410). Podemos concluir que o ALA pode modular o ERE através da via da IRE1/XBP, levando ao aumento das chaperonas (CCT4), o que pode demonstrar um potencial terapêutico do ALA em pacientes obesos. / Currently, obesity is considered a worldwide epidemic. It is associated with chronic inflammation and stress activation of the endoplasmic reticulum (ERE), related to the pathogenesis of various diseases such as type 2 diabetes mellitus, cardiovascular diseases, cancer, hypertension, among others. In this context, studies are needed to find alternatives that improve the inflammatory process. Several studies in humans and animals have already demonstrated the anti-inflammatory properties of omega-3 fatty acid. The objective of our study was to evaluate the effects of alpha-linolenic acid (ALA) supplementation on the metabolism and stress of the endoplasmic reticulum in obese patients. A prospective, randomized, placebo-controlled, double-blind study was conducted. In total, 52 patients were randomized to supplementation with 3 g / day of ALA or placebo, 27 individuals from the omega-3 / ALA group and 25 from the control group. Lipid, glycidic and inflammatory profile were evaluated before and after supplementation. Visceral adipose tissue (TAV) was collected during bariatric surgery after supplementation. The fatty acid profile incorporated in the TAV was evaluated by gas chromatography. Genes were evaluated by real-time PCR. There was no change in serum levels of IL-6 (p = 0.2201), TNF-? (0.7703) and CRP (p = 0.57) after supplementation with ALA, but we observed a decrease in serum Leptin levels (P = 0.0154) and IP-10 (p = 0.0410), inflammatory cytokines, and increase in IL-4 (p = 0.0211), anti-inflammatory cytokine. Significant incorporation of ALA (p = 0.0002), EPA (p <0.0001) and DHA (p = 0.0005) into the TAV was observed. Molecular evaluation showed an increase in the gene expression of XBP1 (p = 0.0013), sXBP1 (p <0.0001), EIF2-? (p = 0.0063), GADD34 (p=0,0117) and CCT4 chaperone (p = 0.0001), decrease in the gene expression of leptin (p = 0.0410) and ATF-6 (p=0,0305) and a tendency to decrease the gene expression of CHOP. We can conclude that ALA can modulate ERE through the IRE1 / XBP, PERK and ATF-6 pathways, leading to increased chaperones (CCT4), which may demonstrate a therapeutic potential of ALA in obese patients.

Atualmente, a obesidade é considerada uma epidemia mundial. Está associada a um estado de inflamação crônica e ativação do estresse do retículo endoplasmático (ERE), relacionados à patogênese de diversas doenças como diabetes mellitus tipo 2, doenças cardiovasculares, câncer, hipertensão, dentre outras. Nesse contexto, são necessários estudos para encontrar alternativas que melhorem o processo inflamatório. Vários estudos em humanos e animais já demostraram as propriedades anti-inflamatórias do ácido graxo ômega-3. O objetivo do nosso trabalho foi avaliar os efeitos da suplementação do ácido alfa-linolênico (ALA) no metabolismo e no estresse do retículo endoplasmático em obesos grau III. Foi conduzido um estudo prospectivo, randomizado, placebo-controlado, duplo-cego. No total, 52 pacientes foram randomizados para a suplementação com 3g/dia de ALA ou placebo, sendo 27 indivíduos do grupo ômega-3/ALA e 25 do grupo controle. Foi avaliado perfil lipídico, glicídico e inflamatório antes e após suplementação. O tecido adiposo visceral (TAV) foi coletado durante cirurgia bariátrica após suplementação. O perfil de ácidos graxos incorporados no TAV foi avaliado por cromatografia gasosa. Os genes foram avaliados através de PCR em tempo real. Não houve alteração nos níveis séricos de IL-6 (p=0,2201), TNF-? (0.7703) e PCR (p=0,57) após suplementação com ômega-3/ALA, porém observamos diminuição nos níveis séricos de Leptina (p=0.0154) e IP-10 (p=0.0410), citocinas inflamatórias, e aumento na IL-4 (p=0,0211), citocina anti-inflamatória. Foi observado incorporação significativa do ALA (p=0,0002), EPA (p<0,0001) e DHA (p=0,0005) no TAV. Avaliação molecular evidenciou um aumento da expressão gênica do XBP1 (p=0,0013), sXBP1 (p<0,0001), EIF2-? (p=0,0063) e da chaperona CCT4 (p=0,0001) e diminuição na expressão gênica da leptina (p=0,0410). Podemos concluir que o ALA pode modular o ERE através da via da IRE1/XBP, levando ao aumento das chaperonas (CCT4), o que pode demonstrar um potencial terapêutico do ALA em pacientes obesos. / Currently, obesity is considered a worldwide epidemic. It is associated with chronic inflammation and stress activation of the endoplasmic reticulum (ERE), related to the pathogenesis of various diseases such as type 2 diabetes mellitus, cardiovascular diseases, cancer, hypertension, among others. In this context, studies are needed to find alternatives that improve the inflammatory process. Several studies in humans and animals have already demonstrated the anti-inflammatory properties of omega-3 fatty acid. The objective of our study was to evaluate the effects of alpha-linolenic acid (ALA) supplementation on the metabolism and stress of the endoplasmic reticulum in obese patients. A prospective, randomized, placebo-controlled, double-blind study was conducted. In total, 52 patients were randomized to supplementation with 3 g / day of ALA or placebo, 27 individuals from the omega-3 / ALA group and 25 from the control group. Lipid, glycidic and inflammatory profile were evaluated before and after supplementation. Visceral adipose tissue (TAV) was collected during bariatric surgery after supplementation. The fatty acid profile incorporated in the TAV was evaluated by gas chromatography. Genes were evaluated by real-time PCR. There was no change in serum levels of IL-6 (p = 0.2201), TNF-? (0.7703) and CRP (p = 0.57) after supplementation with ALA, but we observed a decrease in serum Leptin levels (P = 0.0154) and IP-10 (p = 0.0410), inflammatory cytokines, and increase in IL-4 (p = 0.0211), anti-inflammatory cytokine. Significant incorporation of ALA (p = 0.0002), EPA (p <0.0001) and DHA (p = 0.0005) into the TAV was observed. Molecular evaluation showed an increase in the gene expression of XBP1 (p = 0.0013), sXBP1 (p <0.0001), EIF2-? (p = 0.0063), GADD34 (p=0,0117) and CCT4 chaperone (p = 0.0001), decrease in the gene expression of leptin (p = 0.0410) and ATF-6 (p=0,0305) and a tendency to decrease the gene expression of CHOP. We can conclude that ALA can modulate ERE through the IRE1 / XBP, PERK and ATF-6 pathways, leading to increased chaperones (CCT4), which may demonstrate a therapeutic potential of ALA in obese patients.