Effects of the ITPR1 gene overexpression on Ca²⁺ concentration, lipid content and calcium transport-related genes in duck uterine epithelial cells / 生物工程学报

Inositol 1,4,5-trisphosphate receptor 1 (ITPR1) is an important intracellular channel for releasing Ca²⁺. In order to investigate the effects of the ITPR1 overexpression on Ca²⁺ concentration and lipid content in duck uterine epithelial cells and its effects on calcium transport-related genes, the structural domain of ITPR1 gene of duck was cloned into an eukaryotic expression vector and transfected into duck uterine epithelial cells. The overexpression of the ITPR1 gene, the concentration of Ca²⁺, the lipid content, and the expression of other 6 calcium transport-related genes was determined. The results showed that the concentration of Ca²⁺ in uterine epithelial cells was significantly reduced after transfection (P<0.05), the triglyceride content was significantly increased (P<0.01), and the high-density lipoprotein content was significantly decreased (P<0.01). The correlation analysis results showed that the overexpression of the C-terminal half of the ITPR1 gene was significantly positively correlated with the total cholesterol content (P<0.01), which was significantly positively correlated with the low-density lipoprotein content (P<0.05). The overexpression of the N-terminal half of the ITPR1 gene was significantly positively correlated with the triglyceride content (P<0.01), which was significantly negatively correlated with the concentration of Ca²⁺ (P<0.05). RT-qPCR results of 6 calcium transport-related genes showed that the overexpression of the C-terminal half of the ITPR1 gene significantly inhibited the expression of the IP3R2, VDAC2 and CAV1 genes, and the overexpression of the N-terminal half of the ITPR1 gene significantly promoted the expression of the IP3R3 and CACNA2D1 genes. In conclusion, the ITPR1 gene overexpression can promote Ca²⁺ release in duck uterus epithelial cells, promote the synthesis of triglyceride, low-density lipoprotein and cholesterol, and inhibit the production of high-density lipoprotein, and the ITPR1 gene overexpression affected the expression of all 6 calcium transport-related genes.

Lipid content and cryotolerance of in vitro-produced bovine embryos treated with forskolin before vitrification / Acúmulo de lipídios intracitoplasmáticos, desenvolvimento e criotolerância de embriões bovinos produzidos in vitro e tratados com diferentes concentrações de forskolin antes da vitrificação

The aim of the present study was to evaluate the intracytoplasmic lipid content, development and cryotolerance of in vitro-produced bovine embryos treated with different concentrations of forskolin before vitrification. Embryos were produced from abattoir-derived ovaries and allocated into four groups. In the treatment groups, forskolin was added to the in vitro culture medium on Day 6 and incubated for 24 hours in one of the following concentrations: 2.5µM (Forsk 2.5 group), 5.0µM (Forsk 5.0 group) or 10.0µM (Forsk 10.0 group). Embryos from the control group were cultured without forskolin. On Day 7 of culture, the expanded blastocysts were stained with the lipophilic dye Sudan Black B for determination of the intracytoplasmic lipid content or were cryopreserved via the Vitri-Ingá® procedure. Although there were no significant differences (P>0.05) in the blastocyst rates between the Control group (44.9%) and the other treatments, the embryo production was lower (P<0.05) in Forsk 10.0 group (38.8%) compared to Forsk 2.5 (50.5%) and Forsk 5.0 (54.7%) groups. The intracytoplasmic lipid content (expressed in arbitrary units of pixels) in blastocysts from the Control group (1.00±0.03) was similar (P>0.05) to that found in Forsk 2.5 (0.92±0.03) and Forsk 10.0 groups (1.06±0.03) groups; however the lipid accumulation in blastocysts from Forsk 5.0 group (0.82±0.04) was lower than in the Control group (P<0.05). Based on these results, Forsk 5.0 treatment was tested for cryotolerance and it was observed that the blastocoel re-expansion rate evaluated 24 hours after warming was greater (P<0.05) in Forsk 5.0 group (72.2%) compared to the Control group (46.2%). In conclusion, pre-treatment with forskolin at a concentration of 5.0 µM for 24 hours before vitrification is effective in reducing the intracytoplasmic lipid content and, consequently, improves cryotolerance of IVP bovine embryos.(AU)

Os embriões foram produzidos a partir de ovários obtidos em abatedouro e foram alocados em quatro grupos experimentais. Nos grupos tratados, o forskolin foi adicionado ao meio de cultivo in vitro no dia 6 do cultivo e os embriões foram incubados durante 24 horas com uma das seguintes concentrações: 2,5µM (grupo Forsk 2,5), 5,0µM (grupo Forsk 5,0) ou 10,0µM (grupo Forsk 10,0). Os embriões do grupo controle foram cultivados na ausência de forskolin. No dia 7 do cultivo, os blastocistos expandidos foram corados com o corante lipofílico Sudan Black B para a determinação do teor de lípidos intracitoplasmáticos ou foram criopreservados através do protocolo Vitri-Ingá®. Não foi observada diferença significativa (P>0,05) na taxa de produção de blastocistos entre o grupo Controle (44,9%) e os demais tratamentos, todavia observou-se menor produção de embriões (P<0,05) no grupo Forsk 10,0 (38,8%) em comparação com os grupos Forsk 2,5 (50,5%) e Forsk 5,0 (54,7%). A quantidade de lipídeos intracitoplasmáticos do grupo Controle (1,00±0,03) foi semelhante (P>0,05) a dos grupos Forsk 2,5 (0,92±0,03) e Forsk 10,0 (1,06±0,03); no entanto, o acúmulo de lípidos nos blastocistos do grupo Forsk 5.0 (0,82 ± 0,04) foi menor do que no grupo controle (P<0,05). A partir destes resultados, o grupo Forsk 5,0 foi testado quanto à criotolerância e foi observado que a taxa de re-expansão da blastocele 24 horas após o aquecimento foi maior (P<0,05) no grupo Forsk 5,0 (72,2%) quando comparado ao grupo Controle (46,2%). Em conclusão, o pré-tratamento com forskolin na concentração de 5,0 µM durante 24 horas antes da vitrificação foi eficiente para promover a redução da quantidade de lipídeos intracitoplasmáticos e, consequentemente, melhorou a criotolerância de embriões bovinos produzidos in vitro.(AU)

Chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted by emulsified canola oil / Composição química, propriedades microbiológicas e perfil de ácidos graxos de Salame tipo Italiano com substituição da gordura animal por óleo de canola emulsionado

ABSTRACT: Vegetable oils have been used to substitute pork backfat to improve the fatty acid profile of fermented sausages. The aim of this study was to assess the chemical composition, microbiological properties, and fatty acid profile of Italian-type salami with pork backfat substituted (15% and 30%) by emulsified canola oil. Fat contents decreased while moisture contents increased in Italian-type salami with emulsified canola oil. The growth of lactic acid bacteria in salami was not affected by canola oil and absence of fecal coliforms, coagulase-positive staphylococci, and Salmonella were reported during processing of fermented sausages. Lower levels of saturated fatty acids (SAFAs), higher levels of monounsaturated fatty acids (MUFAs) and polyunsaturated fatty acids (PUFAs) were observed in salami with emulsified canola oil. Together, our results indicated that substituting pork backfat with emulsified canola oil improved the nutritional values of Italian-type salami.

RESUMO: O uso de óleos vegetais como substitutos da gordura animal em embutidos fermentados tem surgido como uma alternativa de produzir produtos cárneos com um melhor perfil lipídico. Assim, o objetivo deste trabalho foi avaliar a composição química, propriedades microbiológicas e o perfil de ácidos graxos de salame tipo Italiano elaborado com a substituição parcial da gordura suína (15 e 30%) por emulsão com óleo de canola. A incorporação do óleo de canola reduziu o teor de gordura e aumentou o teor de umidade dos embutidos fermentados. Em relação às características microbiológicas, a adição de óleo de canola emulsionado não alterou o crescimento das bactérias láticas durante o processamento. Ainda, não foram detectadas contagens de coliformes fecais, staphylococci coagulase positiva e Salmonella. Em relação ao perfil de ácidos graxos, foi possível observar a redução nos teores totais de ácidos graxos saturados (SAFAs) e uma elevação nos valores de ácidos graxos monoinsaturados (MUFAs) e poliinsaturados (PUFAs) nos tratamentos adicionados de óleo de canola. Dessa forma, a substituição da gordura suína por óleo de canola emulsionado em salame tipo Italiano torna-se uma alternativa interessante na produção de produtos cárneos com melhores características nutricionais.

Effects of Sodium Nitrate and Mixotrophic Culture on Biomass and Lipid Production in Hypersaline Microalgae Dunaliella Viridis Teod

To access the potential application of Dunaliella viridis Teod. for biofuel production, the effects of culture media composition on biomass and lipid content of this microalgae were investigated. Measured at the 20 th day, sodium nitrate at 5.0 mM augmented biomass production by 26.5 percent compared to control (1 mM sodium nitrate). Total lipids expressed as µg mL-1 of culture also increased with increase in nitrate concentration up to 5.0 mM sodium nitrate, whereas when expressed on the per cell basis, total lipids stayed relatively constant at most of the tested nitrate concentrations except at 0.5 mM which was 31.4 percent higher compared to 1.0 mM nitrate. At 5.0 mM sodium nitrate, by using 20 g L-1 of glucose in mixotrophic culture of D. viridis, cell number augmented by 36.4 percent compared to the cultures with no added glucose. Llipid content per cell and per mL of culture was increased by 71.4 and 135.1 percent, respectively. Among plant hormones, 10-9 M indole-3- acetic acid (IAA) plus 10 -8 M trans-zeatin riboside led to 22.8 percent higher biomass relative to control (without hormone and at 1.0 mM sodium nitrate). It is concluded that altering the growth conditions of D. viridis can lead to higher cell densities and higher lipids content which can be exploited for biofuel production.

Cultivation of Scenedesmus dimorphus for C/N/P removal and lipid production

Background CO2 emission, water pollution and petroleum shortage are the issues coming with the development of industry. A cost effective system was constructed to fix the CO2 in flue gas (15% CO2), remove nitrogen and phosphorus from manure wastewater and produce biofuels at the same time. The significant cultivation conditions were selected by Plackett-Burman design, and then optimized with central composite design. Results Optimum culture condition was predicted at light intensity of 238 µmol·m- 2·s- 1, TN of 152 mg·L- 1, inoculum density of 0.3 g·L- 1, under which the measured CO2 fixation rate, total nitrogen and phosphorus removing rate, and lipid content were 638.13 mg·L- 1·d- 1, 88.16%, 73.98% and 11.9%, respectively. The lipid content was then enhanced to 24.2% by a nitrogen starvation strategy. Conclusion A cultivation strategy was suggested to achieve effective C/N/P removal from flue gas and manure wastewater, and meanwhile obtained high lipid content from microalgal biomass.

Depositional characteristics of atmospheric polybrominated diphenyl ethers on tree barks

OBJECTIVES: This study was conducted to determine the depositional characteristics of several tree barks, including Ginkgo (Ginkgo biloba), Pine (Pinus densiflora), Platanus (Platanus), and Metasequoia (Metasequoia glyptostroboides). These were used as passive air sampler (PAS) of atmospheric polybrominated diphenyl ethers (PBDEs). METHODS: Tree barks were sampled from the same site. PBDEs were analyzed by highresolution gas chromatography/high-resolution mass spectrometer, and the lipid content was measured using the gravimetric method by n-hexane extraction. RESULTS: Gingko contained the highest lipid content (7.82 mg/g dry), whereas pine (4.85 mg/g dry), Platanus (3.61 mg/g dry), and Metasequoia (0.97 mg/g dry) had relatively lower content. The highest total PBDEs concentration was observed in Metasequoia (83,159.0 pg/g dry), followed by Ginkgo (53,538.4 pg/g dry), Pine (20,266.4 pg/g dry), and Platanus (12,572.0 pg/g dry). There were poor correlations between lipid content and total PBDE concentrations in tree barks (R2=0.1011, p =0.682). Among the PBDE congeners, BDE 206, 207 and 209 were highly brominated PBDEs that are sorbed to particulates in ambient air, which accounted for 90.5% (84.3-95.6%) of the concentration and were therefore identified as the main PBDE congener. The concentrations of particulate PBDEs deposited on tree barks were dependent on morphological characteristics such as surface area or roughness of barks. CONCLUSIONS: Therefore, when using the tree barks as the PAS of the atmospheric PBDEs, samples belonging to same tree species should be collected to reduce errors and to obtain reliable data.

Conteúdo energético das operárias da formiga cortadeira, Atta sexdens / Workers energy content in leaf-cutting ants, Atta sexdens

O conhecimento do conteúdo energético das operárias é essencial para mensurar a disponibilidade energética, responsável pela manutenção e execução de atividades essenciais para o crescimento da colônia de formigas. Mas pouco se conhece sobre o conteúdo energético das operárias e quanto estará disponível para realizar atividades de, por exemplo, escavar do ninho. O presente estudo determinou o teor de lipídeo e o conteúdo energético das operárias de formigas cortadeiras inativas, antes e pós atividade de escavação. Por meio da determinação do teor de lipídeos, pode-se calcular o conteúdo energético das operárias em repouso (paradas) e compará-las com as que escavaram. O teor de lipídeo e conteúdo energético das operárias foram em média 9,1±0,8% e 111,31±54,71J, respectivamente, entretanto, as séries experimentais não diferiram significativamente. Adicionalmente, a taxa do fluxo catabólico, embasado na massa corporal das operárias foi de 14,76±10,11µW. Conclui-se que o recurso energético para a escavação do ninho não é proveniente de reserva de lipídeos corporal e, dessa forma, o conteúdo energético das operárias não alterou antes e pós-atividade de escavação.

The knowledge of the workers energy content is essential to measure the energy availability for maintenance and performance of activities essences for the colony growth. But little is known about the workers energy content, and how much will be available to carry out activities, for example, the excavation of the nest. The present study determined the lipid content and energy content of the worker ants inactive before and after excavation activity. Through the lipids determination, it can be calculated the energy content of workers resting (standing) with those which excavated. The lipid content and energy content of the workers were on average 9.1±0.8% and energy content of the workers was on average 111.31±54.71J, respectively, however, the experimental series did not differ significantly. Additionally, catabolic flux rate based in workers body mass was 14.76±10.11µW. It was concluded that the energy resource for the excavation of the nest is not coming from reserve lipid body, and thus the energy content of the workers did not change before and after excavation activity.

Esforço de escavação e teor de lipídios em rainhas da formiga cortadeira Atta sexdens rubropilosa / Digging effort and lipid content in leaf-cutting ant queens Atta sexdens rubropilosa

As formigas cortadeiras, importantes pragas agrícolas, anualmente realizam a fundação de novas colônias. Para o início de novas colônias, é preciso a escavação de túneis e de câmaras no solo, sendo que, para isso, as rainhas recém fecundadas gastam muita energia, no entanto a quantificação dessa energia gasta é desconhecida. Nossa hipótese é que a reserva lipídica, armazenada no corpo da rainha, seja utilizada durante a escavação do ninho. Testamos essa hipótese comparando o aumento do esforço de escavação das rainhas, as quais foram experimentalmente estimuladas a escavar uma, duas ou três vezes consecutivas, em relação ao controle, ou seja, em relação às rainhas que não escavaram. Foram quantificados a massa corporal e o teor de lipídios das rainhas. Os resultados mostraram que, ao contrário de nossas expectativas iniciais, massa e teor de lípideos não foram afetadas pelo aumento do esforço de escavação do ninho. Conclui-se que a escavação pelas rainhas não afetou os percentuais de lipídios em seu corpo, consequentemente, o conteúdo de energia. Provavelmente, os recursos energéticos para a escavação não se originam de fontes lipídicas, mas de outros, provavelmente carboidratos.

Nest digging spends a lot of energy existing a question about how much energy is required to dig the tunnel and initial chamber. It was hypothesized that the lipid content is used during nest digging. This hypothesis was tested by comparing the consequences of increasing digging effort in queens that were experimentally stimulated to excavate a complete founding nest either once, twice or three times consecutively, compared to control queens that didn't dig. Weight and lipid content of queens were quantified. Results showed that, in contrast with the initial expectations, weight and lipid content were not affected by the increased digging effort in the experimentally-induced successive excavations. It was conclude that the excavation by the queens did not affect the percentages of lipids and consequently the energy content, in their bodies. Probably, energy resources for the excavation wasn't originated from lipid reserves, but from other energy sources, perhaps carbohydrates.

Lipid and energy contents in the bodies of queens of Atta sexdens rubropilosa Forel (Hymenoptera, Formicidae): pre-and post-nuptial flight / Conteúdo de lipídeos e energético das rainhas de Atta sexdens rubropilosa Forel (Hymenoptera, Formicidae): pré e pós-vôo nupcial

The nuptial flight allows males and females to meet and copulate and both need energy to perform this activity. Before leaving the nest, males and females are well nourished and ready to mate. However, little is known about the lipid and energy contents in females before the nuptial flight (virgins) and after it (mated females). In this work we measured lipid concentrations in relation to body weight in these individuals. Our results showed that 16.82% of the bodies of young virgin females one month before mating flight are composed of lipids, contrasting with the 32.62% lipid content in mated females who had not excavated their nest yet, and 32.88% in those who had. The energy content measured for virgin females was 2942.63 J, contrasting with 6110.01 J for queens before excavating the nest and 5677.51 J after excavation. Based on our results, we conclude that the body mass, and therefore the lipid and energy contents in the bodies of Atta sexdens rubropilosa queens double during the last month before the nuptial flight. This energy resource is fundamental to the activities required during the nuptial flight, digging the nest and the founding of the colony.

O vôo nupcial permite que machos e fêmeas se encontrem e copulem. Para isso, tanto o macho quanto a fêmea precisam de energia para efetuar tal atividade. Antes de partir do ninho de origem, machos e fêmeas estão bem nutridos e prontos para o acasalamento. Mas pouco se conhece sobre o teor de lipídeos e conteúdo energético pré-vôo nupcial (fêmeas virgens) e pós-vôo nupcial (fêmeas acasaladas). Nossos resultados indicam que as fêmeas virgens apresentaram 16,82% de lipídeos em seu corpo, enquanto as rainhas (fêmeas acasaladas) que não escavaram 32,62%, e as que escavaram 32,88%. O conteúdo energético foi de 2942,63 J para as fêmeas virgens, 6110,01 J para rainhas sem escavação e 5677,51 J para aquelas que escavaram. Com base nos resultados obtidos, concluí-se que as rainhas de Atta sexdens rubropilosa dobram a sua massa corporal, e consequentemente, o teor de lipídeos e conteúdo energético, um mês antes da revoada. Esse recurso energético é fundamental para as atividades exigidas durante o vôo nupcial, escavação do ninho e a fundação da colônia.

Maintenance of filtering molluscs in aquaria for sub-chronic studies

This work determined the best survival conditions for the clam Mytella guyanensis and the mussel Perna perna in the estuary and sea aquaria respectively over at least 12 days, which could enable their use in the ecotoxicological studies. The aquaria were set up with the appropriate water and sedimen, and allowed to establish for a minimum of one month before adding the organisms. The best survival conditions for M. guyanensis required more time for the aquarium stabilization, addition of inocula and more frequent water changes than for P. perna. The organisms' lipid contents increased and their condition index was maintained indicating the good conditions of the aquaria, hence, their possible use in the sub-chronic studies.

Estudos sobre a dinâmica de contaminação de organismos marinhos devem ser feitos sob condições controladas pelo tempo necessário para que os organismos possam responder à presença do agente contaminante. No entanto, a manutenção de organismos em aquários por determinado período pode ser difícil porque todas as outras variáveis do ambiente precisam ser próximas às condições naturais. Este trabalho determinou as melhores condições de sobrevivência do marisco Mytella guyanensis e do mexilhão Perna perna, respectivamente em aquários de estuário e de mar, por período de até 12 dias. Os aquários foram montados com água e sedimento de estuário ou de mar e estabilizados por, no mínimo, um mês antes da colocação dos respectivos organismos. As melhores condições de sobrevivência de M. guyanensis requisitaram mais tempo de estabilização do aquário, adição de inóculos e trocas de água mais freqüentes do que para os P. perna. Os conteúdos de lipídios aumentaram com o tempo e o índice de condição dos organismos foi mantido, indicando as boas condições dos aquários e, conseqüentemente, a possibilidade de uso em pesquisas ecotoxicológicas.

Effect of Young Barley Leaf on Lipid Contents and Hepatic Lipid-Regulating Enzyme Activities in Mice Fed High-Fat Diet

This study was conducted to investigate the effects of powdered young barley leaf and its water extract on body weight and lipid metabolism in high-fat fed mice. Male mice were divided into normal group, high-fat (HF) group, highfat group supplemented with powdered young barley leaf (HF-YBL) and high-fat group supplemented with water extract of the powdered young barley leaf (HF-WYBL). The powdered young barley leaf or its water extract was added to a standard diet based on 1% dried young barley leaf (1 g YBL/100 diet and 0.28 g WYBL/100 g diet) for 8 weeks. Supplementation of YBL and WYBL significantly reduced body weight and epididymal adipose tissue weight in highfat fed mice. Food intake and daily energy intake were significantly lower in the YBL group than in the HF group. After 8 weeks, plasma triglyceride and cholesterol concentrations were significantly higher in the HF group than in the Normal group; however, both YBL and WYBL significantly lowered those of the high-fat fed mice. The ratio of HDL-cholesterol/ total cholesterol of the YBL and WYBL groups were significantly elevated compared to that of HF group. Both YBL and WYBL significantly increased fecal excretion of triglyceride in high-fat fed mice, whereas they did not affect fecal cholesterol concentration. The triglyceride levels of liver, adipose tissue and heart were significantly lower in the YBL and WYBL groups than in the HF group. Supplementation of WYBL also lowered the kidney triglyceride and heart cholesterol concentrations compared to those of HF group. Hepatic lipid regulating enzyme activities, fatty acid synthase, HMG-CoA reductase and acyl-coenzyme A: cholesterol acyltransferase, were significantly lower in the YBL and WYBL groups than in the HF group. Accordingly, these results suggest that YBL and WYBL improve plasma and organ lipid levels partly by increasing fecal lipid excretion and inhibiting fatty acid and cholesterol biosynthesis in the liver.