Influence of one selected Tisochrysis lutea strain rich in lipids on Crassostrea gigas larval development and biochemical composition

Effects of a remarkably high overall lipid Tisochrysis lutea strain (T+) upon gross biochemical composition, fatty acid (FA), sterol and lipid class composition of Crassostrea gigas larvae were evaluated and compared with a normal strain of Tisochrysis lutea (T) and the diatom Chaetoceros neogracile (Cg). In a first experiment, the influence of different single diets (T, T+ and Cg) and a bispecific diet (TCg) was studied, whereas, effects of monospecific diets (T and T+) and bispecific diets (TCg and T+Cg) were evaluated in a second experiment. The strain T+ was very rich in triglycerides (TAG: 93–95% of total neutral lipids), saturated FA (45%), monounsaturated FA (31–33%) and total fatty acids (4.0–4.7 pg cell−1). Larval oyster survival and growth rate were positively correlated with 18:1n-7 and 20:1n-7, in storage lipids (SL), and negatively related to 14:0, 18:1n-9, 20:1n-9, 20:4n-6 and trans-22-dehydrocholesterol in membrane lipids (ML). Surprisingly, only the essential fatty acid 20:5n-3 in SL was correlated positively with larval survival. Correlations suggest that physiological disruption by overabundance of TAG, FFA and certain fatty acids in larvae fed T+ was largely responsible for the poor performance of these larvae. ‘High-lipid’ strains of microalgae, without regard to qualitative lipid composition, do not always improve bivalve larval performance.

Allocation du carbone et métabolisme azoté chez l'haptophyte Tisochrysis lutea

The aim of this thesis is to improve knowledge on mechanisms involved in the response to nitrogen limitation and in lipid accumulation in the microalgae haptophyte Tisochrysis lutea. The wild type strain and a lipid accumulating mutant strain were grown under different nitrogen limitation and starvation and analyzed by functional genomics. Four genes of high-affinity nitrate/nitrite transporter (Nrt2) were identified and characterized to reveal the mechanisms involved in mineral absorption in this species. Transcriptomes of both strains were sequenced and proteins affected by nitrogen starvation and differentially expressed between the two strains were identified. We so identified the functions regulated by nitrogen deficiency and potentially involved in the accumulation of storage lipids. The responses of both strains to thin variations of nitrogen limitation were studied. The results of high-throughput proteomic analyzes suggest that the lipid-accumulation in the mutant strain is the result of carbon metabolism impacted overall, this spurred on signaling mechanisms. Two proteins have been studied since probably involved in carbon and nitrogen remobilization from amino acids catabolism during nitrogen limitation. This work increases knowledge on haptophytes, and brings assumptions on metabolic key involved in nitrogen limitation and carbon allocation in microalgae.

Exposure to toxic Alexandrium minutum activates the detoxifying and antioxidant systems in gills of the oyster Crassostrea gigas

Harmful algal blooms of Alexandrium spp. dinoflagellates regularly occur in French coastal waters contaminating shellfish. Studies have demonstrated that toxic Alexandrium spp. disrupt behavioural and physiological processes in marine filter-feeders, but molecular modifications triggered by phycotoxins are less well understood. This study analyzed the mRNA levels of 7 genes encoding antioxidant/detoxifying enzymes in gills of Pacific oysters (Crassostrea gigas) exposed to a cultured, toxic strain of A. minutum, a producer of paralytic shellfish toxins (PST) or fed Tisochrysis lutea (T. lutea, formerly Isochrysis sp., clone Tahitian (T. iso)), a non-toxic control diet, in four repeated experiments. Transcript levels of sigma-class glutathione S-transferase (GST), glutathione reductase (GR) and ferritin (Fer) were significantly higher in oysters exposed to A. minutum compared to oysters fed T. lutea. The detoxification pathway based upon glutathione (GSH)-conjugation of toxic compounds (phase II) is likely activated, and catalyzed by GST. This system appeared to be activated in gills probably for the detoxification of PST and/or extra-cellular compounds, produced by A. minutum. GST, GR and Fer can also contribute to antioxidant functions to prevent cellular damage from increased reactive oxygen species (ROS) originating either from A. minutum cells directly, from oyster hemocytes during immune response, or from other gill cells as by-products of detoxification.

Impact of recent coastal development and human activities on Nha Trang Bay, Vietnam : evidence from a Porites lutea geochemical record

Nha Trang Bay (NTB) is located on the Central Vietnam coast, western South China Sea. Recent coastal development of Nha Trang City has raised public concern over an increasing level of pollution within the bay and degradation of nearby coral reefs. In this study, multiple proxies (e.g., trace metals, rare earth elements (REEs), and Y/Ho) recorded in a massive Porites lutea coral colony were used to reconstruct changes in seawater conditions in the NTB from 1995 to 2009. A 14-year record of REEs and other trace metals revealed that the concentrations of terrestrial trace metals have increased dramatically in response to an increase in coastal development projects such as road, port, and resort constructions, port and river dredging, and dumping activities since 2000. The effects of such developmental processes are also evident in changes in REE patterns and Y/Ho ratios through time, suggesting that both parameters are critical proxies for marine pollution.

Steroidogenesis in the desensitized rat corpora-lutea

Earlier workers have observed that in the leydig cell desensitization brings results in addition to down regulation of receptors, in leisons in the steroidogenlc pathway. In the present study immature rats having heavily leutinized ovaries were given 50 iu hCG and the desenasitized CL removed 48h later were used. At that time no change in the 5 3MSD activity and CAMP binding activity(a measure of CAMP dependent protein kinase) was observed.Followlng desensitization however,l)a significent increase in phosphodiestrase activity,ii)a 50% reduction in total mitochondrial cholesterol level, iii)a significant reduction in its ability to utilize cholesterol or hydrolyse its ester and iv)a significant lowering(by 66%)in cholesterol side chain clean age activity(by measuring pregnanalone formed) was observed. Pregnanalone production was restored to normalcy if exogenous cholesterol was added to the mitohondrial preparation. The results suggest that luteal desensitization is due in addition to down regulation of LH receptors, to a marked reduction in available cholesterol pool in the mitochondrial compartment. The increase in phosphodiestrase activity, though probably a secondary effect,might effectively contribute to the overall reduction in the steroid out-put by increasing the catabolism of CAMP.(Aided by grants from ICMR,New Delhi and WHO, Geneva).

Ovarian expression of insulin-like peptide 3 (INSL3) and its receptor (RXFP2) during development of bovine antral follicles and corpora lutea and measurement of circulating INSL3 levels during synchronized estrous cycles

Insulin-like peptide 3 (INSL3), a major product of testicular Leydig cells, is also expressed by the ovary but its functional role remains poorly understood. Here, we quantified expression of INSL3 and its receptor RXFP2 in theca interna (TIC) and granulosa (GC) compartments of developing bovine antral follicles and in corpora lutea (CL). INSL3 and RXFP2 mRNA levels were much higher in TIC than GC and increased progressively during follicle maturation with INSL3 peaking in large (11-18mm) estrogen-active follicles and RXFP2 peaking in 9-10mm follicles before declining in larger (11-18mm) follicles. Expression of both INSL3 and RXFP2 in CL was much lower than in TIC. In situ hybridization and immunohistochemistry confirmed abundant expression of INSL3 mRNA and protein in TIC. These observations indicate follicular TIC rather than CL as the primary site of both INSL3 production and action, implying a predominantly auto-/paracrine role in TIC. To corroborate the above findings, we showed that in vitro exposure of TIC to a luteinizing concentration of LH greatly attenuated expression of both INSL3 and its receptor while increasing progesterone secretion and expression of STAR and CYP11A1. Moreover, in vivo, a significant cyclic variation in plasma INSL3 was observed during synchronized estrous cycles. INSL3 and estradiol-17β followed a similar pattern, both increasing after luteolysis, before falling sharply after the LH surge. Thus, theca-derived INSL3, likely from the dominant pre-ovulatory follicle, is detectable in peripheral blood of cattle and expression is down-regulated during luteinisation induced by the pre-ovulatory LH surge. Collectively, these findings underscore the likely role of INSL3 as an important intrafollicular modulator of TIC function/steroidogenesis, whilst raising doubts about its potential contribution to CL function.

Amino acid, Antioxidant and Ion Profiles of Carpolobia lutea Leaf (Polygalaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

ANTIMICROBIAL ACTIVITY of CARPOLOBIA LUTEA EXTRACTS and FRACTIONS

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Calor específico e densidade da polpa de cajá (Spondias lutea L.) com diferentes concentrações de sólidos solúveis sob baixas temperaturas

A polpa de cajá tem sido exportada da região Nordeste para todo o Brasil, e uma das maneiras de conservá-la é processando-a e armazenado-a na forma congelada. Durante o processamento, existem dois fatores que favorecem a qualidade do produto a ser armazenado: a rapidez com que o produto é congelado e a adição de açúcar como forma de dificultar a ação microbiológica. Na linha de processo, algumas propriedades termofísicas, como o calor específico e a densidade, se alteram com as temperaturas negativas e com o teor de sólidos solúveis totais; dessa forma, para o dimensionamento adequado de máquinas e equipamentos, torna-se necessário conhecer essas propriedades, principalmente quando se operam equipamentos criogênicos, onde as temperaturas podem chegar a -196 ºC. Neste trabalho, determinaram-se a densidade e o calor específico da polpa do cajá com quatro diferentes níveis de sólidos solúveis totais (natural da polpa; 20; 40 e 60%) e temperaturas entre -196 ºC e -18 ºC. A densidade foi obtida pela razão entre a massa da polpa e o seu volume. O volume foi determinado pelo método do deslocamento de massa (água) e o calor específico por meio de balanço de calor entre a massa da polpa, a massa do cilindro que contém a polpa e a massa do gelo. Conclui-se que a densidade da polpa de cajá aumenta em função da redução da temperatura e do aumento do teor de sólidos solúveis totais de 9 para 60 °Brix e que o calor específico da polpa diminui com a diminuição de temperatura e aumenta com a concentração de sólidos solúveis totais.

Antidiarrheal mechanism of Carpolobia lutea leaf fractions in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anti-nociceptive effects of Carpolobia lutea G. Don (Polygalaceae) leaf fractions in animal models

Leaves from Carpolobia lutea (Polygalaceae) were screened to establish the antiulcer ethnomedicinal claim and to quantitatively isolate, elucidate the active compounds by semi-preparative HPLC. The anti-nociceptive effects of Carpolobia lutea (CL) G. Don (Polygalaceae) organic leaf extracts were tested in experimental models in mice. The anti-nociceptive mechanism was determined using tail-flick test, acetic acid-induced abdominal constrictions, formalin-induced hind paw licking and the hot plate test. The fractions (ethanol, ethyl acetate, chloroform, n-hexane) and crude ethyl acetate extract of CL (770 mg/kg, i.p.) produced significant inhibitions of both phases of the formalin-induced pain in mice, a reduction in acetic acid-induced writhing as well as and an elevation of the pain threshold in the hot plate test in mice. The inhibitions were greater to those produced by indomethacin (5 mg/kg, i.p.). Ethyl acetate fraction revealed cinnamic and coumaric acids derivatives, which are described for the first time in literature. These cinnamalglucosides polyphenols characterised from CL may in part account for the pharmacological activities. These findings confirm its ethnomedical use in anti-inflammatory pain and in pains from gastric ulcer-associated symptoms. © 2011 Springer Basel AG.

Development of a topical formulation containing S. Lutea extract: Stability, in vitro studies and cutaneous permeation

Flavonoids have been widely incorporated into cosmetic and dermatological formulations, affording benefits such as antioxidant action, improved skin tone and fewer lines and wrinkles. Brazil has a huge biodiversity, with one of the richest flora in the world, and existing studies justify the quest for greater research efforts in this area. The cajazeira (Spondias lutea L.), a plant of the Anacardiaceae family from tropical America, is widely disseminated in Brazil. This plant was chosen because of the presence of flavonoids that exhibit antioxidant activity. The purpose of this research was to develop a stable topical formulation containing Spondias lutea extract with the aim of preventing skin diseases caused by UV radiation. Hydro ethanolic extract of Spondias lutea fruit was prepared and assayed for its the flavonoids content. The antioxidant activity was estimated by DPPH and superoxide assay. The physicochemical stability and skin permeation of the cream containing 8% (w/w) of extract were assessed. The results showed that the S. lutea extract possessed antioxidant activity, and that it is possible to obtain a stable cosmetic containing the extract, which is able to penetrate the skin. Thus, it is possible to use this extract to produce an anti-aging cosmetic.

Antiulcer effects of Ethyl acetate fraction of Carpolobia lutea leaf

Carpolobia lutea leaves (CLL) (Polygalaceae) were earlier screened and the antiulcer ethnomedicinal claim established. This article seeks to quantitatively isolate, elucidate the active compounds from most active CLL fraction. Fractionation was by semi-preparative HPLC; the active fraction was subjected to radical scavenging assays (RSA) and quantification of the total phenolic content (TPC) were also executed. Results: Ethyl acetate fraction (EAF) was observed to be the most pharmacologically active antiulcer fraction when screened using acute ulcer models induced in rodents. The EAF demonstrated significant (p < 0.05-0.001) antiulcer activity in various in vivo induced ulceration models by reducing the ulcer index and increasing the preventive ratio. The EAF demonstrated > 70% in TPC and < 20 % in RSA. Cinnamic and coumaric acids derivatives were isolated from EAF. Cinnamic acids have been implicated and patented as antiulcer agent. Isolated compounds could in part mediate the observed pharmacological activities which lend credence to its ethnobotanical uses.