Epicardial fat volume is associated with coronary microvascular response in healthy subjects: a pilot study

Epicardial fat (EF) is an active ectopic fat depot, which has been associated with coronary atherosclerosis, and which could early influence endothelial function. We thus investigated the relationship between EF and endothelium-dependent vasoreactivity of the coronary microcirculation, in highly selected healthy volunteers. Myocardial blood flow (MBF) was determined by measuring coronary sinus flow with velocity-encoded cine magnetic resonance imaging (MRI) at 3T. We measured MBF at baseline and in response to sympathetic stimulation by cold pressor testing (CPT) in 30 healthy volunteers with normal left ventricular (LV) function (age 22 ± 4 years, BMI = 21.3 ± 2.8 kg/m(2)). EF volume was volumetrically assessed by manual delineation on short-axis views. CPT was applied by immersing one foot in ice water for 4 min. Mean EF volume was 56 ± 26 ml and mean LV mass 100 ± 28 g. CPT significantly increased heart rate (HR) by 32 ± 19%, systolic blood pressure by 14 ± 10%, and rate-pressure product by 45 ± 25%, P < 0.0001. The increase in HR, reflecting sympathetic stimulation, was not influenced by sex, age or EF volume. CPT induced a decrease in coronary vascular resistance (135 ± 72 vs. 100 ± 42 mm Hg.ml(-1).min.g, P = 0.0006), and a significant increase in MBF (0.81 ± 0.37 vs. 1.24 ± 0.56 ml.min(-1).g(-1), P < 0.0001). Interestingly, we found a significant negative correlation between EF volume and ΔMBF (r= - 0.40, P = 0.03), which remained significant after adjusting for ΔHR. ΔMBF was also associated with adiponectin (r = 0.41, P = 0.046), but not with waist circumference, BMI, C-reactive protein, lipid or glycemic parameters. In multivariate analysis, adiponectin and EF volume remained both independently associated with ΔMBF. A high EF amount is associated with a lower coronary microvascular response, suggesting that EF could early influence endothelial function.     

The effects of water temperature and hormone treatments on circulating LH and ovulation in tench (Tinca tinca)

The effectiveness of three hormone treatments commonly used for artificial reproduction of tench was evaluated under three thermal regimes, with focus on the need for dopamine antagonist. Mature females were divided into nine groups (n?=?8) and gradually exposed, over a 24?h period, to three temperature regimes: cold (18.1?±?0.02?°C), optimal (22.05?±?0.03?°C), and warm (26.3?±?0.01?°C). Each temperature regime comprised three experimental groups injected with one of three hormone treatments: carp pituitary extract (CPE; 3?mg?kg^?1); [D-Arg⁶, Pro⁹, NEt]-sGnRH (10???g?kg^?1); and [D-Arg⁶, Pro⁹, NEt]-sGnRH (10???g?kg^?1)?+?metoclopramide (20?mg?kg^?1) (combined treatment). No differences were found between ovulation induction (ovulation rate????75?%) with sGnRHa alone and with the combined treatment; whereas CPE at cold and warm water temperatures was significantly less effective (P?<?0.05) than above mentioned treatments. Administration of sGnRHa alone induced a gradual increase in luteinizing hormone (LH) levels with LH peaks close to ovulation, in contrast to the immediate LH surge with high LH levels throughout the entire study observed with the combined treatment under all thermal regimes. LH levels induced by GnRHa alone were significantly lower (P?<?0.05) at all temperatures compared to the combined treatment, with the exception of the final sample at 26?°C, when no difference was recorded. Based on these results we recommend the application of sGnRHa without the addition of dopamine antagonist as a reliable method for inducing ovulation in tench under suboptimal temperature conditions.     

Effect of onion-type multilamellar liposomes on Trametes versicolor laccase activity and stability

Trametes versicolor laccase was encapsulated into onion-type, lipid-based multilamellar vesicles (MLVs). When encapsulated, laccase was isolated from the assay medium but was still active once freed from its capsule. The encapsulation efficiency was larger than 65% at 25 °C and 37 °C and decreased to 55% by introducing 140 mM NaCl into the buffered medium (pH = 4.5). MLVs were shown to drastically improve both laccase stability and activity. At 25 °C, laccase activity was doubled in the presence of MLVs. At 37 °C in the salt-free medium, the half-life time of laccase was increased from 2hr 30-65 h without and with MLVs, respectively. This effect was even more pronounced in the salted medium where laccase activity was unchanged for 6 days in the presence of MLVs. These beneficial effects were attributed to the immobilization of laccase onto MLV surface. Laccase activity as well as stability was notably shown to be directly correlated to MLV stability.     

Comparative content of some bioactive compounds in apples,peaches and pears and their influence on lipids and antioxidant capacity in rats

The aim of this study was to compare some bioactive compounds in apples, peaches and pears and their influence on lipids and antioxidant capacity in rats. The content of total polyphenols (g/100g) was 0.23 +/- 0.03; 0.22 +/- 0.03 and 0.68 +/- 0.1 in peeled fruits and 0.48 +/- 0.04, 0.47 +/- 0.04 and 1.2 +/- 0.12 in peels of peaches, pears and apples, respectively. Caffeic, p-coumaric and ferulic acids and the total radical-trapping antioxidative potential (TRAP) values in peeled apples and their peels were significantly higher than in peaches and pears, respectively. Contrarary, no significant differences in the content of dietary fiber among the studied fruits were found. The content of all studied indices in peels was significantly higher than peeled fruits (p < 0.05 ). A good correlation between the total polyphenols and the TRAP values was found in all fruits. Diets supplemented with apples and to a less extent with peaches and pears have improved lipid metabolism and increased the plasma antioxidant potential especially in rats fed with added cholesterol. The highest content of biologically active compounds and the best results in the experiment on rats makes apple preferable for dietary prevention of atherosclerosis and other diseases.     

Comparison of the contents of the main biochemical compounds and the antioxidant activity of some Spanish olive oils as determined by four different radical scavenging tests

The aim of this study was to compare the contents of the main biochemical compounds and the antioxidant capacity of five Spanish olive oils by four different antioxidant tests and to find out the most valuable oil for disease preventing diets. Fatty acids, sterols and individual antioxidant compounds in Arbequina, Hojiblanca, Extra Virgin, Picual and Lampante Spanish olive oils were determined. Antioxidant activities were done as well using different radical scavenging activities: total radical-trapping antioxidative potential by ABAP (TRAP-ABAP), radical scavenging activity by DPPH (RSA-DPPH), antioxidant assay by beta-carotene-linoleate model system (AA-beta-carotene) and total antioxidant status by ABTS (TAA-ABTS). The highest content of all studied antioxidant compounds (353; 329; 4.6 and 2.7 mg/kg for tocopherols, tocotrienols, polyphenols and o-diphenols, respectively) was found in Extra Virgin oil. Also the highest antioxidant capacity was observed in Extra Virgin oil (668 nM/ml; 29.4%; 40.4% and 2.64 mM TE/kg for TRAP-ABAP, RSA-DPPH, AA- beta-carotene and TAA-ABTS, respectively). The correlation between total phenols and antioxidant capacities measured by four methods was very high, but the highest for the beta-carotene (R = 0.9958). In conclusion, the best method for determination of the antioxidant capacity of olive oils is the beta-carotene test. Extra Virgin olive oil has high organoleptic properties and the highest antioxidant activity. The above-mentioned makes this oil a preferable choice for diseases preventing diets.     

A role for the lymphotoxin/LIGHT axis in the pathogenesis of murine collagen-induced arthritis

A lymphotoxin-beta (LTbeta) receptor-Ig fusion protein (LTbetaR-Ig) was used to evaluate the importance of the lymphotoxin/LIGHT axis in the development and perpetuation of arthritis. Prophylactic treatment with the inhibitor protein LTbetaR-Ig blocked the induction of collagen-induced arthritis in mice and adjuvant arthritis in Lewis rats. Treatment of mice with established collagen-induced arthritis reduced the severity of arthritic symptoms and joint tissue damage. However, in a passive model of anti-collagen Ab-triggered arthritis, joint inflammation was not affected by LTbetaR-Ig treatment precluding LT/LIGHT involvement in the very terminal immune complex/complement/FcR-mediated effector phase. Collagen-II and Mycobacterium-specific T cell responses were not impaired, yet there was evidence that the overall response to the mycobacterium was blunted. Serum titers of anti-collagen-II Abs were reduced especially during the late phase of disease. Treatment with LTbetaR-Ig ablated follicular dendritic cell networks in the draining lymph nodes, suggesting that impaired class switching and affinity maturation may have led to a decreased level of pathological autoantibodies. These data are consistent with a model in which the LT/LIGHT axis controls microenvironments in the draining lymph nodes. These environments are critical in shaping the adjuvant-driven initiating events that impact the subsequent quality of the anti-collagen response in the later phases. Consequently, blockade of the LT/LIGHT axis may represent a novel approach to the treatment of autoimmune diseases such as rheumatoid arthritis that involve both T cell and Ab components.     

The effect of (1-->3)-beta-D-glucans, carboxymethylglucan and schizophyllan on human leukocytes in vitro

(1-->3)-beta-D-glucans are known as potent inductors of humoral and cell-mediated immunity in humans and animals. (1-->3)-beta-D-glucans isolated from various sources differ in their chemical structure and physical parameters and consequently in their immunomodulatory potential. In this study the immunomodulatory activity of two (1-->3)-beta-D-glucans schizophyllan (SPG) and carboxymethylglucan (CMG) was determined and compared on human blood leukocytes in vitro. Both SPG and CMG activated blood phagocytes and lymphocytes as demonstrated by increased whole blood production of reactive oxygen species, by increased production of pro-inflammatory cytokines IL-6, IL-8, and TNF-alpha, by increased surface expression of CD69 on lymphocytes, and by altered expression of CD11b and CD62L on polymorphonuclear leukocytes and monocytes. SPG demonstrated a significantly higher potential to stimulate blood phagocytes and production of selected pro-inflammatory cytokines than CMG. The higher potency of SPG to stimulate human blood phagocytes in vitro could be caused by factors such as higher branching frequencies or neutral polymer charge of SPG or different conformation in solution if compared with CMG.     

The extracellular matrix of porcine mature oocytes: Origin,composition and presumptive roles

The extracellular matrix (ECM) of porcine mature oocytes was revealed by transmission electron microscopy (TEM) after treatment with tannic acid and ruthenium red. Present in the perivitelline space (PVS) and on the surface of the zona pellucida (ZP), it appeared to be composed of thin filaments and granules at the interconnections of the filaments, which were interpreted respectively as hyaluronic acid chains and bound proteoglycans. In order to determine whether this material is produced by the corona cells (the same ECM was found also on the surface of the zona pellucida and between cumulus cells) or by the oocyte itself, the synthesis of glycoproteins and glycosaminoglycans was checked by autoradiography on semi-thin and thin sections observed by light and electron microscopy. Immature oocytes within or without cumulus cells, were incubated with L [3H-] fucose or L [3H-] glucosamine – precursors respectively of glycoproteins and hyaluronic acid or hyaluronan (HA) bound to proteoglycans – for various times (with or without chase) and at different stages during in vitro maturation. In the first case, incorporation was found in both cumulus cells and ooplasm (notably in the Golgi area for 3H-fucose) and labeled material accumulated in the ECM of the PVS and of the ZP surface. Labeling in the PVS with both precursors was maximum between metaphase I (MI) and metaphase II (MII) and was partially extracted by hyaluronidase but not by neuraminidase. Tunicamycin, an inhibitor of glycoprotein synthesis, significantly decreased the amount of ³H-fucose labeled molecules in the PVS and increased the incidence of polyspermic penetration during subsequent in vivo fertilization. Since cumulus-free oocytes also secreted ³H-glucosamine containing compounds, both oocyte and cumulus cells probably contribute to the production of the ECM found in the PVS of mature oocytes. ECM and particularly its HA moiety present on both sides of the ZP may constitute a favourable factor for sperm penetration.     

Antioxidative properties of Jaffa sweeties and grapefruit and their influence on lipid metabolism and plasma antioxidative potential in rats

The effective substances (polyphenols, phenolic and ascorbic acids, flavonoids and dietary fibers) and antioxidative activities, using different radical-scavenging tests, were determined for Jaffa sweeties and grapefruit. The antioxidative activities comprised the contributions from polyphenols, phenolic acids, flavonoids and ascorbate components, and were well-correlated with polyphenols and flavonoids. The correlation coefficient between the polyphenols and antioxidative activity varied from 0.73 to 0.99. All applied methods showed that sweeties had higher antioxidative activity than grapefruit. Experiments on laboratory animals show that diets supplemented with sweeties, and to a lesser extent with grapefruit, increased the plasma antioxidative potential and improved the lipid metabolism, especially in the rats fed with added cholesterol. These findings provide additional characterization of the nutritional value of citrus fruits and their influence on the lipid metabolism in rats.     

The Xenopus Xmus101 protein is required for the recruitment of Cdc45 to origins of DNA replication

The initiation of eukaryotic DNA replication involves origin recruitment and activation of the MCM2-7 complex, the putative replicative helicase. Mini-chromosome maintenance (MCM)2-7 recruitment to origins in G1 requires origin recognition complex (ORC), Cdt1, and Cdc6, and activation at G1/S requires MCM10 and the protein kinases Cdc7 and S-Cdk, which together recruit Cdc45, a putative MCM2-7 cofactor required for origin unwinding. Here, we show that the Xenopus BRCA1 COOH terminus repeat-containing Xmus101 protein is required for loading of Cdc45 onto the origin. Xmus101 chromatin association is dependent on ORC, and independent of S-Cdk and MCM2-7. These results define a new factor that is required for Cdc45 loading. Additionally, these findings indicate that the initiation complex assembly pathway bifurcates early, after ORC association with the origin, and that two parallel pathways, one controlled by MCM2-7, and the other by Xmus101, cooperate to load Cdc45 onto the origin.