Enhancement in liver SREBP-1c/PPAR-α ratio and steatosis in obese patients: Correlations with insulin resistance and n-3 long-chain polyunsaturated fatty acid depletion

Sterol receptor element-binding protein-1c (SREBP-1c) and peroxisome proliferator-activated receptor-α (PPAR-α) mRNA expression was assessed in liver as signaling mechanisms associated with steatosis in obese patients. Liver SREBP-1c and PPAR-α mRNA (RT-PCR), fatty acid synthase (FAS) and carnitine palmitoyltransferase-1a (CPT-1a) mRNA (real-time RT-PCR), and n-3 long-chain polyunsaturated fatty acid (LCPUFA)(GLC) contents, plasma adiponectin levels (RIA), and insulin resistance (IR) evolution (HOMA) were evaluated in 11 obese patients who underwent subtotal gastrectomy with gastro-jejunal anastomosis in Roux-en-Y and 8 non-obese subjects who underwent laparoscopic cholecystectomy (controls). Liver SREBP-1c and FAS mRNA levels were 33% and 70% higher than control values (P < 0.05), respectively, whereas those of PPAR-α and CPT-1a were 16% and 65% lower (P < 0.05), respectively, with a significant 62% enhancement in the SREBP-1c/PPAR-α ratio. Liver n-3 LCPUFA levels were 53% lower in obese patients who also showed IR and hipoadiponectinemia over controls (P < 0.05). IR negatively correlated with both the hepatic content of n-3 LCPUFA (r = ? 0.55; P < 0.01) and the plasma levels of adiponectin (r = ? 0.62; P < 0.005). Liver SREBP-1c/PPAR-α ratio and n-3 LCPUFA showed a negative correlation (r = ? 0.48; P < 0.02) and positive associations with either HOMA (r = 0.75; P < 0.0001) or serum insulin levels (r = 0.69; P < 0.001). In conclusion, liver up-regulation of SREBP-1c and down-regulation of PPAR-α occur in obese patients, with enhancement in the SREBP-1c/PPAR-α ratio associated with n-3 LCPUFA depletion and IR, a condition that may favor lipogenesis over FA oxidation thereby leading to steatosis.     

Neuroprotective effects and magnetic resonance imaging of mesenchymal stem cells labeled with SPION in a rat model of Huntington's disease

Bone marrow mesenchymal stem cells (MSC) have been tested and proven effective in some neurodegenerative diseases, but their tracking after transplantation may be challenging. Our group has previously demonstrated the feasibility and biosafety of rat MSC labeling with iron oxide superparamagnetic nanoparticles (SPION). In this study, we investigated the therapeutic potential of SPION-labeled MSC in a rat model of Huntington's disease, a genetic degenerative disease with characteristic deletion of striatal GABAergic neurons. MSC labeled with SPION were injected into the striatum 1h after quinolinic acid injection. FJ-C analysis demonstrated that MSC transplantation significantly decreased the number of degenerating neurons in the damaged striatum 7 days after lesion. In this period, MSC transplantation enhanced the striatal expression of FGF-2 but did not affect subventricular zone proliferation, as demonstrated by Ki67 proliferation assay. In addition, MSC transplantation significantly reduced the ventriculomegaly in the lesioned brain. MRI and histological techniques detected the presence of the SPION-labeled cells at the lesion site. SPION-labeled MSC produced magnetic resonance imaging (MRI) signals that were visible for at least 60 days after transplantation. Our data highlight the potential of adult MSC to reduce brain damage under neurodegenerative diseases and indicate the use of nanoparticles in cell tracking, supporting their potential as valuable tools for cell therapy.     

Oxidative stress, microsomal and peroxisomal fatty acid oxidation in the liver of rats treated with acetone

Parameters of oxidative stress, microsomal cytochrome P450 activity and peroxisomal fatty acid oxidation were studied in liver of rats following acetone (1% v/v) consumption for 7 days. Acetone treatment increased the activity of catalase and decreased the activities of superoxide dismutase (SOD) and glutathione peroxidase (GTPx), but did not significantly modify the liver content of malondialdehyde (MDA) and reduced glutathione. Also, acetone increased the total content of cytochrome P450, the microsomal lauric acid hydroxylation, aminopyrine N-demethylation and the peroxisomal beta-oxidation of palmitoyl CoA. These effects were similar to those found previously in starved and ethanol-treated rats, supporting the hypothesis that ketone bodies would be the common inducer of microsomal and peroxisomal fatty acid oxidation in these metabolic states.     

Three-dimensional structure of the shell plate assembly of the chiton Tonicella marmorea and its biomechanical consequences

This study investigates the three-dimensional structure of the eight plate exoskeletal (shell) assembly of the chiton Tonicella marmorea. X-ray micro-computed tomography and 3D printing elucidate the mechanism of conformational change from a passive (slightly curved, attached to surface) to a defensive (rolled, detached from surface) state of the plate assembly. The passive and defensive conformations exhibited differences in longitudinal curvature index (0.43 vs. 0.70), average plate-to-plate overlap (～62% vs. ～48%), cross-sectional overlap heterogeneity (60-82.5% vs. 0-90%, fourth plate), and plate-to-plate separation distance (100% increase in normalized separation distance between plates 4 and 5), respectively. The plate-to-plate interconnections consist of two rigid plates joined by a compliant, actuating muscle, analogous to a geometrically structured shear lap joint. This work provides an understanding of how T. marmorea achieves the balance between mobility and protection. In the passive state, the morphometry of the plates and plate-to-plate interconnections results in an approximately continuous curvature and constant armor thickness, resulting in limited mobility but maximum protection. In the defensive state, the underlying soft tissues gain protection and the chiton gains mobility through tidal flow, but regions of vulnerability open dorsally, due to the increase in plate-to-plate separation and decrease in plate-to-plate overlap. Lastly, experiments using optical and scanning electron microscopy, mercury porosimetry, and Fourier-transform infrared spectroscopy explore the microstructure and spatial distribution of the six layers within the intermediate plates, the role of multilayering in resisting predatory attacks, and the detection of chitin as a major component of the intra-plate organic matrix and girdle.     

Evidence for gene flow differs from observed dispersal patterns in the Humboldt penguin, Spheniscus humboldti

The Humboldt penguin, once common throughout its range, is today listed as Vulnerable by the IUCN. Mark-recapture and telemetry studies indicate that adult Humboldt penguins are sedentary, suggesting strong genetic differentiation between colonies. We developed genotypes for 336 individuals at 12 microsatellite loci sampled at four different localities spanning the entire range of this species. Results show that long-term gene flow has occurred but appears to be affected by geographic distance as pairwise F _ST comparisons involving the colony at Punta San Juan (Peru) and the two colonies at Algarrobo (central Chile) and Puñihuil (southern Chile) are significant. Bayesian estimates of recent migration rates indicate substantial dispersal among all colonies. Despite the dramatic decline in numbers, we did not observe a bottleneck in any population. Furthermore, we did not detect a founder effect in the recently discovered colony at Puñihuil. As our indirect estimates signal strong gene flow between populations, we suggest that Humboldt penguin colonies need to be managed as a metapopulation rather than as discrete management units.     

Modulation of cytochrome P450 activity in the kidney of rats following long-term red wine exposure

Cytochrome P450 (CYP)-dependent oxidation of lauric acid, p-nitrophenol and ethanol by microsomal fractions of kidney were studied in control rats and in animals given either ethanol, red wine, or alcohol-free red wine for 10 weeks. Ethanol increased the total CYP content and specifically CYP 2E1, as well as p-nitrophenol and ethanol oxidation. The effects of ethanol treatment on the content and activity of CYP 2E1 were attenuated when red wine was administered, while the alcohol-free red wine values were similar to those of the control group. Although lauric acid hydroxylation was decreased by red wine treatment, the content of CYP 4A1 was not influenced by drinking fluids. We conclude that red wine administration attenuates the ethanol-induced enhancement of microsomal activities dependent on CYP 2E1 of rat kidney. Our results suggest that the non-alcoholic constituents of red wine could account for this modulation.     

Copper exposure and potential biomarkers of copper metabolism

Relevant biological effects associated with mild to moderate copper deficiency and copper excess are unknown. It is difficult to identify markers of these early changes because limits of the homeostatic range are still undefined and early changes may represent adaptive responses that do not imply necessarily risk of damage. We report here a series of studies carried out to shed light on the responses within the homeostatic range, by assessing classic parameters of copper status in humans at different copper exposure. In adult healthy volunteers that had an estimated daily intake of 0.9 mg Cu/day (approximately 15 g/kg/d), exposure to additional 50–60 g of copper/kg/day for three months or up to 150 g/kg/d for two months resulted in no significant changes of SOD activity in erythrocytes, of copper concentration (in serum, erythrocytes and mononuclear cells) and of serum ceruloplasmin (ANOVA). Neither were found differences by gender or age. As in previous studies in infants, the non-ceruloplasmin copper fraction was positively correlated to serum copper (r=0.58). Assessing variations on copper absorption, infants supplemented/not supplemented with oral copper (80 ug/kg/14 days), at age 1 and 3 months, showed copper absorption close to 80% at both ages; no effect was observed for age or supplementation, suggesting that either these concentrations do not elicit regulatory mechanisms or that at this age down regulation for copper absorption is not efficient. These studies indicate that in the range of the copper homeostasis area the markers tested are not suitable to detect mild changes (within the homeostatic range) of copper metabolism.     

Biochemical characterization of a thermostable cysteine synthase from Geobacillus stearothermophilus V

The cysK gene encoding a cysteine synthase of Geobacillus stearothermophilus V was overexpressed in E. coli and the recombinant protein was purified and characterized. The enzyme is a thermostable homodimer (32 kDa/monomer) belonging to the beta family of pyridoxal phosphate (PLP)-dependent enzymes. UV-visible spectra showed absorption bands at 279 and 410 nm. The band at 279 nm is due to tyrosine residues as the enzyme lacks tryptophan. The 410 nm band represents absorption of the coenzyme bound as a Schiff base to a lysine residue of the protein. Fluorescence characteristics of CysK's Schiff base were influenced by temperature changes suggesting different local structures at the cofactor binding site. The emission of the Schiff base allowed the determination of binding constants for products at both 20 degrees C and 50 degrees C. At 50 degrees C and in the absence of sulphide the enzyme catalyzes the decomposition of O-acetyl-l-serine to pyruvate and ammonia. At 20 degrees C, however, a stable alpha-aminoacrylate intermediate is formed.     

Biosynthesis of agglomerin A: stereospecific incorporation of pro-R- and pro-S-hydrogens at sn-C-3 of glycerol into the branched C(3) moiety

The biosynthetic origin of the C(3) branched unit of agglomerin A has been investigated. Feeding of sn-(3R)- and sn-(3S)-[3-(2)H]glycerols to Enterobacter agglomerans PB-6042 followed by (2)H NMR analysis of the resulting agglomerin A revealed that pro-R and pro-S hydrogens at sn-C-3 of glycerol were incorporated stereospecifically into 5E and 5Z hydrogens of agglomerin A, respectively. These results imply that the immediate precursor of the C(3) branched unit is not pyruvate, but 1,3-bisphosphoglyceric acid or its biological equivalent.