AICAR Protects Vascular Endothelial Cells from Oxidative Injury Induced by the Long-Term Palmitate Excess

Hyperlipidemia manifested by high blood levels of free fatty acids (FFA) and lipoprotein triglycerides is critical for the progression of type 2 diabetes (T2D) and its cardiovascular complications via vascular endothelial dysfunction. However, attempts to assess high FFA effects in endothelial culture often result in early cell apoptosis that poorly recapitulates a much slower pace of vascular deterioration in vivo and does not provide for the longer-term studies of endothelial lipotoxicity in vitro. Here, we report that palmitate (PA), a typical FFA, does not impair, by itself, endothelial barrier and insulin signaling in human umbilical vein endothelial cells (HUVEC), but increases NO release, reactive oxygen species (ROS) generation, and protein labeling by malondialdehyde (MDA) hallmarking oxidative stress and increased lipid peroxidation. This PA-induced stress eventually resulted in the loss of cell viability coincident with loss of insulin signaling. Supplementation with 5-aminoimidazole-4-carboxamide-riboside (AICAR) increased endothelial AMP-activated protein kinase (AMPK) activity, supported insulin signaling, and prevented the PA-induced increases in NO, ROS, and MDA, thus allowing to maintain HUVEC viability and barrier, and providing the means to study the long-term effects of high FFA levels in endothelial cultures. An upgraded cell-based model reproduces FFA-induced insulin resistance by demonstrating decreased NO production by vascular endothelium.     

Beneficial and Sexually Dimorphic Response to Combined HDAC Inhibitor Valproate and AMPK/SIRT1 Pathway Activator Resveratrol in the Treatment of ALS Mice

Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurodegenerative disorder. There is no cure and current treatments fail to slow the progression of the disease. Epigenetic modulation in the acetylation state of NF-kB RelA and the histone 3 (H3) protein, involved in the development of neurodegeneration, is a drugable target for the class-I histone deacetylases (HDAC) inhibitors, entinostat or valproate, and the AMP-activated kinase (AMPK)-sirtuin 1 pathway activator, resveratrol. In this study, we demonstrated that the combination of valproate and resveratrol can restore the normal acetylation state of RelA in the SOD1(G93A) murine model of ALS, in order to obtain the neuroprotective form of NF-kB. We also investigated the sexually dimorphic development of the disease, as well as the sex-sensibility to the treatment administered. We showed that the combined drugs, which rescued AMPK activation, RelA and the histone 3 acetylation state, reduced the motor deficit and the disease pathology associated with motor neuron loss and microglial reactivity, Brain-Derived Neurotrophic Factor (BDNF) and B-cell lymphoma-extra large (Bcl-xL) level decline. Specifically, vehicle-administered males showed earlier onset and slower progression of the disease when compared to females. The treatment, administered at 50 days of life, postponed the time of onset in the male by 22 days, but not in a significant way in females. Nevertheless, in females, the drugs significantly reduced symptom severity of the later phase of the disease and prolonged the mice’s survival. Only minor beneficial effects were produced in the latter stage in males. Overall, this study shows a beneficial and sexually dimorphic response to valproate and resveratrol treatment in ALS mice.     

Static fatigue of Hi-Nicalon™-S fiber at elevated temperature in air,steam, and silicic acid-saturated steam

A facility for testing SiC fiber tows in static fatigue and creep at elevated temperatures in air and steam was developed. Static fatigue of Hi-Nicalon™-S fibers was investigated at 800°C-1100°C at applied stresses between 115 and 1250 MPa in air, in Si(OH)₄(g)-saturated steam, and in unsaturated steam. Fibers tested in Si(OH)₄(g)-saturated steam and in air had silica scales throughout the test sections, but those tested in unsaturated steam did not develop scales near the steam injection point. Fiber lifetimes in static fatigue were shortest in unsaturated steam, intermediate in Si(OH)₄(g)-saturated steam, and longest in air. Failure strains did not exceed 0.3%. Steady-state strain rates and static fatigue lifetimes are modelled empirically by the Monkman-Grant relationship. Failure mechanisms are discussed.     

“Arbequina” Olive Oil Composition Is Affected by the Application of Regulated Deficit Irrigation during Pit Hardening Stage

Three new regulated deficit irrigation (RDI) treatments were applied to “Arbequina” olive orchards during pit hardening. Oil quality was determined by measuring analytical parameters for olive oil grading, antioxidant activity, total phenol content, fatty acid profile, volatile compounds profile, and sensory analysis. Oils from RDI were classified as “extra virgin olive oil” and their quality was improved due to their higher antioxidant potential (ABTS⁺ [increased ~75%] and DPPH^˙ [increased ~25%] assays) and phenols (increased ~53%) than control. Concentration of total volatile compounds decreased (~27%) but RDI olive oils showed a more balanced profile (alcohols, aldehydes, and esters). Monounsaturated fatty acid content increased (~5%) and atherogenic and thrombogenic indexes decreased (~8.5%) in RDI olive oil. Regarding sensory analysis, RDI provided more balanced oils with higher fruit aroma than control. Other benefits of RDI olive oil, when compared with oil from full irrigated orchards are reduced use of water and improved functional and sensory quality.     

The diastereofacial selectivity of the addition of various 1-metalated 1-heptynes to substituted cyclopentanecarbaldehydes. – The use of 1-titanated 1-heptyne for the construction of the alkyl side chain of Prostaglandins

A highly diastereoselective addition of 1-heptyne to substituted cyclopentanecarbaldehydes is one of the indispensable prerequisites to the construction of the (S)-3-hydroxy-1(E)-octenyl side chain of prostaglandins by palladium(II)-catalyzed [3,3]-sigmatropic rearrangement of allylic acetates. It was found that 1-titanated 1-heptynes afford the Cram product at best with a diastereofacial selectivity of 92.5%. The corresponding lithium and magnesium compounds are less diastereoselective.     

Macrophage-Mediated Melanoma Reduction after HP-NAP Treatment in a Zebrafish Xenograft Model

The Helicobacter pylori Neutrophil Activating Protein (HP-NAP) is endowed with immunomodulatory properties that make it a potential candidate for anticancer therapeutic applications. By activating cytotoxic Th1 responses, HP-NAP inhibits the growth of bladder cancer and enhances the anti-tumor activity of oncolytic viruses in the treatment of metastatic breast cancer and neuroendocrine tumors. The possibility that HP-NAP exerts its anti-tumor effect also by modulating the activity of innate immune cells has not yet been explored. Taking advantage of the zebrafish model, we examined the therapeutic efficacy of HP-NAP against metastatic human melanoma, limiting the observational window to 9 days post-fertilization, well before the maturation of the adaptive immunity. Human melanoma cells were xenotransplanted into zebrafish embryos and tracked in the presence or absence of HP-NAP. The behavior and phenotype of macrophages and the impact of their drug-induced depletion were analyzed exploiting macrophage-expressed transgenes. HP-NAP administration efficiently inhibited tumor growth and metastasis and this was accompanied by strong recruitment of macrophages with a pro-inflammatory profile at the tumor site. The depletion of macrophages almost completely abrogated the ability of HP-NAP to counteract tumor growth. Our findings highlight the pivotal role of activated macrophages in counteracting melanoma growth and support the notion that HP-NAP might become a new biological therapeutic agent for the treatment of metastatic melanomas.     

Mechanical behavior of cold-water fish gelatin gels crosslinked with 1,4-butanediol diglycidyl ether

In this work, chemical crosslinking with 1,4-butanediol diglycidyl ether (BDDGE) is used as strategy to enhance mechanical performance of fish gelatin (FG) gels in order to meet the properties' range of mammalian gelatin physical gels. Joint analysis of free amino groups, swelling ratio, and total soluble material indicates that crosslinking degree increases with increasing FG concentration and it is favored by a 0.2 BDDGE/FG ratio. Increasing crosslinking degree enhances gel indentation strength and shear modulus (μ) while decreases fracture toughness (G_IC). Measured μ and G_IC values lies within the range exhibited by mammalian gelatin physical gels, but the relationship between these parameters is opposite. This is due to the different fracture mechanisms occurring in chemically crosslinked and physical gels.     

Technical note: Serum total protein and immunoglobulin G concentrations in neonatal dairy calves over the first 10 days of age

Efficacy of passive transfer of immunity in young calves is commonly assessed using total serum protein (STP) or serum immunoglobulin G (IgG) concentration tested within the first few days of life. To our knowledge, no research has measured changes in these concentrations over this period to establish an appropriate age range for testing. The aim of this study was to monitor changes in STP and serum IgG concentrations from birth until 10 d of age to provide a basis for recommendations for when passive transfer of immunity in dairy calves can be measured. Concentrations of STP and IgG of 12 calves were measured at 11 time points: at approximately 30 min before colostrum feeding, at 24 h after colostrum feeding, and daily from d 2 to 10 of age. Mean (± standard deviation) STP and IgG concentrations were 4.61 ± 0.3 g/dL and 0.6 ± 0.6 mg/mL at birth, 5.83 ± 0.73 g/dL and 22.2 ± 9.6 mg/mL at 24 h after colostrum feeding, and 5.78 ± 0.52 g/dL and 16.1 ± 7.3 mg/mL at d 10 of age, respectively. The IgG concentration declined over subsequent days relative to IgG measured at 24 h at a rate of approximately 0.69 mg/mL per day, declining by 27.6 ± 6.2% (mean ± SD) on d 10. The concentration of STP did not decrease over time. Concentrations of IgG at 24 h after colostrum feeding were highly correlated with each of the measures of IgG over the 10-d period (r ≥0.97). These correlations were supported by the Bland-Altman plots of agreement between the 24-h sample and subsequent samples. Compared with the reference value at 24 h, STP concentrations were highly correlated on d 2 and 3 (r ≥0.98), highly correlated but variable from d 4 to 9 (r ≥0.88), and lower at d 10 (r = 0.76). These results indicate that calves may be reliably tested for passive transfer of immunity using IgG or STP concentrations up to 9 d of age.     

Identification of polyphenolic compounds in red and white grape varieties grown in R. Macedonia and changes of their content during ripening

The composition of polyphenols in the skin, seed and pulp extracts of the grapes of Vranec and Smederevka varieties, and Merlot and Chardonnay as well, was analyzed by liquid chromatography-diode array detection-mass spectrometry (HPLC-DAD-MS) and MS/MS techniques. Thirty-one phenolic compounds, including anthocyanins, flavonols and flavan-3-ols, as well as phenolic acids derivatives, have been identified in the extracts prepared from the grapes at physiological maturity, by mass spectrometry applying electrospray ionization operated in alternating ion mode and by performing tandem MS experiments in the ion trap mass analyzer. Grapes were analyzed at three different phases: (i) veraison, (ii) physiological ripeness and (iii) late harvest, in order to follow the evolution of polyphenolic content during berry development, applying spectrophotometric methods. Vranec had a higher polyphenolic content compared to Merlot due to the higher levels of total phenolics and flavonoids in the skins and seeds as well as a higher content of flavan-3-ols and anthocyanins in the skins, allowing discriminating the varieties. HPLC-DAD-MS analysis presented significantly higher relative amounts of anthocyanin monoglucosides and p-coumaroylglucosides in Vranec grapes. Smederevka seeds had a higher amount of flavan-3-ols than Chardonnay, while a higher content of total phenolics and flavonoids was found in the skins of the Chardonnay variety. Anthocyanin content in both red varieties increased during the berry ripening, while flavan-3-ols in seeds were mainly accumulated in the veraison phase followed by decrease of the content with ripening.     

Identification of novel pyranoanthocyanins in berry juices

In most fruit and berry products the attractive red color is unstable and easily susceptible to degradation. The colors of strawberry and raspberry juices can be enhanced and stabilized by the addition of cinnamic acids. The aim of this study was to identify the novel anthocyanin products responsible for the improved juice color. The pyranoanthocyanins formed were detected using high-performance liquid chromatography, and the fractions from this were analyzed using a nano-ESI tandem MS technique. The pyranoanthocyanins identified were formed from reactions of the main anthocyanins of strawberry and raspberry juices with ferulic acid or sinapic acid. The new anthocyanin derivatives were the 4-vinylguaiacol and 4-vinylsyringol adducts of pelargonidin and cyanidin. This is the first time that pelargonidin 3-glucoside-based vinylphenol pyranoanthocyanins and pyranoanthocyanins with more complex sugar residues have been found. This is also the first time pyranoanthocyanins have been detected in strawberry and raspberry juices.