New Formulation of a Methylseleno-Aspirin Analog with Anticancer Activity Towards Colon Cancer

Aspirin (ASA) has attracted wide interest of numerous scientists worldwide thanks to its chemopreventive and chemotherapeutic effects, particularly in colorectal cancer (CRC). Incorporation of selenium (Se) atom into ASA has greatly increased their anti-tumoral efficacy in CRC compared with the organic counterparts without the Se functionality, such as the promising antitumoral methylseleno-ASA analog (1a). Nevertheless, the efficacy of compound 1a in cancer cells is compromised due to its poor solubility and volatile nature. Thus, 1a has been formulated with native α-, β- and γ-cyclodextrin (CD), a modified β-CD (hydroxypropyl β-CD, HP-β-CD) and Pluronic F127, all of them non-toxic, biodegradable and FDA approved. Water solubility of 1a is enhanced with β- and HP- β-CDs and Pluronic F127. Compound 1a forms inclusion complexes with the CDs and was incorporated in the hydrophobic core of the F127 micelles. Herein, we evaluated the cytotoxic potential of 1a, alone or formulated with β- and HP- β-CDs or Pluronic F127, against CRC cells. Remarkably, 1a formulations demonstrated more sustained antitumoral activity toward CRC cells. Hence, β-CD, HP-β-CD and Pluronic F127 might be excellent vehicles to improve pharmacological properties of organoselenium compounds with solubility issues and volatile nature.     

Characterization of tributyrin hydrolysis by immobilized lipase on woolen cloth using conventional batch and novel spinning cloth disc reactors

Optimal loading and operating conditions for a new, superior immobilization of amano lipase from Pseudomonas fluorescens on woolen cloth were determined. The optimal enzyme loading was 46.8 mg g dry cloth⁻¹ with activity of 200 U. A batch reactor was used to characterize process conditions important to industrial application of the wool immobilized lipase. The optimal pH for immobilized lipase in tributyrin hydrolysis was 7, slightly lower than that of free lipase (pH 8). The optimal temperature for both free and immobilized lipase was 45 °C. The immobilized lipase was more stable to reuse than some other lipase immobilizations, maintaining 85% of its activity after 6 long term runs and 75.8% of the original activity after storage of 40 weeks at 4 °C. The thermal stability of lipase was improved by 2.4 times after immobilization. The thermal deactivation rate of immobilized lipase followed the Arrhenius law with E_d = 199 kJ mol⁻¹. The Michaelis–Menten constant (K_m) of the lipase increased from 1.63 mM to 4.48 mM after immobilization. The immobilized lipase was also successfully applied for tributyrin hydrolysis in a novel enzyme process intensification technology – the spinning cloth disc reactor (SCDR): conversion increased by around 13% under similar conditions compared to a conventional batch stirred tank reactor. The SCDR is therefore key to exploiting the advantages of the wool immobilized lipase developed in this work.     

Evaluation of the effect of probiotic cultures added to commercial yogurt over a known population of Listeria monocytogenes and Escherichia coli O157:H7

The effect of probiotic cultures over Listeria monocytogenes and Escherichia coli O157:H7 during yogurt storage was evaluated. Two different yogurt brands, one with additional probiotic cultures (Lactobacillus casei and L. acidophilus) were inoculated with known populations (106 UFC/g) of either L. monocytogenes or E. coli O157:H7 in three different times and stored for 32 days at 5 degrees C. Every four days the count of lactic bacteria, the added pathogens and pH was evaluated, according to the methodology described in the Bacteriological Analytical Manual. The pH and lactic bacteria population were constant during the testing period. Yogurt with additional probiotic cultures reduced the population of L. monocytogenes in 8 days, the population of E. coli O157:H7 in 16; yogurt with no additional probiotics took 20 days to reduce L. monocytogenes to non-detectable levels and even after 28 days of storage, E. coli O157:H7 was cultured. In this work, the beneficial effects of additional probiotic cultures in yogurt is confirmed again.     

Scavenging of reactive oxygen and nitrogen species with nanomaterials

Reactive oxygen and nitrogen species (RONS) are essential for normal physiological processes and play important roles in cell signaling, immunity, and tissue homeostasis. However, excess radical species are implicated in the development and augmented pathogenesis of various diseases. Several antioxidants may restore the chemical balance, but their use is limited by disappointing results of clinical trials. Nanoparticles are an attractive therapeutic alternative because they can change the biodistribution profile of antioxidants, and possess intrinsic ability to scavenge RONS. Herein, we review the types of RONS, how they are implicated in several diseases, and the types of nanoparticles with inherent antioxidant capability, their mechanisms of action, and their biological applications.     

FTIR spectroscopy characterization of poly (vinyl alcohol) hydrogel with different hydrolysis degree and chemically crosslinked with glutaraldehyde

In this work, poly (vinyl alcohol) (PVA) hydrogels with different degree of hydrolysis (DH) were prepared by chemical crosslinking with glutaraldehyde (GA). The nanostructure of the resulting hydrogels was investigated by Fourier Transform Infrared Spectroscopy (FTIR) and Synchrotron small-angle X-ray scattering characterization (SAXS). In vitro tests were performed by swelling ratio assays in different pH solutions. The infrared spectra of the crosslinked PVA showed absorption bands of the acetal bridges resulted from the reaction of the GA with the OH groups from PVA. Also the FTIR spectroscopy was used to determine the crystallinity of the PVA film based on the relative intensity of the vibration band at 1141 cm^− 1. The results have showed an increase of hydrogel crystallinity with higher DH of PVA. SAXS patterns have clearly indicated important modifications on the PVA semicrystalline structure when it was crosslinked by GA. The swelling ratio was significantly reduced by chemically crosslinking the PVA network. PVA-derived hydrogel with chemically modified network was found to be pH-sensitive, indicating a high potential to be used in drug delivery polymer system.     

Real-time assessment of fluid flow generated by appendage movements of Daphnia using standing square-wave chronamperometry

Standing square-wave chronoamperometry (SSWCA) was applied to the analysis of the microfluid flow generated by the movement of the appendages of the Crustacea Daphnia. This novel approach provided for the first time real-time assessment and analysis of the breathing rate/fluid flow of individual organisms. An electrochemical tracer was delivered into the fluid inflow of the organism and a carbon fiber microelectrode placed in the fluid outflow's path. The variation of the net concentration/flux of the electroactive tracer, dopamine, at the electrode surface was measured with SSWCA. The observed chronoamperometric peaks (with fine structure) of the outflow are seen as a direct representation of appendage movement and, too, the workings and responses of the organism to its environment, e.g., external stimuli such as food or chemicals. It was concluded that SSWCA follows primarily the variation of the convective component of the Nernst-Plank equation for flux and, to lesser extent, diffusion and migration. In this work, SSWCA can clearly be used to monitor changes in the Daphnia-generated fluid outflow on a different time scale than was previously possible. This new application of SSWCA is faster and likely more accurate than using high-speed video.     

The association of age, sex and helmet use with the risk of death for occupants of two-wheeled motor vehicles involved in traffic crashes in Spain

Background

This study was designed to separate the association of age, sex and helmet use with the risk of death for occupants of two-wheeled motor vehicles (TWMV) involved in crashes into its two theoretical components: severity of the crash and occupant resilience.

Methods

We analyzed the retrospective cohort comprising all 48 016 pairs of drivers and passengers aged 14 years or more in TWMV involved in crashes with victims in Spain from 1993 to 2007 recorded in the Spanish traffic crash registry. The outcome (death or survival), age, sex and helmet use was known for both occupants. Adjusted relative risks (RR) for the association of age, sex and helmet with the risk of death were calculated with Poisson regression models.

Results

Each 1-year increase in age was related with a 3% increase in the risk of death related with lower resilience. The severity-dependent RR of death was 1.84 for male sex and 0.86 for non-helmet use, and the resilience-dependent RR was 0.72 and 2.53, respectively.

Conclusions

The direction and magnitude of the association between age, sex and helmet use and the risk of death of an occupant of a TWMV involved in a crash changed depending on which component of risk was considered: crash severity or occupant resilience. Specifically, female sex and non-helmet use seemed to be associated with crashes of lower severity, but together with increased age they were also related with lower resilience to the energy released in the crash, and therefore with a higher risk of death after adjustment for crash severity. This should be taken into account when assessing the association of individual factors with the risk of death after a crash.     

Stability of the antimicrobial activity of acetogenins from avocado seed,under common food processing conditions,against Clostridium sporogenes vegetative cell growth and endospore germination

Anticlostridial activity of acetogenins from avocado seed was further characterised, and their stability and effectiveness under food processing conditions, and in a model food system, studied for the first time. Isolated AcO‐avocadenyne (1) and AcO‐avocadene (2) showed anticlostridial potential, particularly the latter molecule. Enriched acetogenins extracts, obtained at laboratory scale (EAE) and semi‐commercially (Avosafe^®), presented similar MIC values (3.9–9.8 ppm) and a bacteriostatic effect. Extracts bioactivity showed resistance to heat (≤120 °C), high hydrostatic pressure (HHP; 300–600 MPa, 3–6 min, 25 °C) and salt (≤3% w/v). In addition, the extract was most stable at pH ≥ 7.0 and potency against endospores increased after HHP treatment and exposure to pH 9.5, suggesting a positive effect on solubility or structure of particular acetogenins. In a model food system processed by HHP, acetogenins were retained; however, initial quantities gradually declined by 63% and 32% at 25 and 4 °C, respectively, at the end of the storage period (42 days). Most stable molecules (persediene (4) > persenones > AcO‐avocadene (2)) possess a keto or trans‐enone group at C‐4 in the aliphatic chain, which could support hydrogen donation to surrounding carbon atoms and confer antioxidant activity. Active endospores were completely inhibited by 5000 ppm Avosafe^® in the model food system (37 °C, 72 h) and lower concentrations (500–1000 ppm) resulted in 1–2 log reduction of a 3 log inoculum target. Efficacy information generated in the present work is considered crucial to improve scientific knowledge on spore inhibition properties of avocado acetogenins.     

Effect of addition of Agaricus blazei mushroom residue to milk enriched with Omega‐3 on the prevention of lipid oxidation and bioavailability of bioactive compounds after in vitro gastrointestinal digestion

The residue from a hydroalcoholic extract of the mushroom Agaricus blazei (MAR) was evaluated for phenolic compounds, flavonoids and antioxidant activity. The ability of MAR to slow the oxidation of Omega‐3 resulting from light exposure in milk matrix, and its bioavailability after in vitro digestion was investigated. MAR presented phenolic compounds and flavonoids and showed antioxidant activity. At each concentration, addition of MAR to Omega‐3‐supplemented milk inhibited the production of conjugated dienes and malonaldehyde compared with samples without MAR. The bioavailability assay showed that polyphenols were still present after in vitro digestion and had antioxidant activity.