Tailoring reversible insulin aggregates loaded in electrosprayed arabinoxylan microspheres intended for colon-targeted delivery

Arabinoxylans (AX) covalent gels are little affected by pH changes and fermentable by colonic microbiota, which make them suitable for insulin oral administration and colon-targeted delivery. In this work, the tailoring of insulin aggregates size using glutamic acid and their loading in AX microspheres (2%w/v) prepared by triaxial electrospray is presented. Dynamic light scattering showed that insulin-glutamic acid aggregates are reversible under intestinal conditions. AX microspheres presented a spherical shape, a mean diameter of 233 μm, a heterogeneous microstructure as shown by scanning electron microscopy, and a homogeneous distribution of insulin aggregates as observed by confocal laser scanning microscopy. In this AX-insulin system, 20% of insulin is released under simulated gastrointestinal pH conditions, indicating that most of the insulin stays into the microspheres, available for colonic release. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47960.     

Protease (PrA and PrB) and prolyl and arginyl aminopeptidase activities from Debaryomyces hansenii as a function of growth phase and nutrient sources

The effects of nutrient sources and growth phase of Debaryomyces hansenii on the protease (PrA and PrB) and aminopeptidase (prolyl-[PAP] and arginyl-[AAP] aminopeptidases) activities were investigated. These activities were also monitored during growth on a whole sarcoplasmic muscle protein extract (WSPE) and on an equivalent medium but free of compounds under 10 kDa (SPE>10 kDa). The levels of specific protease and aminopeptidase activities were higher when cells were grown in urea and dipeptides than when grown in either ammonium or free amino acids as nitrogen sources. The level of each aminopeptidase (PAP or AAP) activity was preferentially induced by its own substrate (ProLeu or LysAla), suggesting a role in the utilization of exogenous peptides. Higher specific activities for all proteolytic enzymes were detected when using acetate as carbon source. The time course experiments carried out on urea or sarcoplasmic protein-containing media revealed an increase in all activities during transition and advanced stages of stationary phase of growth. In muscle protein extracts, the absence of low molecular mass nutrients (SPE>10 kDa) initially induced the production of PrA, PrB, and AAP activities, possibly involved in the breakdown of muscle oligopeptides.     

Isothermal DNA amplification coupled with DNA nanosphere-based colorimetric detection

We present a simple, rapid method for detecting short DNA sequences that combines a novel isothermal amplification method (EXPAR) with visual, colorimetric readout based on aggregation of DNA-functionalized gold nanospheres. The reaction is initiated by a trigger oligonucleotide, synthetic in nature for this proof-of-principle study, which is exponentially amplified at 55 degrees C and converted to a universal reporter oligonucleotide capable of bridging two sets of DNA-functionalized gold nanospheres. This reaction provides >10(6)-fold amplification/conversion in under 5 min. When combined with a solution containing DNA nanospheres, the bridging reporter causes nanosphere aggregation. The resulting color change from red to dark purple or blue is enhanced through spotting the solution onto a C18 reversed-phase thin-layer chromatography plate. The reaction can easily be adapted for detection of different trigger oligonucleotides using the same set of DNA nanospheres. It permits detection of as low as 100 fM trigger oligonucleotide in under 10 min total assay time, with minimal reagent consumption and requirement for instrumentation. We expect that combining this simple, versatile assay with trigger generation from a genomic target DNA sequence of interest will be a powerful tool in the development of rapid and simple point-of-care molecular diagnostic applications.     

Explanation-based learning:A problem solving perspective

This article outlines explanation-based learning (EBL) and its role in improving problem solving performance through experience. Unlike inductive systems, which learn by abstracting common properties from multiple examples, EBL systems explain why a particular example is an instance of a concept. The explanations are then converted into operational recognition rules. In essence, the EBL approach is analytical and knowledge-intensive, whereas inductive methods are empirical and knowledge-poor. This article focuses on extensions of the basic EBL method and their integration with the problem solving system. 's EBL method is specifically designed to acquire search control rules that are effective in reducing total search time for complex task domains. Domain-specific search control rules are learned from successful problem solving decisions, costly failures, and unforeseen goal interactions. The ability to specify multiple learning strategies in a declarative manner enables EBL to serve as a general technique for performance improvement. 's EBL method is analyzed, illustrated with several examples and performance results, and compared with other methods for integrating EBL and problem solving.     

Lipoprotein Profile in Immunological Non-Responders PLHIV after Antiretroviral Therapy Initiation

Nuclear magnetic resonance (NMR)-based advanced lipoprotein tests have demonstrated that LDL and HDL particle numbers (LDL-P and HDL-P) are more powerful cardiovascular (CV) risk biomarkers than conventional cholesterol markers. Of interest, in people living with HIV (PLHIV), predictors of preclinical atherosclerosis and vascular dysfunction may be associated with impaired immune function. We previously stated that immunological non-responders (INR) were at higher CV risk than immunological responders (IR) before starting antiretroviral therapy (ART). Using Liposcale^® tests, we characterized the lipoprotein profile from the same cohort of PLHIV at month 12 and month 36 after starting ART, intending to explore what happened with these indicators of CV risk during viral suppression. ART initiation dissipates the differences in lipoprotein-based CV risk markers between INR and IR, and only an increase in the number of HDL-P was found in INR + IR when compared to controls (p = 0.047). Interestingly, CD4⁺ T-cell counts negatively correlated with medium HDL-P concentrations at month 12 in all individuals (ρ = −0.335, p = 0.003). Longitudinal analyses showed an important increase in LDL-P and HDL-P at month 36 when compared to baseline values in both IR and INR. A proper balance between a proatherogenic and atherogenic environment may be related to the reconstitution of CD4⁺ T-cell count in PLHIV.     

Phenolic profiles,antioxidant activity and in vitro antiviral properties of apple pomace

Methanolic and acetonic extracts of apple pomace were evaluated for phenolic profiles, antioxidant properties and antiviral effect against herpes simplex virus type 1 (HSV-1) and 2 (HSV-2). Acetone extraction yielded the higher amounts of phenolic compounds. The extraction method influenced the phenolic composition although antioxidant activity correlated weakly with phenols concentration. Among the polyphenols analysed, quercetin glycosides were the most important family, followed by dihydrochalcones. Apple pomace extracts were able to inhibit both HSV-1 and HSV-2 replication in Vero cells by more than 50%, at non-cytotoxic concentrations. Selectivity indexes (SI) ranged from 9.5 to 12.2.     

Influence of germination on the soluble carbohydrates and dietary fibre fractions in non-conventional legumes

Influence of light exposure during germination on structural and soluble carbohydrates including total dietary fibre (TDF), insoluble dietary fibre (IDF) and soluble (SDF) dietary fibre fractions and also raffinose family oligosaccharides (RFOs) was studied in non-conventional legumes Vigna unguiculata (cowpea), Canavalia ensiformis (jack bean), Stizolobium niveum (mucuna), and Lablab purpureus (dolichos), and compared to a well known and used Glycine max (soybean). Non-conventional legumes were rich in DF, mainly IDF, which represented 93–97% of TDF. It was relevant the proportion of protein that remained associated to the insoluble DF matrix. Non-conventional legumes exhibited important levels of RFOs but their profile was different depending on the tropical seed. The germination of seeds produced changes in the carbohydrate fraction, mainly an increase of TDF in most instances, except for soybean, and a drastic reduction of RFOs, from 98% to 63%, with the corresponding increase in the amounts of total soluble sugars.     

Coculture-inducible bacteriocin activity of Lactobacillus plantarum strain J23 isolated from grape must

Detection and characterization of bacteriocin production by Lactobacillus plantarum strain J23, recovered from a grape must sample in Spain, have been carried out. Bacteriocin activity was degraded by proteolytic enzymes (trypsin, alfa-chymotrypsin, papaine, protease, proteinase K and acid proteases), and it was stable at high temperatures (121 degrees C, 20min), in a wide range of pH (1-12), and after treatment with organic solvents. L. plantarum J23 showed antimicrobial activity against Oenococcus oeni, and a range of Lactobacillus and Pediococcus species. Bacteriocin production was detected in liquid media only when J23 was cocultivated with some inducing bacteria, and induction took place when intact cells or 55 degrees C heated cells of the inducer were cocultivated with J23, but not with their autoclaved cells. Bacteriocin activity of J23 was not induced by high initial J23 inocula, and it was detected in cocultures during the exponential phase. The presence of ethanol or acidic pH in the media reduced bacteriocin production in the cocultures of J23 with the inducing bacteria. The presence of plantaricin-related plnEF and plnJ genes was detected by PCR and sequencing. Nevertheless, negative results were obtained for plnA, plnK, plNC8, plS and plW genes.