Oleanolic acid (OA), a natural component of many plant food and medicinal herbs, is endowed with a wide range of pharmacological properties whose therapeutic potential has only partly been exploited until now. Throughout complex and multifactorial mechanisms, OA exerts beneficial effects against diabetes and metabolic syndrome. It improves insulin response, preserves functionality and survival of β-cells, and protects against diabetes complications. OA may directly modulate enzymes connected to insulin biosynthesis, secretion, and signaling. However, its major contributions appear to be derived from the interaction with important transduction pathways, and many of its effects are consistently related to activation of the transcription factor Nrf2. Doing that, OA induces the expression of antioxidant enzymes and phase II response genes, blocks NF-κB, and represses the polyol pathway, AGEs production, and hyperlipidemia. The management of type 2 diabetes requires an integrated approach, which includes the early intervention to prevent or delay the disease progression, and the use of therapies to control glycemia and lipidemia in its late stages. In this sense, the use of functional foods or drugs containing OA is, undoubtedly, an interesting path.

De novo protein design has been successful in expanding the natural protein repertoire. However, most de novo proteins lack biological function, presenting a major methodological challenge. In vaccinology, the induction of precise antibody responses remains a cornerstone for next-generation vaccines. Here, we present a protein design algorithm called TopoBuilder, with which we engineered epitope-focused immunogens displaying complex structural motifs. In both mice and nonhuman primates, cocktails of three de novo–designed immunogens induced robust neutralizing responses against the respiratory syncytial virus. Furthermore, the immunogens refocused preexisting antibody responses toward defined neutralization epitopes. Overall, our design approach opens the possibility of targeting specific epitopes for the development of vaccines and therapeutic antibodies and, more generally, will be applicable to the design of de novo proteins displaying complex functional motifs.

This work establishes a new procedure for the extraction and analysis of pentacyclic triterpenes, with which fruits and leaves from three Spanish olive cultivars ("Picual", "Hojiblanca", and "Arbequina") has been studied. The leaf contains important amounts of oleanolic acid (3.0-3.5% DW), followed by significant concentrations of maslinic acid and minor levels of ursolic acid, erythrodiol, and uvaol. The abundance and profile of triterpenoids change during the leaf ontogeny. In the fruit, triterpenes are exclusively located in the epicarp at concentrations 30-fold lower than that in the leaf. Maslinic acid is the main triterpenoid, only accompanied of oleanolic acid. Along the ripening the levels of these triterpenes decreased. All the analyzed leaves and fruits come from the same agricultural estate, with identical climate and culturing conditions. For this reason, the found differences could majorly be attributable to the genetic factors of the olive cultivars.

The genetic characterization of four previously reported mutants of human respiratory syncytial (RS) virus resistant to monoclonal antibody 63G is described. Sequences of the G protein genes were obtained from: (i) mRNA derived cDNA recombinants, (ii) direct mRNA sequencing and (iii) amplified vRNA derived cDNAs. The results obtained indicate that the original escape mutants, recovered from individual plaques, contained heterogeneous viral populations. This heterogeneity affected the number of adenosine residues present after nucleotides 588 or 623 of the G protein gene. Mutant viruses recovered after a second plaque purification step generated homogeneous sequences but contained single adenosine insertions or deletions at those two sites compared with the Long sequence. These genetic alterations introduced frameshift changes which are reflected in both the antigenic and structural properties of the mutant G proteins. The origin and importance of frameshift mutations in the RS virus G protein gene are discussed.

Monoclonal antibodies directed against the glycoproteins of human respiratory syncytial virus were used in competitive enzyme-linked immunosorbent assays for topological mapping of epitopes. Whereas epitopes of the F glycoprotein could be ascribed to five nonoverlapping antigenic sites, anti-G antibodies recognized unique epitopes, many of whose competition profiles overlapped extensively. Variant viruses selected with a neutralizing (47F) anti-F antibody lost the binding for only 47F and 49F antibodies, which mapped in the same antigenic area. In contrast, viruses selected with an anti-G antibody lost the capacity to bind most of the anti-G antibodies, and their G protein was not recognized by an anti-virus antiserum, indicating major changes in the antigenic structure of the G molecule. Finally, we found great antigenic variation of the G protein among viral isolates. This occurred even within viruses of the same subtype with only limited divergence of amino acid sequence between strains. All of these data indicate marked differences in the antigenic organization of the G and F glycoproteins of respiratory syncytial virus; we discuss these differences in terms of the chemical structure of the glycoproteins.

A total of 124 methicillin-resistant Staphylococcus aureus (MRSA) isolates were ascertained at the University Hospital of the Canary Islands between January 1997 and April 2000. Genotyping included pulsed-field gel electrophoresis (PFGE) (SmaI digestion) and PCR-restriction fragment length polymorphism (PCR-RFLP) analysis for the coagulase (coa) and protein A (spa) genes. Antibiotic resistance was the main phenotypic marker correlated with genotyping results. Three main PFGE types were detected: A (with 12 subtypes), B (with 2 subtypes), and C. PFGE type A1 was the most commonly found (61% of isolates) and the one responsible for all the epidemic outbreaks. Other genetics markers used (coa and spa RFLPs) were significantly correlated with the PFGE types detected (P < 0.001). These PCR-RFLP assays were useful as molecular markers for a quick, preliminary study of MRSA outbreaks.

a b s t r a c tThe present work evaluates the potential of hazelnut kernels as a source of antioxidants to be incorporated into new products. First, the effects of extraction conditions on the isolation of hazelnut kernels' total phenols and antioxidants were evaluated. Six conditions, involving different solvents (water, methanol and aqueous acetone) and contact times, were studied. The highest total phenol contents were obtained with boiling water for 30 min, 44.3 ± 7.7 mg GAE/g extract , and 80% (v/v) aqueous acetone solution for 24 h, 36.2 ± 8.8 mg GAE/g extract . Increasing the contact time for the acetonic extractions did not improve the total phenols content. Regarding antioxidant activity, the highest DPPH-scavenging effect value was obtained with 80% (v/v) aqueous acetone for 24 h with an effective concentration (EC 50 ) equal to 1.12 ± 0.07 mg/mL. When other nuts -walnuts, almonds, pine nuts and peanuts -were extracted under this condition, only walnut extract exhibited higher phenol content (268 ± 32 mg GAE/g extract ), antioxidant activity as measured by reducing power (EC 50 = 0.091 ± 0.015 mg/mL) and free radical scavenging capacity (DPPH assay) (EC 50 = 0.060 ± 0.010 mg/mL) than hazelnut extract. The present work demonstrates that some nuts might be a natural source of bioactive compounds that can be incorporated into new health-related products or be substitutes of synthetic compounds of questionable safety, promoting human health and reducing disease risks.

The antioxidant activity and chemical composition of essential oils and methanolic extracts of twenty Spanish Thymus mastichina L. populations were studied. Both essential oils and methanolic extracts possessed antioxidant properties. However, the total phenol contents of the methanolic extracts varied between 2.90 and 9.15 mg GAE/g extract and the EC 25 values of DPPH free radical scavenging activity between 0.90 and 3.45 mg/mL for the methanolic extracts and 78e241 mg/mL for essential oils, these showing low antioxidant potential. Actually, in essential oils the main compound determined was the 1,8-cineole (56.8e69.6%), whereas thymol, g-terpinene, terpinolene and geraniol (species with considerable DPPH scavenging activity) were observed in low amounts. Concerning methanolic extracts, rosmarinic acid was the most abundant polyphenol (1.70e9.85 mg/g), followed by methoxysalicylic acid, apigenin, kaempferol and luteolin.