Development of Methotrexate Complexes Endowed with New Biological Properties Envisioned for Musculoskeletal Regeneration in Rheumatoid Arthritis Environments

Methotrexate (MTX) administration is the gold standard treatment for rheumatoid arthritis (RA), but its effects are limited to preventing the progression of the disease. Therefore, effective regenerative therapies for damaged tissues are still to be developed. In this regard, MTX complexes of general molecular formula M(MTX)·xH₂O, where M = Sr, Zn, or Mg, were synthesized and physicochemically characterized by TGA, XRD, NMR, ATR–FTIR, and EDAX spectroscopies. Characterization results demonstrated the coordination between the different cations and MTX via two monodentate bonds with the carboxylate groups of MTX. Cation complexation provided MTX with new bioactive properties such as increasing the deposition of glycosaminoglycans (GAGs) and alternative anti-inflammatory capacities, without compromising the immunosuppressant properties of MTX on macrophages. Lastly, these new complexes were loaded into spray-dried chitosan microparticles as a proof of concept that they can be encapsulated and further delivered in situ in RA-affected joints, envisioning them as a suitable alternative to oral MTX therapy.     

AMD-Like Substrate Causes Epithelial Mesenchymal Transition in iPSC-Derived Retinal Pigment Epithelial Cells Wild Type but Not C3-Knockout

The Bruch’s membrane (BrM) is a five-layered extracellular matrix (ECM) that supports the retinal pigment epithelium (RPE). Normal age-related changes in the BrM may lead to RPE cell damage and ultimately to the onset and progression of age-related macular degeneration (AMD), which is the most common cause of visual loss among the elderly. A role for the complement system in AMD pathology has been established, but the disease mechanisms are poorly understood, which hampers the design of efficient therapies to treat millions of patients. In an effort to identify the mechanisms that lead from normal aging to pathology, we have developed a cell-based model using complement deficient human induced pluripotent stem cell (iPSC)-derived RPE cells cultured on an AMD-like ECM that mimics BrM. The data present evidence that changes in the ECM result in loss of differentiation and promote epithelial mesenchymal transition (EMT) of healthy RPE cells. This pathological process is mediated by complement activation and involves the formation of a randomly oriented collagen meshwork that drives the dedifferentiation of the RPE monolayer. Genetic ablation of complement component 3 has a protective effect against EMT but does not prevent the abnormal deposition of collagens. These findings offer new insights into the sequence of events that initiate AMD and may guide the design of efficient therapies to treat this disease with unmet medical needs.     

Designing antimicrobial bioactive glass materials with embedded metal ions synthesized by the sol–gel method

Bioactive glasses (SiO₂–P₂O₅–CaO) having tailored concentrations of different biocide metal ions (copper or silver) were produced by the sol–gel method. All the particles release phosphorous ions when immersed in water and simulated body fluid (SBF). Moreover, a surface layer of polycrystalline hydroxy-carbonate apatite was formed on the particle surfaces after 10 day immersion in SBF as confirmed by X-ray diffraction and scanning electron microscopy (SEM) showing the bioactive materials. Samples with embedded either copper or silver ions were able to further release the biocide ions with a release rate that depends on the metal embedded and the dissolution medium: water or SBF. This biocide ion release from the samples explains the antimicrobial effect of our active particles against Escherichia coli DH5α ampicillin-resistant (Gram-negative) and Streptococcus mutans (Gram-positive) as determined by the Minimum Bactericidal Concentration (MBC) method. The antimicrobial behavior of the particles depends on the bacteria and the biocide ion used. Noteworthy, although samples with copper are able to release more metal ion than samples with silver, they present higher MBC showing the high effect of silver against these bacteria.     

Design and Characterization of Corn Starch Edible Films Including Beeswax and Natural Antimicrobials

The effectiveness of edible films (EFs) used as coatings to maintain the quality and safety of fresh produce for long time depends on their functional properties characterization. This study was aimed to design and evaluate physicochemical, barrier, mechanical, and antimicrobial properties of EFs based on corn starch (acetylated cross-linked (ACLS) or oxidized (OS)), micro-emulsified beeswax (BW, 0–1 % w/w), and two natural antimicrobials (lauric arginate (LAE, 400–4000 mg/L) and natamycin (NAT, 80–800 mg/L)). EFs based on ACLS or OS made with 1 % BW microemulsion produced homogeneous EFs surface and did not show changes in thickness or opacity. Water vapor permeability (WVP, 0.57 ± 0.04 g mm m^?2 h^?1 kPa^?1 for ACLS, and 0.56 ± 0.05 g mm m^?2 h^?1 kPa^?1 for OS) was reduced; tensile strength (TS, 51.48 ± 5.92 MPa for ACLS, and 40.96 ± 4.98 MPa for OS), and elastic modulus (EM, 211.30 ± 7.85 MPa for ACLS, and 203.50 ± 5.35 MPa for OS) were decreased, whereas elongation at break (E, 4.59 ± 1.11 % for ACLS, and 4.76 ± 4.98 % for OS) increased. The additive effect showed by the combination of natural antimicrobials (2000 mg/L of LAE plus 400 mg/L of NAT) incorporated into EFs with 1 % BW completely inhibited Rhizopus stolonifer, Colletotrichum gloeosporioides, Botrytis cinerea, and Salmonella Saintpaul. These properties of corn starch EFs used as coatings represent an excellent alternative to extend the shelf life of fresh produce.     

Polyphenolic compositions of Basque natural ciders: A chemometric study

Polyphenolic compositions of Basque natural ciders were determined by high-performance liquid chromatography, with diode array detection following thiolysis, in order to differentiate ciders according to the geographical origin of the main raw material used for their elaboration. Fifty percent of the apples used for cidermaking in the Basque Country are imported from France or Galicia (N.W. Spain); this gives beverages of different chemical compositions and sensory qualities. A data set, consisting of 64 cider samples and 33 measured variables, was evaluated using multivariate chemometric techniques. A preliminary study of data structure was performed by cluster analysis and principal component analysis. Different classification systems for the two categories were obtained on the basis of the chemical data by applying several supervised pattern recognition procedures, such as linear discriminant analysis (LDA), K-nearest neighbours (KNN), soft independent modelling of class analogy (SIMCA), and multilayer feed-forward artificial neural networks (MLF-ANN). KNN, SIMCA and the MLF neural network provided complementary results: KNN allowed the correct classification of almost all the ciders of the Galician category, SIMCA provided a model for the ciders of the French category that excluded all ciders made with Galician apples (50% of raw material), and the neural network achieved a level of hits for the classification of the ciders obtained from French apples (50% of raw material) above 95%. Polyphenolic profiles of the ciders provide enough information to develop classification rules for identifying ciders according to the geographical origin of the raw material used for cidermaking.     

Health-promoting activities of ultra-filtered okara protein hydrolysates released by in vitro gastrointestinal digestion: identification of active peptide from soybean lipoxygenase

Okara, a major by-product of the soymilk industry, which is rich in proteins, could possibly release under physiological conditions potential bioactive peptides. Thus, an okara protein isolate was digested sequentially with pepsin and pancreatin for c.a. 4 h. On the basis of its relatively high degree of hydrolysis and antioxidant activities (power reduction and radical scavenging activity), the okara protein hydrolysates at the end of the in vitro digestion were fractionated by ultra-filtration and the obtained ultra-filtered fractions were further tested for angiotensin-converting enzyme inhibition and multifunctional antioxidant activities. In the <1 kDa molecular weight cutoff ultra-fraction the amino acid sequence, TIIPLPV, of a peptide from soybean lipoxygenase-1 with a calculated mass 751.48 Da was identified using LC-ESI-MS/MS techniques. The hydrophobic amino acids present in this peptide, particularly Val at terminal position, could likely be associated with the relatively high health-promoting attributes tested. This study evidenced that the consumption of okara protein may exert health benefits on the basis of the bioavailability and bioactivity of the identified peptide.     

Effect of Freezing and Frozen Storage on Chemical and Microbiological Characteristics in Sheep Milk Cheese

ABSTRACT: Most chemical components and water activity of an the Los Pedroches' brand ewes' milk cheese were not altered by frozen storage. Lactic acid concentration and pH were significantly different (p < 0.05) in control cheeses and those kept in frozen storage for 3 and 6 months from those stored for 9 months. Proteolysis continued slowly during frozen storage, with significantly higher rates of non protein nitrogen and amino acid nitrogen (p < 0.05) present at the end of the storage period. Counts of microorganisms, except for enterococci, tended to decrease during frozen storage. Freezing rates did not affect the chemical and microbiological characteristics studied.     

Antioxidant activity of seed polyphenols in fifteen wild Lathyrus species from South Spain

Antioxidant activity of seed phenolics was studied in the following Lathyrus species: Lathyrus hirsutus, Lathyrus filiformis, Lathyrus sativus, Lathyrus cicera, Lathyrus angulatus, Lathyrus sphaericus, Lathyrus annuus, Lathyrus clymenum, Lathyrus pratensis, Lathyrus ochrus, Lathyrus aphaca, Lathyrus latifolius, Lathyrus setifolius, Lathyrus tingitanus and Lathyrus amphicarpos. Phenolic contents ranged from 3.8 mg/g meal in L. setifolius to 29.2 mg/g meal in L. sphaericus. In general, non-cultivated Lathyrus species contained higher phenolic contents than cultivated ones. A negative correlation between seed size and phenolic contents was observed and was related to the higher proportion of hulls in the smaller seeds. L. annuus possessed phenolics with highest specific antioxidant activity. These phenolics were more than two times more antioxidant than equivalent amounts of phenolics extracted from commercial chickpea, lupin or soy. On the other hand, L. aphaca possessed the highest antioxidant activity per mg of flour extract. This antioxidant activity was twice that observed in same amounts of extracted flours from commercial chickpea, lupin or soy. Results show that studied Lathyrus species are rich in phenolic compounds with higher antioxidant activity than phenolics of widely consumed legumes such as soy, chickpea or lupin. In conclusion, Lathyrus may represent an interesting source of phenolic compounds with high antioxidant activity that may be useful as natural antioxidants and contribute to revalorize the cultivation of these legumes.