Fermentation with Lactic Acid Bacteria for Bean Flour Improvement: Experimental Study and Molecular Modeling as Complementary Tools.

Pulses are considered superfoods for the future world due to their properties, but they require processing to reduce antinutritional factors (ANFs) and increase bioactivity. In this study, bean flour (Phaseolus vulgaris L.) was fermented under different conditions (addition of Lactiplantibacillus plantarum CRL 2211 and/or Weissella paramesenteroides CRL 2182, temperature, time and dough yield) to improve its nutri-functional quality. Fermentation for 24 h at 37 °C with the mixed starter increased the lactic acid bacteria (LAB) population, acidity, polyphenol content (TPC) and ANF removal more than spontaneous fermentation. Statistical and rep-PCR analysis showed that fermentation was mainly conducted by Lp. plantarum CRL 2211. Metabolic modeling revealed potential cross-feeding between Lp. plantarum and W. paramesenteroides, while the molecular docking and dynamic simulation of LAB tannases and proteinases involved in ANF removal revealed their chemical affinity to gallocatechin and trypsin inhibitors. Fermentation was better than soaking, germination and cooking for enhancing bean flour properties: it increased the free amino acids content by 50% by releasing glutamine, glutamic acid, arginine, leucine and lysine and modified TPC by increasing gallic acid and decreasing caffeic, ferulic and vanillic acids and quercetin-3-glucoside. The combination of experimental and simulation data may help us to understand fermentation processes and to design products with desirable features.

In vitro and in vivo digestibility of prebiotic galactooligosacharides synthesized by ß-galactosidase from Lactobacillus delbruecki subsp. bulgaricus CRL450.

BACKGROUND: The general assumption that prebiotics reach the colon without any alterations has been challenged. Some in vitro and in vivo studies have demonstrated that 'non-digestible' oligosaccharides are digested to different degrees depending on their structural composition. In the present study, we compared different methods aiming to assess the digestibility of oligosaccharides synthesized by ß-galactosidase (ß-gal) of Lactobacillus delbruecki subsp. bulgaricus CRL450 (CRL450-ß-gal) from lactose, lactulose and lactitol. RESULTS: In the simulated gastrointestinal fluid method, no changes were observed. However, the oligosaccharides synthesized by CRL450-ß-gal were partially hydrolyzed in vitro, depending on their structure and composition, with rat small intestinal extract (RSIE) and small intestinal brush-border membrane vesicles (BBMV) from pig. Digestion of some oligosaccharides increased when mixtures were fed to C57BL/6 mice used as in vivo model; however, lactulose-oligosaccharides were the most resistant to the physiological conditions of mice. In general ß (1→6) linked products showed higher resistance compared to ß (1→3) oligosaccharides. CONCLUSION: In vitro digestion methods, without disaccharidases, may underestimate the importance of carbohydrates hydrolysis in the small intestine. Although BVMM and RSIE digestion assays are appropriate in vitro methods for these studies, in vivo studies remain the most reliable for understanding what actually happens in the digestion of oligosaccharides. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Subcutaneous Application of a Gelatin/Hyaluronic Acid Hydrogel Induces the Production of Skin Extracellular Matrix.

The development of injectable hydrogels with natural biopolymers such as gelatin (Ge) and hyaluronic acid (Ha) is widely performed due to their biocompatibility and biodegradability. The combination of both polymers crosslinked with N-Ethyl-N'-(3-dimethyl aminopropyl) carbodiimide hydrochloride (EDC) can be used as an innovative dermal filler that stimulates fibroblast activity and increases skin elasticity and tightness. Thus, crosslinked Ge/Ha hydrogels with different concentrations of EDC were administered subcutaneously to test their efficacy in young and old rats. At higher EDC concentrations, the viscosity decreases while the particle size of the hydrogels increases. At all concentrations of EDC, amino and carboxyl groups are present. The histological analysis shows an acute inflammatory response, which disappears seven days after application. At one and three months post-treatment, no remains of the hydrogels are found, and the number of fibroblasts increases in all groups in comparison with the control. In addition, the elastic modulus of the skin increases after three months of treatment. Because EDC-crosslinked Ge/Ha hydrogels are biocompatible and induce increased skin tension, fibroblast proliferation, and de novo extracellular matrix production, we propose their use as a treatment to attenuate wrinkles and expression lines.

Role of LTB4 and nitric oxide in the gastroprotective effect of Prosthechea karwinskii leaves extract in the indomethacin-induced gastric injury in the rat.

Gastric injury is mainly described by inflammation of the gastric epithelium. Recently, our group of work demonstrated that Prosthechea karwinskii leaves extract induces both an in vitro antioxidative action and an in vivo gastroprotective effect in a rat. However, the molecules involved in the gastroprotective action by Prosthechea karwinskii are not known. Thus, the aim of this study is to determine whether Prosthechea karwinskii extract modifies anti-inflammatory and antioxidative biomarkers in an in vivo rat model of indomethacin-induced gastric injury. Rats were orally administered with indomethacin and Prosthechea karwinskii leaf extract. Our results suggest that the gastroprotective effect of Prosthechea karwinskii leaf extract is related to the reduction in leukocyte infiltration and antioxidative action in a model of indomethacin-induced gastric injury. Further studies are warranted to investigate the role of the compounds identified in the gastroprotective action of Prosthechea karwinskii leaves extract.

Gelatin/Hyaluronic Acid Scaffold Coupled to CpG and MAGE-A5 as a Treatment against Murine Melanoma.

The half-time of cells and molecules used in immunotherapy is limited. Scaffolds-based immunotherapy against cancer may increase the half-life of the molecules and also support the migration and activation of leukocytes in situ. For this purpose, the use of gelatin (Ge)/hyaluronic acid (HA) scaffolds coupled to CpG and the tumor antigen MAGE-A5 is proposed. Ge and HA are components of the extracellular matrix that stimulate cell adhesion and activation of leucocytes; CpG can promote dendritic cell maturation, and MAGE-A5 a specific antitumor response. C57BL/6 mice were treated with Ge/HA/scaffolds coupled to MAGE-A5 and/or CpG and then challenged with the B16-F10 melanoma cell line. Survival, tumor growth rate and the immune response induced by the scaffolds were analyzed. Ge/HA/CpG and Ge/HA/MAGE-A5 mediated dendritic cell maturation and macrophage activation, increased survival, and decreased the tumor growth rate and a tumor parenchyma with abundant cell death areas and abundant tumor cells with melanin granules. Only the scaffolds coupled to MAGE-A5 induced the activation of CD8 T cells. In conclusion, Ge/HA scaffolds coupled to CpG or MAGE-A5, but not the mixture, can induce a successful immune response capable of promoting tumor cell clearance and increased survival.

Validation of the traditional medicinal use of a Mexican endemic orchid (Prosthechea karwinskii) through UPLC-ESI-qTOF-MS/MS characterization of its bioactive compounds.

Ethnopharmacological relevance: The orchid Prosthechea karwinskii is a medicinal orchid in Oaxaca, Mexico, used to treat diabetes, cough, wounds, and burns, prevent miscarriage and assist in labor. Each part of the plant (leaves, pseudobulbs, or flowers) is used by healers for certain treatment conditions, indicating that each part has different biocompounds with specific pharmacological activity. Aim of the study: To characterize the biocompounds in extracts from leaves, pseudobulbs, and flowers of P. karwinskii and evaluate their ROS inhibition capacity to associate it with medicinal uses. Materials and methods: The compounds present in extracts from leaves, pseudobulbs, and flowers of P. karwinskii were identified by UPLC-ESI-qTOF-MS/MS. The chemical differentiation of each extract was tested by principal component analysis (PCA) using compound intensity values. For each extract, total phenol and flavonoid contents were quantified. Their antioxidant capacity was evaluated ex vivo by inhibition of ROS with DCFH-DA and in vitro with DPPH radical. Results: Based on the PCA, it was observed that some compounds were completely separated from others according to the correlation that they presented. The compounds common to all three plant parts were quinic, malic, succinic, azelaic, and pinellic acids. Among the compounds identified, two were exclusive to leaves, four to pseudobulbs, and ten to flowers. Some of the identified compounds have well-known antioxidant activity. The leaves had the highest content of total phenols and flavonoids, and the highest in vitro and ex vivo antioxidant capacity. A strong correlation was observed between phenol and flavonoid contents, and antioxidant capacity ex vivo and in vitro. Conclusions: It was found that the bioactive compounds and antioxidant capacity of each part of the plant were associated with its traditional medicinal use. A pharmacological potential was also found in P. karwinskii for further biological studies because of the type of compounds it contained.

Immunomodulatory Properties of Masticadienonic Acid and 3α-Hydroxy Masticadienoic Acid in Dendritic Cells.

Dendritic cells are antigen-presenting cells, which identify and process pathogens to subsequently activate specific T lymphocytes. To regulate the immune responses, DCs have to mature by the recognition of TLR ligands, TNFα or IFNγ. These ligands have been used as adjuvants to activate DCs in situ or in vitro, with toxic effects. It has been shown that some molecules affect the immune system, e.g., Masticadienonic acid (MDA) and 3α-hydroxy masticadienoic acid (3α-OH MDA) triterpenes naturally occurring in several medicinal plants, since they activate the nitric oxide synthase in macrophages and induce T lymphocyte proliferation. The DCs maturation induced by MDA or 3a-OH MDA was determined by incubating these cells with MDA or 3α-OH MDA, and their phenotype was afterwards analyzed. The results showed that only 3α-OH MDA was able to induce DCs maturation. When mice with melanoma were inoculated with DCs/3α-OH MDA, a decreased tumor growth rate was observed along with an extended cell death area within tumors compared to mice treated with DCs incubated with MDA. In conclusion, it is proposed that 3α-OH MDA may be an immunostimulant molecule. Conversely, it is proposed that MDA may be a molecule with anti-inflammatory properties.

Fermentation of chickpea flour with selected lactic acid bacteria for improving its nutritional and functional properties.

AIMS: To improve the nutri-functional quality of chickpea flour by fermentation with selected lactic acid bacteria (LAB) to formulate functional legume-derived products. METHODS AND RESULTS: A Randomized Complete Block Design was carried out to assess the influence of experimental conditions (presence/absence of Lactiplantibacillus plantarum CRL2211 and/or Weissella paramesenteroides CRL2182, temperature, time and dough yield) on LAB population, acidification, antinutritional factors and total phenolic contents (TPCs) of chickpea flour. Fermentation with both strains for 24 h at 37°C produced an increase in LAB (up to 8.9 log CFU/g), acidity (final pH 4.06), TPC (525.00 mg GAE/100 g) and tannin and trypsin inhibitor removal (28.80 mg GAE/100 g and 1.60 mg/g, respectively) higher than the spontaneously fermented doughs. RAPD and Rep-PCR analysis revealed that fermentation was dominated by L. plantarum CRL2211. Molecular docking and dynamics simulations were useful to explain LAB enzyme behaviour during fermentation highlighting the chemical affinity of LAB tannases and proteinases to gallocatechin and trypsin inhibitors. Compared with other processing methods, fermentation was better than soaking, germination and cooking for increasing the techno-functional properties of chickpea flour. Fermented doughs were applied to the manufacture of crackers that contained 81% more TPC and 64% more antioxidant activity than controls. CONCLUSIONS: Fermentation for 24 h at 37°C with selected autochthonous LAB was the best method for improving the quality of chickpea flour and derived crackers type cookies. SIGNIFICANCE AND IMPACT OF STUDY: Chickpea is suitable for the development of novel functional foods. Fermentation with selected LAB would improve the final product quality and bioactivity. The combination of experimental and simulation approaches can lead to a better understanding of the fermentation processes to enhance the properties of a food matrix.

Epicutaneous Administration of 17ß-Estradiol Induces Langerhans Cells Depletion.

BACKGROUND: Langerhans cells (LC) number and function in mouse vaginal mucosa are affected by 17ß-estradiol (E2) application; nonetheless, its effect on epidermal LC has not been studied. The purpose of this study was to evaluate the effect of topical administration of E2 on the number, phenotype, and migratory ability of LC in mouse skin. METHODS: Ears of adult CD1 male mice were topically treated once with several doses. Immunohistochemical staining for CD207 and TUNEL staining were performed. LC migration to lymph nodes and the effect on the expression of costimulatory molecules on cultured dendritic cells (DC) were also evaluated. RESULTS: E2 decreased the number of CD207+ LC in a dose-dependent manner. One hour after treatment, 1 and 10 µg/mL E2 significantly reduced the LC number by 21% and 26%, respectively, after two hours, the reduction was 23% and 41%, respectively. After 48 hours, LC recovered, and after 96 hours of treatment, the CD207+/MHCII+ DC numbers were increased in regional lymph nodes. However, CD86 and CD40 molecules were expressed at lower levels than in positive control. The TUNEL assay did not show apoptotic cells. Furthermore, in cultured DC, E2 promoted a decrease in CD40 and CD86 expression and an increase in CD273, CD274, MHCII, and CCR7. CONCLUSIONS: The topical administration of E2 induced a transitory local diminution of LC population and a tolerogenic phenotype. This decrease in epidermal LC suggests that E2 may affect skin immune responses, inducing an inhibitory response, which should be considered when prescribing topical E2 medications.

Three-Dimensional Porous Scaffolds Derived from Bovine Cancellous Bone Matrix Promote Osteoinduction, Osteoconduction, and Osteogenesis.

The use of three-dimensional porous scaffolds derived from decellularized extracellular matrix (ECM) is increasing for functional repair and regeneration of injured bone tissue. Because these scaffolds retain their native structures and bioactive molecules, in addition to showing low immunogenicity and good biodegradability, they can promote tissue repair and regeneration. Nonetheless, imitating these features in synthetic materials represents a challenging task. Furthermore, due to the complexity of bone tissue, different processes are necessary to maintain these characteristics. We present a novel approach using decellularized ECM material derived from bovine cancellous bone by demineralization, decellularization, and hydrolysis of collagen to obtain a three-dimensional porous scaffold. This study demonstrates that the three-dimensional porous scaffold obtained from bovine bone retained its osteoconductive and osteoinductive properties and presented osteogenic potential when seeded with human Wharton's jelly mesenchymal stromal cells (hWJ-MSCs). Based on its characteristics, the scaffold described in this work potentially represents a therapeutic strategy for bone repair.

Bioactive compounds from Prosthechea karwinskii decrease obesity, insulin resistance, pro-inflammatory status, and cardiovascular risk in Wistar rats with metabolic syndrome.

ETHNOPHARMACOLOGICAL RELEVANCE: The orchid Prosthechea karwinskii is a species endemic to Mexico, which is used in traditional medicine to lower glucose levels in patients with diabetes, and to treat inflammation-related problems. Recent studies have shown that this orchids can reduce glucose, cholesterol, and triglyceride levels in Wistar rats, which were previously induced to have metabolic syndrome (MS). AIM OF THE STUDY: To evaluate the effect of P. karwinskii leaves extract on the components of metabolic syndrome: obesity, insulin resistance, pro-inflammatory status, and cardiovascular risk in a Wistar rat model, and to identify the bioactive compounds in the extract. MATERIALS AND METHODS: UPLC-ESI-qTOF-MS/MS was used to identify the compounds present in the extract. MS was induced in Wistar rats through administration of a 40% sucrose diet for 20 weeks. The rats were divided into five groups that received different treatments for 4 weeks: one group without any treatment, one group receiving metformin (200 mg/kg p.o.), and three groups receiving different doses of P. karwinskii leaves extract (100, 200, and 300 mg/kg p.o.). The animals' body weights were recorded weekly, and at the end of the experiment, they were sacrificed after fasting for 18 h to determine the levels of glucose, insulin, insulin resistance index, total cholesterol, triglycerides, and adiponectin in the serum, as well as levels of TNF-α and HS-CRP in the serum and liver homogenates. The abdominal and pericardial fat weights were also recorded. RESULTS: The main bioactive compounds of the extract were quinic acid, neochlorogenic acid, chlorogenic acid, rutin, kaempferol-3-o-ß-rutinoside, and embelin, known to exhibit MS-related bioactivity. Oral administration of P. karwinskii leaves extract at a dose of 300 mg/kg decreased weight gain, abdominal and pericardial fat deposits, and insulin resistance. At the end of the treatment, levels of triglycerides, TNF-α, HS-CRP, and adiponectin returned to levels similar to normal. CONCLUSION: P. karwinskii extract (300 mg/kg) had an anti-obesity effect, decreased insulin resistance, pro-inflammatory status, and cardiovascular risk in rats with induced MS by increasing adiponectin levels and decreasing TNF-α and HS-CRP levels. The compounds identified in the extract could be responsible for these effects, acting alone or in synergy, as several compounds in the extract are known to have MS-related bioactivity. The foliar extract of P. karwinskii has potential as an effective alternative to a cocktail of drugs used to treat problems associated with MS.

Cellulose-Chitosan-Nanohydroxyapatite Hybrid Composites by One-Pot Synthesis for Biomedical Applications.

The development of organic-inorganic hybrid materials deserves special interest for bone tissue engineering applications, where materials must have properties that induce the survival and activation of cells derived from the mesenchyme. In this work, four bio-nanocomposites based on cellulose and variable content of chitosan, from 15 to 50 w% based on cellulose, with nanohydroxyapatite and ß-Glycerophosphate as cross-linking agent were synthesized by simplified and low-energy-demanding solvent exchange method to determine the best ratio of chitosan to cellulose matrix. This study analyzes the metabolic activity and survival of human dermal fibroblast cells cultivated in four bio-nanocomposites based on cellulose and the variable content of chitosan. The biocompatibility was tested by the in vitro cytotoxicity assays Live/Dead and PrestoBlue. In addition, the composites were characterized by FTIR, XRD and SEM. The results have shown that the vibration bands of ß-Glycerophosphate have prevailed over the other components bands, while new diffraction planes have emerged from the interaction between the cross-linking agent and the biopolymers. The bio-nanocomposite micrographs have shown no surface porosity as purposely designed. On the other hand, cell death and detachment were observed when the composites of 1 and 0.1 w/v% were used. However, the composite containing 10 w% chitosan, against the sum of cellulose and ß-Glycerophosphate, has shown less cell death and detachment when used at 0.01 w/v%, making it suitable for more in vitro studies in bone tissue engineering, as a promising economical biomaterial.

A comparison of pesticide residues in soils from two highly technified agricultural valleys in northwestern Mexico.

A pesticide characterization is presented for two highly technified valleys in northwest Mexico: Culiacan Valley (CV) in Sinaloa and Yaqui Valley (YV) in Sonora. Approximately 250,000 kg of active ingredients are used every year, half of which are considered highly hazardous pesticides. Legacy pesticides are still present in the soils of these valleys. The aim of the present study was to identify and quantify a wide variety of pesticides in soils and correlate their concentrations with historical and current use. Agricultural soils from both valleys were sampled and analyzed using accelerated solvent extraction and subsequent quantification by gas chromatography with selective detectors. The most frequently detected pesticides (mean, µg g-1) in CV were organochlorines (0.1967), organophosphates (0.0928), synthetic pyrethroids (0.2565), organonitrogen (0.0552), and miscellaneous pesticides (0.1851). In YV, the most frequently detected pesticides were organochlorines (0.8607), organophosphates (0.0001), synthetic pyrethroids (0.0124), and miscellaneous pesticides (0.0009). The pesticides were more diverse in CV compared to those of YV, which was based on the types of crops produced. Both locations presented highly hazardous pesticides, including concentrations above the action levels established by the Canadian Soil Quality Guide. A follow-up risk assessment is recommended to assess potential effects.

Prebiotic galactooligosaccharides production from lactose and lactulose by Lactobacillus delbrueckii subsp. bulgaricus CRL450.

Galactooligosaccharides (GOS) are useful dietary ingredients recognized worldwide as prebiotics. In the present study, we evaluated the ß-galactosidase (ß-gal) activity of a panel of lactic acid bacteria (LAB) in order to select strains for the synthesis of oligosaccharides from lactose (GOS) and lactulose (OsLu) with a potential prebiotic effect. Fifteen strains out of 20 were able to grow on lactose and showed ß-gal activities between 0.03 and 2.06 U mg-1, whereas eleven were able to synthesize GOS. Lactobacillus delbrueckii subsp. bulgaricus CRL450, the strain with the highest ß-gal activity, synthesized a maximum of 41.3% GOS and 21.0% OsLu from lactose and lactulose, respectively, with ß-(1 → 6) and secondary ß-(1 → 3) linkages. When these compounds were tested without purifying, as carbon sources for the development of recognized probiotics and the producer strain, high growth was observed compared to non-prebiotic sugars like glucose and lactose. When the purified oligosaccharides were tested, the bacterial growth decreased, but the microorganisms displayed metabolic activity evidenced by the consumption of carbohydrates and the production of lactic acid. Additionally, the purified oligosaccharides demonstrated a bifidogenic effect. The obtained results support the potential of L. delbrueckii subsp. bulgaricus CRL450 for the production of the prebiotics GOS and OsLu and encourage the optimization of their synthesis for the design of new functional food ingredients.

Tuberculosis y síndrome inflamatorio de reconstitución inmunológica. Caso pediátrico / Tuberculosis and immune reconstitution inflammatory syndrome. Pediatric case

La tuberculosis es una enfermedad muy frecuente en nuestro medio. A pesar de que la detección precoz y el tratamiento adecuado logran la curación en la mayoría de los pacientes, la dificultad en el diagnóstico, el abandono del tratamiento y la aparición de resistencia a los fármacos tradicionales generan que, en la actualidad, continúe siendo un importante problema de salud pública. En la Argentina, la tasa de morbilidad es de 25/100 000 habitantes, con un leve aumento en la mortalidad.Se presenta el caso de una paciente pediátrica con tuberculosis, que tuvo múltiples complicaciones asociadas a la enfermedad y a su tratamiento, entre las cuales se incluye el síndrome in-flamatorio de reconstitución inmunológica, también conocido como reacción paradojal al tratamiento antituberculoso. Este representa una consecuencia clínica adversa al restablecimien-to de la inmunidad en el paciente que padece una infección sistémica grave, como la tuberculosis miliar.

Tuberculosis is a very frequent disease in our environment. Although early detection and adequate treatment achieve cure in most patients, the difficulty in diagnosis, the abandonment of treatment and the appearance of resistance to traditional drugs generate that at present it continues to represent an im-portant public health problem. In Argentina, the morbidity rate is 25/100,000 inhabitants, with a slight increase in mortality.We present the case of a pediatric patient with tuberculosis and multiple complications associated with the disease and its treatment. One of these complications was the immune re-constitution inflammatory syndrome or paradoxical reaction to antituberculosis treatment. It represents an adverse clinical con-sequence of the restoration of immunity in the patient suffering from a serious systemic infection such as miliary tuberculosis.

[Tuberculosis and immune reconstitution inflammatory syndrome. Pediatric case]. / Tuberculosis y síndrome inflamatorio de reconstitución inmunológica. Caso pediátrico.

Tuberculosis is a very frequent disease in our environment. Although early detection and adequate treatment achieve cure in most patients, the difficulty in diagnosis, the abandonment of treatment and the appearance of resistance to traditional drugs generate that at present it continues to represent an important public health problem. In Argentina, the morbidity rate is 25/100,000 inhabitants, with a slight increase in mortality. We present the case of a pediatric patient with tuberculosis and multiple complications associated with the disease and its treatment. One of these complications was the immune reconstitution inflammatory syndrome or paradoxical reaction to antituberculosis treatment. It represents an adverse clinical consequence of the restoration of immunity in the patient suffering from a serious systemic infection such as miliary tuberculosis.

La tuberculosis es una enfermedad muy frecuente en nuestro medio. A pesar de que la detección precoz y el tratamiento adecuado logran la curación en la mayoría de los pacientes, la dificultad en el diagnóstico, el abandono del tratamiento y la aparición de resistencia a los fármacos tradicionales generan que, en la actualidad, continúe siendo un importante problema de salud pública. En la Argentina, la tasa de morbilidad es de 25/100 000 habitantes, con un leve aumento en la mortalidad. Se presenta el caso de una paciente pediátrica con tuberculosis, que tuvo múltiples complicaciones asociadas a la enfermedad y a su tratamiento, entre las cuales se incluye el síndrome inflamatorio de reconstitución inmunológica, también conocido como reacción paradojal al tratamiento antituberculoso. Este representa una consecuencia clínica adversa al restablecimiento de la inmunidad en el paciente que padece una infección sistémica grave, como la tuberculosis miliar.

Prosthechea karwinskii, an orchid used as traditional medicine, exerts anti-inflammatory activity and inhibits ROS.

ETHNOPHARMACOLOGICAL RELEVANCE: Prosthechea karwinskii (Mart.) J.M.H. Shaw is a Mexican orchid used in traditional medicine by some indigenous communities to treat issues related to inflammation (cough, wounds, burns, and diabetes). Pharmacological research of this orchid could validate its therapeutic uses and demonstrate its potential for treating other health conditions of high prevalence in Mexico, including those associated with oxidative stress such as diabetes, cancer, atherosclerosis, and hypertension as well as inflammation. AIM OF THE STUDY: The leaf extract from P. karwinskii was examined to identify its compounds and elucidate its inhibitory effect on reactive oxygen species as well as its anti-inflammatory activity and gastroprotective effects in an animal model. MATERIALS AND METHODS: Compounds were identified via ultra-high-performance liquid chromatography coupled with electrospray ionization with quadrupole time of flight-mass spectrometry. Inhibition of reactive oxygen species was determined ex vivo in peripheral blood mononuclear cells with 2',7'-dichlorodihydrofluorescein diacetate. The anti-inflammatory activity was assessed using a carrageenan-induced paw edema model in Wistar rats; nitric oxide and tumor necrosis factor alpha levels were quantified. The gastroprotective effect was evaluated in Wistar rats with indomethacin-induced gastric injury. RESULTS: Nine compounds were identified in the P. karwinskii leaf extract. Most compounds, such as quinic acid, malic acid, neochlorogenic acid, chlorogenic acid, rutin, embelin, pinellic acid, and azelaic acid, were reported to exhibit antioxidant and/or anti-inflammatory activity. The extract was also found to inhibit reactive oxygen species in the ex vivo model. Unlike other anti-inflammatory drugs, the extract exerted a dual effect: anti-inflammatory activity and protection of the gastric mucosa. The results showed that the extract could significantly inhibit the release of nitric oxide without a dose-response relationship. CONCLUSION: P. karwinskii leaf extract inhibited reactive oxygen species and exerted an anti-inflammatory effect. Moreover, this extract did not induce gastric damage in the animals. The bioactivity of the species was found to support its use in traditional medicine. This orchid could be used to treat inflammatory diseases without causing the side effects associated with nonsteroidal anti-inflammatory drugs. It can also be employed to treat other pathological conditions associated with oxidative stress. The findings herein form the basis for the future discovery of natural products that may serve as safe alternative therapies for inflammatory disorders.

Influence of the PLGA/gelatin ratio on the physical, chemical and biological properties of electrospun scaffolds for wound dressings.

Chronic wounds are a global health problem, and their treatments are difficult and long lasting. The development of medical devices through tissue engineering has been conducted to heal this type of wound. In this study, it was demonstrated that the combination of natural and synthetic polymers, such as poly (D-L lactide-co-glycolide) (PLGA) and gelatin (Ge), were useful for constructing scaffolds for wound healing. The aim of this study was to evaluate the influence of different PLGA/gelatin ratios (9:1, 7:3 and 5:5 (v/v)) on the physical, chemical and biological properties of electrospun scaffolds for wound dressings. These PLGA/Ge scaffolds had randomly oriented fibers with smooth surfaces and exhibited distances between fibers of less than 10 µm. The 7:3 and 5:5 PLGA/Ge scaffolds showed higher swelling, hydrophilicity and degradation rates than pure PLGA and 9:1 (v/v) PLGA/Ge scaffolds. Young's moduli of the scaffolds were 72 ± 10, 48 ± 6, 58 ± 6 and 6 ± 1 MPa for the pure PLGA scaffold and the 9:1, 7:3 and 5:5 (v/v) PLGA/Ge scaffolds, respectively. Mesenchymal stem cells (MSCs) seeded on all the PLGA/Ge scaffolds were viable, and the cells were attached to the fibers at the different analyzed timepoints. The most significant proliferation rate was observed for cells on the 7:3 PLGA/Ge scaffolds. Biocompatibility analysis showed that all the scaffolds produced inflammation at the first week postimplantation; however, the 7:3 and 5:5 (v/v) PLGA/Ge scaffolds were degraded completely, and there was no inflammatory reaction observed at the fourth week after implantation. In contrast, the 9:1 PLGA/Ge scaffolds persisted in the tissue for more than four weeks; however, at the eighth week, no traces of the scaffolds were found. In conclusion, the scaffolds with the 7:3 PLGA/Ge ratio showed suitable physical, chemical and biological properties for applications in chronic wound treatments.

Toxicity of silver nanoparticles in mouse bone marrow-derived dendritic cells: Implications for phenotype.

Silver nanoparticles (AgNP) are one of the most studied nanoparticles due to their anti-bacterial, -fungal, -viral, -parasitic, and -inflammatory properties. This raises the need to evaluate the toxicity and biological effects of AgNP in the immune system in order to develop new safer biomedical products. In this study, an AgNP formulation currently approved for veterinary applications was applied to mouse bone marrow-derived dendritic cells (BMDC), considered important antigen-presenting cells of the immune system, to evaluate cytotoxicity, genotoxicity, and any significant influence on expression of cellular markers associated with BMDC phenotype and maturation status. The results showed that after 12 h of AgNP exposure, a significant decrease in BMDC viability occurred at the highest concentration tested (1.0 µg AgNP/ml) and at lower doses, the cells maintained membrane integrity and metabolic activity. DNA damage was not significant with any AgNP level aside from the 1.0 µg AgNP/ml level. Regarding phenotype, no differences in expression of CD40 (co-stimulatory molecule highly present in mature BMDC) or in CD273 (a marker for inhibitory T-cell response) were observed. The current results showed that the toxicity of this AgNP formulation was dose-related. The findings also suggest BMDC could maintain structural conservation of co-stimulatory/co-inhibitory surface molecules after 12 h of exposure to this AgNP. This work represents the first step in identifying the toxic effects of this AgNP formulation on dendritic cells.

Differential adhesion and fibrinolytic activity of mesenchymal stem cells from human bone marrow, placenta, and Wharton's jelly cultured in a fibrin hydrogel.

Mesenchymal stem cells isolated from different tissues should share associated markers and the capability to differentiate to mesodermal lineages. However, their behavior varies in specific microenvironments. Herein, adhesion and fibrinolytic activity of mesenchymal stem cells from placenta, bone marrow, and Wharton's jelly were evaluated in fibrin hydrogels prepared with nonpurified blood plasma and compared with two-dimensional cultures. Despite the source, mesenchymal stem cells adhered through focal adhesions positive for vinculin and integrin αV in two dimensions, while focal adhesions could not be detected in fibrin hydrogels. Moreover, some cells could not spread and stay rounded. The proportions of elongated and round phenotypes varied, with placenta mesenchymal stem cells having the lowest percentage of elongated cells (~10%). Mesenchymal stem cells degraded fibrin at distinct rates, and placenta mesenchymal stem cells had the strongest fibrinolytic activity, which was achieved principally through the plasminogen-plasmin axis. These findings might have clinical implications in tissue engineering and wound healing therapy.