Effect of conA-unbound proteins from Melipona beecheii honey on the formation of Pseudomonas aeruginosa ATCC 27853 biofilm.

Pseudomonas aeruginosa is an opportunistic bacterium that can form a biofilm with the ability to colonize different surfaces and for increasing resistance to antibiotics. An alternative to solve this problem may be the use of non-glucose/mannose glycosylated proteins from Melipona beecheii honey, which are capable of inhibiting the growth of this pathogen. In this work, the antibiofilm activity of the conA-unbound protein fraction (F1) from M. beecheii was evaluated. The crude protein extract (CPE) and the F1 fraction inhibited the P. aeruginosa biofilm growth above 80% at 4 and 1.3 µg/mL, respectively. These proteins affected the structure of the biofilm, as well as fleQ and fleR gene expressions involved in the formation and regulation of the P. aeruginosa biofilm. The results demonstrated that the F1 fraction proteins of M. beecheii honey inhibit and affect the formation of the P. aeruginosa biofilm.

Tailoring Heat Transfer and Bactericidal Response in Multifunctional Cotton Composites.

Through the execution of scientific innovations, "smart materials" are shaping the future of technology by interacting and responding to changes in our environment. To make this a successful reality, proper component selection, synthesis procedures, and functional active agents must converge in practical and resource-efficient procedures to lay the foundations for a profitable and sustainable industry. Here we show how the reaction time, temperature, and surface stabilizer concentration impact the most promising functional properties in a cotton-based fabric coated with silver nanoparticles (AgNPs@cotton), i.e., the thermal and bactericidal response. The coating quality was characterized and linked to the selected synthesis parameters and correlated by a parallel description of "proof of concept" experiments for the differential heat transfer (conversion and dissipation properties) and the bactericidal response tested against reference bacteria and natural bacterial populations (from a beach, cenote, and swamp of the Yucatan Peninsula). The quantification of functional responses allowed us to establish the relationship between (i) the size and shape of the AgNPs, (ii) the collective response of their agglomerates, and (iii) the thermal barrier role of a surface modifier as PVP. The procedures and evaluations in this work enable a spectrum of synthesis coordinates that facilitate the formulation of application-modulated fabrics, with grounded examples reflected in "smart packaging", "smart clothing", and "smart dressing".

Antimicrobial and antioxidant activity of proteins isolated from Melipona beecheii honey.

Proteins from Melipona beecheii honey were purified by concanavalin A (conA) affinity chromatography and eluted with a stepwise glucose gradient into fractions named F2-F5. The conA-unbound fraction (F1) was further separated by molecular exclusion into fractions named MbF1-1,2 and MbF1-3. All fractions were evaluated for antibacterial activity against foodborne pathogens and antioxidant capacity. F1 fraction possessed highest antimicrobial activity against S. aureus, L. monocytogenes, S. Typhimurium, E. coli and P. aeruginosa with MIC's 1.4 ± 0.2, 15 ± 1, 39 ± 2, 1 ± 0.1, and 75 ± 2 µg/mL, respectively. F1, MbF1-1,2 and MbF1-3 had bactericidal effect except against P. aeruginosa. When the antioxidant capacity of the fractions was determined, F2 had the highest antioxidant activity measured by DPPH radical scavenging activity (IC50 = 2.4 ± 0.4 µg/µL) and reducing power of Fe(III) (IC50 = 1.8 ± 0.2 µg/µL). We provide evidence that M. beecheii honey proteins possess broad spectrum antibacterial and antioxidant activity, the latter probably through their reducing agent and free radical scavenger properties.

Electrospun and co-electrospun biopolymer nanofibers for skin wounds on diabetic patients: an overview.

Wound healing treatment in diabetic patients worldwide represents around 2.1 trillion dollars to global health sectors. This is because of the complications presented in the wound healing process of skin ulcers, such as a lack of macrophage and fibroblast growth factors (TGF-ß1 and PDGF, respectively) that are both needed for extracellular matrix (ECM) synthesis. Therefore, there is a need for research on new and cost-effective materials to enable ECM synthesis. Such materials include co-electrospun nanofibers used as wound dressings, since they have a similar morphology to the ECM, and therefore, possess the advantage of using different materials to accelerate the wound healing process. Co-electrospun nanofibers have a unique structural configuration, formed by a core and a shell. This configuration allows the protection and gradual liberation of healing agent compounds, which could be included in the core. Some of the materials used in nanofibers are polymers, including natural compounds, such as chitosan (which has been proven to possess antimicrobial and therapeutic activity) and gelatin (for its cell growth, adhesion, and organisational capacity in the wound healing process). Synthetics such as polyvinyl-alcohol (PVA) (mainly as a co-spinning agent to chitosan) can also be used. Another bioactive compound that can be used to enhance the wound healing process is eugenol, a terpenoid present in different medicinal plant tissues that have scarring properties. Therefore, the present review analyses the potential use of co-electrospun nanofibers, with chitosan-PVA-eugenol in the core and gelatin in the shell as a wound dressing for diabetic skin ulcers.

Effects of phenolic and protein extracts from Melipona beecheii honey on pathogenic strains of Escherichia coli and Staphylococcus aureus.

Antimicrobial effects of Melipona beecheii honey have been attributed to diverse factors, in this sense, certain components such as proteins and phenolics could explain relevant aspects of its antimicrobial activity. The aim of this study was to evaluate the antibacterial activity of phenolic and protein extracts from M. beecheii honey against two bacterial pathogens: Staphylococcus aureus and Escherichia coli. With respect to phenolic content, HPLC analysis allowed the identification of phenolic acids like chlorogenic acid, caffeic acid, and flavonoids like catechin, myricetin, quercetin and apigenin. On the other hand, seven bands with molecular weight from 7.6 to 95 kDa were detected in protein extract by SDS-PAGE system. It was determined the antibacterial activity of both extracts, with MICs lower than 145 µg/mL and 60 µg/mL for the phenolic and protein extracts respectively. These results indicate that phenolic and protein components of M. beecheii honey contribute significantly to the antibacterial activity.

Partial characterization of ethanolic extract of Melipona beecheii propolis and in vitro evaluation of its antifungal activity

ABSTRACT The ethanolic extract of Melipona beecheii Bennett 1831, propolis from Yucatán Mexico was evaluated in vitro for the determination of its phenolic compound content, antioxidant capacity and antifungal activity. The results were compared against those of the ethanolic extract of Apis mellifera propolis. The total phenolic content, flavonoid content and flavanones-dihydroflavonols content were assessed by colorimetric methods. The antifungal activity was assessed in vitro against Candida albicans. For the ethanolic extract of M. beecheii propolis; total phenolic content, was 263.25 ± 8.78 µg/ml, total flavonoid content was 768 ± 204 µg/ml and flavanones-dihydroflavonols content was 335.42 ± 15.08 µg/ml. For antioxidant activity assessed as DPPH scavenging and iron reducing power ethanolic extract of M. beecheii propolis reported IC50 of 32.47 and 1.60 µg/ml of gallic acid equivalent respectively. Regarding antifungal activity against C. albicans, the minimal inhibitory concentration and minimal fungicidal concentration for ethanolic extract of M. beecheii propolis were 1.62 ± 0.33 and 2.50 ± 0.22 µg/ml of dry extract; For both minimal inhibitory concentration and minimal fungicidal concentration, ethanolic extract of M. beecheii propolis required 30% less concentration of dry extract than ethanol extract of A. mellifera propolis to exert the same antifungal actions against C. albicans. To the best of our knowledge, this is the first report about the flavonoid and flavanones-dihydroflavonols content of M. beecheii propolis. Due to the lack of information available about the stingless bee's honeycomb products, the study and conservation of endemic honeybees should remain as an active focus of research.

Pentacyclic triterpenes and other constituents in propolis extract from Melipona beecheii collected in Yucatan, México

ABSTRACT Thirteen pentacyclic triterpenes, methyl 3-oxours-12-en-23-oate, marsformosanone, taraxerone, β-amyrenone, α-amyrenone, lupenone, 24-methylencycloartan-3-one, moretenol acetate, β-amyrin acetate, germanicol acetate, 24-methylencycloartanyl acetate, β-amyrin, and α-amyrin were identified in a chloroform-methanol propolis extract from Melipona beecheii. Additionally, were identified in this propolis, hexadecanoic acid, methyl ester, octadecanoic acid, methyl ester and 1-triacontanol. The purification of the propolis extract was carried out using different chromatographic techniques, including vacuum liquid chromatography, gravity column chromatography and gel filtration chromatography Sephadex LH-20. The identification of the metabolites was performed using mass spectrometry.

Antioxidant, antihypertensive, anti-hyperglycemic, and antimicrobial activity of aqueous extracts from twelve native plants of the Yucatan coast.

Looking for a biotechnical potential, aqueous extracts of leaves of 12 native species used in the Mayan traditional medicine of the coastal dune and mangrove of Yucatan (Mexico) were selected to evaluate their biological activities. Rhizophora mangle and Manilkara zapota showed the highest free radical scavenging activity (3.94 ± 0.19 and 6.42 ± 0.32 µg/mL, respectively), and the highest antihypertensive activity was obtained from Solanum donianum (0.38 µg/mL). The anti-hyperglycemic activity of these species was also tested; the highest activities were registered with R. mangle. The antimicrobial activity of Malvaviscus arboreus, S. donianum, M. zapota, and R. mangle at 10% (w/v) was positive against six human pathogenic bacteria and Bonellia macrocarpa against one pathogenic fungus. Solanum donianum, M. zapota, B. macrocarpa, and R. mangle were positive against two pathogenic plant fungi. These results show that the aqueous extracts of five native plants of the Yucatan coast have potential as antioxidants, ACE inhibitors, α-amylase and α-glucosidase inhibitors, and as antimicrobials, which make their exploration for utilization in the agricultural and pharmaceutical industries a possibility.

Encapsulation of vegetable oils as source of omega-3 fatty acids for enriched functional foods.

Polyunsaturated omega-3 fatty acids (PUFAs), a functional component present in vegetable oils, are generally recognized as being beneficial to health. Omega-3 PUFAs are rich in double bonds and unsaturated in nature; this attribute makes them highly susceptible to lipid oxidation and unfit for incorporation into long shelf life foods. The microencapsulation of oils in a polymeric matrix (mainly polysaccharides) offers the possibility of controlled release of the lipophilic functional ingredient and can be useful for the supplementation of foods with PUFAs. The present paper provides a literature review of different vegetable sources of omega-3 fatty acids, the functional effects of omega-3 fatty acids, different microencapsulation methods that can possibly be used for the encapsulation of oils, the properties of vegetable oil microcapsules, the effect of encapsulation on oxidation stability and fatty acid composition of vegetable oils, and the incorporation of long-chain omega-3 polyunsaturated fatty acids in foods.

[ANTIOXIDANT POTENTIAL OF MELIPONA BEECHEII HONEY AND ITS RELATIONSHIP TO HEALTH: A REVIEW]. / POTENCIAL ANTIOXIDANTE DE LA MIEL DE MELIPONA BEECHEII Y SU RELACIÓN CON LA SALUD: UNA REVISIÓN.

The present article provides a literature review about the biological potential of Melipona beecheii. The objective is to project some tendecies in research about nutraceutical aspects related to the bioactive compounds presents in the honey of this stingless bee species, known for its medicinal properties traditional, in the Yucatan Peninsula. Currently, there is strong evidence that M. beecheii honey has bioactive compounds such as proteins, flavonoids and polyphenols with high antioxidant activity. The scientific evidence allows to propose to the honey of stingless bee species as a potential alternative for the obtention of bioactive compounds with antioxidant activity in the Yucatan Peninsula and natural food being proposed to reduce some diseases associated with stress oxidative physiological human cells. However, there is still information that explains such antioxidant activity, therefore, according to the literature reviewed, sees the need to address nutraceuticals and functional aspects correlated with the bioactive compounds present in this honey bee.

El presente artículo provee una revisión de literatura sobre el potencial biológico de la miel de Melipona beecheii. El objetivo es proyectar algunas tendencias en investigación sobre los aspectos nutracéuticos relacionados con los compuestos bioactivos presentes en la miel de esta especie de abeja sin aguijón, reconocida por sus características medicinales tradicionales, en la Península de Yucatán. Actualmente existe una evidencia sólida que demuestra que la miel de M. beecheii posee compuestos bioactivos tales como proteínas, flavonoides y polifenoles, con alta actividad antioxidante. La evidencia científica obtenida permite proponer a la miel de esta especie de abeja sin aguijón como alternativa para la obtención de compuestos bioactivos con actividad antioxidante en la Península de Yucatán, y ser propuesto como alimento natural para reducir algunos tipos de cáncer asociados al estrés oxidativo de las células fisiológicas del ser humano. Sin embargo, aún falta información que explique dicha actividad antioxidante; por lo tanto, de acuerdo con la literatura revisada, se ve la necesidad de abordar aspectos nutracéuticos y funcionales en correlación con los compuestos bioactivos presentes en esta miel de abeja.

Efficient transformation of Mycosphaerella fijiensis by underwater shock waves.

Black leaf streak disease, also known as black Sigatoka, causes dramatic losses in production of banana and plantains fruits. The disease is caused by the pathogenic fungus Mycosphaerella fijiensis (anamorph Pseudocercospora fijiensis; Mycosphaerellaceae). Genetic transformation of M. fijiensis would allow a better understanding of molecular basis of pathogenicity and design novel approaches to control the infection caused by this pathogen. However, transformation of this fungus has not been easy. We report here a protocol for genetic transformation of M. fijiensis employing underwater shock waves and intact conidia. The recombinant strains recovered showed genetic stability over >10 generations. The frequency of transformation obtained was between 75 and 150 times higher than the efficiency reported in the only article published on transformation of M. fijiensis using spheroplasts. This improvement allowed the use of a thousand times less cells than the amount employed before, avoiding the need for cumbersome successive batch cultures. Our protocol is simple, highly efficient, fast and reproducible and together with the available genomes of M. fijiensis and Musa acuminata, it offers new possibilities to study the diverse mechanisms of pathogenesis of the fungus.

Potencial antioxidante de la miel de Melipona beecheii y su relación con la salud: una revisión / Antioxidant potential of melipona beecheii honey and its relationship to health: a review

El presente artículo provee una revisión de literatura sobre el potencial biológico de la miel de Melipona beecheii. El objetivo es proyectar algunas tendencias en investigación sobre los aspectos nutracéuticos relacionados con los compuestos bioactivos presentes en la miel de esta especie de abeja sin aguijón, reconocida por sus características medicinales tradicionales, en la Península de Yucatán. Actualmente existe una evidencia sólida que demuestra que la miel de M. beecheii posee compuestos bioactivos tales como proteínas, flavonoides y polifenoles, con alta actividad antioxidante. La evidencia científica obtenida permite proponer a la miel de esta especie de abeja sin aguijón como alternativa para la obtención de compuestos bioactivos con actividad antioxidante en la Península de Yucatán, y ser propuesto como alimento natural para reducir algunos tipos de cáncer asociados al estrés oxidativo de las células fisiológicas del ser humano. Sin embargo, aún falta información que explique dicha actividad antioxidante; por lo tanto, de acuerdo con la literatura revisada, se ve la necesidad de abordar aspectos nutracéuticos y funcionales en correlación con los compuestos bioactivos presentes en esta miel de abeja (AU)

The present article provides a literature review about the biological potential of Melipona beecheii. The objective is to project some tendecies in research about nutraceutical aspects related to the bioactive compounds presents in the honey of this stingless bee species, known for its medicinal properties traditional, in the Yucatan Peninsula. Currently, there is strong evidence that M. beecheii honey has bioactive compounds such as proteins, flavonoids and polyphenols with high antioxidant activity. The scientific evidence allows to propose to the honey of stingless bee species as a potential alternative for the obtention of bioactive compounds with antioxidant activity in the Yucatan Peninsula and natural food being proposed to reduce some diseases associated with stress oxidative physiological human cells. However, there is still information that explains such antioxidant activity, therefore, according to the literature reviewed, sees the need to address nutraceuticals and functional aspects correlated with the bioactive compounds present in this honey bee (AU)

Recombinant expression of four oxidoreductases in Phanerochaete chrysosporium improves degradation of phenolic and non-phenolic substrates.

Phanerochaete chrysosporium belongs to a group of lignin-degrading fungi that secretes various oxidoreductive enzymes, including lignin peroxidase (LiP) and manganese peroxidase (MnP). Previously, we demonstrated that the heterologous expression of a versatile peroxidase (VP) in P. chrysosporium recombinant strains is possible. However, the production of laccases (Lac) in this fungus has not been completely demonstrated and remains controversial. In order to investigate if the co-expression of Lac and VP in P. chrysosporium would improve the degradation of phenolic and non-phenolic substrates, we tested the constitutive co-expression of the lacIIIb gene from Trametes versicolor and the vpl2 gene from Pleurotus eryngii, and also the endogenous genes mnp1 and lipH8 by shock wave mediated transformation. The co-overexpression of peroxidases and laccases was improved up to five-fold as compared with wild type species. Transformant strains showed a broad spectrum in phenolic/non-phenolic biotransformation and a high percentage in synthetic dye decolorization in comparison with the parental strain. Our results show that the four enzymes can be constitutively expressed in a single transformant of P. chrysosporium in minimal medium. These data offer new possibilities for an easy and efficient co-expression of laccases and peroxidases in suitable basidiomycete species.

Electrophoresis characterisation of protein as a method to establish the entomological origin of stingless bee honeys.

Increasing production of stingless-bee honey and the prospect of broader marker for natural and organic products indicate the need to establish parameters to determinate the entomological origin and authenticity of honey. In this research, honeys of Apis mellifera, Melipona beecheii and Trigona spp. were collected in Yucatan, Mexico. Stingless-bee honeys contained more water and less total sugars and reducing sugars. SDS-PAGE patterns show distinctive bands for each kind of honey. The SDS-PAGE pattern of A. mellifera proteins honey showed three bands with molecular weights between 10.2 and 74.8kDa, there were five proteins bands in M. beecheii honey with molecular weights between 6.1 and 97.0kDa and nine for Trigona spp. proteins between 9.3 and 86.7kDa. Conventional physicochemical parameters along with electrophoresis profiles of stingless-bee honeys proteins could be an alternative for determination of entomological origin.

Antioxidant compounds, antioxidant activity and phenolic content in peel from three tropical fruits from Yucatan, Mexico.

The aim of this study was to determine the antioxidant compounds, antioxidant activity and content of individual phenolic compounds of freeze-dried peel from three tropical fruits grown in Yucatan, México: purple star apple (Chrysophyllum cainito L.), yellow cashew and red cashew (Anacardium occidentale). The freeze-dried peels were good source of antioxidant compounds. ABTS and DPPH values in the peel from each fruit were 3050.95-3322.31 µM Trolox/100g dry weight (DW) or 890.19-970.01 mg of vitamin C/100 g DW, and 1579.04-1680.90 µM Trolox/100 g DW or 340.18-362.18 mg of vitamin C/100 g DW, respectively. Six phenolic compounds were identified in the peel from the tropical fruits studied: ferulic, caffeic, sinapic, gallic, ellagic and myricetin. This study demonstrated that freeze-dried peels from purple star apple, yellow cashew and red cashew, could serve as potential sources of antioxidants for use in food and pharmaceutical industries.

Tandem shock waves to enhance genetic transformation of Aspergillus niger.

Filamentous fungi are used in several industries and in academia to produce antibiotics, metabolites, proteins and pharmaceutical compounds. The development of valuable strains usually requires the insertion of recombinant deoxyribonucleic acid; however, the protocols to transfer DNA to fungal cells are highly inefficient. Recently, underwater shock waves were successfully used to genetically transform filamentous fungi. The purpose of this research was to demonstrate that the efficiency of transformation can be improved significantly by enhancing acoustic cavitation using tandem (dual-pulse) shock waves. Results revealed that tandem pressure pulses, generated at a delay of 300 µs, increased the transformation efficiency of Aspergillus niger up to 84% in comparison with conventional (single-pulse) shock waves. This methodology may also be useful to obtain new strains required in basic research and biotechnology.

Determination of some physicochemical characteristics, bioactive compounds and antioxidant activity of tropical fruits from Yucatan, Mexico.

The aim to the study was to determine the physicochemical composition, bioactive compounds and antioxidant activity of fruits from Yucatan, Mexico such as star apple, cashew, mombin, mamey sapote, white sapote, sugar apple, sapodilla, dragon fruit, nance, ilama, custard apple, mamoncillo and black sapote. The physicochemical characteristics were different between fruits and were good sources of bioactive compounds. The edible part with the highest values of antioxidant activity were mamoncillo, star apple, mombin, cashew, white sapote, ilama, custard apple, sugar apple, and nance. Total soluble phenols content showed a correlation with antioxidant activity by ABTS (R=0.52, P⩽0.05) and DPPH (R=0.43, P⩽0.05). A high correlation was obtained between the two assays (ABTS and DPPH) used to measure antioxidant activity in the tropical fruit species under study (R=0.82, P⩽0.05). The results show promising perspectives for the exploitation and use of tropical fruits studied with significant levels of nutrients and antioxidant activity.

A novel and highly efficient method for genetic transformation of fungi employing shock waves.

Genetic transformation of filamentous fungi is an essential tool in many areas such as biotechnology, medicine, phytopathology and genetics. However, available protocols to transform fungi are inefficient, laborious and have low reproducibility. We report the use of underwater shock waves as a novel method to transform filamentous fungi. An experimental piezoelectric shock wave generator was designed to expose fungal conidia to heterologous DNA. The device was successfully tested in Aspergillus niger, Fusarium oxysporum, Trichoderma reesei and Phanerochaete chrysosporium. The transformation frequency per number of conidia was between two and four orders of magnitude higher in comparison to previously published methods. For example, the frequency of transformation in A. niger was improved up to 5400-fold as compared with Agrobacterium protocols. Transformation was verified by expression of the green fluorescent protein, PCR and Southern blot. Our method offers new possibilities for fast, easy and efficient genetic manipulation of diverse fungal species.