Toxicity Evaluation of a Polyphenolic Extract from Flourensia cernua DC through Artemia Lethality Assay, Hemolytic Activity, and Acute Oral Test.

Flourensia cernua DC, commonly known as hojasen or tarbush, is a medicinal plant used in arid regions due to its therapeutic properties, especially in the treatment of gastrointestinal disorders. This study aimed to assess the toxicity of a polyphenolic extract obtained from F. cernua. This research involved both in vitro (hemolytic and brine shrimp assay) and in vivo tests (acute oral toxicity) to determine the safety profile of this extract. The extract was obtained through a novel ultrasound-microwave extraction and purified by ion-exchange chromatography. Analysis of the polyphenolic extract revealed a rich composition of flavonoids and hydroxycinnamic acids, mainly apigenin glycosides. In toxicity tests, the polyphenols did not exhibit toxicity towards Artemia salina at a concentration of 1 mg/ml. Furthermore, incubation at 500 µg/ml for 4 hours showed a slight toxic effect on erythrocytes. In the acute oral toxicity test in mice, doses of 300 mg/kg and 2000 mg/kg did not result in animal mortality, indicating that the LD50 exceeds 2000 mg/kg. However, the higher dose induced signs of toxicity, including lethargy, drowsiness, piloerection, and a significant decrease in weight during the initial two days postadministration of the polyphenolic extract. In addition, histological analysis suggested potential kidney damage at the 2000 mg/kg dose. According to OECD guidelines, while the extract can be classified as category 5 (low acute toxicity) due to the absence of mortality at 2000 mg/kg, the observed signs of toxicity should be considered in the overall risk assessment. These findings highlight the potential of F. cernua in pharmaceutical and nutraceutical applications due to its high polyphenolic content. However, further investigations are necessary to explore the specific effects of the compounds present in the extract. In addition, continuous evaluation of its long-term toxicity is essential to fully understand the extract's safety profile and efficacy.

Grape Pomace Polyphenols Reduce Acute Inflammatory Response Induced by Carrageenan in a Murine Model.

Grape pomace (GP), a by-product of wine production, contains bioactive polyphenols with potential health benefits. This study investigates the anti-inflammatory properties of a polyphenolic fraction derived from GP, obtained by ultrasound-microwave hybrid extraction and purified using ion-exchange chromatography. In the inflammation model, mice were divided into six groups: intact, carrageenan, indomethacin, and three GP polyphenols treatment groups. Paw edema was induced by subplantar injection of carrageenan, and the GP polyphenols were administered intraperitoneally at doses of 10, 20, and 40 mg/kg. The anti-inflammatory effect was evaluated by measuring paw volume, and expression of inflammatory markers: cyclooxygenase-2 (COX-2), myeloperoxidase (MPO), and cytokines (IL-1ß and IL-6), along with lipid peroxidation levels. The GP polyphenols significantly reduced paw edema and expression levels of COX-2, MPO, and cytokines in a dose-dependent manner effect, with the highest dose showing the greatest reduction. Additionally, lipid peroxidation levels were also decreased by GP polyphenols treatment at doses of 10 and 20 mg/kg. These findings suggest that ultrasound-microwave extraction combined with amberlite purification proved to be effective in obtaining a polyphenolic-rich fraction from GP. Thus, GP polyphenols may serve as a natural anti-inflammatory and antioxidant agent for treating inflammation and oxidative stress-related diseases.

Antiproliferative and cytotoxic effects of polyphenols from pomegranate peel and coffee pulp on cancer cells.

Pomegranate peel (PP) and coffee pulp (CP) are by-products of the food industry that can cause environmental pollution if not handled adequately. These by-products contain a significant amount of polyphenolic compounds which have antioxidant and possibly anticancer properties. We investigated the antiproliferative and cytotoxic activities of polyphenols from PP, CP and a 50-50% mixture of both against HeLa, A549, MDA-MB and Hek-293 cells. The total phenolic content of the PP and CP extracts was determined by high performance liquid chromatography/electrospray ionisation/mass spectrometry, and the antiproliferative and cytotoxic potentials were evaluated by MTT (3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide) and Lactate Dehydogenase assays, respectively. Results showed antiproliferative and cytotoxic effects of polyphenols from PP and CP when administered at different concentrations or mixtures on HeLa, A549 and MDA-MB cells. No significant antiproliferative effects were observed on Hek-293 cells treated under similar conditions. These results suggest the potential of PP and CP polyphenols, individually or in combination, to modulate biological mechanisms involved in cervical, breast and lung cancer.

Polyphenolic characterisation and antiprotozoal effect of extracts obtained by maceration, ultrasound, microwave and ultrasound/microwave of Porophyllum ruderale (Jacq.) Cass.

Porophyllum ruderale (Jacq.) Cass. (Asteraceae) has antiprotozoal properties and contains extractable phenolic compounds by the maceration method (M). However, new extraction proposals such as ultrasound (U), microwaves (MW), and ultrasound/microwaves (U/MW) have emerged to optimise yields, but it is unknown if these methods modify effectiveness. Therefore, the study consisted of extracting the aerial part of P. ruderale with ethanol using the M, U, MW and U/MW methods to study its composition by RP-HPLC-ESI-MS, its total polyphenol content and its effect against Entamoeba histolytica. The study showed that U, MW and U/MW did not modify the extraction yield compared to M, but they did change the composition and the total polyphenol content. All extracts contain phloretin, caffeic acid 4-O-glucoside, todolactol A, quercetin 3-O-glucoside, quercetin 3-O-rhamnoside, luteolin and 3,7-dimethylquercetin, and affected the growth of E. histolytica. However, M and U extracts were the most effective at 5 mg/mL.

Tubulin as a potential molecular target for resveratrol in Giardia lamblia trophozoites, in vitro and in silico approaches.

Giardia lamblia is a globally distributed protozoan parasite that causes intestinal disease. Recently, there is an increase in refractory cases of giardiasis to chemotherapeutic agents, and drugs available cause side effects that may limit its use or cause therapeutic non-compliance. Therefore, search for alternative and less harmful drugs to treat giardiasis is an important task. In this sense, resveratrol (RSV) is a polyphenol with a wide range of pharmacological effects such as antimicrobial, anticarcinogenic and antioxidant. The aim of this study was to evaluate the effects of RSV on Giardia lamblia trophozoites in vitro and in silico, focusing on tubulin affectation, a major protein of the Giardia cytoskeleton which participates in relevant processes for cell survival. In vitro determinations showed that RSV inhibits parasite growth and adherence, causes morphological changes, and induces apoptosis-like cell death through tubulin alterations demonstrated by immunolocalization and Western blot assays. Bioinformatic analysis by molecular docking suggested that RSV binds to Giardia tubulin interface heterodimer, sharing binding residues to those reported with depolymerization inhibitors. These findings suggest that RSV affects microtubular dynamics and make it an interesting compound to study for its safety and antigiardiasic potential.

Antifungal In Vitro Activity of Phoradendron sp. Extracts on Fungal Isolates from Tomato Crop.

Synthetic chemicals are mainly used for the control of fungal diseases in tomato, causing the phytopathogens to generate resistance to the chemical active ingredient, with a consequent risk to human health and the environment. The use of plant extracts is an option for the control of these diseases, which is why the main objective of this research was to study an alternative biocontrol strategy for the management of plant diseases caused by fungi through obtaining polyphenol extracts from mistletoe plants growing on three different tree species-mesquite (Prosopis glandulosa), cedar (Cedrus), and oak (Quercus), which contain flavones, anthocyanins, and luteolin. The overall chemical structure of the obtained plant extracts was investigated by RP-HPLC-ESI-MS liquid chromatography. The antifungal effect of these extracts was examined. The target phytopathogenic fungi were isolated from tomato plantations located in Altamira, Tamaulipas, Mexico. The microorganisms were characterized by classical and molecular methods and identified as Alternaria alternata, Fusarium oxysporum, Fusarium sp., and Rhizoctonia solani.

Anticancer Effect of Pomegranate Peel Polyphenols against Cervical Cancer.

Polyphenols are a broad group of bioactive phytochemicals with powerful antioxidant, anti-inflammatory, immunomodulatory, and antiviral activities. Numerous studies have demonstrated that polyphenol extracts obtained from natural sources can be used for the prevention and treatment of cancer. Pomegranate peel extract is an excellent source of polyphenols, such as punicalagin, punicalin, ellagic acid, and caffeic acid, among others. These phenolic compounds have antineoplastic activity in in vitro models of cervical cancer through the regulation of cellular redox balance, induction of apoptosis, cell cycle arrest, and modulation of different signaling pathways. The current review summarizes recent data from scientific reports that address the anticancer activity of the predominant polyphenol compounds present in PPE and their different mechanisms of action in cervical cancer models.

Polyphenolic extract from Punica granatum peel causes cytoskeleton-related damage on Giardia lamblia trophozoites in vitro.

Background: Diarrheal diseases caused by protozoa have a great impact on human health around the world. Giardia lamblia is one of the most common flagellates in the intestinal tract. Factors such as adverse effects to first-line drugs or the appearance of drug-resistant strains, make it necessary to identify new treatment alternatives. Agroindustry waste, like pomegranate peel, are a source of phenolic compounds, which possess antiparasitic activities. In vivo studies demonstrated antigiardiasic potential by reducing cyst shedding and protecting intestinal cells; however, they did not identify the compounds or elucidate any mechanism of action in the parasite. The objective of this study is to identify potential molecular targets and to test the in vitro effects of polyphenols from Punica granatum on Giardia lamblia. Methods: The in vitro antigiardial potential of polyphenolic extract from pomegranate peel (Punica granatum L.) obtained using microwave-ultrasound methodology was evaluated on Giardia lamblia trophozoites. Extract phytochemical identification was performed by HPLC/MS analysis. The effect of polyphenolic extract on growth and adhesion capacity was determined by parasite kinetics; morphological damage was evaluated by SEM, alteration on α-tubulin expression and distribution were analyzed by western blot and immunofluorescence, respectively. Results: The pomegranate peel extract showed the presence of ellagitannins (punicalin and punicalagin, galloyl-dihexahydroxydiphenoyl-hexoside), flavones (luteolin), and ellagic acid, that caused an inhibitory effect on growth and adhesion capacity, particularly on cells treated with 200 µg/mL, where growth inhibition of 74.36%, trophozoite adherence inhibition of 46.8% and IC50 of 179 µg/mL at 48 h were demonstrated. The most important findings were that the extract alters α-tubulin expression and distribution in Giardia trophozoites in a concentration-independent manner. Also, an increase in α-tubulin expression at 200 µg/mL was observed in western blot and diffuse or incomplete immunolabeling pattern, especially in ventral disk. In addition, the extract caused elongation, disturbance of normal shape, irregularities in the membrane, and flagella abnormalities. Discussion: The pomegranate peel extract affects Giardia trophozoites in vitro. The damage is related to the cytoskeleton, due to expression and distribution alterations in α-tubulin, particularly in the ventral disk, a primordial structure for adhesion and pathogenesis. Microtubule impairment could explain morphological changes, and inhibition of adhesion capacity and growth. Besides, this is the first report that suggests that ellagic acid, punicalin, punicalagin and luteolin could be interactioning with the rich-tubulin cytoskeleton of Giardia. Further investigations are needed in order to elucidate the mechanisms of action of the isolated compounds and propose a potential drug alternative for the giardiasis treatment.

Recovery of Bioactive Ellagitannins by Ultrasound/Microwave-Assisted Extraction from Mexican Rambutan Peel (Nephelium lappaceum L.).

Rambutan (Nephelium lappaceum L.) is a tropical fruit from Asia which has become the main target of many studies involving polyphenolic analysis. Mexico produces over 8 million tons per year of rambutan, generating a huge amount of agro-industrial waste since only the pulp is used and the peel, which comprises around 45% of the fruit's weight, is left behind. This waste can later be used in the recovery of polyphenolic fractions. In this work, emerging technologies such as microwave, ultrasound, and the hybridization of both were tested in the extraction of phenolic compounds from Mexican rambutan peel. The results show that the hybrid technology extraction yielded the highest polyphenolic content (176.38 mg GAE/g of dry rambutan peel). The HPLC/MS/ESI analysis revealed three majoritarian compounds: geraniin, corilagin, and ellagic acid. These compounds explain the excellent results for the biological assays, namely antioxidant activity evaluated by the DPPH, ABTS, and LOI (Lipid oxidation inhibition) assays that exhibited great antioxidant capacity with IC50 values of 0.098, 0.335, and 0.034 mg/mL respectively, as well as prebiotic activity demonstrated by a µMax (maximum growth) of 0.203 for Lactobacillus paracasei. Lastly, these compounds have shown no hemolytic activity, opening the door for the elaboration of different products in the food, cosmetic, and pharmaceutical industries.

Co-microencapsulation: a promising multi-approach technique for enhancement of functional properties.

Co-microencapsulation is a growing technique in the food industry because it is a technique that, under the same fundamentals of microencapsulation, allows the generation of microcapsules with a longer shelf life, using a smaller number of encapsulating materials and a smaller amount of active compounds, while having a greater beneficial activity. This responds to consumer demand for higher quality foods that limit the use of ingredients with low nutritional content and provide beneficial health effects, such as probiotics, prebiotics, vitamins, fatty acids, and compounds with antioxidant activity. The combination of two or more active compounds that achieve a synergy between them and between the encapsulating materials offers an advantage over the well-known microencapsulation. Among the main active compounds used in this process are probiotics, prebiotics, fatty acids, and polyphenols, the main combination being that of probiotics with one of the other active compounds that enhances their benefits. The present review discusses the advantages and disadvantages of the different encapsulating materials and techniques used to obtain co-microencapsulants, where the main result is a higher survival of probiotics, higher stability of the active compounds and a more controlled release, which can lead to the generation of new foods, food supplements, or therapeutic foods for the treatment of common ailments.

Purshia plicata Triggers and Regulates Proteins Related to Apoptosis in HeLa Cancer Cells.

Cervical cancer represents a public health problem, develops resistance to traditional therapies and cost-of-treatment is high. These disadvantages have led to the search for alternative bioactive-compound-based therapies. Said bioactive compounds include phenolic compounds, flavonoids, and tannins. The present study aimed to evaluate the therapeutic effect of a P. plicata extract on the HeLa cell line. Viability and apoptosis assays were run on the two cell lines treated with the extract. The peptides, up- and down-expressed in both cell lines, were identified by PDQuest analysis software and high-performance liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS). Our results show that a 500 mg/L treatment deregulated cell viability, with different apoptotic morphologies observed which are associated with the presence of bio-compounds, which up- and down-regulated the peptides. In conclusion, P. plicata regulates proteins associated with apoptosis in HeLa cancer cells.

Insecticidal capacity of polyphenolic seed compounds from neem (Azadirachta indica) on Spodoptera frugiperda (J. E. Smith) larvae.

The objective of this study was to evaluate the insecticidal activity of the polyphenolic compounds found in neem on S. frugiperda larvae. Three neem extracts (1:12 (m/v) with 70% ethanol, 1:12 (m/v) with 0% ethanol (only water), and 1:4 (m/v) with 0% ethanol) were employed. Subsequently, the extraction of phytochemical compounds of each extract was performed using ultrasound and microwave technologies simultaneously. The compound characterization was performed by HPLC-mass. In addition, the insecticidal evaluation of the neem extract was performed against S. frugiperda of the second-stage larvae. The extracts were applied by spraying the larvae according to each bioassay. Results showed that the extract obtained with a 1:12 (m/v) relationship and 70% ethanol was effective for the control of S. frugiperda larvae. In this extract, the predominant organic compound families were: methoxyflavones, flavonols, hydroxycoumarins, anthocyanins, methoxycinnamic acid, and alkylflavones. Phytochemical compounds obtained from neem seeds with environmentally friendly solvents and alternative technologies (ultrasound and microwave) have potent insecticidal activity against S. frugiperda larvae.

Green Bean, Pea and Mesquite Whole Pod Flours Nutritional and Functional Properties and Their Effect on Sourdough Bread.

In this study, proximal composition, mineral analysis, polyphenolic compounds identification, and antioxidant and functional activities were determined in green bean (GBF), mesquite (MF), and pea (PF) flours. Different mixtures of legume flour and wheat flour for bread elaboration were determined by a simplex-centroid design. After that, the proximal composition, color, specific volume, polyphenol content, antioxidant activities, and functional properties of the different breads were evaluated. While GBF and PF have a higher protein content (41-47%), MF has a significant fiber content (19.9%) as well as a higher polyphenol content (474.77 mg GAE/g) and antioxidant capacities. It was possible to identify Ca, K, and Mg and caffeic and enolic acids in the flours. The legume-wheat mixtures affected the fiber, protein content, and the physical properties of bread. Bread with MF contained more fiber; meanwhile, PF and GBF benefit the protein content. With MF, the specific bread volume only decreased by 7%. These legume flours have the potential to increase the nutritional value of bakery goods.

Punicalin and ellagic acid from pomegranate peel extract facilitate apoptotic behaviour in the Hela cell line.

Polyphenols may be an effective therapy for both the prevention and treatment of cancer. Previous studies have found that these compounds may inactive Hela cells, which may even be converted into a normal cells post-treatment. The present study extracted phenolic compounds from pomegranate peel, with the polyphenols then purified using different solvents and identified by means of high-performance liquid chromatography-tandem mass spectrometry (HPLC/MS). Once the phenolic compounds had been purified, we evaluated their cytotoxic effects on both the Hela and NIH-3T3 cell lines, on which an apoptosis assay was also carried out. Additionally, apoptosis assay was carried out on Hela and NIH-3T3. Lastly, the proteome profile was analysed via two-dimensional gel electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS). We isolated and then purified punicalagin and ellagic acid (EA) from pomegranate peel, with both compounds likely to have a cytotoxic effect on Hela and NIH-3T3. However, this effect depends on both concentration and exposure time. Results obtained using a Cayman commercial assay kit suggests that punicalin and EA regulate the apoptosis on the Hela and NIH-3T3 cell lines. Finally, we observed that polyphenols compounds regulate the expression of proteins related to apoptosis. In conclusion, punicalin and EA have a cytotoxic effect on Hela and, furthermore, reactive the apoptotic pathway in this cell.

Classical and New Pharmaceutical Uses of Bacterial Penicillin G Acylase.

BACKGROUND: ß-lactam antibiotics are the most used worldwide for the treatment of bacterial infections. The consumption of these classes of drugs is high, and it is increasing around the world. To date, the best way to produce them is using penicillin G Acylase (PGA) as a biocatalyst. OBJECTIVE: This manuscript offers an overview of the most recent advances in the current tools to improve the activity of the PGA and its pharmaceutical application. RESULTS: Several microorganisms produce PGA, but some bacterial strains represent the primary source of this enzyme. The activity of bacterial PGA depends on its adequate expression and carbon or nitrogen source, as well as a specific pH or temperature depending on the nature of the PGA. Additionally, the PGA activity can be enhanced by immobilizing it to a solid support to recycle it for a prolonged time. Likewise, PGAs more stable and with higher activity are obtained from bacterial hosts genetically modified. CONCLUSION: PGA is used to produce b-lactam antibiotics. However, this enzyme has pharmaceutical potential to be used to obtain critical molecules for the synthesis of anti-tumor, antiplatelet, antiemetic, antidepressive, anti-retroviral, antioxidant, and antimutagenic drugs.

Compuestos fenólicos de Tagetes lucida Cav. con efecto antibacteriano por daños a la membrana / Phenolic compounds of Tagetes lucida Cav. with antibacterial effect due to membrane damage

Tagetes lucida Cav. (Asteraceae = Compositae) se usa para tratar infecciones estomacales. El estudio se centró en evaluar la composición y el efecto antimicrobiano de un extracto de T. lucida Cav. La planta se extrajo con etanol al 10% p/v, y la composición del extracto se analizó por Rp-HPLC-MS. El efecto antibacteriano se evaluó contra Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa y Salmonella choleraesuis utilizando métodos de difusión por disco, microdilución y bioautografía. Los ensayos de sytox y cometa fueron utilizados para evaluar el mecanismo de acción. De esta forma, se detectaron nueve compuestos fenólicos en el extracto de T. lucida. El extracto exhibió actividad solo en S. aureus (MIC de 4.000 mg/ml). La bioautografía reveló que los compuestos fenólicos podrían actuar sinérgicamente. Las pruebas de sytox y cometa mostraron una acción antibacteriana del extracto sobre la membrana bacteriana y el ADN de esta cepa bacteriana.

Tagetes lucida Cav. (Asteraceae=Compositae) is used for treating stomach infections. The study focused on evaluating the composition and antimicrobial effect of an extract of T. lucida Cav. The plant extracted with ethanol at 10% w/v, and the extract composition analyzed by Rp-HPLC-MS. The antibacterial effect was evaluated against Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Salmonella choleraesuis using disk diffusion, microdilution and bioautography methods. The sytox and comet assays were used to evaluate the mechanism of action. In this way, nine phenolic compounds were detected in the extract of T. lucida. The extract exhibited activity only on S. aureus (MIC of 4.000 mg/ml). The bioautography revealed that the phenolic compounds could act synergistically. The sytox and comet tests showed an antibacterial action of the extract on the bacterial membrane and DNA of this bacterial strain.

Native yeasts for alternative utilization of overripe mango pulp for ethanol production / Levaduras nativas para la utilización alternativa de pulpa de mango senescente para la producción de etanol

Mango fruits (Mangifera indica L.) are highly perishable, causing postharvest losses and producing agroindustrial waste. In the present work, native yeasts were used to evaluate ethanol production in overripe mango pulp. The two isolated strains showed similar sequences in the 18S rDNA region corresponding to Kluyveromyces marxianus, being different to the data reported in the NCBI database. Values of up to 5% ethanol (w/v) were obtained at the end of fermentation, showing a productivity of 4g/l/day, a yield of up to 49% of ethanol and a process efficiency of 80%. These results represent a viable option for using the surplus production and all the fruits that have suffered mechanical injury that are not marketable and are considered as agroindustrial waste, thus achieving greater income and less postharvest losses.

Las frutas de mango (Mangifera indica L.) son altamente perecederas, lo cual causa pérdidas poscosecha y produce desechos agroindustriales. En el presente trabajo, se utilizaron 2 levaduras nativas para evaluar la producción de etanol en pulpa de mango senescente. Las 2 cepas aisladas mostraron similitud en la región 18S ADNr, correspondiente a Kluyveromyces marxianus, la cual es diferente a lo reportado en la base de datos del NCBI. Se obtuvieron valores de hasta el 6% de etanol (v/v) al final de la fermentación, con una productividad de hasta 4g/l/día, un rendimiento de hasta 49% de etanol y una eficiencia en el proceso fermentativo del 80%. Esto representa una opción viable para utilizar excedentes de producción o frutos que han sufrido daño mecánico y no son comercializables, al lograr más ingresos y menos pérdida poscosecha.

Exploring the Degradation of Gallotannins Catalyzed by Tannase Produced by Aspergillus niger GH1 for Ellagic Acid Production in Submerged and Solid-State Fermentation.

Due to great interest on producing bioactive compounds for functional foods and biopharmaceuticals, it is important to explore the microbial degradation of potential sources of target biomolecules. Gallotannins are polyphenols present in nature, an example of them is tannic acid which is susceptible to enzymatic hydrolysis. This hydrolysis is performed by tannase or tannin acyl hydrolase, releasing in this way, biomolecules with high-added value. In the present study, chemical profiles obtained after fungal degradation of tannic acid under two bioprocesses (submerged fermentation (SmF) and solid state fermentation (SSF)) were determined. In both fermentation systems (SmF and SSF), Aspergillus niger GH1 strain and tannic acid as a sole carbon source and inducer were used (the presence of tannic acid promotes production of enzyme tannase). In case of SSF, polyurethane foam (PUF) was used like as support of fermentation; culture medium only was used in case of submerged fermentation. Fermentation processes were monitored during 72 h; samples were taken kinetically every 8 h; and all extracts obtained were partially purified to obtain polyphenolic fraction and then were analyzed by liquid chromatography-mass spectrometry (LC-MS). Molecules like gallic acid and n-galloyl glucose were identified as intermediates in degradation of tannic acid; during SSF was identified ellagic acid production. The results obtained in this study will contribute to biotechnological production of ellagic acid.

Búsqueda de biomarcadores a partir de péptidos expresados en biopsias de cáncer de mama / Search for biomarkers from peptides expressed in breast cancer biopsies / Busca de biomarcadores a partir de peptídeos expressos em biópsias câncer de mama

El cáncer de mama (CM) es una de las principales causas de muerte en México. Se ha observado un incremento en la incidencia de éste en mujeres de 15-29 años. A fin de comprender las causas en el desarrollo del CM, se pretendió buscar la asociación entre los genes/enfermedad empleando técnicas de Biología Molecular. Se analizaron, por genómica funcional, 50 biopsias frescas de pacientes con CM (BFCM), 50 biopsias embebidas en parafina de CM (BEPCM) y 10 biopsias frescas de pacientes con sospecha de CM (BFSC), obtenidas de mujeres que residen en Coahuila, México. Las muestras proteicas se cuantificaron y se resolvieron en geles de poliacrilamida dodecil sulfato de sodio (SDS-PAGE) y en dos dimensiones (2-DE). El perfil proteico de las BFCM, BEPCM versus BFSC mostró diferencias entre las bandas peptídicas observadas en los geles. Aquellos péptidos que se diferenciaron por su expresión fueron analizados por cromatografía líquida acoplada a masas en tándem (LC/ MS/MS). Las huellas peptídicas obtenidas, a su vez, se analizaron por medio del banco de genes (PubMed). Se encontraron, en las muestras de cáncer, proteínas asociadas a migración celular, supresión de tumores, estrés oxidativo y choque térmico. Por último, estos hallazgos se confirmaron empleando inmuno-electro transferencia o Western blot (WB) con anticuerpos contra vimentina.

Breast cancer (BC) is one of the leading causes of death in Mexico. Moreover, BC is the main cause of death in women between 15-29 years old in northern Mexico. Proteomic techniques have been used in order to achieve a better understanding of the genes involved in the development of BC. The proteins in BC extracted from 50 fresh breast cancer tissues (FBCT), 50 paraffin embedded breast cancer tissues (PEBCT) and 10 biopsies from women suspected of cancer (SC), residing in Coahuila, Mexico were analyzed in this paper. The quantity of protein extracted was similar in both samples FBCT and PEBCT. However, protein quality was lower in PEBCT than FBCT. Subsequently, these proteins were resolved in SDS-PAGE and 2DE. Differences were noticed in protein profile and all those suspect proteins were analyzed by LC/MS/MS. Amino acidic fingerprint allowed for the identification of peptides associated with a) cell migration, b) tumor suppression, c) oxidative stress or heat shock.

O câncer da mama (CM) é uma das principais causas de morte no México. Observou-se um aumento na incidência desse câncer em mulheres entre os 15-29 anos de idade. Para compreender as causas do desenvolvimento de CM, visou-se encontrar a associação entre os genes/doença utilizando técnicas de Biologia molecular. Analisaram-se por genômica funcional, 50 biópsias frescas de pacientes com CM (BFCM), 50 biópsias embebidas em parafina (BEPCM) e 10 biópsias frescas de pacientes com suspeita de CM (BFSC), obtidas de mulheres residentes em Coahuila, México. As amostras de proteínas foram quantificadas e separadas em géis de poliacrilamida dodecil sulfato de sódio (SDS-PAGE) e em duas dimensões (2-DE). O perfil proteico das BFCM, BEPCM comparado com BFSC mostrou diferenças entre as bandas peptídicas observadas nos géis. Esses peptídeos que diferem em sua expressão foram analisados por cromatografia líquida acoplada a massas em tandem (LC/MS/MS). As pegadas peptídicas obtidas, por sua vez, foram analisadas utilizando o banco de genes (PubMed). Verificaram-se nas amostras de câncer, proteínas associadas à migração celular, supressão de tumores, estresse oxidativo e choque térmico. Finalmente, estes achados foram confirmados utilizando a imuno-eletro transferência ou Western Blot (WB) com anticorpos contra vimentina.

Efecto de las diferentes fuentes de polifenoles sobre la eficiencia de liberación de ácido elágico por Aspergillus niger / Effect of different polyphenol sources on the efficiency of ellagic acid release by Aspergillus niger

Fungal hydrolysis of ellagitannins produces hexahydroxydiphenic acid, which is considered an intermediate molecule in ellagic acid release. Ellagic acid has important and desirable beneficial health properties. The aim of this work was to identify the effect of different sources of ellagitannins on the efficiency of ellagic acid release by Aspergillus niger. Three strains of A. niger (GH1, PSH and HT4) were assessed for ellagic acid release from different polyphenol sources: cranberry, creosote bush, and pomegranate used as substrate. Polyurethane foam was used as support for solid-state culture in column reactors. Ellagitannase activity was measured for each of the treatments. Ellagic acid was quantified by high performance liquid chromatography. When pomegranate polyphenols were used, a maximum value of ellagic acid (350.21 mg/g) was reached with A. niger HT4 in solid-state culture. The highest amount of ellagitannase (5176.81 U/l) was obtained at 8 h of culture when cranberry polyphenols and strain A. niger PSH were used. Results demonstrated the effect of different polyphenol sources and A. niger strains on ellagic acid release. It was observed that the best source for releasing ellagic acid was pomegranate polyphenols and A. niger HT4 strain, which has the ability to degrade these compounds for obtaining a potent bioactive molecule such as ellagic acid.

La hidrólisis fúngica de los elagitaninos produce ácido hexahidroxidifénico, considerado como una molécula intermedia en la liberación de ácido elágico. El ácido elágico tiene importantes y deseables propiedades benéficas para la salud humana. El objetivo de este trabajo fue identificar el efecto de la fuente de elagitaninos sobre la eficiente liberación de ácido elágico por Aspergillus niger. La liberación de ácido elágico se realizó con tres cepas de A. niger (GH1, PSH y HT4) en presencia de diferentes fuentes de polifenoles (arándano, gobernadora y granada), usadas como sustrato. Se empleó espuma de poliuretano como soporte para el cultivo en estado sólido en reactores en columna. Se midió la actividad elagitanasa a cada uno de los tratamientos. El ácido elágico liberado se cuantificó por cromatografía líquida de alta resolución. Cuando se utilizaron los polifenoles de granada, se alcanzó un valor máximo de 350,21 mg/g de ácido elágico con A. niger HT4 en cultivo en estado sólido. La mayor actividad elagitanasa (5176.81 U/l) se obtuvo a 8 h de cultivo cuando se usaron los polifenoles de arándano como sustrato y A. niger PSH. Los resultados demostraron el efecto que tiene la fuente de polifenoles y la cepa de A. niger en la liberación de ácido elágico. Se observó que la mejor fuente para la liberación de ácido elágico fueron los polifenoles de granada y que la cepa A. niger HT4 posee la habilidad de degradar estos compuestos para la obtención de potentes moléculas bioactivas, como el ácido elágico.