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.

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.

Association of the Vascular Endothelial Growth Factor Gene Polymorphism +936 C/T with Diabetic Neuropathy in Patients with Type 2 Diabetes Mellitus.

BACKGROUND: Peripheral neuropathy is one of the most common late complications of diabetes. Vascular endothelial growth factor (VEGF) gene polymorphisms have been associated with the development of peripheral neuropathy in different populations of patients with type 2 diabetes mellitus (DM2). OBJECTIVE: To analyze the prevalence of the +936 C/T VEGF gene polymorphism among patients with DM2 with and without peripheral neuropathy. STUDY DESIGN AND METHODOLOGY: 218 unrelated DM2 patients, 90 with and 128 without peripheral neuropathy were genotyped for the +936 C/T VEGF gene polymorphism using PCR amplification followed by restriction length polymorphism analysis. RESULTS: The CC homozygous VEGF+936 C/T (rs3025039) was the predominant genotype in DM2 patients with peripheral neuropathy, whereas the predominant genotype in patients without neuropathy was the heterozygous C/T. No statistical association was found between genotype distribution and the presence of neuropathy (p = 0.063). The distribution of the genotypes according to the dominant (CC vs. CT + TT) and recessive (TT vs. CT + CC) models showed that the homozygous CC and TT genotypes, respectively, are not risk factors for neuropathy. The CT genotype conferred a protective effect as seen in the over-dominant model (CT vs. CC + TT) (OR = 0.52; 95% CI = 0.300-0.90; p = 0.019). CONCLUSION: We conclude that the VEGF+936 C/T (rs3025039) gene polymorphisms are related to peripheral neuropathy in Mexican DM2 patients, with the heterozygous genotype potentially conferring a protective effect.

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.

Gallic acid decreases hepatitis C virus expression through its antioxidant capacity.

Gallic acid (GA) is a natural phenolic compound that possesses various biological effects, including antioxidant, anti-inflammatory, antibiotic, anticancer, antiviral and cardiovascular protection activities. In addition, numerous studies have reported that antioxidants possess antiviral activities. Hepatitis C virus (HCV) is one of the most important causes of chronic liver diseases worldwide, but until recently, only a small number of antiviral agents had been developed against HCV. Therefore, the present study investigated whether GA exhibits an anti-HCV activity. The effects of GA on HCV expression were examined using a subgenomic HCV replicon cell culture system that expressed HCV nonstructural proteins (NSs). In addition, GA cytotoxicity was evaluated at concentrations between 100-600 mg/ml using an MTT assay. Huh-7 replicon cells were incubated with 300 mg/ml GA for different times, and the HCV-RNA and protein levels were measured by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Pyrrolidine dithiocarbamate (PDTC) was used as an antioxidant control and reactive oxygen species (ROS) production was measured during the exposure. The results indicated that GA did not produce a statistically significant cytotoxicity in parental and HCV replicon cells. Furthermore, GA downregulated the expression levels of NS5A-HCV protein (~55%) and HCV-RNA (~50%) in a time-dependent manner compared with the levels in untreated cells. Notably, GA treatment decreased ROS production at the early time points of exposure in cells expressing HCV proteins. Similar results were obtained upon PDTC exposure. These findings suggest that the antioxidant capacity of GA may be involved in the downregulation of HCV replication in hepatoma cells.

A novel arrangement of Cys residues in a paralytic peptide of Conus cancellatus (jr. syn.: Conus austini), a worm-hunting snail from the Gulf of Mexico.

The present study details the purification, the amino acid sequence determination, and a preliminary characterization of the biological effects in mice of a new conotoxin from the venom of Conus cancellatus (jr. syn.: Conus austini), a worm-hunting cone snail collected in the western Gulf of Mexico (Mexico). The 23-amino acid peptide, called as25a, is characterized by the sequence pattern CX1CX2CX8CX1CCX5, which is, for conotoxins, a new arrangement of six cysteines (framework XXV) that form three disulfide bridges. The primary structure (CKCPSCNFNDVTENCKCCIFRQP*; *, amidated C-terminus; calculated monoisotopic mass, 2644.09Da) was established by automated Edman degradation after reduction and alkylation, and MALDI-TOF and ESI mass spectrometry (monoisotopic mass, 2644.12/2644.08Da). Upon intracranial injection in mice, the purified peptide provokes paralysis of the hind limbs and death with a dose of 240 pmol (~0.635 µg, ~24.9 ng/g). In addition, a post-translational variant of this peptide (as25b) was identified and determined to contain two hydroxyproline residues. These peptides may represent a novel conotoxin gene superfamily.

In vitro and in vivo biocompatibility of apatite-coated magnetite nanoparticles for cancer therapy.

The aim of this study was to determine the biocompatibility and potential toxicity of apatite-coated magnetite nanoparticles. The in vitro biocompatibility with human red blood cells was evaluated, not hemolytic effects were found at concentrations lower than 3 mg/ml. For the in vivo study, Balb/c mice were used. The animals were injected intravenously or intraperitoneally, the doses ranged from 100 to 2,500 mg/Kg. All the injected animals showed normal kidney and liver function. No significant changes were found in the body weight, the organs weight and the iron levels in liver due to the administration. In conclusion, apatite-coated magnetite nanoparticles did not induce any abnormal clinical signs in the laboratory animals. The results demonstrated that apatite-coated magnetite nanoparticles of 8 ± 2 nm in size did not have hemolytic effect in human erythrocytes and did not cause apparent toxicity in Balb/c mice under the experimental conditions of this study.