Special Section : Medical and Pharmaceutical Biotechnology

 

1-Phenyl-1-Propanol Obtained by Reduction of Propiophenone with Nocardia corallina B-276

María Teresa Lara Carvajal, Herminia Inés Pérez Méndez, Aida Solís Oba, Norberto Manjarrez Alvarez, Liliana Hernández Vázquez, Myrna Solís Oba.

  • Abstract

    Currently the production of flavoring agents, agrochemicals and pharmaceutical products has become important, through the development of environmentally friendly biotechnological processes to obtain enantiomerically pure compounds by enzymes or microorganisms. 1-phenyl-1-prophanol is an important chiral reagent used as a flavoring, in perfumery and precursor of chiral complex compounds in organic chemistry, with a high commercial value, so in this investigation enantioselective bioreduction of propiophenone to 1-phenyl-1-propanol had been studied with whole cells of Nocardia corallina B-276 in phosphate buffer at pH 7.0 with a substrate:wet cells ratio (m/m) 1:500; compounds were characterized by IR and 1H NMR and 13C NMR, analytical methods were developed by GC to determine percentage conversion and by HPLC to determine enantiomeric ratio. According to the results it was observed that the bioreduction of propiophenone to 1-phenyl-1-propanol with Nocardia corallina is pH dependent; the optimum pH of the supernatant of the liquid fermentation for the conversion of the ketone to the R-enantiomer is at pH 5.67 in 72 h.

 

Implementation of an Intelligent Controller into a Robotic Device for Rehabilitation Intended for Lower Extremity Paralysis Patients

Gerson Figueroa-Flores, Alfredo Leal-Naranjo, Christopher René Torres-San Miguel, Guillermo Urriolagoitia-Sosa, Guillermo Urriolagoitia-Calderón

  • Abstract

    This paper presents the design of an intelligent controller for an inverted pendulum system. First briefly describe the most recent and current research on this topic, specifically in the last 10 years. Then a series of definitions which aims to delve the reader to understand more in detail the literature of this research will be proposed. Once and bases studied issue is known we proceed to develop the mathematical model of the system and then convert the equations of motion both transfer function as state variables, eventually stability analysis will be developed in open loop the two methods mentioned above, in order to carry out a digital simulation by blocks through Matlab Simulink©. Is presented through graphs, the behavior of the system variables under study. Then based on the results obtained above will proceed to design different fuzzy controllers, in order that only one of these is the one that achieves optimal response to obtain a good system stability. Also presented through graphs, the state variables of the system after being controlled by the fuzzy controller designed. To check its stability analysis is presented, from the graphical method of phase plane. Finally obtained the desired response, an application of the inverted pendulum system focused on the biomechanics, a robot which will help the rehabilitation of patients with lower limb paralysis will be designed, the application will be virtually simulated to verify proper operation.

 

Effect of Metatera Extract on Wound Closure

Adriana Martínez Cuazitl, Julia Antonia Ceja de León, María del Consuelo Gómez

García, Blanca Rosa Buenrostro Espinosa, Eduardo San Martín Martínez, Francisco Javier Paz Bermudez, Mario García Solis,David Guillermo Pérez Ishiwara1.

  • Abstract

    Wounds represent a major health problem, in terms of morbidity and mortality,the primary goals of wound treatment are the rapid wound closure and a functional scar.There are medicinal plants used empirically for treatment of wounds such as metatera plant (Scrophulariaceae family). Our group demonstrated in vitro that metatera extract (ME) induces important events for wound closure as proliferation, migration and differentiation of fibroblast. The main objective of this work was to determinate the wound closure effect induced by metatera extract in an excisional in vivo model. Metatera extract included in a hydrogel (ME-H) was optimized for dairy topical application during 15 days. We analyzed the wound closure rate effect, the histopathology changes and the total collagen content in tissues treated with ME-H. The morphometric analysis showed that ME clearly improve wound closure. Moreover, histopathology changes induced by this extract suggest that ME induced different cellular events, such as fibroblast maturation and fibroblast and collagen fibers arrangement. Our results supported theethnobotanic use of this plant for wound care in Mexico.

 

Chronic Prostatitis / Chronic Pelvic Pain Syndrome:Histological Evidences of treatment with anImmunotherapeutic Agent in a Murine Model 

Pérez-Alvarado Carlos Manuel, Ma del Consuelo Gómez García, Miguel Reyes López,Mario García Solís, Elizabeth Pérez Hernández, Carlos Adolfo Pérez de la Mora, Bulmaro Esteban Correa Meza, Virginia Sánchez Monroy, Francisco Paz Bermudez8 and David Guillermo Pérez Ishiwara.

  • Abstract

    Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) has a strongautoimmune component and is the most important type of prostatitis in worldwide. Hereusing a modification of the murine model of experimental autoimmune prostatitis (EAP), we showed that besides an infiltrate of inflammatory cells in the prostatic tissue, animals developed benign prostatic hyperplasia. Different doses of immune regulatory agent were administrated to experimental animals, observing that the dialyzable leukocyte extract (DLE) decreased the infiltrate of inflammatory and mast cells.

 

Bioreduction of Phenyl Ketones for the Obtention of Enantiopure Alcohols

Cervantes Fadia, Solís Aída, Martínez Rosa, Pérez Herminia, Manjarrez Norberto, Hernández Liliana, Solís Myrna.

  • Abstract

    The demand for chiral intermediate compounds with high enantiomeric purity has increased markedly in industries. Chiral secondary alcohols are frequently required as important intermediates for the introduction of chiral center into the pharmaceuticals, flavor, aroma and agricultural chemicals, and specialty materials. Biocatalytic asymmetric reductions can offer highly selective reactions, environmentally benign processes, and energy-effective operations and thus of great interest. Alcohol dehydrogenases (ADH), are a class of nicotinamide dependent oxidoreductases, which can catalyze the reduction of carbonyl compounds to produce alcohols. Three different sources of oxidoreductase were tested (alberjon, alubia and ayocote) for the biotransformation of acetophenone (1a) and propiophenone (1b) to their respective alcohols (2a,2b), with alberjon as enzyme source, the reduction of both ketones was similar. With alubia and ayocote bean the enzyme was sensitive to the chain size of the ketone, the reduction of 1a was almost the double than that of 1b this could be because a steric effect. With regard to the ee, there was not a correlation between the chain size and the ee, it was more dependent on the enzyme source. Alberjon, 2a 71%, 2b (10%); alubia, 2b had a 58% ee, 28% for 2a; ayocote beans, 2a 38% ee and 29% ee for 2b.

 

Aqueous Crude Extract of Montanoa tomentosa Exerts Anxiolytic-Like Effects in Female Rats with Long-Term Absence of Ovarian Hormones

Juan Francisco Rodríguez-Landa, Magdalena Guadalupe Rodríguez-Santiago, María de Jesus Rovirosa-Hernández, Francisco García-Orduña, Miguel Carro-Juárez

  • Abstract

    Natural or “surgical” menopause is characterized by a reduced concentration of steroid hormones, which is associated with a major incidence of vasomotor symptoms, vaginal dryness, osteoporosis, cognitive deterioration, hot flashes, irritability, anxiety and mood swings, among others. Hormonal and pharmacological therapies have been used to treat those alterations, but in some woman produce side effects limiting their long-term use. Therefore new studies to found alternative therapies for the management of those alterations in women are needed. In the Mexican traditional medicine there are some plants as Montanoa tomentosa recommended to prevent and treat women´s illness, including emotional and mood swings, but specific studies on the effects produced by this plant on anxiety associated with low concentration of steroid hormones have not available. Therefore in the present study the aim was to evaluate the effect of the aqueous extract of M. tomentosa in rats with long-term absence of ovarian hormones induced by ovariectomy. Three concentration of M. tomentosa extract (12.5, 25 and 50 mg/kg) were evaluated in rats subjected to elevated plus maze and open field tests and comparisons against a control group and diazepam as reference of an anxiolytic drug were described. In the elevated plus maze only 50 mg/kg of M. tomentosa extract, alike to diazepam, increased the time spent and the number of entries into the open arms, respect to control group. Similar to diazepam, in the open field test none of the evaluated treatments produced significant changes in crossing and rearing, but 50 mg/kg of M. tomentosa extract increase time spent in grooming behavior as compared with control group. In conclusion M. tomentosa extract produces an anxiolyticlike effect in rats with long-term absence of ovarian hormones. Findings partially support the traditio

 

Cytotoxic Effects of Sábila (Aloe Vera): Commercial Products as Food Supplement

Zaira López, Peter Knauth, Guadalupe Avalos-Navarro, Virgilio Bocanegra-García, José A. Ramírez, Rosalía Reynoso-Camacho and Gildardo Rivera

  • Abstract

    Aloe vera is used intensively in traditional medicine because it’s widely number of therapeutic properties, and due to that it is used as food supplement. Although it is well known that the main class of compounds responsible for aloe-induced toxicity is anthraquinones, it seems to be that their concentrations are reduced in commercial products. To our knowledge, few studies concerning to the toxicity of aloe-based product have been documented. This lack of data suggested us to evaluate the effect of two different commercial products on cell viability, which were 200x Dehydrated Inner Leaf Powder (ILP) and 100x Dehydrated Whole Leaf Powder (WLP). The HPLC analysis revealed two peaks in each product; aloin (5.82 ppm for ILP and 4.75 ppm for WLP) and aloe-emodin (5.73 ppm for ILP and 1.68 ppm for WLP) indicating low concentrations compared with other studies. For cytotoxic analysis, both products were dissolved in cell culture medium getting final concentrations of 0.5x, 1x, 5x, and 10x; at pH 7.3 and 50 μg/ml gentamicine. Cells were analyzed by cell membrane integrity (LDH-release), metabolic activity (WST-1), functional lysosomes (NRU) and apoptosis induction (Annexin V/PI). The results indicated that both products are not cytotoxic at the recommended concentration of 1x, which it is used as food supplement; this could be explained due to the low anthraquinones concentration. The cytotoxic effects were observed from 5 up to 10x fold over-concentrated; a possible reason is that the toxic effect might be caused by mechanical stress by the fibres. For another side, no apoptotic effect was detected at concentrations up to 10x with bothproducts. This work was a short cytotoxicity study in a classical cancer cell line (HeLa), weare considering to do further toxicological evaluations with other commercial products in order to know the risk for to use them as food supplements 

 

Essential Oil from Poly-phytopharm Herbs AttenuateManifestations of Influenza Illness Mice Model

Hernández Alarcón Rita Xochitl, María del Consuelo Gómez Garcia, Elizabeth PérezHernández, Roberto González González, Nury Pérez Hernández, Virginia Sánchez Monroy Olivia Medel Flores, Elvia Pérez Soto and David Guillermo Pérez

Ishiwara1,2

  • Abstract

    Influenza caused by influenza a virus, is an acute self-limited respiratory illness, its severe form is characterized by an intense inflammatory response, pulmonary infiltrates, vasodilator shock and high mortality. Treatment of disease is mainly focus to the use of a limited set of anti-viral drugs. To search for the development of new potential drugs for the treatment of influenza disease, here we report the therapeutic effects of an essential oil obtained from a poliphytopharmacological formulation (PEO), based on cinnamon, guava, propolis, and menthol on influenza A murine model. Animals administered prophylactically with PEO showed less severe clinical symptoms. Histophatologically, there were no evidence of pulmonary parenchymal consolidation and in contrast to non-treated animals, the lung inflammatory process was very limited. 

 

Generation And Preliminary Characterization of Murine IgM Monoclonal Antibodies against the N-terminal β-strand Of A Fibrillogenic Light Chain Variable Domain

Olga Mata Ruiz, Yolanda Medina Flores, Miying Dessire Gómez Cervantes1, Diana Reynada Castillo, Robin Axel Ruiz Zamora, Francisco Javier Rodríguez Álvarez, Karen Cortes Sarabia, Luis del Pozo Yauner.

  • Abstract

    Based on findings of previous studies, we have identified the N-terminal segment of the light chains variable domains as a structurally important region for the mechanism of amyloid fibril assembling. Thus, we have suggested that this region could be target for monoclonal antibodies (MAb) with the potential for inhibiting the amyloid aggregation of the light chains, and/or suitable for the inmunohistological detection of the AL deposits. In this work, we report the generation and preliminary characterization of a group of murine monoclonal antibodies against the N-terminal β strand of the fibrillogenic variable domain protein 6aJL2. The immunization of the laboratory animals (three female Balb/c mice 4-6 weeks old) was performed by intraperitoneal inoculation of a mix composed by the synthetic peptide Asn1-Lys17 (50 μg per animal) homogenized with the adjuvant TiterMax® Gold (Sigma-Aldrich Cat No. T2684). The generation of the hybridomas was accomplished by the conventional methodology, using murine myeloma cells X63Ag8.653. The hybridoma screening and antibody characterization were performed with two different methodologies: indirect ELISA and dot-blot. In both techniques, the antibody recognition was evaluated against the synthetic peptide Asn1-Lys17 and the recombinant protein 6aJL2, from which the peptide originated. Ten different hybridomes were selected by the capacity to produce monoclonal antibodies (MAb) that bind the peptide Asn1-Lys17. Interestingly, all the MAb were of IgM class, as determined by indirect ELISA. The preliminary characterization indicates that all the antibodies recognize the 6aJL2 protein in the fibrillar state, but not in the native conformation.

 

Antioxidant Capacity of Prosthechea karwinskii (Orchidaceae) Extracts Obtained by Sonication

Alejandra Rojas-Olivos, Rodolfo Solano-Gómez, Manuel Jiménez-Estrada, Blanca Yakelina Silva-Valeriano, Luicita Lagunez-Rivera.

  • Abstract

    Prosthechea karwnskii is a valuable ornamental orchid in Oaxaca, Mexico. In the Mixteca region, this plant has uses in traditional medicine as well as in religious ceremonies. The presence of phenolic compounds with antioxidant potential in this orchid has been reported, although not determined. Considering the advantages of the use of sonication in secondary metabolite extraction from medicinal plants, hydroethanolics and hydroethanolics extracts were obtained from each part of this orchid (pseudobulb, leaf and flower) using sonication pretreatment in order to determine antioxidant capacity. The results indicated that both hydroethanolic (IC50: 3.52 ± 0.078 mg / ml) and hydromethanolic (IC50: 8.88 ± 1.826 mg / ml) leaf extracts had a higher antioxidant capacity than pseudobulbs and flower extracts. Leaf extracts also exhibited inhibition in the breast cancer cell line (MCF-7). The results demonstrated the antioxidant capacity of this orchid and its potential for application in assessments related to oxidative stress diseases.

 

Histopathological And Clinical Analysis Of Balb/C Mice Infected With Mouse-Adapted Pandemic Influenza A/H1N1 Virus Using Different Viral Doses

Elvia Pérez Soto, Mario García Solís, Carlos Cabello Gutiérrez, María del Consuelo Gómez García, Carlos Adolfo Pérez de la Mora, Bulmaro Esteban Correa Meza, Pablo Muriel, Guillermo Pérez Ishiwara.

  • Abstract

    Influenza A is a serious respiratory illness that can be debilitating and may cause complications leading to hospitalization and death. Pathology has been associated with severe immune response in the lung and is the main cause of death among patients with viral pneumonia caused by pandemic (H1N1) 2009, producing acute respiratory distress syndrome (ARDS) with diffuse alveolar damage (DAD). Here, we examined serial pathological changes in the lungs of the mice infected with mouse-adapted pandemic influenza 2009 A/H1N1 virus using different viral doses. Correlating the clinical signs and the histopathological changes in the different experimental groups, we identified specific viral doses causing from a slight pneumonia to ARDS/DAD. In the group infected with 8 hemagglutinating unit (HA), we found edema, lymphocytic infiltrates, acute interstitial pneumonitis and fibrosis. However, in the group infected with 16 HA, severe interstitial pneumonitis, alveolar collapse, hemorrhage, septal congestion and dense fibrosis was developed as reported in humans. The infection with 16 HA is an ideal, fast and efficient model to evaluate immunomodulatory drugs that diminished the ARDS and DAD.

 

Assessment of the Antiviral Effect of a CysLT1 Receptor Inhibitor on the Infection rate of Dengue Virus 2 In Vero Cells

Aldo Segura-Cabrera,2J.S. Salas-Benito,Carlos A. García-Pérez, Mónica Ascención De Nova-Ocampo and Mario A. Rodríguez-Pérez

  • Abstract

    Dengue (DENV) is the most important arbovirosis worldwide, afflicting humans mainly of the tropical and subtropical developing regions. Neither effective DENV vaccines nor antiviral agents are currently available. The DENV polyprotein precursor is cleaved by both host proteases and the two-component virus protease NS2B-NS3. This complex protease is accounted for when designing DENV antivirals. A structure-based virtual screening protocol was designed to identify a promising set of leads against the catalytic domain in active form of DENV NS2B-NS3 protease. The docking results were subjected to a consensus scoring analysis. Thus, six docked poses were ranked among the top 50 compounds, which were used to molecular dynamics simulation and free-energy calculation. One of those leads resulted to be a FDA-approved CysLT1 receptor inhibitor. To confirm the inhibitory activity of the candidate, it was assayed in vitro with the PlateliaTM Dengue NS1 ELISA (BioRad) and using DENV 2 (NG strain) and Vero cells. A reduction in the infection of 29% was seen at a concentration of 80 μg/ml but this did not reach statistical significance (p>0.05) when using a completely random ANOVA with post hoc mean comparison Bonferroni-test. However a highly significant (p <0.01) reduction of 36% was noted at 160 μg/ml.

 

Western Blot to Diagnose Invasive Amoebiasis. A Biotechnological Development

M.S. Flores, G. Mena,K. Arévalo, G. Maldonado and L. Galán.

  • Abstract

    Entamoeba histolytica infects 50 million persons worldwide and cause 50,000 to 100,000 deaths yearly. Intestinal Invasive amoebiasis induces dysentery with trophozoites and cysts in feces; the amoebic liver abscess is the most frequent manifestation of extra intestinal amoebiasis. Parasites detected in feces diagnose intestinal amoebiasis, but patients with amoebic liver abscess frequently do not release amoebas in stool. Amoebic liver abscess is diagnosed by ultrasound or computed tomography and serological tests. Here we present a biotechnological development to diagnose invasive amoebiasis using preserved antigenic molecules without enzymatic inhibitors. The amoebic antigenic extract was resolved by SDS-PAGE and transferred to nitrocellulose membranes. Each membrane strip was incubated with sera from patients with amoebic liver abscess; sera from patients with intestinal amoebiasis or sera from clinically healthy volunteers. We identified six antigenic amoebic immunodominant bands, recognized exclusively by the antibodies from patients with ALA, but not recognized by the antibodies present in the sera of healthy people without amoebiasis living in endemic zones, neither sera from individuals with intestinal amoebiasis. These antigenic bands constitute a WB pattern to diagnose invasive amoebiasis. The WB diagnostic test to invasive amoebiasis is useful in amoebiasis endemic zone. The project was supported by PAICYT, UANL.

 

Molecular Effectors Induced By Metatera Extract During Skin Wound Healing In a in vivo Model

Adriana Martínez Cuazitl; Consuelo Gómez Garcia; Virginia Sánchez Monroy; Eduardo San Martin Martinez; Francisco Javier Paz Bermudez Mario García Solis, David Guillermo Pérez Ishiwara.

  • Abstract

    Wound healing is regulated by several molecular interactions, characterized by three phases. In vitro, we have found that extract of metatera plant (ME) (Scrophulariaceae family) induce fibroblast differentiation, migration, proliferation and adhesion. The main objective of this work was determinated the molecular effectors and signaling pathways induced by ME plant in a in vivo model, and the correlation with morphometric and histopathological changes. Positive control and ME were evaluated in excisional in vivo wound model. We analyze morphometric and histopatological changes at 3, 5 and 7 days post wounding. Moreover, we investigated by immunohistochemistry the expression of molecular effectors induced by ME. Our results showed that the inflammation process and fibroblast migration were regulated by TGF􀀀1 in ME groups. Additionally, ME regulates key molecules related with proliferation and differentiation fibroblast, like PCNA, 􀀀-SMA and TGF􀀀1. It also promotes the expression of TGF􀀀3 and matrix components, such as collagen type I, stimulating neoformation,  aturation, arrangement and horizontal alignment of collagen fibers. Our results demonstrated that metatera extract regulated the expression of key molecules that control several molecular and cellular events in the three phases of healing process, accelerating the wound closure in excisional in vivo model. 

 

Total Carotenoid Content of Actinobacteria Isolated from Marine Sediment of the Gulf of México

Lucía Ortega Cabello, Herminia I. Pérez Méndez, Luis A. Maldonado Manjarrez, Julia Cassani Hernández, Erika T. Quintana Cano, Norberto Manjarrez Alvarez

  • Abstract

    Carotenoids are organic pigments that are synthesized by photosynthetic organisms such as bacteria. These types of molecules have important applications due to their chemopreventive activity and as color providers to different products. Carotenoid production from microbial sources have been important because is an eco-friendly, stable and high productivity technique. The aim of the present research was to compare total carotenoid content between five actinobacteria strains (named JOC-4, JOC-5, JOC-7, JOC-8 and JOC-9) isolated from the Gulf of México. Strains were grown on plates of GYEA medium during ten days at 28ºC, followed by fermentation at 28ºC, 150 rpm for six days, biomass and supernatant were lyophilized and carotenoid extraction was performed using a mixture of dichloromethane:methanol:acetone (1:1:2), total carotenoids were calculated with UV-Vis spectrometry, 450 nm. To determine significant differences, an anidated experiment and a Scheffè test were performed. A significant difference in carotenoid content was determined between biomass and the strains with an F value of 21.29 and 11.19, respectively (p < 0.05). The Scheffé test indicated that biomass from strains JOC-5 (25.37±3.02 μg/mL) and JOC-8 (39.55±4.55 μg/mL) presented the highest carotenoid content and the less variation. While JOC-4, JOC-7 and JOC-9 presented high variation in carotenoid content.

 

Comparison of Carotenoid Production of Two Carotenogenic Strains in GYEA Media Supplemented with Iron and Copper salts

Lucía Ortega Cabello, Herminia I. Pérez Méndez, Luis A. Maldonado Manjarrez, Julia Cassani Hernández, Erika T. Quintana Cano, Norberto Manjarrez Alvarez

  • Abstract

    Microbial production of carotenoids can be influenced by some factors, among these factors, the presence of mineral salts can help the microorganism to produce a higher quantity of carotenoids. The aim of the present research was to compare carotenoid production of two actinobacteria strains (JOC-5 and JOC-8) in GYEA alone and with FeCl2, FeSO4 and CuSO4. Strains were grown on plates of GYEA alone and with salts at concentrations ranging from 0.025-1.00%, 28ºC, followed by fermentation, 28 ºC, 150 rpm, biomass was lyophilized; carotenoid extraction was performed with a mixture of dichloromethane: methanol: acetone (1:1:2), followed by quantification with UV-Vis spectrometry, 450 nm. A one-way ANOVA was performed. Optimal growth was observed at concentrations of 0.025% within ten days. A significant difference in carotenoid content was determined between the different salts with an F value for JOC-5 and JOC-8 of 19.36 and 7.61 respectively (p < 0.05). Carotenoid production is affected in JOC-5 by the presence of FeCl2 and CuSO4, and JOC-8 with FeSO4 and CuSO4. While JOC-5 with FeSO4 and JOC-8 with FeCl2 yielded a similar amount carotenoid compared to GYEA alone. Smaller quantities of FeSO4 and FeCl2 with respective strain could enhance total carotenoid content.

 

Evaluation of Antagonistic Activityin Strains of Micromonospora sp. Isolated from the Gulf of California Against Candida albicans

Omar E. Valencia Ledezma, Herminia I. Pérez Méndez, Luis A. Maldonado Manjarrez, Erika T. Quintana Cano, Norberto Manjarrez Alvarez

  • Abstract

    The marine ecosystem represents a rich source of metabolites of pharmacological interest, among these organisms, the actinomycetes, particularly the genus Micromonospora have shown to have a high biotechnological value. Antagonist activity was determined using 12 strains of Micromonospora sp. (M001-M012) against Candida albicans ATCC 10231 were grown in medium (GYM) employing various methods: a) agar disk diffusion, b) double layer, c) against disk drive, d) confrontation dual type, e) modified technique of Fleming, selection of antagonistic strains was assessed by determining zones of inhibition. Streptomyces sp. was used as positive control. Antagonism was positive after 60 days of growth and the technique that allowed the best measure of inhibition percentage was the modified technique of Fleming as this allows diffusion of secondary metabolites. According to the assay, the strains that showed a high inhibition percentage greater than 90% were M007, M009, M010 and M012, with an average of 3.23±0.17 mm of inhibition. A positive antagonism at least by four different Micromonospora species against Candida albicans 10231 was observed. This indicates that fungal agents are produced and that are active against Candida albicans.

 

Determination Of Energetic Content For 3,5-Dimethylisoxazole Like Antilipolityc Drug

E. Adriana Camarillo, Henoc Flores and Gastón Perdomo

  • Abstract

    Adipose tissue is the body's largest energy reservoir and a major source of metabolic fuel. In periods of food supply, energy is stored as triglycerides. The Lipolysis is defined as the hydrolytic cleavage of ester bonds in triglycerides, resulting in the generation of fatty acids and glycerol. The rate of lipolysis is regulated by lipolytic and antilipolytic hormones, they reduces the free fatty acid levels in the blood stream; insulin is the most potent antilipolytic hormone known. 3, 5-dimethylisoxazole has been investigated on lipid and carbohydrate metabolism and it showed an antilipolityc effect. Thermodynamic properties acquire growing importance in studies of biologically active substances. The biothermodynamics is being actively developed now. Therefore, it seems particularly important to study thermodynamic properties for this compound. The purpose of this work is to report experimental measurements of thermochemical properties of 3, 5-dimethylisoxazole. The energy of combustion was determined using a combustion static bomb calorimeter. The molar energy of combustion for the compound in the condensate phase at T = 298.15 K was −(2900.02 ± 1.48) kJ⋅mol-1. From this value, the corresponding standard molar enthalpy of formation was obtained as −(66.08 ± 1.69) kJ⋅mol-1.

 

Carboxymethylation of Exocellular β-(1→6)-D-Glucan (Lasiodiplodan) to Enhance Biological Potential

*Carlos R.M. Malfatti , Francini Y. Kagimura, Thais Theis, Mário A. A. da Cunha

  • Abstract

    Many fungal β-glucans have biological activities associated with their structure and molecular weight. The study aimed the chemical modification the lasiodiplodan by carboxymethylation seeking to increase their solubility and therefore enhance its biological action. A chemical modification was confirmed by Fourier Transform Infrared Spectroscopy (FTIR) and the Degree of Substitution (DS) was determined by neutralization titration. Antioxidant capacity was assessed by DPPH and ABTS radicals scavenging assay and FRAP assay. FT-IR spectrum of no modified and derivative biomolecule were similar, exhibiting the typical signals of polysaccharides in the region from 4000 cm-1 and 500 cm-1.The carboxymethylation was confirmed by two new bands of strong absorption in the region 1604 cm-1(COO-) and 1421 cm-1 (COO-). The substitution degree of the derivatized biomolecule was 1.2 and the solubility in water increased by around 65%. The antioxidant activity was increased by derivatization. At concentrations of 5 mg/L the ABTS radical scavenging potential increased from 3.0% (no modified molecule) to 9.7% (derivatized molecule) and the DPPH radical scavenging ability increased from 1% to 7.5%. Ferric ion reducing antioxidant power was also increased (15%). The carboxymethylation contributed to increase its solubility in water which may contribute to greater antioxidant action.

 

Atomistic Study Of The Interaction PPAR -gamma With EPA: A Computational Study

Adriana Morales Martínez, Francisca Pérez-Severiano & Absalom Zamorano Carrillo

  • Abstract

    PPARs (Peroxisome Proliferator Activated Receptors) are members of the nuclear receptor family that can be activated by ligands endogenous (fatty acids) or exogenous (thiazolidinediones, fibrates). There are three isotypes of PPARs (PPARα, PPARβ/δ and PPARɣ) studied as therapeutic targets in different diseases, mainly those characterized for high oxidative stress levels. PPARβ/δ and PPARɣ are highly expressed in brain and their activation by ligands could increase the level of protecting molecules against tissular damage; however, a wider description of the mechanisms is necessary. Two ligands, DHA (docosahexaenoic acid) and EPA (eicosapentaenoic acid) are fatty acids that can activate PPARs. Herein, we describe atomistically the affinity of PPARɣ with EPA and DHA by computational simulations; in particular, Docking was performed in Autodock Vina and the results were analyzed by MOE software. Nine conformers were obtained, and the top affinity was -7.1 Kcal/mol for both DHA and EPA, sharing the same ligand binding domain. DHA and EPA interact with four polar residues of PPAR-ɣ arm I (Ser289, His323, His449, and Tyr473). Thus, the affinity of EPA and DHA with PPAR-ɣ are highly similar. Hydrogen bonds might be provided by polar residues and the carboxylate groups

 

Determination of the Parameters of the Bergman´s Model for Diabetic Mice Treated with Ibervillea sonorae

N.R. Budar-Aleman, A.A. Sotelo de, M.G. Ramírez-Sotelo, A.I. Cabrera-Llanos

  • Abstract

    This article shows the estimation of Bergman's minimal model parameters for glucose-insulin interaction in three states: evolution of the concentration of insulin, evolution of glucose and glucose/insulin interaction. First, the dynamics of glucose measured in mice that were induced diabetes mellitus type 2 experimentally, which were treated with extracts of Ibervillea sonorae root in a range of 100-400 mg/kg, likewise, the estimate for healthy mice is shown and the different parameter values obtained are compared with each other showing how they influence the dynamics of Bergman's model. The values of the glucose/insulin interaction where obtained in vivo by blood samples of the mice every hour. Showing the evolution and variability of the estimate is shown in graphical form; also, the estimation error is quantified by performance curves, which were associated to the energy that is implied in the difference between the values of the parameters obtained and the data obtained in the kinematics of the mice. The results indicate the estimated level reached, where the upper value of the performance curve was 3.0128, while the lower was 0.1978, with standard error values between 1.55% and 10.47%.

 

Portable Measurament System of Athletic Performance in Aerobic Phase with Adquisition of LabVIEW Platform and ATmega Controllers

E. Sanchez-Velarde, O. Rico-Asención, M.G. Ramírez-Sotelo, A.I. Cabrera-Llanos

  • Abstract

    In this work, a remote monitoring system for measuring heart rate and deployment of an ECG signal is presented. This system uses a pulse sensor plug-and-play type placed on the lobe of the patient to obtain a signal, which is represented by squares; each symmetric pulse width is the width of the plethysmographic signal obtained by the sensor. And, an electrocardiograph developed for obtaining a derivative (positive, negative, and reference electrode), a filtering step to obtain the complete signal through the basic vector comprising Eithoven triangle. Both signals were transmitted via a ZigBee protocol radio frequency, using two Xbee modules series S1, data were received through a ATMEGA328P, later, by the LabVIEW platform, and anthropometric data of the individual, to monitor the heart rate, continued deployment of the 2 signals obtained presented along a routine of aerobic exercise and a history of heart and indicator signal obtained this in order to observe their status. A series of measurements of heart rate and ECG signal for individuals undergoing physical activity of different intensity is presented

 

Inhibition Of Dopachrome Synthesis By Mistletoe Extracts (Cladocolea loniceroides)

Frida Paredes Ruiz, María José Serrano Maldonado, Ivette González Palma, Jennifer López Sánchez, Edith Ponce Alquicira, Jorge Soriano Santos

  • Abstract

    Mistletoe is a parasitic plant which grows attached to and within the branches of trees and shrubs. Mistletoe impact is rather negative causing great tree mortality. The Mexican mistletoe,Cladocolea loniceroides contains different polyphenols, which are an excellent natural antioxidants resource from this plant. Various tropical vegetables with high antioxidant concentrations have been known for their anti-tyrosinase activity which could be potentially prepared for skin-whitening preparations. The objective of this study was to assess tyrosinase inhibitory activity of mistletoe extracts through the quantification of dopachrome. The green and ripe fruits, leaf and stem were extracted with five solvents including hexane, chloroform, ethyl acetate, methanol, ethanol and water. The results showed that the water extract by ripe fruit, exerted a considerable level of in vitro mushroom tyrosinase inhibition (IC50=1.1 mg/mL) compared to a positive control of hydroquinone (IC50=0.048 mg/mL). After evaluating the antioxidant activity by DPPH and ABTS radicals scavenging activity, it was founded that the water extract by ripe fruit had the highest antioxidant activity in both methods (EC50=1.36 mg/mL and 0.29 mg/mL, respectively), also showed non-competitive inhibition (Ki´=0.22 mg/mL). So, this extract has potential as skin-whitening agent, but further study is needed to identify the phytochemical compounds with anti-tyrosinase activity

 

Phytochemical Characterization of K. flammea and Its Effect On The Biological Neoplastic Events In Prostate Cancer

Iván Arias-González; Cynthia Ordaz-Pichardo

  • Abstract

    Despite decades of research, the treatment and management of malignant tumors still remain a formidable challenge for public health. Medicinal plant drug discovery continues to provide new and important leads against various pharmacological targets including cancer. Kalanchoe flammea is a native plant of Mexico that has shown anticancer activity against prostate cancer. In vitro treatment of androgen-insensitive (PC-3) prostate cancer cells with an ethyl acetate extract of K. flammea (Kf-AcOEt) reduced cell viability and no significant effect shown in non-neoplastic human prostate epithelial cells (PrEC) under identical conditions. Also, Kf-AcOEt induced cell death in a time-dependent manner and shown different biochemical hallmarks of apoptosis like activation of caspase-9, -3 and release of cytochrome C from mitochondria into the cytosol, proteolysis of PARP-1, downregulation of antiapoptotic protein Bcl-2, ROS production and DNA fragmentation. Moreover, Kf-AcOEt causes cell cycle arrest in the S phase. In vivo studies indicate that Kf-AcOEt inhibit growth of prostate tumors in mouse xenograft model. These data suggest that Kf-AcOEt may contain metabolites that may serve as a lead compounds for targeting so far incurable androgen-insensitive prostate cancers, which could have potential as a future chemotherapeutic agents.

 

Effect of an Analogue of Praeruptorin A in Streptozotocin-Induced Diabetic Rats

Elix Alberto Dominguez Mendoza, Jorge Cornejo-Garrido, Eleuterio Burgueño Tapia, Cynthia Ordaz-Pichardo

  • Abstract

    Diabetes is a chronic metabolic disorder that includes various alterations in carbohydrate, fat and protein metabolism. It is characterized by hyperglycemia resulting from absolute or partial deficiency in insulin secretion, insulin resistance, or both. In Mexico, there are more than 300 plants traditionally used for the treatment of diabetes, one of these plants is Arracacia tolucensis known by the common name of "Acocotillo". In our laboratory, different extracts of A. tolucensis were evaluated in a diabetic rat model. AcOEt extract had the highest effect, showing a blood glucose of 158 mg/dL ± 25 of compared with, the diabetic group 361 ± 27 mg/dL (difference of 203 mg/dL). The presence of 2 major metabolites were found, Praeruptorin A and 4-hidroxiangeoloilvisaminol. We synthesize an analogye of praeruptorin A (PAA) with hypoglycemic activity at a concentration of 15 mg/kg showing a blood glucose of 128 ± 6.4 mg/dL compared with the diabetic group 327.5 ± 23.8 mg/dL (difference of 198.4 mg/dL).The effect of the analogue was very similar to Glibenclamide or Metformin, compounds widely used in the clinic for the treatment of diabetes. It’s of our interest to evaluate the mechanism of action that caused this hypoglycemic activity.

 

Glucosylation of Naringin and Rutin Catalyzed by CGTase from Thermoanaerobacter sp.

Violeta Campis, Edmundo Castillo, Liliana Hernández, Julia Cassani

  • Abstract

    The flavonoids have proved their importance as natural antioxidants in addition to the prevention of cardiovascular diseases and some kinds of cancer. This work describes the methodology to obtain more soluble glucosylated products of Naringin (flavanone) and Rutin (flavonol) by biocatalytic reactions using ciclodextrin glucosyl transferase (CGTase), which catalyzes the transfer of glucose to acceptors as flavonoids. The glucosylation reaction of Naringin with CGTase from Thermoanaerobacter sp. (0.182 U/mL) and soluble starch was incubated at 50°C with stirring for 14 days; two products (N2 and N3) were observed by thin layer chromatography (TLC), the same reaction conditions were applied to Rutin glucosylation with β-Ciclodextrin (βCD) for 3 days, a unique product (R2) was observed by TLC. The extraction of the products was performed by preparative TLC and N3 was purified in 45.3 and N2 8.2% yield, R2 was purified in 38.7% yield. The reaction conditions of the substrates in presence of CGTase resulted favorable to obtain glucosylated products of Naringin and Rutin to help increase their solubility

 

Anticancer Activity of Arantho on a Rodent Breast Cancer Model

Cynthia Carolina Estanislao Gómez, David Guillermo Pérez Ishiwara, Francisco Javier Paz Bermúdez, Abraham Aquino Carreño, Jesús Gabriel Silva Escobedo, María del Consuelo Gómez García.

  • Abstract

    Breast cancer represents the highest mortality rate worldwide. Chemotherapy is the most common treatment which uses a combination of different anticancer drugs. However, these drugs produce side effects and resistance; so, new research is necessary to find new natural compounds that can reduce side effects. Particularly Arantho is a plant used in traditional medicine and it induced cytotoxicity in breast cancer cells. So, the objective of the work was to evaluate the anticancer activity of the essential oil of Arantho (EO) on in vivo breast cancer model. The rat breast cancer model was induced by NMU carcinogen, which can generate ductal adenocarcinomas. Different groups were established, each one received a single intraperitoneal dose of 50 mg NMU/kg; Group 1 was the negative control and Groups 2 to 5 received doses of 30, 50, 100 and 150 mg/kg of the EO, respectively. Results indicated that 100 mg/kg were the most effective, because it reduced tumor size comparing with the control group. Histopathologic analysis demonstrated the presence of ductal adenocarcinomas similar in each group. However, liver, kidney and mammary gland tissues demonstrated differences between groups. In conclusion, the EO of Arantho showed an important antitumoral activity on these model.

 

Synthesis of Xanthones with Antioxidant Activity

Aarón Mendieta Daniel Zarate, Víctor Soto, María del Carmen Cruz, Fabiola Jiménez, Asiel Tabarez and Joaquín Tamariz

  • Abstract

    In the search for new antioxidant agents, the present study was focused on the synthesis of a series of diversely substituted xanthones and on their in vitro evaluation in their antioxidant properties. This evaluation was based on the scavenging activity method using the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. Among the evaluated xanthones, we found that those compounds having a methoxy group and a methyl acetate fragment in their structures exhibited a good antioxidant activity.

 

Synthesis of Xanthones with Antioxidant Activity

Aarón Mendieta, Daniel Zarate, Víctor Soto, María del Carmen Cruz, Fabiola Jiménez, Asiel Tabarez and Joaquín Tamariz

  • Abstract

    In the search for new antioxidant agents, the present study was focused on the synthesis of a series of diversely substituted xanthones and on their in vitro evaluation in their antioxidant properties. This evaluation was based on the scavenging activity method using the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. Among the evaluated xanthones, we found that those compounds having a methoxy group and a methyl acetate fragment in their structures exhibited a good antioxidant activity.

 

Perezone Productiom by Adventitious Roots of Acoutria hebeclada DC

Ma. Del Carmen García Méndez, Patricia Castillo España, Irene Perea Arango, Alfonso Leija Salas, Laura Álvarez, Silvia Marquina, José de Jesús Arellano García

  • Abstract

    Acourtia hebeclada DC and others species of the Acourtia genus had been used since prehispanic times to prepare a beverage to cope with stomachache. Commonly known as pipitzáhuac, this medicinal plant produces and accumulates in its roots high amounts of the sesquiterpene quinone known as perezone; a crystalline substance with a deep orange color, easily extracted from roots of several Acourtia species. Perezone can be used as pigment and to produce other pigments. Pharmacologically perezone has been shown to have cytotoxic activity and may be a protectionist for reperfusion damage. The aim of this project was the establishment of a protocol for obtaining adventitious root cultures to produce perezone. Leaves, roots and stem nodes from 30 days old in vitro cultured seedlings of A. hebeclada were used as explants. Nine treatments were established on MS solid medium, amended with three different concentrations naphthaleneacetic acid and 6-benzylaminopurine. Adventitious roots were obtained with treatments M2 and M3 in leaf and root explants. Analysis of perezone production by different lines of adventitious roots was carried out by thin layer chromatography and by GC-mass spectrometry. Further work should be done to establish a process of perezone production from the root lines obtained.

 

Clinical Study Of Patients With Pre-Malignant Lesions Of Cervical Carcinoma: Therapeutic Evaluation Of Dialyzable Leukocyte Extract Of Crocodile

Acosta Ríos María del Pilar ,Sauer Ramírez Eduardo, García Solís Mario, Consuelo Gómez García, Castro Muñoz Leonardo, Correa Meza Bulmaro, Pérez De la Mora Carlos, Ocadiz Delgado Rodolfo, Pérez Ishiwara David Guillermo

  • Abstract

    Cervical Cancer (CaCu) is the consequence of premalignant lesions of progressive evolution. The human papillomavirus (HPV) that has been implicated in this pathology. The Immunotherapy with dialyzable leukocyte extract (DLE), increases the activation of monocytes, macrophages and NK cells. 54 women with low-grade dysplasia were divided into two groups, the control group (Placebo) and DLE group. Histopathological analysis of biopsies shows the koilocytes with loss of cytoplasmic maturation in the lower third of the epithelium. In the remaining two thirds also was observed atypical nuclei and sometimes acanthosis, parakeratosis was observed and dyskeratosis. The viral genotype analysis done by PCR showed that all patients have HPV infections predominantly 31, 16 and 18. Comparative studies colposcopy before and after therapeutic intervention, showed a reduction in cervical lesions in patients treated with DLE. Comparative histopathological analysis showed decreased leukocyte infiltration and decreased cervicitis. Immunohistochemical analysis showed decreased expression of PCNA in the cervical epithelium of patients treated with the DLE. Low expression of PCNA after DLE treatment suggests decrease of proliferation of infected cells by HPV.

 

In vitro Binding Effect Of Potential Inhibitors Identified By Docking To The Nickel Response Regulator (NikR) From Helicobacter Pylori

Ponciano García, Wendy Cruz, Rubén Cantú, Arturo Rojo, Aldo Segura, Yolanda López and Xianwu Guo

  • Abstract

    NikR protein is a regulator responsible for controlling the expression of certain genes such as ureA and ureB in Helicobacter pylori. The proteins ureA and ureB are the structural subunits of urease, an enzyme important for the H. pylori adaptation in low pH niche as an essential factor for living in human stomach. Thus, NikR is a good candidate for medicine targeting. Previously, we reported a molecular docking between X-ray structure of H. pylori NikR and a chemical database, and some potential inhibitors have been identified. The NikR from H. pylori strain 26695 was cloned into the plasmid pProExHTb for expression in Escherichia coli BL21 (DE3). Native H. pylori NikR with hexa-histidine tag was purified. In native conditions, HpNiKR was a tetrameric molecule as seen by N–PAGE and by size-exclusion chromatography, and confirmed by MALDI mass spectrometry. The ureA promoter fragment for HpNikR binding experiments was constructed via incubation by two synthesized complementary single oligonucleotide chains. The EMSAs competition assays showed the disassociation from DNA promoter–Holo-HpNikR complex as the addition of two compounds in range of 250-300 μM. The in vitro experiments showed that two compounds can bind to HpNikR, affecting the protein recognition with promoter of ureA.

 

TLR-9 and CD1d Expression in Mexican Patients with Human Papilloma Virus Infection

Leonardo Josué Castro Muñoz, Eduardo Sauer Ramirez, Pilar Acosta Ruiz, Mario García Solís, Virgina Sánchez Monroy, Consuelo Gómez García, Rodolfo Ocádiz Delgado, Olivia Medel Flores, David Guillermo Pérez Ishiwara

  • Abstract

    During the early stages of HPV infection, the components of the innate immune response are the first line of defense. Receptors as the TLR-9 and cells as the NKT, which are activated specifically by CD1d receptor, are able to identify and to eliminate infected cells; these molecules are also important components for the correct activation of adaptive immune response against virus. In this respect, there are few studies determining the role of innate immune response in early stages of HPV infection. Therefore, the objective of this study was to determine the expression of TLR-9 and CD1d in samples from Mexican patients with cervical intraepithelial neoplasia grade I (CIN-I), positive to HPV infection. Results from multiplex PCR demonstrated that 11 patients showed multiple infections, 10 presented infection with HPV-31, the most common among the high risk type, while 4 presented only low risk 11 type. On the other hand, TLR-9 and CD1d that are expressed in the basal epithelium of non-infected tissue, were detected in the cytoplasm of the upper epithelium, while no expression was found in cells of the basal layer, suggesting a diminished activity of the innate immune response during HPV viral infection.

 

Characterization and In Vitro Evaluation ofBiopolymer/Nano-Hydroxyapatite Composites for Bone Tissue Regeneration

Georgina Carbajal-De-la-Torre, Marco Aurelio Pardo-Galván, Ana Beatriz Martínez-Valencia, Tania Saluén Gaytán-Compeán, Ana Edith Higareda-Mendoza

  • Abstract

    The purpose of this study was to evaluate the in vitro cell biocompatibility of composites consisting of polyurethane (PU) and nano-hydroxyapatite (nHAp). MC3T3-E1 mouse calvarial preosteoblasts were cultured on the surface of the PU/nHAp composites. Cell proliferation, cytotoxicity, and osteoblast mineralization of seeded MC3T3-E1 were investigated. Cell expression of osteogenic markers such as type 1 collagen (COL-1), bone sialoprotein (BSP), and osteocalcin (OCN) were examined by reverse transcription polymerase chain reaction. Cell viability and total cellularity were similar between the different combinations of PU/nHAp tested. However, expression of osteogenic markers and mineralization of native bone tissue were significantly greater when osteoblasts were cultured on PU/nHAp composites compared to control. The results demonstrate that one of the composites presented improved osteoinductive, physicochemical and mechanical properties without additional effects on cell viability or proliferation.

 

Cytotoxic and α-Glucosidase Inhibitory Activity from Eucidaris thouarsi (Echinoidea) and Mithrodia bradleyi (Asteroidea)

Francisco Vargas-Betancourt, Claudia J. Hernández-Guerrero, Antonio Nieto Camacho, Ma. Teresa Ramírez Apan and Bárbara González Acosta.

  • Abstract

    The cytotoxic and α-glucosidase inhibitory activity from organic extracts and fractions of sea urchin Eucidaris thouarsi and starfish Mithrodia bradleyi of Punta Arena de La Ventana, B. C. S., Mexico was evaluated. The cytotoxic activity of extracts and fractions (50 μg mL-1) toward different cancer cell lines U251, PC-3, K562, HCT-15, MCF-7 and SKLU were determined using the protein-binding dye sulforhodamine B assay in microculture to measure cell growth. Cell viability was determined by MTT assays against murine cell line J774. The inhibitory activity of α-glucosidase (Type 1) of extracts and fractions assay was determined at different concentrations (1, 10 and 100 μg mL-1). The fraction F1 of M. bradleyi showed strong cytotoxicity but low cell viability against the cell line J774. While ether extract and fractions F2 and F4 of E. thouarsi showed a high and selective cytotoxicity against cancer cell lines. The fractions F3 and F4 of E. thouarsi, viscera extract and fraction F2 of M. bradleyi showed strong α-glucosidase inhibitory activity similar to quercetin, which is a potent inhibitor used as reference. Eucidaris thouarsi and Mithrodia bradleyi displayed potential cytotoxic and α-glucosidase inhibitory activity. Thus these organisms are a good source for the identification of active compounds.

 

Methylation Analysis of Genes for the T-type Voltage-Dependent Calcium Channels in Cervical Cancer Derived Cell Lines

Katherine Campaña, Pedro Chavez, Javier Camacho and Efraín Garrido

  • Abstract

    Cervical cancer is the second cause of death in women worldwide. A positive correlation between increased gene methylation and increased pathological changes exists during the pathogenesis of cervical cancer. The intracellular calcium control allows orderly cell cycle progression, and play a vital role in growing and cell proliferation. T-type calcium channels play a fundamental role in the regulation of calcium in non-excitable cells. The expression of the T-type calcium channels isoforms has been reported in a wide range of cells including cancer cells. Our study has been aimed to evaluate the methylation in the promoter and regulatory regions of two genes encoding the main isoforms of T-type calcium channels: Cav 3.1 and Cav 3.2, and its correlation with their expression in cervical-cancer cells. Previously, it was reported no methylation of the T-type calcium channels in normal breast epithelial tissue, bone marrow, colon mucosa and placenta tissue suggesting that Cav 3.1 gene methylation is cancer specific. Using standard RT-PCR, the mRNA expression of the calcium channels was verified. Cav 3.1 mRNA was detected in CALO, while Cav 3.2 mRNA was detected in C-33A. After bisulfite DNA conversion the percentage of methylation was established, and correlated with the expression of the channels

 

Anxiolytic and Sedative Effects of Psidium Sartorianum (Guayabilla)

Hilda Rangel Rocha, Kristopher Goroztieta García, Emma Lilia Hernández Sánchez, Eva Aguirre Hernández, Hortensia Rosas Acevedo

  • Abstract

    Psidium sartorianum (Myrtaceae) is commonly known as Arrayan, guayabilla or guayabillo, is a tree up to 20 m in height, its bark is smooth and clear, and is apparent in fine layers. The leaves are ovate and occasionally obovate, lanceolate or elliptic. The flowers are white and the fruits are globose and yellow. This plant is mainly used for the treatment of digestive disorders; such as diarrhea and dysentery. We now describe the sedative effects of extracts of leaves of this species. A significant reduction in the exploratory activity of mice was produced in the extracts at dose of 300 mg/kg (hole board model) In the test for anxiety behavior, a significant increase in the time spent in the open-side arms was observed for ethyl acetate extract (100 mg/kg). For other hand hexane, ethyl acetate and methanol extracts (300 mg/kg) induce a significant increase in the sleeping time in comparison to the control group.

 

Bacterial Contamination Control with Antimicrobial Peptides in Bovine Semen Cryopreservation

Y. Sánchez-Acosta, K. Ramírez, S. Orduz and O. Camargo

  • Abstract

    Cryopreserved semen must remain free of microorganisms to be used in artificial insemination in a safe way. Despite the sanitary controls, many cryopreserved semen straws are contaminated with microorganism, some of which display antibiotic resistance. Therefore, this technology involves the risk of spreading pathogens, threatening the health of inseminated females. Our objective was to explore antimicrobial peptides for bacterial control in bovine semen cryopreservation. Using microdilution susceptibility test, we assessed the potency of seven dermaseptin S4 analogues, including three newly designed: S4(5-28)a, K20S4(5-28) and K4S4(1-15), against Escherichia coli, Staphylococcus aureus and aerobic bacteria isolated from frozen-thawed bovine semen, belonging to the genera Bacillus, Staphylococcus and gram positive cocci. K4S4, S4 (5-28) a, K20S4 (5-28) and K4S4 (1-15) a, with a net charge of +5, were the most potent peptides. S4 (5-28) and K4S4 (1-15), with a net charge of +4, had lower activity. K4S4 (1-16) a, that possess increased charge (+6) but is less hydrophobic than K4S4 (1-15) a (0.43 and 0.52, respectively), had less activity. Modifying the charge-hydrophobicity relationships of the peptides, resulted in short analogues that displayed high activity in the micromolar range (<10μM) against problematic bacteria in semen contamination. These results suggest a potential for development of peptide-based antibiotics applicable in bovine semen cryopreservation.

 

Modeling Nucleoprotein Rabies Virus

Grecia C. Tovar-Rojas, Fernando Bastida-González, Paola B. Zárate-Segura 

  • Abstract

    Rabies virus (RABV) infects mammals causing acute encephalomyelitis (Lyssavirus: Rhabdoviridae: Mononegavirales). RABV genome consists of a single non segmented negative-strain RNA that encodes the five structural proteins: glycoprotein (G), the matrix protein (M), phosphoprotein (P), viral RNA-dependent RNA polymerase (L) and nucleoprotein (N). The N protein contains 450 amino acids highly conserved. In 2006 the crystal structure of nucleoprotein rabies virus (RABV N) was described, however this crystal lacks the description of loops. For construction RABV N model by homology and refinement loops was usedMODELLER 9.12 program. The 3D model of RABV N obtained was evaluated stereochemical quality and reliability with Ramachandran PLOT and Errat server, showing 78% favorable residues and Z-score 58.6% respectively. Subsequently strongest interations between aminoacids in nucleoprotein model were analyzed with PDBsum server, which were E20-R254, D4-R254, N11-E266, K417-E392 and E403-H369; these were compared with 2GTT interactions showing that E20-R254 and N11-E266 are importants due to these are present in both models. The interactions E20-R254 (polar-acid and polar-basic) and N11-E266 (polar-uncharged and polar-acid) is relevant due to that are the main contacts between neighboring protomers.

 

Characterization Of The Folding Thermodynamics And Kinetics Of Fibrillogenesis Of Mutants Of 6ajl2 Protein At Glu13 And Lys17

Francisco J. Rodríguez Álvarez , Gerardo Vázquez del Angel, Sergio Garay, Adrián Ochoa Leyva, Julio I. Pérez Carreón, Jorge Meléndez Zajgla, Luis del Pozo Yauner

  • Abstract

    It has been reported that somatic mutations disrupting key interactions of the native structure, such as salt bridges, can modulate the amyloidogenic potential of the protein. The crystal structure analysis of the recombinant variable domain protein 6aJL2 suggests that residues Glu13 and Lys17, located in the A and B β strands, are involved in a salt bridge. This study was aimed to evaluate the effect of mutations in the aforementioned residues on the folding thermodynamic and fibrillogenesis kinetic of 6aJL2 protein. Single and double mutants of 6aJL2 protein by Gly, Ala, Gln and Leu at positions 13 and 17 were generated by conventional PCR. In vitro fibrillogenesis of the proteins was performed on 500 μg/ml sample dilutions in PBS pH 7.4. The samples, in triplicate, were incubated at 37 ° C with constant orbital stirring of 1000 rpm, the formation of amyloid aggregates was determined by the tioflavin T assay. The folding thermodynamic of the mutants was determined by reversible unfolding with urea. 6aJL2 mutations which replace E13 and K17 with smaller residues, such as Gly and Ala, increased the thermodynamic stability. The fibrillogenic kinetic of the proteins is influenced by the structure of the replacing amino acid side chain.

 

Investigating The Molecular Basis Of Peptide Ser26-Arg39 Amyloidogenesis

Robin Axel Ruiz Zamora, Adrián Ochoa Leyva, Julio Isael Pérez Carreón, Jorge Meléndez Zajgla, Luis del Pozo Yauner

  • Abstract

    This work was aimed to identify the residues determining the fibrillogenic potential of the fragment Ser26-Arg39 of the amyloidogenic light chain variable domain 6aJL2. The contribution of each residue was determined by a Pro-scanning mutagenesis analysis. For this purpose, we evaluated the in vitro fibrillogenesis of the wild-type peptide and sixteen different single-mutants by Pro in a 150 μM dilution in PBS pH 7.4 plus 2.0 M guanidine hydrochloride. It was observed an increase of the ThT fluorescence in the Wild-type peptide Ser26-Arg39, indicative of amyloid fibril formation, that was accompanied by the decrease of the intrinsic fluorescence of the soluble fraction, at the end of the incubation. By other hand, the point-mutations by Pro at positions from Ile30 to Tyr36 suppressed nearly totally the aggregation capacity of the peptide, as indicate the very low values of ThT fluorescence and the high intrinsic fluorescence of the soluble fraction. We conclude that the region determining the amyloidogenic potential of peptide Ser26-Arg39 spans from Ile30 to Tyr36. This finding suggests that this region may be part of the structural core of the amyloid fibril formed by the intact protein, playing a key role in the mechanism of fibril self-assembling.

 

Arantho (Rutaceae): a medicinal mexican plant against breast cancer

Alejandro Arroyo Román, Erik Ocaranza Sánchez, David Guillermo Pérez Ishiwara, María Guadalupe García Meza, Cynthia Carolina Estanislao Gómez, Nury Pérez Hernández, María del Consuelo Gómez García

  • Abstract

    Breast cancer is the most common malignancy in women worldwide representing the leading cause of death. There are several therapeutic methods for these illness as the chemotherapy; which includes the administration of drugs. However, these treatment affect normal cells causing severe side effects in patients. In addition to the low diversity of drugs on the market leads to the search for new drugs to treat this condition. Medicinal plants have chemical components of great importance to the discovery of new active compound. Mexico is one of the countries with the greatest diversity of plants in the world after China and the use of plants for medical purposes has been a tradition. In this work we identified a mexican plant (Arantho) that is collected for medical purposes by the local people of El Cardonal province, in Hidalgo State located in the center of Mexico. Arantho belongs to the Rutaceae family. People use the leaves to prepare infusions and drink the tea to treat breast cancer. Besides, the plant is dried and stored in unfavorable seasons in order to use them later. The study of the potential cytotoxic properties of Arantho against breast cancer is currently performed in cellular lines.

 

Development of Nanostructured Titanium Dioxide Reservoirs Including Poorly Soluble Antiepileptic Drugs For Intranasal Application

Cecilia Zavala-Tecuapetla, Tessy María López Goerne

  • Abstract

    Nanostructured controlled delivery systems are an innovative approach to the treatment of diseases such as epilepsy. We focus on developing nanostructured delivery system where low solubility antiepileptic drugs (AEDs) can be encapsulated and later released. Nanostructured reservoirs were synthesized by the sol-gel method using as precursor titanium n-butoxide. AEDs were encapsulated by adding them during the gelation step. The reservoirs obtained (empty and containing the AED) were characterized by means of several physical techniques. It was shown that the poorly soluble AEDs can be encapsulated in titanium dioxide reservoir. As a result of the encapsulation process, the structure of the drug is not chemically altered. The reservoirs containing AEDs exhibit higher surface areas and a heterogeneous morphology formed by irregular aggregates which are spherical in shape and are approximately 0.30-0.48 μm in diameter. The in vitro release kinetics for the AEDs evaluated, showed two distinct phases of release: an initial rapid release and subsequent slow sustained release. Intranasal administration of these titanium dioxide reservoirs could promote the release of poorly permeable AEDs to the blood brain barrier and  ould act directly on the brain decreasing the frequency or severity of seizures in epileptic patients.

 

Silencing of White Spot Syndrome Virus (WSSV) Gene Vp281 By dsRNA and its Antiviral Effect in Shrimp (Litopenaeus Vannamei)

José Socorro Rodríguez Navarro , Píndaro Álvarez Ruiz, Carina Gámez Jiménez, Gabriela López Cervantes

  • Abstract

    The white spot syndrome virus (WSSV) has caused huge losses to the shrimp industry. It has been shown that silencing of viral genes using RNA interference (RNAi) decreases viral replication. We evaluated the effect of silencing the gene 281 of WSSV using double stranded RNA (dsRNA) to trigger RNAi in Litopeneaus vannamei. Shrimp were collected from a commercial farm. Treatments were separated by dsRNA dose (1, 2 or 4 μg/shirmp) and dsRNA size (215, 394 or 843 bp). The dsRNA of each fragment was synthesized with a commercial kit. 10 shrimp were injected with dsRNA. After two days, were challenged against WSSV. Mortality was recorded twice a day. At the end of experiments, WSSV presence was confirmed by PCR.No antiviral effect was observed in the treatments with the dsRNA fragments of 215 or 395 bp, regardless of dsRNA dose. These results suggest that silencing of VP281 gene does not protect against WSSV infection. However, the treatment with the largest dsRNA (843 pb) and the higher dsRNA dose, achieved some survivors. As a conclusion, we showed that antiviral effect induced by RNAi depends of size and quantity of dsRNA supplied to shrimp.