Verbascoside production in long-term Buddleja cordata Kunth cell suspension cultures.

Previously, our group reported the establishment of a white callus cell line of Buddleja cordata Kunth that is a high producer of the secondary metabolite, verbascoside (VB, also named acteoside), under suspension culture conditions. Here, we present experimental evidence of the sustained ability of that cellular line to grow and produce high amounts of VB for 5 years of continuous culture. Cellular line profiles were determined at the early (at the beginning) and late stages (at the end of 5 years of continuous subculturing) by analyzing relevant parameters of culture growth, i.e., specific growth rate [µ], doubling time [dt], and growth index [GI], as well as VB production. Late-stage cultures exhibited a 61% faster growth rate than early-stage subcultures, and 25 and 3% lower doubling time and growth index. The extents of growth phases were found to be different. Similar amounts of biomass were found (9.5 g and 9.4 g L-1). Verbascoside production increased parallel to cell growth; maximal yield level occurred in the mid-exponential phase and lasted until the end of the stationary phase (i.e., from the 15th to the 25th day and from the 9th to the 21st day for the early and late stages, correspondingly). The content of VB was higher in the late-stage culture (1.43 ± 0945 g L-1) than in the early-stage culture (1.21 ± 0.0286 g L-1). Productivity values point out the potential use of B. cordata cell line in the biotechnological production of VB and for research focused on the biochemistry of secondary metabolism.

[Helicobacter pylori detected in atheroma plaque]. / Detección de Helicobacter pylori en placas de ateroma.

INTRODUCTION: In general, infection and chronic inflammation have been implied as etiologic agents for atheroesclerosis and in particular coronary illness (CI). Several studies have correlated the infection of Helicobacter pylori with CI, especially with virulent strains (lineage Cag A). OBJECTIVE: Demonstrate the immunohistochemical presence of H. Pylori in atheroscletotic plaques obtained from endarterectomy of different vascular regions. MATERIAL AND METHODS: 34 atherosclerotic plaques of different vascular areas were studied, (25 men and 9 women). The tissues were fixed with 10% neutral buffered-formalin and decalcifying in formic acid 5% was used when necessary. The tissue sections were included in paraffin, cut and colored with H&E and subjected to Immunohistochemistry (IHC) of H. Pylori. Briefly, tissues were deparaffinized and thermally treated with a citrate-based solution of antigenic retrieval (ImmunoDNA Retriever with Citrate, BIO SB, Santa Barbara, CA) using a water bath at 95 degrees C for 1 hour. The IHC was conducted using a high sensitivity Biotin-Streptavidin-HRP-DAB IHC system (ImmunoDetector HRP/DAB, BIO SB). The microscopic observation evaluated the presence of mononuclear inflammatory cells and the identification of the bacteria in the wall or the vascular lumen. RESULTS: Of the 34 cases studied 14 were positive, where one could identify the bacillus in their different forms (41,17%) associated with chronic inflammation.

Detección de Helicobacter Pylori en placas de ateroma / Elicobacter Pilori detected in atheroma plague

Introducción: la infección y la inflamación crónica han sido implicadas como agentes etiológicos para la ateraesclerasis (ATE). Varios estudios han relacionado a la infección por H. Pylori (HP) con la EC, especialmente con los linajes mas virulentos (linaje Cag A). Objetivo: demostrar la presencia del HP en placas de endarterectomías, utilizando una técnica Inmunohistoquimica (lHQ) específica que revela una reacción Ag-Ac mediante un cromógeno. Material y Métodos: se estudiaron 34 placas ATE de distintos territorios vasculares. Se fijaron en formol descalcificándolas en ácido fórmico según necesidad. Fueron incluidos en parafina, cortados y coloreados con H-E y técnicas de IHQ específicas para HP. Luego fueron desparafinados y tratados térmicamente con una solución de recuperación antigénica (lnmuno DNA Retriever with Citrate) utilizando olla a presión. La IHQ se efectuó con un sistema de alta sensibilidad Biotina-Estreptavidina-Peroxidasa-DAB). La observación morfológica evaluó células inflamatorias mononucleares y la identificación de la bacteria en la pared o la luz vascular. Resultados: de los 34 casos estudiados, en 14 se pudo identificar el bacilo en sus diferentes formas (41,17%), asociado a signos de inflamación crónica. Conclusión: el HP estuvo presente en un número sustancial de lesiones ATE y se asoció con inflamación. Estudios recientes sugieren que la presencia de Hp, demostrada por técnicas de IHQ, potenciaría los FR para ATE, induciendo una respuesta celular inflamatoria crónica por irritación persistente de la pared arterial.

Introduction: In general, infection and chronic inflammation have be en implied as etiologic agents for atherosclerosis and in particular coronary illness (CI). Several studies have correlated the infection of Helicobacter pylori with CI, especially with virulent strains (lineage Cag A). Objective: Demonstrate the immunohistochemical presence of H. Pylori in atherosclerotic plaques obtained from endarterectomy of different vascular regions. Material and methods: 34 atherosclerotic plaques of different vascular areas were studied, (25 men and 9 women). The tissues were fixed with 10% neutral buffered-formalin and decalcifying in formic acid 5% was used when necessary. The tissue sections were included in paraffin, cut and colored with H&E and subjected to Immunohistochemistry (lHC) of H.Pylori. Briefly, tissues were deparaffinized and thermally treated with a citrate-based solution of antigenic retrieval (lmmunoDNA Retriever with Citrate, BlO SB, Santa Barbara, CA) using a water bath at 95°C for 1 hour. The IHC was conducted using a high sensitivity BiotinStreptavidin-HRP-DAB IHC system (lmmunoDetector HRPIDAB, BlO SB). The microscopic observation evaluated the' presence of mononuclear inflammatory cells and the identification of the bacteria in the wall or the vascular lumen. Results: Of the 34 cases studied 14 were positive, where one could identify the bacillus in their different forms (41, 17%) associated with chronic inflammation.

Winery solid residue revalorization into oil and antioxidant with nutraceutical properties by an enzyme assisted process.

Revalorization of the winery industry residue, grape seed is studied for the production of an oil and defatted meal with nutraceutical properties. Conventional grape seed oil extraction process is carried out by pressing at high temperature affecting the product quality. Oil extraction by cold pressing improves product quality, but it gives a low oil yield. Oil extracted is increased at the pressing stage, when an enzymatic pre-treatment is incorporated in to the conventional process. The yield is determined by determining the residual oil in the pressed cake. Using an enzymatic treatment during 9 hours at 45 degrees C and 50% of moisture, with a mixture of two commercial enzymes grape seed oil extraction yield by cold pressing is raised up to 72%, being a 59.4% increment in comparison to the yield obtained by the control, without enzymes. The defatted meal by enzimatic assisted process improves its phenolic compounds between 2 and 4 times, depending on the conditions of phenolics extraction in comparison to the control samples.

Thermal inactivation of immobilized penicillin acylase in the presence of substrate and products.

Inactivation of immobilized penicillin acylase has been studied in the presence of substrate (penicillin G) and products (phenylacetic acid and 6-aminopenicillanic acid), under the hypothesis that substances which interact with the enzyme molecule during catalysis will have an effect on enzyme stability. The kinetics of immobilized penicillin acylase inactivation was a multistage process, decay constants being evaluated for the free-enzyme and enzyme complexes, from whose values modulation factors were determined for the effectors in each enzyme complex at each stage. 6-Aminopenicillanic acid and penicillin G stabilized the enzyme in the first stage of decay. Modulation factors in that stage were 0.96 for penicillin G and 0.98 for 6-aminopenicillanic acid. Phenylacetic acid increased the rate of inactivation in both stages, modulating factors being -2.31 and -2.23, respectively. Modulation factors influence enzyme performance in a reactor and are useful parameters for a proper evaluation.

Using metaphors in therapy: dichos and Latino clients.

The clinical literature has given increased attention to the importance of culturally appropriate interventions with diverse populations. The use of metaphor as a tool in psychotherapy has become increasingly salient. Metaphor can also be used in work with Latino clients through the incorporation of dichos, or sayings, that exist in Mexican American and other Latino cultures. These folk sayings exhibit cultural beliefs and ideals embedded in figurative language that describe the human condition. The Spanish language has crystallized human situations, frailties, and anecdotes in the dichos used in everyday conversations. These dichos offer the clinician culturally viable tools for mitigating resistance, enhancing motivation, or reframing problems. Importantly, dichos provide an ambience that contributes to culturally sensitive treatment.

[Immobilization of microbial lactase]. / Inmovilización de lactasa microbiana.

Lactase (beta-D galactoside-galactohydrolase, E.C.3.2.1.23) is a relevant enzyme to the dairy industry as it modifies undesirable functional and nutritional properties derived from the lactose content in milk and dairies, and as a way of recovering or upgrading cheese whey. This latter aspect has been considered to develop an enzyme catalyst suitable for the continuous hydrolysis of whey permeate. The selection of enzyme and support and the immobilization procedure has been reported previously. Results obtained in the immobilization of fungal lactase on activated chitin have prompted us to scale-up the procedure, a system being developed in which the enzyme is immobilized within the reactor (in situ). Results are presented for the in situ immobilization of lactase with and without recirculation of the reagents. Previous procedure was reproduced, although moderate profiles of activity were generated through the catalyst bed which were not eliminated by recirculation. Packed bed reactors with immobilized lactase were operated at varying flowrates and lactose concentrations, results being compared, in terms of substrate conversion and reactor productivity, with a theoretical model based on the corresponding kinetic expression and ideal flow regime. Deviations are significant at high flowrates which is attributed to backmixing and channeling through the catalyst bed. The model fits reasonably well at low flowrates and high feed substrate concentration. Productivity was 58 g of glucose/l.h at 40 ml/h of 120 g/l of lactose. Stability of the immobilized lactase was assessed in long-term reactor operation with whey permeate (35 g/l of lactose) at 40 degrees C and pH 4.0. Operational half-life was 120 days.

Disparate effects of dietary fatty acids on activity of 5'-nucleotidase of rat liver plasma membrane.

The effects of incorporation of dietary n-3 polyunsaturated fatty acids (PUFA) into rat liver plasma membrane on the activity of 5'-nucleotidase (EC 3.1.3.5) was studied. The membrane phospholipids from rats fed a diet containing 10% by weight menhaden oil (MO) for 3 wk contained more n-3 PUFA and less n-6 PUFA in phospholipids classes, i.e., 24% and 65% less linoleic and arachidonic acid in phosphatidylcholine, than in rats fed 10% coconut oil (CNO) diets. The specific activity of 5'-nucleotidase in n-3 PUFA-enriched hepatic plasma membranes was 1.6- to 2-fold higher than that in rats fed CNO or corn oil (CO). Lineweaver-Burk plots for 5'-nucleotidase in liver plasma membranes isolated from rats fed MO and CNO showed no significant differences in Km values but the Vmax was increased by 67% in MO-fed rats. Arrhenius plots showed a break point in 5'-nucleotidase activity at 28.3 degrees C and 30.8 degrees C in plasma membranes from MO- and CNO-fed rats, respectively. The implications of this in the generation of adenosine and its possible impact on physiological functions are discussed.

[Organization of family labor and fertility in rural Mexico]. / Organizacion del trabajo familiar y fecundidad en el Mexico rural.

PIP: The relationship between the organization of a family's work force and fertility in rural Mexico is assessed. The data are from the 1981 Rural Family Planning Survey, which covered approximately 8,000 households. Four groups are examined separately: farmers, agricultural wage earners, self-employed, and nonagricultural wage earners. Consideration is given both to fertility differentials among these groups and to differences in levels of contraceptive practice. (SUMMARY IN ENG)^ieng