Economic analysis of Uricase production under uncertainty: Contrast of chromatographic purification and aqueous two-phase extraction (with and without PEG recycle)

Uricase is the enzyme responsible for the breakdown of uric acid, the key molecule leading to gout in humans, into allantoin, but it is absent in humans. It has been produced as a PEGylated pharmaceutical where the purification is performed through three sequential chromatographic columns. More recently an aqueous two-phase system (ATPS) was reported that could recover Uricase with high yield and purity. Although the use of ATPS can decrease cost and time, it also generates a large amount of waste. The ability, therefore, to recycle key components of ATPS is of interest. Economic modelling is a powerful tool that allows the bioprocess engineer to compare possible outcomes and find areas where further research or optimization might be required without recourse to extensive experiments and time. This research provides an economic analysis using the commercial software BioSolve of the strategies for Uricase production: chromatographic and ATPS, and includes a third bioprocess that utilises material recycling. The key parameters that affect the process the most were located via a sensitivity analysis and evaluated with a Monte Carlo analysis. Results show that ATPS is far less expensive than chromatography, but that there is an area where the cost of production of both bioprocesses overlap. Furthermore, recycling doesn't impact the cost of production. This study serves to provide a framework for the economic analysis of Uricase production using alternative techniques. This article is protected by copyright. All rights reserved

Efecto tóxico in vivo e in vitro de los extractos polares de nueve plantas utilizadas en medicina tradicional.

Los vegetales son fuente de metabolitos secundarios que pudieran ser sustancias biológicamente activas, pero también podemos encontrar compuestos con toxicidad de leve a aguda como: terpenos, esteroides, cumarinas, alcaloide y flavonoides. Estos últimos tienen la propiedad de causar de formación, fragilidad osmótica y agregación en eritrocitos (E). El objetivo de este estudio es determinar efecto tóxico in vivo e in vitro de los extractos acuoso (A) y metanólico (M) de nueve plantas utilizadas en medicina tradicional mexicana. Se realizó la caracterización e identificación de compuestos fitoquímicos por pruebas colorimétricas y por HPLC, DL50 con el ensayo de Artemia salina, presencia de:TBARS, H2O2, eriptosis por marcaje con Annexina V-FITC y VCM en E humanos de donador sano, citotoxicidad en línea celular VERO, daño histológico y bioquímico en ratas macho Wistar e inducción de CYP3A4. El rendimiento de las plantas fue de 1.20 a 17%. Los cromatogramas por HPLC muestran componentes similares a los estándares utilizados. En el análisis por Citometria de flujo la exposición de los E a los extractos causó la inducción de eriptosis en un % similar al control positivo. En peroxidación lipídica sobre E se encontró que para TBARS/MDA los extractos A y M observaron una media por encima de lo establecido como normal. En la cuantificación de H2O2 en los E tratados con los extractos M, mostraron una media de 2.6 µM, y los A una media de 15.0 µM en comparación con los E sin tratamientos de 4.0 µM y con el valor citotóxico de ≥ a 50µM. En A. salina los extractos que mostraron toxicidad fueron Tillandsia recurvata, Sphaeralcea angustifolia y Gnaphallium oxyphyllum. En células VERO, T. recurvata y Loeselia mexicana presentaron citotoxicidad. El extracto de Calea ternifolia, mostró cambios histológicos en riñón en ratas macho Wistar. Los extractos de las plantas presentaron inhibición en la enzima CYP3A4 en baculosomas humanos. En conclusión los componentes fitoquímicos de C. ternifolia, Prosopis glandulosa, L. mexicana, Eryngium heterophyllum, G. oxyphyllum, Casuarina equisetifolia, T. recurvata, S. angustifolia y Tecoma stans con propiedades medicinales también inducen eriptosis, peroxiación lipídica e incremento de H2O2, efecto tóxico y citotóxico e inducen la expresión de CYP3A4. ABSTRACT The vegetables represent a source of secondary metabolites. Not only we can find biologically active substances, but also we can find compounds with high and low toxicity. For instance, terpenes, steroids, coumarin, alkaloids and flavonoids. The last compounds, can cause deformation, osmotic fragility and erythrocyte aggregation. The objective of this research is to determine the toxic effect, in vivo e in vitro, from the aqueous and methanol extract of nine plants used in the traditional mexican medicine. The following tests were done: the characterization and identification of phytochemical compounds by colorimetric test and HPLC, DL50 using Artemia salina model, presence of TBARS, H2O2 and eryptosis by Annexin V-FITC and VCM (in E humans and healthy donor), cytotoxicity on Vero cell lines, Histological and biochemical damage in male Wistar rats and CYP3A4 induction. The performance of the plants was from 1.20 to 17%. The HPLC's chromatograms showed similar components to the standards. The analysis, via flow cytometry, showed that the exposition from erythrocytes to extracts causes an induction of eryptosis in a similar percentage to the positive control. As a result, in the erythrocytes' lipid peroxidation, for TBARS/MDA, was observed that the A and M extracts had a median above the established value (normal value). In the H2O2 quantification, from the erythrocytes treated with M extracts, was showed a median of 2.6 µM. In the other hand, the erythrocytes treated with A extracts, showed a median of 15.0 µM. In comparison with the erythrocytes without treatment which showed a median of 4.0 µM, and a cytotoxicity value greater than or equal to 50 µM. Using Artemia salina, in fact, the extracts which showed toxicity were: Tillandsia recurvata, Sphaeralcea angustifolia y Gnaphallium oxyphyllum. By comparison, in VERO cells, the extracts which showed cytotoxicity were: Tillandsia recurvata y Loeselia mexicana. Furthermore, the Calea ternifolia's extract showed histological changes in the male Wistar rats' kidney. In addition, the plants' extracts showed inhibition in the CYP3A4 enzyme, as well as in human baculosomes. All things considered, the Calea ternifolia's phytochemical components, Prosopis glandulosa, Loeselia mexicana, Eryngium heterophyllum, Gnaphallium oxyphyllum, Casuarina equisetifolia, Tillandsia recurvata, Sphaeralcea angustifolia y Tecoma stans, with medical properties, also induces eryptosis as well as lipid peroxidation, increased of H2O2, toxic and cytotoxic effect and the expression of CYP3A4

Genotype Imputation to Improve the Cost-Efficiency of Genomic Selection in Farmed Atlantic Salmon

Genomic selection uses genome-wide marker information to predict breeding values for traits of economic interest, and is more accurate than pedigree-based methods. The development of high density SNP arrays for Atlantic salmon has enabled genomic selection in selective breeding programmes, alongside high-resolution association mapping of the genetic basis of complex traits. However, in sibling testing schemes typical of salmon breeding programmes, trait records are available on many thousands of fish with close relationships to the selection candidates. Therefore, routine high density SNP genotyping may be prohibitively expensive. One means to reducing genotyping cost is the use of genotype imputation, where selected key animals (e.g. breeding programme parents) are genotyped at high density, and the majority of individuals (e.g. performance tested fish and selection candidates) are genotyped at much lower density, followed by imputation to high density. The main objectives of the current study were to assess the feasibility and accuracy of genotype imputation in the context of a salmon breeding programme. The specific aims were: (i) to measure the accuracy of genotype imputation using medium (25 K) and high (78 K) density mapped SNP panels, by masking varying proportions of the genotypes and assessing the correlation between the imputed genotypes and the true genotypes; and (ii) to assess the efficacy of imputed genotype data in genomic prediction of key performance traits (sea lice resistance and body weight). Imputation accuracies of up to 0.90 were observed using the simple two-generation pedigree dataset, and moderately high accuracy (0.83) was possible even with very low density SNP data (~250 SNPs). The performance of genomic prediction using imputed genotype data was comparable to using true genotype data, and both were superior to pedigree-based prediction. These results demonstrate that the genotype imputation approach used in this study can provide a cost-effective method for generating robust genome-wide SNP data for genomic prediction in Atlantic salmon. Genotype imputation approaches are likely to form a critical component of cost-efficient genomic selection programmes to improve economically important traits in aquaculture

SEED ABORTION IN NATURALLY POLLINATED FLOWERS OF MEXICAN NATIVE PLANTS OF \u3ci\u3ePhaseolus coccineus\u3c/i\u3e L.

INTRODUCTION: Phaseolus coccineus L. plants require insects or hummingbirds to pollinate their flowers and set pods. The number of pods that can be produced by a plant is set by the number of flowers while the number of seeds is set by the number of ovules within the flowers (Stephenson, 1981). The arrest of the development of the seed after its partial differentiation, - seed abortion-, also determines the number of seeds per plant. The objective of this work is to determine the percentage of seed abortion per plant of two Mexican native varieties of Phaseolus coccineus L. MATERIALS AND METHODS: Two varieties (accessions no. 8446 and 8448) were selected from the Mexican bean collection of the Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP). Seeds harvested in November 2014, were sown in pots on April 5, 2015 and seedlings were transplanted outdoors on April 20, distant 2 meter apart. The plants were of indeterminate growth, climbing type. There were a natural occurrence of bees, bumblebees, and hummingbirds pollinators during the flowering period. The experiment was a complete randomized design, with two treatments (varieties), five replications (one plant per replication) and five sampling dates (Oct. 14, Nov. 4, Nov. 25, Dec. 9, and Dec. 16). At each sampling date, the mature pods per plant were harvested and opened. The following data were registered: a) the number of normal seeds per pod; b) the number of aborted seeds per pod including early abortions detected with the stereoscopic microscope. The sum of (a) and (b) = c, which represented the potential number of seeds per POD in each sampling date. It was evident at this point that practically all the pods in a variety had the same potential number of seeds. Therefore, the potential number of seeds per PLANT in each sampling date represented by Y = c*n, where n represents the number of pods per plant in each sampling date. Following when applicable, the similar procedure for seed abortion: Z = total number of aborted seeds per plant in each sampling date. The percentage of seed abortion (Z/Y)*100 (total number of aborted seeds per plant in each sampling date/potential number of seeds per plant in each sampling date)

SEED ABORTION IN NATURALLY POLLINATED FLOWERS OF MEXICAN NATIVE PLANTS OF \u3ci\u3ePhaseolus coccineus\u3c/i\u3e L.

INTRODUCTION: Phaseolus coccineus L. plants require insects or hummingbirds to pollinate their flowers and set pods. The number of pods that can be produced by a plant is set by the number of flowers while the number of seeds is set by the number of ovules within the flowers (Stephenson, 1981). The arrest of the development of the seed after its partial differentiation, - seed abortion-, also determines the number of seeds per plant. The objective of this work is to determine the percentage of seed abortion per plant of two Mexican native varieties of Phaseolus coccineus L. MATERIALS AND METHODS: Two varieties (accessions no. 8446 and 8448) were selected from the Mexican bean collection of the Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (INIFAP). Seeds harvested in November 2014, were sown in pots on April 5, 2015 and seedlings were transplanted outdoors on April 20, distant 2 meter apart. The plants were of indeterminate growth, climbing type. There were a natural occurrence of bees, bumblebees, and hummingbirds pollinators during the flowering period. The experiment was a complete randomized design, with two treatments (varieties), five replications (one plant per replication) and five sampling dates (Oct. 14, Nov. 4, Nov. 25, Dec. 9, and Dec. 16). At each sampling date, the mature pods per plant were harvested and opened. The following data were registered: a) the number of normal seeds per pod; b) the number of aborted seeds per pod including early abortions detected with the stereoscopic microscope. The sum of (a) and (b) = c, which represented the potential number of seeds per POD in each sampling date. It was evident at this point that practically all the pods in a variety had the same potential number of seeds. Therefore, the potential number of seeds per PLANT in each sampling date represented by Y = c*n, where n represents the number of pods per plant in each sampling date. Following when applicable, the similar procedure for seed abortion: Z = total number of aborted seeds per plant in each sampling date. The percentage of seed abortion (Z/Y)*100 (total number of aborted seeds per plant in each sampling date/potential number of seeds per plant in each sampling date)

1-(2-Chlorobenzyloxy)-3-[1,2,3]triazol-1-yl-propan-2-ol Derivatives: Synthesis, Characterization, and DFT-Based Descriptors Analysis

A novel series of 1-(2-chlorobenzyloxy)-3-[1,2,3]triazol-1-yl-propan-2-ol derivatives was designed and synthesized using copper catalyzed alkyne-azide cycloaddition in the key step. Theoretical investigation of molecular and electronic properties by means of global and local reactivity indexes of the synthetized compounds was carried out, using DFT (Density Functional Theory) at PBEPBE/6-31++G∗∗ levelCONACY

Two-Swim Operators in the Modified Bacterial Foraging Algorithm for the Optimal Synthesis of Four-Bar Mechanisms

This paper presents two-swim operators to be added to the chemotaxis process of the modified bacterial foraging optimization algorithm to solve three instances of the synthesis of four-bar planar mechanisms. One swim favors exploration while the second one promotes fine movements in the neighborhood of each bacterium. The combined effect of the new operators looks to increase the production of better solutions during the search. As a consequence, the ability of the algorithm to escape from local optimum solutions is enhanced. The algorithm is tested through four experiments and its results are compared against two BFOA-based algorithms and also against a differential evolution algorithm designed for mechanical design problems. The overall results indicate that the proposed algorithm outperforms other BFOA-based approaches and finds highly competitive mechanisms, with a single set of parameter values and with less evaluations in the first synthesis problem, with respect to those mechanisms obtained by the differential evolution algorithm, which needed a parameter fine-tuning process for each optimization problem

Desarrollo de la planta de Agave tequilana mediante la tecnica de fertigacion en tamaulipas : Development of the Agave tequilana plant with fertigation in tamaulipas

Con el objetivo de estudiar las características agronómicas y la acumulación de nutrimentos en la materia seca (MS) durante un ciclo completo de la planta de agave, en un experimento establecido en el sur de Tamaulipas se plantaron hijuelos de agave en tres condiciones de manejo del fertilizante: a) En temporal y sin fertilizante, Testigo Absoluto (TA); b) Fertigación, además de la precipitación, recibieron agua y nutrimentos (N, P, K, Ca, Mg, y micronutrimentos) mediante un sistema de riego por goteo; y c) Fertilización de base (FB)+Fertigación, adicional a lo recibido en el tratamiento anterior, se aplicó a las plantas de agave FB con N, P, K y micronutrimentos. Cada tratamiento se estudió por triplicado en un diseño de bloques al azar. Durante el estudio se realizaron ocho evaluaciones destructivas utilizando una planta por unidad experimental; septiembre del 2004, febrero del 2005, octubre del 2005, 2006, 2007, 2008, 2009 y 2010. Las plantas desarrolladas con Fertigación y FB+Fertigación, en promedio superaron en 17 y 76 % la altura y número de hojas registradas en la planta del TA, respectivamente. En relación con la biomasa, se estimó que estas mismas plantas produjeron 491.4 y 456.1 t ha-1 en la planta total, y 218.1 y 252.4 t ha-1 en la piña, respectivamente. En plantas del TA la biomasa total fue 97 t ha-1 y 46.5 t ha-1 en la piña. La concentración de azucares reductores (AR) en la piña no fue modificada por el manejo con fertilizante (P≥0.05), su concentración fluctuó entre 26.8 y 29.0 %. La concentración de nutrimentos en la MS, tampoco fue modificada con el fertilizante (P≥0.05); en la MS de plantas desarrolladas con Fertigación y Fertigación+FB se registró la mayor cantidad de nutrimentos, los valores medios, fueron 182.7 g de N, 60.1 g de P, 165.1 g de K, 816.4 g de Ca y 54.1 g de Mg, los cuales fueron superiores y significativamente diferentes (P≤0.05) a los registrados en la MS de la planta del TA. Independientemente del manejo de la fertilización, la cantidad de nutrimentos determinados en la MS presentó el siguiente orden Ca>K>N>Mg>P