Yield Of Final Product Research Articles

A Closed-tube Loop-Mediated Isothermal Amplification Assay for the Visual Endpoint Detection of Brucella spp. and Mycobacterium avium subsp. paratuberculosis.

LAMP (loop-mediated isothermal amplification) is an isothermal nucleic acid amplification technique that is characterized by its efficiency, rapidity, high yield of final product, robustness, sensitivity, and specificity, with the blueprint that it can be implemented in laboratories of low technological complexity. Despite the conceptual complexity underlying the mechanistic basis for the nucleic acid amplification, the technique is simple to use and the amplification and detection can be carried out in just one step. In this chapter, we present a protocol based on LAMP for the rapid identification of isolates of Brucella spp. and Mycobacterium avium subsp. paratuberculosis, two major bacterial pathogens in veterinary medicine.

Sludge from paper mill effluent treatment as raw material to produce carbon adsorbents: An alternative waste management strategy

Pulp and paper industry produces massive amounts of sludge from wastewater treatment, which constitute an enormous environmental challenge. A possible management option is the conversion of sludge into carbon-based adsorbents to be applied in water remediation. For such utilization it is important to investigate if sludge is a consistent raw material originating reproducible final materials (either over time or from different manufacturing processes), which is the main goal of this work. For that purpose, different primary (PS) and biological sludge (BS) batches from two factories with different operation modes were sampled and subjected to pyrolysis (P materials) and to pyrolysis followed by acid washing (PW materials). All the materials were characterized by proximate analysis, total organic carbon (TOC) and inorganic carbon (IC), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) and N2 adsorption isotherms (specific surface area (SBET)and porosity determination). Sludge from the two factories proved to have distinct physicochemical properties, mainly in what concerns IC. After pyrolysis, the washing step was essential to reduce IC and to considerably increase SBET, yet with high impact in the final production yield. Among the materials here produced, PW materials from PS were those having the highest SBET values (387–488 m2 g−1). Overall, it was found that precursors from different factories might originate final materials with distinct characteristics, being essential to take into account this source of variability when considering paper mill sludge as a raw material. Nevertheless, for PS, low variability was found between batches, which points out to the reliability of such residues to be used as precursors of carbon adsorbents.

Effect of harvest date on growth, production and quality of honeybush (Cyclopia genistoides and C. subternata)

Cyclopia spp., used for production of honeybush herbal tea, is endemic to specific climatic zones in the Western and Eastern Cape of South Africa. Most of the plant material is harvested from wild plants in mountainous areas. Production and market development of this popular herbal tea is unsustainable without increased commercial production. Information on cultivation requirements, growth habit and production potential is essential to ensure a sustainable industry. The effect of harvest date on growth, production and tea quality was evaluated in non-irrigated commercial plantations of Cyclopiagenistoides and Cyclopia subternata. Cyclopia genistoides bushes, harvested annually, did not recover the height of un-harvested bushes. Fastest C. genistoides bush regrowth and highest dry mass yield occurred after February harvests, with September harvests resulting in the highest sieve quality and mangiferin content (only the second annual harvest). For the “fermented” product, its hot water soluble solids (as indication of cup strength) and total polyphenol content, tended to increase with years of harvest. The mangiferin content of “unfermented” (green) C. genistoides plant material increased with years of harvest. Cyclopia subternata bushes harvested during August and September recovered the fastest in height. The August and May harvests delivered the highest fresh mass yield, while harvesting during August delivered significantly more of the finer processed tea (<1400μm) and less of the coarse fraction (>2800μm) than a September, February or May harvest. Annual harvesting is encouraged, but harvesting should not take place during flowering (August to September), because the flowers add very little to the final product yield. Based on 10,000 bushes/ha, an estimated annual production of 8.5 or 10.4 tons/ha of C. genistoides and C. subternata (fresh plant material), respectively, can be achieved.

Development of palm-based neopentyl glycol diester as dielectric fluid and its thermal aging performance

The potential of palm-based neopentyl glycol diester as dielectric insulating fluid was investigated. The details of the transesterification of high oleic palm oil methyl ester (POME) with neopentyl glycol (NPG) with the final product yield of more than 90 wt% of NPG diester were discussed. The thermal aging performance of NPG diester was compared with conventional mineral insulating oil at 90, 110 and 130 °C. This paper focused mainly in the effects of aging to chemical, physical and electrical properties of NPG diester. Apart from being fully biodegradable and non-toxic, the synthesized diesters exhibited high flashpoint and the breakdown voltage was comparable to mineral oil. The result indicated that throughout the aging period, NPG diester exhibited lower acid value than mineral oil and no significant change in viscosity was observed. The study on mechanical properties of insulating paper aged in NPG diester shows higher tensile strength than paper aged in mineral oil. The synthesized esters have shown great potential to be used as transformer oil.

Abstract 3634: Simultaneous isolation of genomic DNA and total RNA using the Qiagen AllPrep® method

Abstract Introduction: The purification of genomic DNA (gDNA) and total RNA (RNA) from various specimen types using a single sample is currently in high demand in genomic settings due to the limitation of available biological material. There is an increased need for simultaneous isolation of nucleic acids in cancer genetics for use in downstream applications such as genotyping, microarrays or next generation sequencing. The Qiagen AllPrep® technique provides a comprehensive solution for extraction of gDNA and RNA from a single specimen. In this study, we tested the AllPrep® kits for concurrent isolation of gDNA and RNA from five different sample types. Materials and Methods: Genomic DNA and RNA were extracted simultaneously according to Qiagen AllPrep® manufacturer's instructions from five specimen types: bone marrow mononuclear cells (BMMC), cryopreserved cultured cell lines, whole blood, fresh frozen and FFPE samples. Quantity and quality of both nucleic acids were assessed by the NanoDrop 2000 and Qubit fluorometer 2.0. Purified gDNA from BMMC, fresh frozen, whole blood and FFPE samples was further evaluated by library preparation, targeted hybridization capture and sequencing on an Illumina NextSeq500. FastQ files were submitted to a Clinical Genome Analytics (CGA) pipeline for quality metrics analysis. Results: The AllPrep® technique resulted in effective isolation of high quality gDNA and RNA (Table1). All set wet lab and sequencing QC metrics were met for each sample type. Conclusion: We have successfully verified the AllPrep® method can be used for simultaneous extraction of both gDNA and RNA using the same input material without compromising the yield or purity of both final products. This simple procedure based on spin column technology not only gave the advantage of generating high quality results using limited specimen resources, but also decreased the hands on time by 3.5 hours and cost when compared to extraction done with two separate kits. Table 1.Represents average QC results and SD for each specimen type. **n = 8 for sequencing metricsgenomic DNAgenomic DNAtotal RNAtotal RNASample TypeOD260/280Total Yield (ng)OD260/280Total Yield (ng)Mean Target Coverage% DuplicationBMMC (n = 3)1.853033.31.9413855206.34%Whole Blood (n = 2)1.818936.51.881335566.96.47%Fresh Frozen (n = 2)1.69312752.0432375345.44.02%FFPE Specimen (n = 16)1.665133.41.9410524.1**748.2**6.10%Cultured Cell Lines (n = 5)1.8442981.9810489N/AN/ATotal Average (n = 28)1.736898.141.9510443624.75.92%Standard Deviation (SD).249180.90.110616.2228.30.013% Citation Format: Aleksandra Ras, Samantha Helm, Kevin Kelly, Vanessa Spotlow, Guruprasad Ananda, Gregory Tsongalis. Simultaneous isolation of genomic DNA and total RNA using the Qiagen AllPrep® method. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3634.

Equilibrium energy and technological characteristics of plasma synthesis of titanium nitride, carbide, and carbonitride from titanium tetrachloride

Equilibrium energy and technological characteristics of the plasma synthesis of titanium nitride, carbide, and carbonitride, in particular, the composition and yields of final products, energy consumption upon their production, and the required enthalpy of a plasma flow, are calculated for the following reactions: TiCl4 + xH2 + yN2 (synthesis of TiN), TiCl4 + xH2 + zC m H n (synthesis of TiC), and TiCl4 + xH2 + y 1N2 + z 1C m H n (synthesis of TiCN). The calculations are carried out for the temperature range from 500 to 4000 K, at the total pressure of 0.1 MPa in the system, and with the following molar ratios of reagents: x = 8, x = 20; y = 0.5, y = 5, y = 20; z = 0.9, z = 1.0, z = 1.1; y 1 = 5; z 1 = 0.4, z 1 = 0.5, z 1 = 0.6.

A convenient and efficient method for the synthesis of new 2-(4-amino substituted benzilidine) indanone derivatives

An efficient and improved process for the synthesis of novel 1-indanones derivatives has been developed. In the first step of the presented new process, 4-bromobenzaldehyde reacts with 5, 6-dimethoxy-2, 3-dihydro-1H-inden-1-one by N-arylation of corresponding compounds with a variety of aliphatic and aromatic amines to produce a novel category of 1-indanones as the final step. Comparison of this newly designed method with conventional methods show that our method is much more efficient in producing a higher total yield. This innovative synthesis methodology offers several advantages, among which the consumption of inexpensive reagents and the resulting higher total yields of final products are of its remarkable points.

Theoretical justification of the possibility of cryocooling of a solid target at irradiation with proton beam from cyclotron C18

The thermal physics processes of solid target cooling at proton beam irradiation in С18 cyclotron were calculated. The calculations were made with the help of finite-elements method using ANSYS program for beams with nonuniform density of particle distribution in the beam profile. It was shown that the radiation efficiency and conditions of target cooling essentially depend on distributions of particles in the beam and on efficient size of beam. A principal possibility of the cryocooling of target is shown that would essentially increase the radiation efficiency and the yield of final product–the medical isotope 99mTc. The developed calculation method may be of use also for other processes of target irradiation with charged particle beams.

Influence of Linker Length Variations on the Biomass-Degrading Performance of Heat-Active Enzyme Chimeras.

Plant cell walls are composed of complex polysaccharides such as cellulose and hemicellulose. In order to efficiently hydrolyze cellulose, the synergistic action of several cellulases is required. Some anaerobic cellulolytic bacteria form multienzyme complexes, namely cellulosomes, while other microorganisms produce a portfolio of diverse enzymes that work in synergistic fashion. Molecular biological methods can mimic such effects through the generation of artificial bi- or multifunctional fusion enzymes. Endoglucanase and β-glucosidase from extremely thermophilic anaerobic bacteria Fervidobacterium gondwanense and Fervidobacterium islandicum, respectively, were fused end-to-end in an approach to optimize polysaccharide degradation. Both enzymes are optimally active at 90 °C and pH 6.0-7.0 representing excellent candidates for fusion experiments. The direct linkage of both enzymes led to an increased activity toward the substrate specific for β-glucosidase, but to a decreased activity of endoglucanase. However, these enzyme chimeras were superior over 1:1 mixtures of individual enzymes, because combined activities resulted in a higher final product yield. Therefore, such fusion enzymes exhibit promising features for application in industrial bioethanol production processes.

The effect of controlling the drying distortion of laminas on the production yield of cross-laminated timber (CLT) using Larix kaempferi wood

The objective of this study was to evaluate the effect of lamina distortion control on the production yield of cross-laminated timber (CLT) using domestic larch (Larix kaempferi) wood. The measured production yield of CLT was 27 %. The yields based on log volume of sawing, planing, cross-cutting, and finish planing of CLT were 49, 66, 96, and 86 %, respectively. The yield of sawing and planing can be improved by controlling distortion during drying. Using an experimental enforced-loading apparatus, the cup and twist of laminas were reduced by 50 and 90 %, respectively. To analyze the effect of a reduction in sawing dimensions on the final production yield of CLT, an analysis of a geometrically applied sawing pattern of logs was used to predict the sawing yield and the number of laminas after sawing. The geometrical analysis showed that a change in the sawing dimensions affected the production yield of CLT. If the width and thickness of the sawing dimensions can be reduced by 5 mm by decreasing the distortion of lamina during drying, the sawing yield was unchanged (49 %), but it was expected that the number of laminas after sawing would increase from 278 to 303. It was also predicted that the production yield of CLT would increase from 27 to 32 %.

Manufacture of an Ultrafine-Grained TiN–Cu Composition Using an Erosion-Type Coaxial Hybrid Magnetoplasma Accelerator

It is shown that a TiN+Cu powder mixture could be manufactured using a combined barel of a coaxial magnetoplasma accelerator. The method is proved to ensure a wide-range regulation over the copper-to-titanium nitride ratio in the final product yield.

Cord blood processing by a novel filtration system.

ObjectivesAvailability of cord blood (CB) processing has been limited by the need for electrically aided centrifugal techniques, which often produce only low final cell product yield. Here, we describe development and characterization of a novel filter device aimed at allowing CB processing, using gentle gravity‐led flow.Materials and methods CB was processed with a novel filter device (CellEffic CB, consisting of non‐woven fabric), without any centrifugation. Cells were harvested by flushing the filter with either HES or physiological saline solution (SALINE). Differential cell counts and viability analysis, combined with Fluorescence‐Activated Cell Sorting (FACS) (total nucleated cells [TNC], mononuclear cells [MNC], CD45+ CD34+ cells, hematopoietic precursor cells [HPCs]) and clonogenic assay, were employed for analysis of CB pre‐ and post‐processing, and after freeze/thawing.ResultsProcessing using the novel filter yielded high quality RBC depletion while maintaining good recovery of TNC, MNC, CD34+, HPCs and colony forming unit (CFU) output. The filter performed equally well using HES or SALINE. Gravity‐led flow provided gentle cell movement and protection of the stem cell compartment. Post‐thaw CFU output was maintained particularly, an important indicator for CB banking.ConclusionsGeographical limitations of CB transplantation and banking have required a non‐electrical, non‐centrifugal solution. This novel filter CellEffic CB device revealed rapid yet gentle cell processing while maintaining the stem/progenitor cell compartment required for both haematological and regenerative medicine therapies.

Soluble expression, purification and functional characterization of a coil peptide composed of a positively charged and hydrophobic motif.

A de novo heterodimeric coiled-coil system formed by the association of two synthetic peptides, the Ecoil and Kcoil, has been previously designed and proven to be an excellent and versatile tool for various biotechnology applications. However, based on the challenges encountered during its chemical synthesis, the Kcoil peptide has been designated as a "difficult peptide". In this study, we explore the expression of the Kcoil peptide by a bacterial system as well as its subsequent purification. The maximum expression level was observed when the peptide was fused to thioredoxin and the optimized purification process consisted of three chromatographic steps: immobilized-metal affinity chromatography followed by cation-exchange chromatography and, finally, a reverse-phase high-performance liquid chromatography. This entire process led to a final volumetric production yield of 1.5mg of pure Kcoil peptide per liter of bacterial culture, which represents a significant step towards the cost-effective production and application of coiled-coil motifs. Our results thus demonstrate for the first time that bacterial production is a viable alternative to the chemical synthesis of de novo designed coil peptides.

Methane and VFA production in anaerobic digestion of rice straw under dry, semi-dry and wet conditions during start-up phase

ABSTRACTThis work investigates the start-up phase of anaerobic digestion in wet, semi-dry and dry conditions of rice straw analysing the role of volatile fatty acid (VFA) production on process kinetics. Methane production yields and biodegradation kinetics in reactors operated under wet semi-dry and dry conditions were investigated. The experimental results showed a reduction in the specific final methane production yield of 57% and 63% in, respectively, semi-dry (TS = 14.8%) and dry (TS = 23.4%) conditions compared to wet (TS = 4.8%) conditions. The total VFA concentration and speciation are proposed as indicators of process development at different total solids content. High VFA concentrations were found in dry conditions, with a maximum total VFA concentration of 2110 mg/kg in dry conditions, 930 mg/kg in semi-dry conditions and 180 mg/kg in wet conditions.

Estimates of heritabilities and genetic correlations of raw flesh quality traits in a reared gilthead sea bream (Sparus aurata L.) population sourced from broodstocks along the Spanish coasts

Abstract In gilthead sea bream, flesh quality traits such as body composition and texture directly influence yield of final product and consumer preferences so they should be considered in the breeding goal. However, strategies that involve the development of selection schemes for these traits of economic interest are scarce. Taking into account these circumstances, in this study the effect of the origin of the broodstock on the major flesh quality traits was analyzed at harvest size (690 days post-hatching) and genetic parameters (heritabilities and genetic correlations) were estimated as well as their correlations with harvest weight. For this purpose, a population of farmed gilthead sea bream was obtained from three broodstock of different geographical origins along the Spanish coast [Cantabrian Sea (CAN), the Atlantic Ocean (ATL) and Mediterranean Sea (MED)]. Parental assignments between breeders and their offspring were carried out a posteriori using a microsatellite multiplex (SMsa1). In the offspring, raw flesh composition (muscular collagen, fat, moisture and protein contents) were determined (n = 700). Textural parameters (hardness, cohesiveness and derived traits) were measured in an industry relevant number of offspring (n = 890) for the first time in this species. The origin had an effect on muscular fat content as well as on hardness and derived textural parameters. Fish from MED showed the lowest fat percentage and those from CAN the highest values for textural parameters. Differences among origins could be explained through their different genetic backgrounds. However, the effect of the origin could be affected by environmental conditions in the initial facilities, where each origin was reared separately, and by genotype x environment interactions. Heritabilities were medium for muscular fat (0.31 ± 0.08) and moisture (0.24 ± 0.07). The genetic correlation between them was very high and negative (− 0.99 ± 0.02). Selection for harvest weight may lead to an increase in fillet fat percentage due to the genetic correlation between the two traits (0.29 ± 0.14). Hardness showed a medium heritability (0.21 ± 0.06) and an unfavourable negative genetic correlation with harvest weight (− 0.34 ± 0.14). All findings reported in this study should be relevant for the establishment of successful breeding programs in aquaculture of this species. Statement of relevance This study proves the importance of the acquisition of a stock in sea bream since the studied origins of the broodstock has an effect on muscular fat content and textural parameters. Moreover, genetic parameters are estimated for this traits which are scarce in this species.

Purification optimization for a recombinant single-chain variable fragment against type 1 insulin-like growth factor receptor (IGF-1R) by using design of experiment (DoE)

Design of experiment (DoE) is a statistics-based technique for experimental design that could overcome the shortcomings of traditional one-factor-at-a-time (OFAT) approach for protein purification optimization. In this study, a DoE approach was applied for optimizing purification of a recombinant single-chain variable fragment (scFv) against type 1 insulin-like growth factor receptor (IGF-1R) expressed in Escherichia coli. In first capture step using Capto L, a 2-level fractional factorial analysis and successively a central composite circumscribed (CCC) design were used to identify the optimal elution conditions. Two main effects, pH and trehalose, were identified, and high recovery (above 95%) and low aggregates ratio (below 10%) were achieved at the pH range from 2.9 to 3.0 with 32–35% (w/v) trehalose added. In the second step using cation exchange chromatography, an initial screening of media and elution pH and a following CCC design were performed, whereby the optimal selectivity of the scFv was obtained on Capto S at pH near 6.0, and the optimal conditions for fulfilling high DBC and purity were identified as pH range of 5.9–6.1 and loading conductivity range of 5–12.5mS/cm. Upon a further gel filtration, the final purified scFv with a purity of 98% was obtained. Finally, the optimized conditions were verified by a 20-fold scale-up experiment. The purities and yields of intermediate and final products all fell within the regions predicted by DoE approach, suggesting the robustness of the optimized conditions. We proposed that the DoE approach described here is also applicable in production of other recombinant antibody constructs.

ProducingAspergillus tubingensisCTM507 Glucose oxidase by Solid state fermentation versus submerged fermentation: process optimization and enzyme stability by an intermediary metabolite in relation with diauxic growth

BACKGROUND The variability of glucose oxidase (GOD) by Aspergillus tubingensis CTM507 in respect of titer under both submerged (SmF) and solid-state fermentation (SSF) was assessed in relation to growth and substrate consumption. L27 Taguchi experimental design was applied to determine optimum culture conditions for GOD production in SSF. RESULTS Kinetic results showed that fungal growth is a diauxic process which used glucose and gluconic acid as substrates, respectively, and that SmF is suitable for fungi growth. However, SSF led to higher GOD activity (170.59 U mL−1 against 43.73 U mL−1 by SmF). Moreover, GOD stability and liberation were related to the presence and levels of glucose, gluconic acid and to a lesser degree, lactose and arabinose. Optimization of culture media of multi-agro resources led to a final production yield of about 73103 U g−1 of substrates which corresponded to 10 and 84-fold higher than the preliminary SFF and SmF results, respectively. These conditions were validated experimentally and revealed an enhanced GOD yield of 74%. CONCLUSION These findings help an understanding of the phenomena of supremacy of SSF vs SmF to produce functional biomolecules. They also define an SSF process for an important enzyme production at low cost with interesting properties for industrial application. © 2015 Society of Chemical Industry

A numerical study on biomass fast pyrolysis process: A comparison between full lumped modeling and hybrid modeling combined with CFD

This study focuses on process modeling and simulation of a biomass fast pyrolysis system. For the simulation of the biomass fast pyrolysis process, two types of simulation models were developed: lumped model and hybrid model. Employing the above models, the effect of reaction temperature on reaction rate and final product yields were analyzed. It was found that the hybrid model exhibited a peculiar characteristic of displaying multiphase reacting flow occurring in fluidized bed. This behavior of hybrid model could have attributed for the difference in product yields of the models (hybrid and lumped). For the yields of the tar and NCG, hybrid model prediction was very consistent with the experimental results than the lumped model. However, for char yield, the results of both the lumped model and the hybrid model were close to that of experimental results.

Vplyv aplikovaného dusíka na odber vybraných makroelementov a obsah vitamínu C pri pekinskej kapuste

Pot trial was aimed to investigate an effect of fertilizer nitrogen on the content of vitamin C and offtake of certain macroelements by Brassica pekinensis. The experiment was realized in the years 2000 and 2001 in pots containing 10 kg of loamy brownsoil. Optimalization of NPK rates contributed to the increase of N, P, K, Mg and S offtake by the yield of final product in comparison with unfertilized control. The most marked increase offtake of nitrogen (by 14,6%) was obtained when fertilizer N was applied in the diveded form (80% N prior to seeding + 20% N in initial phase of heads formation, nitrogen applied as DAM 390). Positive effect of full NPK nutrition on the vitamin C content was determined. The highest content of vitamin C (629.0 mg.kg-1 of fresh mass) was determined in treatment where nitrogen was applied in the divided rate.

Reduction of invasive bacteria in ethanol fermentations using bacteriophages.

Invasive Lactobacillus bacteria inhibit ethanol fermentations and reduce final product yields. Due to the emergence of antibiotic resistant strains of Lactobacillus spp., alternative disinfection strategies are needed for ethanol fermentations. The feasibility of using the bacteriophage (phage) 8014-B2 to control Lactobacillus plantarum in ethanol fermentations by Saccharomyces cerevisiae was investigated. In 48 h media-based shake flask fermentations, phages achieved greater than 3-log inactivation of L. plantarum, protected final ethanol yields, and maintained yeast viability. The phage-based bacterial disinfection rates depended on both the initial phage and bacterial concentrations. Furthermore, a simple set of kinetic equations was used to model the yeast, bacteria, phage, reducing sugars, and ethanol concentrations over the course of 48 h, and the various kinetic parameters were determined. Taken together, these results demonstrate the applicability of phages to reduce L. plantarum contamination and to protect final product yields in media-based fermentations.