Barley—Botany,production, harvesting,processing, utilization and economics

Barley, probably the oldest cultivated cereal, is widely grown in cooler areas of the world. The annual world production of nearly two and a half billion bushels exceeds that of rye but is less than that of rice, wheat, corn and oats, respectively. Most of the annual 300 million dollar crop of the U.S. is fed to livestock, but about one- third is manufactured into malt.     

A rapid and robust liquid chromatography/tandem mass spectrometry method for simultaneous analysis of anti-tuberculosis drugs—Ethambutol and pyrazinamide in human plasma

Ethambutol and pyrazinamide are two first-line anti-tuberculosis drugs. Though they are normally combined for the treatment, their highly different polarity complicates simultaneous liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis of these two drugs in human plasma with decent peak shape and retention. Here we report a rapid and robust LC/MS/MS method for the simultaneous determination of these two drugs in human plasma. Human plasma samples, together with the isotopically labeled internal standards were extracted using protein precipitation, and then separated on a Chromolith SpeedROD RP-18e column and detected with mass spectrometry. The mobile phase is 0.1% trifluoroacetic acid in water and 0.1% trifluoroacetic acid in methanol. Addition of trifluoroacetic acid in the mobile phases was found to be able to improve peak shape as well as to increase the retention of ethambutol, thus being able to analyze these two drugs at the same time with both drugs having decent peak shape and enough retention on a C₁₈ column. An atmospheric pressure chemical ionization interface was chosen to reduce ion suppression from sample matrix components and provide high sensitivity. The standard curve range was 10.0–5000 ng/mL for ethambutol and 50.0–25,000 ng/mL for pyrazinamide using a plasma sample volume of 50.0 μL. This method has a very short run time of 3.8 min. The method has been fully validated, and <15% relative standard deviation was obtained for both analytes.     

A general-purpose system for the rapid acquisition and computer processing of optical and scanning electron microscope images—II. Software

The design and implementation of software for the operation of a general-purpose optical and electron microscope image processing system is described. The software is a group of programs, controlled by a command-line interpreter called IPR (Image PRocessing). The interpreter may be used interactively, or groups of commands may be issued indirectly after placing them in files. Programs for specialized image processing applications may obtain the services of the system's memory-resident programs, through the same interprogram communication methods that are used by the command-line interpreter.     

Simple,sensitive and rapid HPLC–MS/MS method for the determination of cepharanthine in human plasma

A rapid, sensitive and specific method for the determination of cepharanthine in human plasma using high performance liquid chromatography coupled with tandem mass spectrometry (HPLC–MS/MS) was described. Cepharanthine and the internal standard (I.S.), telmisartan, were extracted from human plasma by methanol to precipitate the protein. A centrifuged upper layer was then evaporated and reconstituted with 100 μL methanol. Chromatographic separation was performed on an AGILENT XDB-C₈ column (150 mm × 2.1 mm, 5.0 μm, Agilent, USA) using a gradient mobile phase with 1 mmol/L ammonium acetate in water with 0.05% formic acid and methanol. Detection and quantitation was performed by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the positive ion mode. The most intense [M+H]⁺ MRM transition of cepharanthine at m/z 607.3 → 365.3 was used for quantitation and the transition at m/z 515.5 → 276.4 was used to monitor telmisartan. The calibration curve was linear within the concentration range of 0.5–200.0 ng/mL (r = 0.9994). The limit of quantification (LOQ) was 0.5 ng/mL. The extraction recovery was above 81.1%. The accuracy was higher than 92.3%. The intra- and inter-day precisions were less than 9.66%. The method was accurate, sensitive and simple and was successfully applied to a pharmacokinetic study after single intravenous administration of 50 mg cepharanthine in 12 healthy Chinese volunteers.     

A rapid and simple method for the isolation of S100a0 (αα) protein by high-performance liquid chromatography

A rapid and simple method to isolate S100a₀ protein from the mixture of bovine S100 protein (S100a₀, S100a and S100b) is described. The S100 mixture purified from bovine brain was applied to an anion-exchange column, equilibrated with 50 mM Tris HC1 buffer (pH 8.0) in a high-performance liquid chromatography (HPLC) system. S100a₀, S100a and S100b proteins could be eluted separately from the column, which were identified by the immunoassay method, by the Tris-HC1 buffer containing a linear concentration gradient (0.25–0.4) M of NaCl. Immunoreactive S100a₀ protein was found in two peak fractions, and each S100a₀ fraction could be isolated (S100a₀-1 and S100a₀-2). Both fractions of S100a₀ protein showed a single band at the same position on acrylamide gel electrophoresis, and eluted in a single peak in the same fractions upon gel-filtration column chromatography. There was no significant difference in the amino acid composition between the two S100a₀ fractions. Since the S100a₀-1 fraction aged for several months at 4°C in the presence of 0.1% NaN₃ was found to contain four protein peaks including the fraction corresponding to the S100a₀-2 fraction, the difference between the two S100a₀ fractions is probably due to some modification of amino acid residues in the molecule, which may occur both in vivo and in vitro.     

A toolbox approach for the rapid evaluation of multi-step enzymatic syntheses comprising a ‘mix and match’ E. coli expression system with microscale experimentation

Abstract

This work describes an experimental ‘toolbox’ for the rapid evaluation and optimisation of multi-step enzymatic syntheses comprising a ‘mix and match’ E. coli-based expression system and automated microwell scale experimentation. The approach is illustrated with a de novo designed pathway for the synthesis of optically pure amino alcohols using the enzymes transketolase (TK) and transaminase (TAm) to catalyze asymmetric carbon-carbon bond formation and selective chiral amine group addition respectively. The E. coli expression system, based on two compatible plasmids, enables pairs of enzymes from previously engineered and cloned TK and TAm libraries to be evaluated for the sequential conversion of different initial substrates. This is complemented by the microwell experimentation which enables efficient investigation of different biocatalyst forms, use of different amine donors and substrate feeding strategies. Using this experimental ‘toolbox’, one-pot syntheses of the diastereoisomers (2S,3S)-2-aminopentane-1,3-diol (APD) and (2S,3R)-2-amino-1,3,4-butanetriol (ABT) were designed and performed, which gave final product yields of 90% mol/mol for APD and 87% mol/mol for ABT (relative to the initial TK substrates) within 25 hours. For the synthesis of APD, the E coli TK mutant D469E was paired with the TAm from Chromobacterium violaceum 2025 while for ABT synthesis the wild-type E. coli TK exhibited the highest specific activity and ee( enantiomeric excess) of >95%. For both reactions, whole-cell forms of the TK-TAm biocatalyst performed better than cell lysates while isopropylamine (IPA) was a preferable amine donor than methylbenzylamine (MBA) since side reactions with the initial TK substrates were avoided. The available libraries of TK and TAm enzymes and scalable nature of the microwell data suggest this ‘toolbox’ provides an efficient approach to early stage bioconversion process design in the chemical and pharmaceutical sectors.     

Assessment of ecosystem services at the national level in Germany—Illustration of the concept and the development of indicators by way of the example wood provision

The EU Biodiversity Strategy stipulates in Target 2, Action 5 that the member states must map and assess the state of the ecosystems and their services and promote the integration into the reporting systems at the EU and national level by 2020. Therefore indicators for capturing and assessing ecosystem services (ES) are needed. In this paper we report for which ES class types currently ES indicators are being developed for Germany in the context of an ongoing research project. Additionally, we provide the indicator specifications, which are based on underlying framework concept. By way of the example of the provisioning service ‘raw wood production’ and the development of the main-indicator ‘annual wood accrual’ and six sub-indicators, we illustrate the concrete procedure, including discussion of results and target values. The indicators for the ES wood provision are not only suitable for an exemplary illustration of procedure, data selection and data basis in Germany. Furthermore, it shows that indicators for provisioning ES can eminently conflict with biodiversity and other ES.     

Spirula—a window to the embryonic development of ammonoids? Morphological and molecular indications for a palaeontological hypothesis

Nautilus is not suitable as a model organism to infer biological functions, embryonic development, or mode of life in ammonoids. A brief review of the available morphological data is given and molecular data are added to discuss the usefulness of Spirula as a biological proxy for ammonoids. Indeed, there are many morphological hints indicating that Spirula could be a useful model organism for approaching the embryonic development of ammonoids. The molecular data seem to support this hypothesis. However, a universal model character of Spirula cannot be detected as, e.g., the mode of feeding probably differs between Spirula and ammonoids.     

Integrated conservation and development project life cycles in the Annapurna Conservation Area, Nepal: Is development overpowering conservation?

The merits of integrated conservation and development projects (ICDPs), which aim to provide development incentives to citizens in return for conservation behaviors, have long been debated in the literature. Some of the most common critiques suggest that conservation activities tend to be strongly overpowered by development activities. We studied this assertion through participant observation and archival analysis of five Conservation Area Management Committees (CAMCs) in the Annapurna Conservation Area (ACA), Nepal. Committee activities were categorized as conservation activities (policy development and conservation implementation), development activities (infrastructure, health care, education, economic development, and sanitation), or activities related to institutional strengthening (administrative development and capacity building activities). Greater longevity of each ICDP was associated with greater conservation activity in relation to development activities. Project life cycles progressed from a focus on development activities in their early stages, through a transitional period of institutional strengthening, and toward a longer-term focus that roughly balanced conservation and development activities. Results suggest that the ICDP concept, as practiced in ACA, has been successful at building capacity for and interest in conservation amongst local communities. However, success has come over a period of nearly a decade, suggesting that prior conclusions about ICDP failures may have been based on unrealistic expectations of the time needed to influence behavioral changes in target populations.     

Technology outlook for real-time quality attribute and process parameter monitoring in biopharmaceutical development—A review

Real-time monitoring of bioprocesses by the integration of analytics at critical unit operations is one of the paramount necessities for quality by design manufacturing and real-time release (RTR) of biopharmaceuticals. A well-defined process analytical technology (PAT) roadmap enables the monitoring of critical process parameters and quality attributes at appropriate unit operations to develop an analytical paradigm that is capable of providing real-time data. We believe a comprehensive PAT roadmap should entail not only integration of analytical tools into the bioprocess but also should address automated-data piping, analysis, aggregation, visualization, and smart utility of data for advanced-data analytics such as machine and deep learning for holistic process understanding. In this review, we discuss a broad spectrum of PAT technologies spanning from vibrational spectroscopy, multivariate data analysis, multiattribute chromatography, mass spectrometry, sensors, and automated-sampling technologies. We also provide insights, based on our experience in clinical and commercial manufacturing, into data automation, data visualization, and smart utility of data for advanced-analytics in PAT. This review is catered for a broad audience, including those new to the field to those well versed in applying these technologies. The article is also intended to give some insight into the strategies we have undertaken to implement PAT tools in biologics process development with the vision of realizing RTR testing in biomanufacturing and to meet regulatory expectations.     

Cost-effective and rapid (3 h) method for combined extraction of DNA,RNA and protein from a single tea leaf sample for genomic and proteomic analysis

This is the first report about the simultaneous extraction of nucleic acids and proteins from tea leaf tissue. Using the present protocol, the DNA, RNA and protein were simultaneously isolated from a single tea leaf sample. The method also maintained the quality and quantity of the isolated biomolecules. The method is cost-effective and takes only 3 h to isolate the starting molecules (DNA, RNA and protein) of central dogma of biology. It was also demonstrated that the isolated DNA, RNA and protein could be successfully used for genomics and proteomic analysis in tea plant which was verified by performing marker study, gene cloning, cDNA preparation, gene expression study and 2-DE.     

The potential of a natural biopolymeric flocculant, ε-poly-l-lysine,for harvesting Chlorella ellipsoidea and its sustainability perspectives for cost and toxicity

Bioprocess and Biosystems Engineering - The successful production of microalgal biomass requires the precise coordination of many different steps. Cell harvesting is a central process in all...     

Farm development at the Arctic cereal limit in northern Norway—continuity and discontinuities

The vegetational development of two neighbouring farm sites at the Arctic cereal limit was investigated by pollen analysis and ¹⁴C-dates. One farm was small and one larger and both had a long habitation history according to archaeological data. Temporary cereal growing at the minor farm may have been introduced about 3100 cal b.p. and was discontinued before 2550 cal b.p. From 1700–1600 cal b.p. Hordeum was cultivated at both farms. At the larger farm there was continuous settlement from c. 2250 cal b.p., but no evidence of cereal growing during the first c. 600 years after the establishment of the farm. It is suggested that changes in the natural conditions, especially paludification, caused a low-activity/abandonment phase at the smaller farm from c. 2550 to c. 1700 cal b.p. A regional abandonment period about 1250 cal b.p. (in the 7th century a.d.) affected the smaller farm in particular.     

Characterization of a rapid and reliable method for iodide biomonitoring in serum and urine based on ion chromatography–ICP-mass spectrometry

An appropriate and controlled supply of thyroid hormones is vital for proper body function. In turn, an appropriate synthesis of T3 and T4 in the thyroid gland is dependent on a sufficient and balanced iodide concentration in blood serum. Due to widespread iodine deficiency or some cases of iodine over exposure, iodide biomonitoring in serum is important and it is that biomonitoring approach being closest to the bioavailable I⁻ supply for the thyroid gland. Therefore, this paper describes a biomonitoring method for iodide determination in serum based on ion chromatography–inductively coupled plasma mass spectrometry (IC–ICP-MS). Since in literature only very few data are available on iodide in serum but many in urine the method is also extended to I⁻ monitoring in urine. The method was additionally designed to have short analysis time (8 min) for increased sample throughput, good precision in serial measurement (serum: 4.86%; urine: 1.4%), and day-to-day determination (serum: 5.7%; urine: 2.28%), high accuracy (serum: 105%; urine: 101%) and good recovery (serum: 102%; urine: 99%) even in matrix-rich samples at low I⁻ concentration. Also, investigations were performed to elucidate whether internal standardization during chromatography, sample preparation for protein-matrix removal or matrix-matched calibration are advantageous for analytical performance. Finally, limits of detection (3σ) of 0.12 μg/L or 0.05 μg/L (serum or urine) and limit of quantification (10σ) of 0.39 μg/L or 0.17 μg/L (serum or urine) were achieved.