Flow cytometric monitoring of influenza A virus infection in MDCK cells during vaccine production

Background  

In cell culture-based influenza vaccine production the monitoring of virus titres and cell physiology during infection is of great importance for process characterisation and optimisation. While conventional virus quantification methods give only virus titres in the culture broth, data obtained by fluorescence labelling of intracellular virus proteins provide additional information on infection dynamics. Flow cytometry represents a valuable tool to investigate the influences of cultivation conditions and process variations on virus replication and virus yields.     

Environmental assessment of cod (<Emphasis Type="BoldItalic">Gadus morhua</Emphasis>) from autoline fisheries

Purpose  

The main purpose of this study has been to document the environmental performance of products based on autoline-caught cod and the distribution of environmental impacts in the value chain from fishing to retail. Another aim has been to document the performed environmental improvement analyses.     

Effect of different allocation methods on LCA results of products from wild-caught fish and on the use of such results

Purpose  

The purpose of this study has been to investigate the effect of different allocation methods on life cycle assessment (LCA) results of products derived from line-caught cod and the consequences of applying these methods considering the main aims of this case study. These aims were for internal improvement work and communication of results to the market.     

Two storage hexamerins from the beet armyworm <Emphasis Type="Italic">Spodoptera exigua</Emphasis>: Cloning,characterization and the effect of gene silencing on survival

Background  

In insects, hemocyanin superfamily proteins accumulate apparently to serve as sources of amino acids during metamorphosis, reproduction and development. Storage hexamerins are important members of the hemocyanin superfamily. Although insects possess storage hexamerins, very little is known about the character and specific functions of hexamerin 1 and storage protein 1 in insect development.     

Genomic signatures of local directional selection in a high gene flow marine organism; the Atlantic cod (Gadus morhua)

Background  

Marine fishes have been shown to display low levels of genetic structuring and associated high levels of gene flow, suggesting shallow evolutionary trajectories and, possibly, limited or lacking adaptive divergence among local populations. We investigated variation in 98 gene-associated single nucleotide polymorphisms (SNPs) for evidence of selection in local populations of Atlantic cod (Gadus morhua L.) across the species distribution.     

Mathematical modeling of the dynamic storage of iron in ferritin

Background  

Iron is essential for the maintenance of basic cellular processes. In the regulation of its cellular levels, ferritin acts as the main intracellular iron storage protein. In this work we present a mathematical model for the dynamics of iron storage in ferritin during the process of intestinal iron absorption. A set of differential equations were established considering kinetic expressions for the main reactions and mass balances for ferritin, iron and a discrete population of ferritin species defined by their respective iron content.     

Single cell oils of the cold-adapted oleaginous yeast <Emphasis Type="Italic">Rhodotorula glacialis</Emphasis> DBVPG 4785

Background  

The production of microbial lipids has attracted considerable interest during the past decade since they can be successfully used to produce biodiesel by catalyzed transesterification with short chain alcohols. Certain yeast species, including several psychrophilic isolates, are oleaginous and accumulate lipids from 20 to 70% of biomass under appropriate cultivation conditions. Among them, Rhodotorula glacialis is a psychrophilic basidiomycetous species capable to accumulate intracellular lipids.     

Life cycle assessment of the environmental performance of conventional and organic methods of open field pepper cultivation system

Purpose

As the scale of the organic cultivation sector keeps increasing, there is growing demand for reliable data on organic agriculture and its effect on the environment. Conventional agriculture uses chemical fertilizers and pesticides, whilst organic cultivation mainly relies on crop rotation and organic fertilizers. The aim of this work is to quantify and compare the environmental sustainability of typical conventional and organic pepper cultivation systems.

Methods

Two open field pepper cultivations, both located in the Anthemountas basin, Northern Greece, are selected as case studies. Life cycle assessment (LCA) is used to quantify the overall environmental footprint and identify particular environmental weaknesses (i.e. unsustainable practices) of each cultivation system. Results are analysed at both midpoint and endpoint levels in order to obtain a comprehensive overview of the environmental sustainability of each system. Attributional LCA (ALCA) is employed to identify emissions associated with the life cycles of the two systems. Results are presented for problem-oriented (midpoint) and damage-oriented (endpoint) approaches, using ReCiPe impact assessment.

Results and discussion

At midpoint level, conventional cultivation exhibits about threefold higher environmental impact on freshwater eutrophication, than organic cultivation. This arises from the extensive use of nitrogen and phosphorus-based fertilizers, with consequent direct emissions to the environment. The remaining impact categories are mainly affected by irrigation, with associated indirect emissions linked to electricity production. At endpoint level, the main hotspots identified for conventional cultivation are irrigation and fertilizing, due to intensive use of chemical fertilizers and (to a lesser degree) pesticides. For organic pepper cultivation, the main environmental hotspots are irrigation, machinery use, and manure loading and spreading processes. Of these, the highest score for irrigation derives from the heavy electricity consumption required for groundwater pumping associated with the fossil-fuel-dependent Greek electricity mix.

Conclusions

Organic and conventional cultivation systems have similar total environmental impacts per unit of product, with organic cultivation achieving lower environmental impacts in ‘freshwater eutrophication’, ‘climate change’, ‘terrestrial acidification’ and ‘marine eutrophication’ categories. Conventional cultivation has a significantly greater effect on the freshwater eutrophication impact category, due to phosphate emissions arising from application of chemical fertilizers.

     

Environmental assessment of black locust (<Emphasis Type="BoldItalic">Robinia pseudoacacia</Emphasis> L.)-based ethanol as potential transport fuel

Purpose  

Lignocellulosic ethanol has received special research interest, driven by concerns over high fuel prices, security of energy supplies, global climate change as well as the search of opportunities for rural economic development. A well-to-wheel analysis was conducted for ethanol obtained from black locust (Robinia pseudoacacia L.) by means of the life cycle assessment (LCA) methodology. This study assesses the environmental profile of using ethanol in mixtures E10 and E85 as transport fuel in comparison with conventional gasoline (CG). In addition, the best model of black locust cultivation was analysed under an environmental point of view.     

Lysis of tubercle bacilli in fresh and stored sputum specimens: implications for diagnosing tuberculosis in stored and paucibacillary specimens by PCR

Background  

Nucleic acid amplification techniques are being used increasingly in diagnosing tuberculosis. In developing countries clinical samples are often stored for subsequent analysis since molecular tests are conducted at only a limited number of laboratories. This study was conducted to assess the speed at which mycobacteria undergo autolysis and free DNA is detected in the supernatant during low-temperature storage.     

Long-term maintenance of in vitro cultured honeybee (Apis mellifera) embryonic cells

Background  

In vitro cultivation of cells allows novel investigation of in vivo- mechanisms and is a helpful tool in developmental biology, biochemistry and functional genomics. Numerous cell lines of insect species, e.g., silkworm and mosquito, have been reported. However, this is not the case for successful long-term cultivation of cells in honeybees.     

A fed‐batch based cultivation mode in Escherichia coli results in improved specific activity of a novel chimeric‐truncated form of tissue plasminogen activator

Aims

A novel chimeric‐truncated form of tissue‐type plasminogen activator (t‐PA) with improved fibrin affinity and resistance to PAI was successfully produced in CHO expression system during our previous studies. Considering advantages of prokaryotic expression systems, the aim in this study was to produce the novel protein in Escherichia coli (BL21) strain and compare the protein potency in batch and fed‐batch processes.

Methods and Results

The expression cassette for the novel t‐PA was prepared in pET‐28a(+). The E. coli expression procedure was compared in traditional batch and newly developed fed batch, EnBase^® Flo system. The protein was purified in soluble format, and potency results were identified using Chromolize t‐PA Assay Kit. The fed‐batch fermentation mode, coupled with a Ni‐NTA affinity purification procedure under native condition, resulted in higher amounts of soluble protein, and about a 30% of improvement in the specific activity of the resulted recombinant protein (46·66 IU mg^?1) compared to traditional batch mode (35·8 IU mg^?1).

Conclusions

Considering the undeniable advantages of expression in the prokaryotic expression systems such as E. coli for recombinant protein production, applying alternative methods of cultivation is a promising approach. In this study, fed‐batch cultivation methods showed the potential to replace miss‐folded formats of protein with proper folded, soluble form with improved potency.

Significance and Impact of the Study

Escherichia coli expression of recombinant proteins still counts for nearly 40% of marketed biopharmaceuticals. The major drawback of this system is the lack of appropriate post‐translational modifications, which may cause potency loss/decline. Therefore, applying alternative methods of cultivation as investigated here is a promising approach to overcome potency decrease problem in this protein production system.     

Lighting during grow-out and <Emphasis Type="Italic">Salmonella</Emphasis> in broiler flocks

Background  

Lighting is used during conventional broiler grow-out to modify bird behaviour to reach the goals of production and improve bird welfare. The protocols for lighting intensity vary. In a field study, we evaluated if the lighting practices impact the burden of Salmonella in broiler flocks.     

The sustainability of communicative packaging concepts in the food supply chain. A case study: part 2. Life cycle costing and sustainability assessment

Purpose  

This paper is the second part of a two-paper series dealing with the sustainability evaluation of a new communicative packaging concept. The communicative packaging concept includes a device that allows changing the expiry date of the product as function of temperature during transport and storage: a flexible best-before-date (FBBD). Such device was analysed in a consumer unit consisting of a nanoclay-based polylactic acid tray filled with pork chops.     

A mitogenomic approach to the taxonomy of pollocks: <Emphasis Type="Italic">Theragra chalcogramma</Emphasis> and <Emphasis Type="Italic">T. finnmarchica</Emphasis> represent one single species

Background  

The walleye pollock (Theragra chalcogramma) and Norwegian pollock (T. finnmarchica) are confined to the North Pacific and North Atlantic Oceans, respectively, and considered as distinct species within the family Gadidae. We have determined the complete mtDNA nucleotide sequence of two specimens of Norwegian pollock and compared the sequences to that of 10 specimens of walleye pollock representing stocks from the Sea of Japan and the Bering Sea, 2 specimens of Atlantic cod (Gadus morhua), and 2 specimens of haddock (Melanogrammus aeglefinus).     

Utilizing high-throughput experimentation to enhance specific productivity of an <Emphasis Type="Italic">E.coli</Emphasis>T7 expression system by phosphate limitation

Background  

The specific productivity of cultivation processes can be optimized, amongst others, by using genetic engineering of strains, choice of suitable host/vector systems or process optimization (e.g. choosing the right induction time). A further possibility is to reduce biomass buildup in favor of an enhanced product formation, e.g. by limiting secondary substrates in the medium, such as phosphate. However, with conventional techniques (e.g. small scale cultivations in shake flasks), it is very tedious to establish optimal conditions for cell growth and protein expression, as the start of protein expression (induction time) and the degree of phosphate limitation have to be determined in numerous concerted, manually conducted experiments.     

Comparing nutritional value and yield as functional units in the environmental assessment of horticultural production with organic or mineral fertilization

Background, aim, and scope  

We report the environmental assessment of the cultivation cycle of cauliflower (Brassica oleracea L. var. botrytis), chosen due to its high levels of natural bioactive compounds, using different fertilization practices. The functional units used during the impact assessment were linked with the quantity produced, considering different units of commercialization, or with the cauliflower quality, considering its antioxidant compounds content. Although nutrient content has been described and used as a possible functional unit, using antioxidant compounds as a functional unit has not previously been published.     

Genomic organization and gene expression of the multiple globins in Atlantic cod: conservation of globin-flanking genes in chordates infers the origin of the vertebrate globin clusters

Background  

The vertebrate globin genes encoding the α- and β-subunits of the tetrameric hemoglobins are clustered at two unlinked loci. The highly conserved linear order of the genes flanking the hemoglobins provides a strong anchor for inferring common ancestry of the globin clusters. In fish, the number of α-β-linked globin genes varies considerably between different sublineages and seems to be related to prevailing physico-chemical conditions. Draft sequences of the Atlantic cod genome enabled us to determine the genomic organization of the globin repertoire in this marine species that copes with fluctuating environments of the temperate and Arctic regions.