Making metabolism accessible and meaningful: is the definition of a central metabolic dogma within reach?

Intermediary metabolism, a dominant research area before the emergence of molecular biology, is attracting renewed interest for fundamental and applied reasons as documented here. Nonetheless, the field may appear to be a thicket precluding entry to all but the most determined. Here we present a metabolic overview that makes this important and fascinating area accessible to a broad range of the molecular biological and biotechnological communities that are being attracted to biological problems crying out for metabolic solutions. This is accomplished by identifying seven key concepts, a so-called metabolic central dogma, that provide a core understanding analogous to the “Central Dogma of Molecular Biology” which focused upon maintenance and flow of genetic information.     

Optimization of cell culture conditions for production of biologically active proteins

We investigated the basic technology of cell culture conditions for production of useful substances such as cytokines, and related proteins produced by Namalwa cells. Namalwa cells (Klein, 1972), human B lymphoblastoid cells, were used for large scale production of alpha-interferon (Klein, 1979). Namalwa KJM-1, a subline of Namalwa cells, adapted to serum- and albumin-free medium, can grow at a high density above 1 × 10⁷ cells/ml in suspension mode by the use of a perfusion culture system, Biofermenter^?, containing a cone-type cell-sedimentation column as cell separator (Sato, 1983).Several kinds of cytokine cDNA can be introduced and expressed in Namalwa KJM-1 cells (Miyaji, 1990a,b,c). Some of these were produced in large quantities by use of a gene amplification method with dhfr (Miyaji, 1990c), even though the Namalwa KJM-1 cells contained endogenous dhfr genes. For stable production of the target protein, Namalwa KJM-1 cells are very useful host cells, because they have no effective endogenous protease activity in the conditioned medium.Using Biofermenter with micro-silicone fibers and a dialysis system, the specific productivity of the target proteins was not depressed at a high cell density.     

Molecular phylogeny of the family Coccidae (Hemiptera,Coccomorpha), with a discussion of their waxy ovisacs

Coccidae is one of the major families of scale insects, with many species considered to be serious agricultural or horticultural pests. However, the phylogenetic relationships among coccid subfamilies, tribes and genera are poorly understood because existing hypotheses are based on morphological characters and cladistic analyses. Here, we present the first molecular phylogeny of the family Coccidae based on DNA fragments of a mitochondrial gene (COI), nuclear ribosomal RNA genes (18S and 28S), and elongation factor-1α (EF-1α). We found that some genera (Coccus Linnaeus and Pulvinaria Targioni Tozzetti), tribes (Coccini, Paralecaniini, Pulvinariini and Saissetiini) and subfamilies (Coccinae and Filippiinae) within the family are nonmonophyletic. Formation of a waxy ovisac and the distribution and structures of ventral tubular ducts have been used to define the tribe Pulvinariini morphologically; however, these were found to be homoplastic traits based on ancestral state reconstruction. Accordingly, we propose a new classification of certain groups as follows: (i) the Paralecaniini is raised to subfamily rank, Paralecaniinae stat.n. , except that Neosaissetia Tao, Wong & Chang is retained as a member of Coccinae; (ii) Megapulvinaria Yang and Pulvinarisca Borchsenius are transferred from Coccinae to Pulvinariscinae stat.n. ; and (iii) Metaceronema Takahashi is transferred from Filippiinae to Pulvinariscinae stat.n. We provide amended diagnoses for the newly proposed subfamilies.     

Animal Welfare in Relation to Standards in Organic Farming

The new EU-regulations on organic farming (1804/1999) are also influencing the animal welfare. A lot of positive regulations is to find, but also regulations that seen to mind more about the general public and customer and their view on organic farming, than the health and welfare of the animals.The paper specially focus on the impact of the regulations and the recommendations that phytotherapeutic essences and homeopathic products take precedence over the so called chemically-synthesised allopatic veterinary medical products, and that the use of the same is prohibited for preventive treatments. Key questions here are the lack of scientific evidence concerning homeopathy in animals, and that Swedish veterinarians are not allowed to work with homeopathy.Differences in interpretation of the regulations between animal owners and veterinarians will also be discussed. What is a disease that needs treatment? Who is to decide about the treatment? Parasitic infections are discussed as an illustrative example.Other consequences of the regulations concerning the animal welfare are problems in certain geographical zones, for instance subarctic areas where necessary crops are impossible to grow.Animal transports and splitting mother-offspring are briefly discussed as future problems to be handled in the regulations, and the paper ends by presenting the need of regulated herd health control programs in organic husbandry, which can detect and focus on welfare and production problems. The organic movement is not static, and must not be so.     

Reduced Medication in Organic Farming with Emphasis on Organic Dairy Production

Medication is an important focus area in organic animal husbandry. The combination of goals relating to improved animal welfare and reduced use of chemicals in general creates a common wish to reduce medication. Based on data from current Danish research projects in organic dairy farming, one specific organic medication pattern or policy cannot be described. The disease treatment pattern is influenced by many factors, e.g. the interaction with colleagues, veterinarians and agricultural advisors. No significant difference could be found with regard to incidence of mastitis treatments or somatic cell counts in 27 organic and 57 conventional herds. A marked tendency to shorter treatment periods in relation to mastitis treatments was described for organic farms in comparison with conventional farms (1.9 days versus 3.2 days (5 organic and 7 conventional herds)). In a study of development of health advisory service in organic herds, the dialogue between farmer, veterinarian and agricultural cattle advisor changed the treatment pattern markedly during a period of 6 months. Among important future challenges for veterinarians in organic farming is pointed at the constructive, open, and critical interaction with the single organic farmer as well as the organic animal husbandry system in general.     

Enhancement of cell growth by intracellularly expressed herpes simplex virus thymidine kinase

A recombinant cell line (NIH3T3:pLtkSN) was made by infecting parental cells (NIH3T3) with a recombinant retrovirus (pLtkSN) encoding herpes simplex virus thymidine kinase (HSVtk) gene. The cells expressing HSVtk (NIH3T3:pLtkSN) grew 2.3 times more than the parental cells (NIH3T3) in Dulbecco's Modified Eagles Media containing 10% (v/v) horse serum. The NIH3T3:pLtkSN cells also showed a significant enhancement in the maximal cell concentration and the specific growth rate even at 2.5% serum concentration. The specific O2 uptake rate of NIH3T3 was 2.1 times greater than that of NIH3T3:pLtkSN. Under both O2-limited and O2-unlimited conditions, it appears that HSVtk plays an important role in enhancing the growth characteristics of animal cells.