Induction and prevention of chondrocyte hypertrophy in culture

Primary chondrocytes from whole chick embryo sterna can be maintained in suspension culture stabilized with agarose for extended periods of time. In the absence of FBS, the cells remain viable only when seeded at high densities. They do not proliferate at a high rate but they deposit extracellular matrix with fibrils resembling those of authentic embryonic cartilage in their appearance and collagen composition. The cells exhibit many morphological and biochemical characteristics of resting chondrocytes and they do not produce collagen X, a marker for hypertrophic cartilage undergoing endochondral ossification. At low density, cells survive in culture without FBS when the media are conditioned by chondrocytes grown at high density. Thus, resting cartilage cells in agarose cultures can produce factors required for their own viability. Addition of FBS to the culture media leads to profound changes in the phenotype of chondrocytes seeded at low density. Cells form colonies at a high rate and assume properties of hypertrophic cells, including the synthesis of collagen X. They extensively deposit extracellular matrix resembling more closely that of adult rather than embryonic cartilage.     

Development of PCR markers linked to resistance to wheat streak mosaic virus in wheat

Wheat streak mosaic virus (WSMV), vectored by the wheat curl mite (Acer tulipae), is an important disease of wheat (Triticum aestivum L.) in the North American Great Plains. Resistant varieties have not been developed for two primary reasons. First, useful sources of resistance have not been available, and second, field screening for virus resistance is laborious and beyond the scope of most breeding programs. The first problem may have been overcome by the development of resistance to both the mite and the virus by the introgression of resistance genes from wild relatives of wheat. To help address the second problem, we have developed polymerase chain reaction (PCR) markers linked to the WSMV resistance gene Wsm1. Wsm1 is contained on a translocated segment from Agropyron intermedium. One sequence-tagged-site (STS) primer set (WG232) and one RAPD marker were found to be linked to the translocation containing Wsm1. The diagnostic RAPD band was cloned and sequenced to allow the design of specific PCR primers. The PCR primers should be useful for transferring Wsm1 into locally adapted cultivars.This is Journal Series No. J-4041 of the Montana Agricultural Experiment Station     

The herpes simplex virus 1 origin binding protein: a DNA helicase.

A recombinant herpes simplex 1 origin binding protein, the product of the herpes UL9 gene, has been overexpressed in mammalian cells and purified to near homogeneity. The origin binding protein shows DNA-dependent nucleoside 5'-triphosphatase and DNA helicase activities in addition to its origin binding activity. The ability to hydrolyze nucleoside 5'-triphosphates is influenced strongly by the structure and sequence of the DNA cofactor. The properties of the recombinant origin binding protein are identical to those of the protein synthesized in herpes simplex 1-infected mammalian cells.     

Staining of hyaluronan in rat cerebellum with a hyaluronectin-antihyaluronectin immune complex

Summary The presence of hyaluronan was studied histochemically in the adult rat cerebellum. We used the hyaluronectin-antihyaluronectin immune complex technique based on the high affinity of hyaluronectin for hyaluronan. The immune complex was prepared with hyaluronectin from a human brain extract and an anti-hyaluronectin monoclonal antibody, which does not react with rat hyaluronectin. This is a specific probe for detecting hyaluronan in rat tissues without any reaction for tissue hyaluronectin.Hyaluronan was found at the nodes of Ranvier, in the perineuronal microenvironment of the deep nuclei and at the Purkinje cells surrounding the initial segment of the axon. It was located at the same places as hyaluronectin, in areas specialized in ion exchanges and neurotransmission. This suggests that the hyaluronectin-hyaluronan complex could be involved in these processes. The immune complex technique with anti-hyaluronectin monoclonal antibody thus seems to be a specific and valuable tool for investigations of the distribution of hyaluronan in the rat cerebellum.     

Amino acid sequence of the N-terminal non-collagenous segment of dermatosparactic sheep procollagen type I.

The non-collagenous N-terminal segment of type I procollagen from dermatosparactic sheep skin was isolated in the form of the peptide Col 1 from a collagenase digest of the protein. The peptide has a blocked N-terminus, which was identified as pyrrolid-2-one-5-carboxylic acid. Appropriate overlapping fragments were prepared from reduced and alkylated peptide Col 1 by cleavage with trypsin at lysine, arginine and S-aminoethyl-cysteine residues and by cleavage with staphylococcal proteinase at glutamate residues. Amino acid sequence analysis of these fragments by Edman degradation and mass spectrometry established the whole sequence of peptide Col 1 except for a peptide junction (7--8) and a single Asx residue (44), and demonstrated that peptide Col 1 consists of 98 amino acid residues. The N-terminal portion of peptide Col 1 (86 residues) shows an irregular distribution of glycine, whereas the C-terminal portion (12 residues) possesses the triplet structure Gly-Xy and is apparently derived from the precursor-specific collagenous domain of procollagen. The central region of the peptide contains ten cysteine residues located between positions 18 and 73 and shows alternating polar and hydrophobic sequence elements. The regions adjacent to the cysteine-rich portion have a hydrophilic nature and are abundant in glutamic acid. The data are consistent with previous physicochemical and immunological evidence that distinct regions at the N- and C-termini of the non-collagenous domain possess a less rigid conformation than does the central portion of the molecule.     

Sensitivity to reinforcement in successive and delayed discriminations

Pigeons' responses were recorded in successive 15-s subintervals of 60-s components of several multiple variable-interval schedules of food reinforcement. In the standard multiple schedule or successive discrimination, discriminative stimuli were present throughout each component. In the delayed discrimination or memory procedure, red or green stimuli were present in the first 15 s of components and were followed by a white stimulus for the remainder of both components. Ratios of responses in the first 15 s of the two components, where discriminative stimuli were present, were sensitive to ratios of reinforcers obtained in the two components, to the same extent in both multiple and memory procedures. In both procedures, sensitivity to reinforcement decreased systematically over component subintervals, but to a greater extent in the memory procedure where discriminative stimuli were absent. The reduction in sensitivity with time since presentation of prior discriminative stimuli in the memory procedure was therefore influenced by two main factors: delayed stimulus control by the discriminative stimuli presented earlier in the component, and a decrease in sensitivity to reinforcement with increasing time since component alternation.     

Supply versus use designs of environmental extensions in input–output analysis: Conceptual and empirical implications for the case of energy

Input–output analysis is one of the central methodological pillars of industrial ecology. However, the literature that discusses different structures of environmental extensions (EEs), that is, the scope of physical flows and their attribution to sectors in the monetary input–output table (MIOT), remains fragmented. This article investigates the conceptual and empirical implications of applying two different but frequently used designs of EEs, using the case of energy accounting, where one represents energy supply while the other energy use in the economy. We derive both extensions from an official energy supply–use dataset and apply them to the same single‐region input–output (SRIO) model of Austria, thereby isolating the effect that stems from the decision for the extension design. We also crosscheck the SRIO results with energy footprints from the global multi‐regional input–output (GMRIO) dataset EXIOBASE. Our results show that the ranking of footprints of final demand categories (e.g., household and export) is sensitive to the extension design and that product‐level results can vary by several orders of magnitude. The GMRIO‐based comparison further reveals that for a few countries the supply‐extension result can be twice the size of the use‐extension footprint (e.g., Australia and Norway). We propose a graph approach to provide a generalized framework to disclosing the design of EEs. We discuss the conceptual differences between the two extension designs by applying analogies to hybrid life‐cycle assessment and conclude that our findings are relevant for monitoring of energy efficiency and emission reduction targets and corporate footprint accounting.