Gibberellin like Substances in Parts of Developing Barley Grain

In husks, the activity of gibberellin-like substances extracted with aqueous methanol (M-“free” GAs) showed a maximum on the 9th day after pollination. In developing embryos, M-“free” GAs showed no biological activity, whereas biological active component(s) were obtained when the embryos were extracted with Tris buffer. The “free” GAs found in the buffer homogenates (B-“free” GAs) of developing embryos showed a maximum of activity on the 33rd day after pollination. Bound GAs recovered from the precipitated protein of the buffer homogenate (“Protein-bound” GAs) were found in embryo and endosperm. Developing endosperm generally contains the major amount of the extractable gibberellin-like substances. In this tissue, the amount of all examined fractions (M-“free” GAs, B-“free” GAs and “protein-bound” GAs) increased after pollination to reach a maximum on the 21st day, before decreasing to a minimum at grain maturity. Moreover, the curves for dry weight increase and gibberellin like substances follow a remarkably similar course, with the latter reaching its maximim slightly earlier than the former one. This result indicates that gibberellines may participate in the regulation of the accumulation process in the endosperm of barley grain.     

A primary phosphorus‐deficient skeletal phenotype in juvenile Atlantic salmon Salmo salar: the uncoupling of bone formation and mineralization

To understand the effect of low dietary phosphorus (P) intake on the vertebral column of Atlantic salmon Salmo salar, a primary P deficiency was induced in post‐smolts. The dietary P provision was reduced by 50% for a period of 10 weeks under controlled conditions. The animal's skeleton was subsequently analysed by radiology, histological examination, histochemical detection of minerals in bones and scales and chemical mineral analysis. This is the first account of how a primary P deficiency affects the skeleton in S. salar at the cellular and at the micro‐anatomical level. Animals that received the P‐deficient diet displayed known signs of P deficiency including reduced growth and soft, pliable opercula. Bone and scale mineral content decreased by c. 50%. On radiographs, vertebral bodies appear small, undersized and with enlarged intervertebral spaces. Contrary to the X‐ray‐based diagnosis, the histological examination revealed that vertebral bodies had a regular size and regular internal bone structures; intervertebral spaces were not enlarged. Bone matrix formation was continuous and uninterrupted, albeit without traces of mineralization. Likewise, scale growth continues with regular annuli formation, but new scale matrix remains without minerals. The 10 week long experiment generated a homogeneous osteomalacia of vertebral bodies without apparent induction of skeletal malformations. The experiment shows that bone formation and bone mineralization are, to a large degree, independent processes in the fish examined. Therefore, a deficit in mineralization must not be the only cause of the alterations of the vertebral bone structure observed in farmed S. salar. It is discussed how the observed uncoupling of bone formation and mineralization helps to better diagnose, understand and prevent P deficiency‐related malformations in farmed S. salar.     

Protein glycosylation mutants of procyclic Trypanosoma brucei: defects in the asparagine-glycosylation pathway

We employed a genetic approach to study protein glycosylation in the procyclic form of the parasite Trypanosoma brucei. Two different mutant parasites, ConA 1-1 and ConA 4-1, were isolated from mutagenized cultures by selecting cells which resisted killing or agglutination by concanavalin A. Both mutant cells show reduced concanavalin A binding. However, the mutants have different phenotypes, as indicated by the fact that ConA 1-1 binds to wheat germ agglutinin but ConA 4-1 and wild type do not. A blot probed with concanavalin A revealed that many proteins in both mutants lost the ability to bind this lectin, and the blots resembled one of wild type membrane proteins treated with PNGase F. This finding suggested that the mutants had altered asparagine- linked glycosylation. This conclusion was confirmed by studies on a flagellar protein (Fla1) and procyclic acidic repetitive protein (PARP). Structural analysis indicated that the N- glycan of wild type PARP is exclusively Man5GlcNAc2 whereas that in both mutants is predominantly a hybrid type with a terminal N- acetyllactosamine. The occupancy of the PARP glycosylation site in ConA 4-1 was much lower than that in ConA 1-1. These mutants will be useful for studying trypanosome glycosylation mechanisms and function.      

Intravenous chloralose is a safe anesthetic for longitudinal use in beagle puppies

Chloralose is an intravenous anesthetic which preserves vagal and central baroreceptor reflexes, thus rendering it useful for physiologic research. However, chloralose is recommended for terminal experiments only, due to concerns relating to long-term toxicity. We investigated the safety of chloralose in longitudinal pulmonary function studies in beagle puppies. Twelve puppies received chloralose anesthesia repeatedly (8-12 times per dog) between the ages of 80 and 300 days. Constant anesthetic depth was maintained reliably throughout the course of the experiments. Recovery lasted approximately 4 hours in each experiment and occurred in four definable stages. Following recovery, the puppies exhibited normal health and growth as compared with other colony animals. There was no biochemical evidence of acute renal, hepatic, pancreatic or cardiac toxicity prior to and immediately after anesthesia, and no evidence of chronic toxicity following completion of the study protocol, after a total cumulative dose of 1.18 g/kg chloralose. These studies demonstrate that intravenous chloralose is a safe anesthetic for longitudinal use.     

Studies on the toxicity and binding kinetics of abrin in normal and Epstein-Barr virus-transformed lymphocyte culture-I. Experimental results - 4

The effects of treatment with varying doses of abrin, a D-galactose binding lectin, on DNA and protein synthesis of normal and Epstein-Barr virus (EBV)-transformed lymphocytes have been previously investigated. Using data on EBV-transformed lymphocyte cell density as a function of both time and dose of abrin, the authors introduced the concept of self- and cross-coupling metabolic variables as a means of understanding how abrin affected DNA and protein uptake. In this paper, the self-coupling constant is studied in more detail and the relationship between DNA and protein synthesis is further expanded. We find that there is a significant linear relationship between DNA and protein synthesis in normal lymphocyte culture as measured by abrin interaction in the culture. We further find that there is a much stronger relationship between these variables in EBV-transformed lymphocyte culture. This relationship is further examined, and possible analytic equations are expressed.     

The distribution of fibronectin in developing zebrafish (Danio rerio) cartilage

The extracellular matrix (ECM) plays a complex and vital role throughout the process of cartilage formation. Fibronectin is a large ECM glycoprotein with an important role in various developmental processes, including skeletogenesis. Taking advantage of the known sequence of cartilage development in zebrafish and using an immunohistochemical stain for collagen type II to identify differentiation phase cartilage, we evaluate the distribution of fibronectin in various cartilaginous elements of the zebrafish (elements of the splanchnocranium, and of the dorsal, caudal, pelvic and pectoral fins). Contrary to what is observed in tetrapods, our data on zebrafish indicate the apparent lack of fibronectin during the condensation phase of cartilage development. This lack is possibly linked to the high developmental rate of the zebrafish and the small size of the condensations, which brings different needs for the extracellular environment to ensure cell survival. Furthermore, the fin disk cartilage of the pectoral fin develops an ECM with a strong fibronectin signal, whereas other cartilage elements show only a weak fibronectin signal in early differentiation, which gradually disappears. Thus, the pectoral fin disk cartilage is unique not only because of its specific way of development (subdivision of a continuous plate into four elements, the proximal radials), but also because of its strong fibronectin‐positive ECM.     

Mechanistic model of fuel selection in the muscle

Fuel selection in human muscle is key to explaining insulin resistance. In obesity and type 2 diabetes mellitus, there is an increased content of lipid within and around muscle fibers. Changes in muscle fuel partitioning of lipid, between oxidation and storage of fat, contribute to the accumulation of intramuscular triglycerides and to the pathogenesis of both obesity and type 2 diabetes mellitus. A mathematical model of the aggregated metabolism in skeletal muscle was developed and the effects of fuel selection for lean and obese individuals under fasting conditions, insulin-stimulated conditions, and oscillating insulin conditions were examined. Model results were complementary to prior observations that elevated lipid oxidation during insulin-stimulated conditions is correlated with insulin resistance. The model also adequately simulated metabolic inflexibility between fat and glucose oxidation in the obese individual. A novel sensitivity analysis indicated the strong interaction effects of parameters of glucose and lipid oxidation pathways on the variables of each pathway.