Modulation by pterins of the phosphorylation and phenylalanine activation of phenylalanine 4-mono-oxygenase.

The interaction between phenylalanine 4-mono-oxygenase and analogues of the natural cofactor (6R)-tetrahydrobiopterin [(6R)-BH4] was studied. The rate of cyclic AMP-dependent phosphorylation of phenylalanine 4-mono-oxygenase was inhibited only by those pterins [(6R)-BH4, (6S)-BH4 and 7,8-dihydrobiopterin (BH2)] that were able to decrease the potency and efficiency of phenylalanine as an allosteric activator of the hydroxylase. Since BH2 lacks cofactor activity, this was not required to modulate either the phosphorylation or the phenylalanine-activation of the hydroxylase. Half-maximal inhibition of the phosphorylation was observed at 1.9 microM-(6R)-BH4, 9 microM-(6S)-BH4 and 17 microM-BH2. Competition experiments indicated that all three pterins acted through binding to the cofactor site of the hydroxylase. Since the phosphorylation site and the cofactor binding site are known to reside, respectively, in the N- and C-terminal domains of the hydroxylase, the pterins were able to induce an interdomain conformational change. BH2, whose dihydroxypropyl group is not subject to epimerization, and (6S)-BH4 both inhibited the phosphorylation less efficiently than did the (6R)-epimer of BH4. Pterins with different spatial arrangements of the dihydroxypropyl side chain thus appeared to elicit different conformations of the phosphorylation site. The hydroxylase reaction showed a higher apparent Km for (6S)-BH4 than for (6R)-BH4 both when the native and the phenylalanine-activated enzyme were tested. For the activated enzyme Vmax was 40% lower with the (6S)-epimer than the (6R)-epimer, also when the more rapid enzyme inactivation occurring with the former cofactor was taken into account.     

Is hypothermia necessary for the winter survival of the GoldcrestRegulus regulus?

Summary The Goldcrest is an interesting species for studies of physiological and behavioural adaptions to cold, since it is the smallest species present in Europe. Although many small birds have to lower their metabolic rate and enter hypothermia in order to make their energy reserves last throughout the night, our measurements of oxygen consumption of five Goldcrests suggest that these birds may be able to balance their nocturnal energy budget during the winter at normothermic body temperatures, provided that the available food supply is adequate. These results are in agreement with those of another small bird, the Common Bushtit, which, like the Goldcrest, reduce nocturnal heat loss by contact roosting. Since the energy saved by hypothermia decreases with decreasing ambient temperature and also decreasing body weight, this strategy might not be such an advantage for the smallest birds living at very low ambient temperatures.

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Zusammenfassung Am Wintergoldhähnchen als dem kleinsten europäischen Vogel interessieren besonders seine physiologischen und Verhaltens-Anpassungen an niedere Temperaturen. Viele Kleinvögel senken ihre Stoffwechselaktivität und verfallen in Hypothermie, um mit ihren Energie-Reserven während der langen Winternächte auszukommen. Unsere Untersuchungen über den O₂-Verbrauch von 5 Wintergoldhähnchen legen jedoch nahe, daß sich das nächtliche Energie-Budget auch im Tiefwinter auf Normalniveau bewegt und daß sie bei ausreichendem Nahrungsangebot ihre normale Körpertemperatur auch nachts beibehalten. Ähnliche Befunde liegen von den vergleichbaren kleinen amerikanischen Buschmeisen vor, die wie Goldhähnchen den nächtlichen Wärmeverlust durch Schlafen auf engem Gefiederkontakt reduzieren. Es ist anzunehmen, daß die Hypothermie als Überwinterungsstrategie für unsere kleinsten Vögel garnicht so vorteilhaft wäre: Die durch Hypothermie eingesparte Energie geht mit abnehmender Umgebungstemperatur und mit geringerer Körpergröße wieder verloren.

     

Lymphatic absorption and transport of retinol and vitamin D-3 from rat intestine Evidence for different pathways

The lymphatic absorption and transport of retinol and vitamin D-3 from rat intestine has been studied. When rats were cannulated in the intestinal lymph duct and given an intraduodenal bolus of [³H]retinol and ¹⁴C-labelled vitamin D-3, ¹⁴C-labeled vitamin D-3 appeared later in the intestinal lymph than [³H]retinol and the rate of absorption of vitamin D-3 was still maximal at a time when that of retinol had declined. Both vitamins were absorbed via the lymphatic route in association with chylomicrons. Almost all the retinol was esterified, while vitamin D-3 appeared in the chylomicrons as free vitamin D-3. In vitro incubations and in vivo studies using hepatectomized and normal rats showed that the retinyl ester was a relatively nonexchangeable component of the chylomicrons and their remnants. Hence, all the vitamin A followed the remnants in their clearance from plasma. In contrast, significant amounts of vitamin D-3 were transferred from the chylomicrons to other plasma fractions. Therefore, only a fraction of this vitamin may be removed in association with the chylomicron remnants.     

Discrimination between Weaned and Unweaned Atlantic Cod (Gadus morhua) in Capture-Based Aquaculture (CBA) by X-Ray Imaging and Radio-Frequency Metal Detector

The aim of this study was to investigate the feasibility of two detection methods for use in discrimination and sorting of adult Atlantic cod (about 2 kg) in the small scale capture-based aquaculture (CBA). Presently, there is no established method for discrimination of weaned and unweaned cod in CBA. Generally, 60–70% of the wild-caught cod in the CBA are weaned into commercial dry feed. To increase profitability for the fish farmers, unweaned cod must be separated from the stock, meaning the fish must be sorted into two groups – unweaned and weaned from moist feed. The challenges with handling of large numbers of fish in cages, defined the limits of the applied technology. As a result, a working model was established, focusing on implementing different marking materials added to the fish feed, and different technology for detecting the feed presence in the fish gut. X-ray imaging in two modes (planar and dual energy band) and sensitive radio-frequency metal detection were the detection methods that were chosen for the investigations. Both methods were tested in laboratory conditions using dead fish with marked feed inserted into the gut cavity. In particular, the sensitive radio-frequency metal detection method with carbonyl powder showed very promising results in detection of marked feed. Results show also that Dual energy band X-ray imaging may have potential for prediction of fat content in the feed. Based on the investigations it can be concluded that both X-ray imaging and sensitive radio-frequency metal detector technology have the potential for detecting cod having consumed marked feed. These are all technologies that may be adapted to large scale handling of fish from fish cages. Thus, it may be possible to discriminate between unweaned and weaned cod in a large scale grading situation. Based on the results of this study, a suggestion for evaluation of concept for in-situ sorting system is presented.     

Oxidative damage to purines in DNA: role of mammalian Ogg1

DNA damage caused by reactive oxygen species is ubiquitous to all living organisms. More than 60 different base lesions have been identified, and the majority of these are removed via the base excision repair pathway. This pathway appears to represent a highly conserved and ancient mechanism of defence counteracting spontaneous DNA decay. In this review, we describe in more detail the Ogg1 enzyme and its conserved action of removing the oxidised base, 7,8-dihydro-8-oxoguanine (oxo(8)G). Recent updates include the cancer-prone ogg1/myh double knockout mouse and an elegant study which looks at the ability of hOgg1 to distinguish between the mutagenic lesion, oxo(8)G, and the vast majority of normal bases.     

Lumican is a major small leucine-rich proteoglycan (SLRP) in Atlantic cod (Gadus morhua L.) skeletal muscle

Knowledge on fish matrix biology is important to ensure optimal fish -quality, -growth and -health in aquaculture. The aquaculture industry face major challenges related to matrix biology, such as inflammations and malformations. Atlantic cod skeletal muscle was investigated for collagen I, decorin, biglycan, and lumican expression and distribution by real-time PCR, immunohistochemical staining and Western blotting. Immunohistochemical staining and Western immunoblotting were also performed using antibodies against glycosaminoglycan side chains of these proteoglycans, in addition to fibromodulin. Real-time PCR showed highest mRNA expression of lumican and collagen I. Collagen I and proteoglycan immunohistochemical staining revealed distinct thread-like structures in the myocommata, with the exception of fibromodulin, which stained in dense structures embedded in the myocommata. Chondroitinase AC-generated epitopes stained more limited than cABC-generated epitopes, indicating a stronger presence of dermatan sulfate than chondroitin sulfate in cod muscle. Lumican and keratan sulfate distribution patterns were strong and ubiquitous in endomysia and myocommata. Western blots revealed similar SLRPs sizes in cod as are known from mammals. Staining of chondroitin/dermatan sulfate epitopes in Western blots were similar in molecular size to those of decorin and biglycan, whereas staining of keratan sulfate epitopes coincided with expected molecular sizes of lumican and fibromodulin. In conclusion, lumican was a major proteoglycan in cod muscle with ubiquitous distribution overlapping with keratan sulfate. Other leucine-rich proteoglycans were also present in cod muscle, and Western blot using antibodies developed for mammalian species showed cross reactivity with fish, demonstrating similar structures and molecular weights as in mammals.     

What Stakeholders Think About Marine Protected Areas: Case Studies from Spain

Marine protected areas (MPAs) are often met with reluctance by affected stakeholders, and in some instances outright objection. Some argue that this is due to insufficient understanding of the functions of MPAs. Others suggest that it could be because of a perception that they are losing more than they are gaining. It is also possible that stakeholders are generally supportive of the idea but think that the MPA should be located elsewhere. We argue that it is images people have about what the MPA is and does that determine how they react. Drawing from three MPA case studies in Spain, we illustrate the importance of critical examination of stakeholders’ images and what they imply for the governance of MPAs.     

An assessment of swinger techniques for the playground swing oscillatory motion

Much attention has been devoted to how playground swing amplitudes are built up by swinger techniques, i.e. body actions. However, very little attention has been given to the requirements that such swinger techniques place on the swinger himself. The purpose of this study was to find out whether different swinger techniques yield significantly different maximum torques, endurance and coordinative skills, and also to identify preferable techniques. We modelled the seated swinger as a rigid dumbbell and compared three different techniques. A series of computer simulations were run with each technique, testing the performance with different body rotational speeds, delayed onset of body rotation and different body mass distributions, as swing amplitudes were brought up towards 90°. One technique was found to be extremely sensitive to the timing of body actions, limiting swing amplitudes to 50° and 8° when body action was delayed by 0.03 and 0.3?s, respectively. Two other more robust techniques reached 90° even with the largest of these delays, although more time (and endurance) was needed. However, these two methods also differed with respect to maximum torque and endurance, and none was preferable in both these aspects, being dependent on the swinger goals and abilities.