Phytoplankton primary production and nutrients in the Oosterschelde (The Netherlands) during the pre-barrier period 1980–1984

Phytoplankton primary production, nutrient concentrations and turbidity were monitored at three stations in the Oosterschelde during 1980–1984 as part of an ecosystem study.From comparisons of dissolved nutrient ratios with the nutrient requirements of phytoplankton, and of ambient nutrient concentrations with half-saturation constants for nutrient uptake by natural phytoplankton populations it was concluded that silicate was a limiting nutrient for diatoms after the spring bloom until the end of the summer. Dissolved inorganic nitrogen and phosphate were not considered to be limiting to phytoplankton growth.In general, the phytoplankton growing season started during the first fortnight of April and ended at the end of September. Column production in the whole Oosterschelde varied between 201 and 540 g C m^–2 yr^–1 and was, on average, 25% higher in the western part than in the eastern part. Basin production in the Oosterschelde varied between 120 and 466 g C m^–2 yr^–1 and was, on average, 55% higher in the western part than in the eastern part; this difference could be explained by differences in the ratio of euphotic depth to mean depth of the compartments.Estimated carbon-specific growth rates in the eastern part varied between < 0.1 and 3 d^–1 and in the western part between < 0.1 and 1 d^–1. This difference could be explained by the great differences in depth of the compartments. Carbon-specific growth rates are discussed in relation to phytoplankton loss rates. It is suggested that in the eastern part sedimentation must be an important sink for phytoplankton.Communication no. 473 of the Delta Institute for Hydrobiological Research, Yerseke, The Netherlands.     

Fluorescence investigation of the sex steroid binding protein of rabbit serum: steroid binding and subunit dissociation

The relationship between steroid binding and protein subunit interactions of rabbit sex steroid binding protein (rSBP) has been studied by steady-state and time-resolved fluorescence spectroscopy. The high-affinity (Ka approximately 10(8) M-1 at 4 degrees C), fluorescent estrogen d-1,3,5(10),6,8-estrapentaene-3,17 beta-diol [dihydroequilenin (DHE)] was used as a fluorescent probe of the steroid-binding site. Perturbation of the binding site with guanidinium chloride (Gdm.Cl) was monitored by changes in the steady-state fluorescence anisotropy of DHE as well as by changes in fluorescence quenching of DHE with acrylamide. The results of acrylamide quenching at 11 degrees C show that, while between 0 and 1 M Gdm.Cl the steroid-binding site is completely shielded from bulk solvent, there is decreased DHE binding. To study the subunit-subunit interactions, rSBP was covalently labeled with dansyl chloride in the presence of saturating 5 alpha-dihydrotestosterone (DHT), which yielded a dansyl-conjugated protein that retained full steroid-binding activity. The protein subunit perturbation was monitored by changes in the steady-state fluorescence anisotropy of the dansyl group. At 11 degrees C, the dansyl anisotropy perturbation, reflecting changes in global and segmental motions of the dimer protein, occurs at concentrations of Gdm.Cl above 1 M. The Gdm.Cl titration in the presence of steroids with equilibrium association constants less than 10(8) M-1 shows a plateau near 3 M Gdm.Cl at 11 degrees C; at this Gdm.Cl concentration, no DHE is bound. No plateau is observed at 21 degrees C. At higher Gdm.Cl concentrations, the dansyl fluorescence anisotropy decreases further and shows no steroid dependence. Recovery of steroid-binding activity (assayed by saturation binding with [3H]DHT), under renaturation conditions, is dependent on both steroid concentration and affinity. Both unlabeled and dansyl-labeled protein recovery the same amount of activity, and according to fluorescence anisotropy, dansyl-labeled rSBP re-forms a dimer upon dilution below 1 M or removal of Gdm.Cl. From the steroid requirement for recovery of steroid-binding activity, it appears that a conformational template is required for the dimeric protein to re-form a steroid-binding site with native-like properties.     

Isolation and characterization of active ribosomal subunits from human placenta.

We have developed a method for the large-scale isolation of active ribosomal subunits from human placenta. The technique involves incubating crude ribosomes for 15 min at 37 degrees C with 0.2 mM puromycin in 50 mM Tris-HCl buffer, pH 7.6, 500 mM KCl and 3 mM MgCl2 followed by centrifugation at 5 degrees C in a BXV zonal rotor using an equivolumetric sucrose gradient in the same buffer, upon which 80--90% of all ribosomes are dissociated into subunits. The purified subunits differ in their chemical composition, the 60-S particle containing no more than 36% protein whereas the 40-S subunit consists of 43% protein. In poly(U)-directed protein synthesis, tested in a completely homologous cell-free system, one recombined couple polymerizes at 37 degrees C 12 to 17 phenylalanine residues at an initial rate of 0.7 residues per minute. However, free 80-S ribosomes obtained by puromycin treatment of the crude ribosomes and reassociation of the subunits without prior isolation, have an even higher incorporating activity (20--25 mol phenylalanine/mol of ribosome). At least 55% of the subunits were estimated to actively participate in the polyphenylalanine synthesis.     

Observation and quantitation of metal-binding sites in the sex steroid-binding protein of human and rabbit sera using the luminescent probe terbium

The first direct evidence for specific metal-binding sites in pure human and pure rabbit sex steroid-binding protein (SBP) is obtained using the luminescent lanthanide terbium. Terbium, a probe for calcium sites in proteins, provided protection of the SBP steroid-binding activity in diluted human serum samples equivalent to that provided by calcium. Pure SBP, first treated with ethylenediaminetetraacetate, was dialyzed against buffer containing TbCl₃. After gel filtration to remove nonspecifically bound terbium, the protein was denatured in urea. The amount of protein-bound terbium was determined by luminescence enhancement of the lanthanide using the chelator dipicolinate, yielding four metal-binding sites per mole of dimer protein from both species.     

Efficacy of different types of aerobic exercise in fibromyalgia syndrome: a systematic review and meta-analysis of randomised controlled trials

Introduction  

The efficacy and the optimal type and volume of aerobic exercise (AE) in fibromyalgia syndrome (FMS) are not established. We therefore assessed the efficacy of different types and volumes of AE in FMS.     

Systemic leakage of 15 micron radioactive microspheres over a period of 24 hours, in awake late-pregnant guinea pigs

The present study was designed to determine the reliability of regional blood flow measurements with 15 microns radioactive microspheres, when 24 h have elapsed between reference sampling and sacrifice. The study was performed in 12 chronically-instrumented late-pregnant guinea pigs. The fraction of microspheres recovered from the lungs was consistently higher, by about 2% of the cardiac output after such a 24-h period, as compared to microsphere experiments performed immediately prior to sacrifice. This finding suggests dislodgement of radioactive particles from the systemic circulation between the time of completion of reference sampling and the subsequent 24 h. No correlation could be demonstrated between the change in lung fraction and the change in any organ fraction in particular. Therefore, it is highly unlikely that the microspheres accumulated in the lungs over this period originate from the placenta or any other specific organ. It is concluded that in the awake late-pregnant guinea pig, the 2% loss of microspheres from the systemic circulation during the 24 h following reference sampling, does not invalidate the cardiac output distribution derived.