Influence of different energy drains on the interrelationship between the rate of respiration,proton-motive force and adenine nucleotide patterns in isolated mitochondria

The respiration of rat liver mitochondria was stimulated by three different ways of energy drain: (a) partial uncoupling (equivalent to direct collapse of the proton-motive force), (b) intramitochondrial utilization of ATP for citrulline synthesis, and (c) extramitochondrial utilization of ATP for glucose phosphorylation. At identical rates of respiration, the intramitochondrial ATP: ADP ratios were the same in all three systems. Furthermore, the proton-motive force was the same in partially uncoupled mitochondria and in the presence of hexokinase plus glucose up to a respiration rate amounting to about 60% of that of the fully active state. However, external ATP: ADP ratios were considerably different in various systems at comparable rates of oxygen uptake, being the lowest under conditions when ATP was being utilized externally. On this basis, it is concluded that the respiratory rate is controlled directly by the proton-motive force and the mitochondrial ATP-synthesizing system operates under near-equilibrium conditions with respect to the membrane energy state parameters. However, a disequilibrium exists at the step of the transport of ATP from mitochondria to the external (cytoplasmic) compartment.     

Homeostasis of the protonmotive force in phosphorylating mitochondria

The relationship between the respiration rate and the magnitude of the electrochemical proton potential (Δμ_H⁺) in rat liver mitochondria was investigated. (1) Under the active-state conditions, the action of inhibitors of either phosphorylation (oligomycin) or respiration (rotenone, malonate) on the respiration and Δμ_H⁺ was measured. Both inhibitors diminished the respiration, whereas rotenone resulted in a decrease of Δμ_H⁺, and oligomycin produced an increase of this potential. The effect of the inhibitors was much more pronounced on the respiration rate than on Δμ_H⁺; for example, the excess of oligomycin produced a 90% inhibition of the respiration while Δμ_H⁺ was changed only by 9%. (2) Under the resting-state conditions, small concentrations of the uncoupler stimulated the respiration while changing Δμ_H⁺ to a relatively small extent. The uncoupler concentrations which doubled and tripled the respiration rate produced only 5 and 9% decrease of Δμ_H⁺, respectively. (3) The present results enabled us to propose a model describing the interrelationship between respiration and Δμ_H⁺.     

Metals in molluscs and associated bottom sediments of the southern baltic

The concentration of Zn, Cu, Pb, Cd, Ni, Co, Ag, Mn, Fe, Ca, Mg, K and Na in molluscsMacoma balthica, Mya arenaria, Cardium glaucum, Mytilus edulis andAstarte borealis from the southern Baltic was determined. The surface sediments and ferromanganese concretions associated with the molluscs were also analysed for concentration of these metals. Species- and regiondependent differences in the metal levels of the organisms were observed. The properties of molluscs analysed which have a tendency toward elevated biological tolerance of selected trace metals were specified. The interelement relationship between metal concentrations in the soft tissue and the shell was estimated and was discussed.     

Distal duplication 14q: Report of three cases and further delineation of the syndrome

Summary Three cases of distal duplication 14q are presented. The first two cases are cousins in a kindred segregating a balanced translocation t(14;18)(q31;q23). The third case resulted from a maternal translocation t(14;18)(q24;p11). By review of these cases and those previously reported, a distal duplication 14q syndrome is further delineated. Common features include postnatal growth retardation, mental retardation, hypotonia, microcephaly, slanted palpebral fissures, ocular hypertelorism, sparse eyelashes and eyebrows, nasal dysmorphism, tented lip, micrognathia, posteriorly rotated ears, and minor skeletal anomalies.     

Platelet derived growth factor BB is a ligand for dermatan sulfate chain(s) of small matrix proteoglycans from normal and fibrosis affected fascia

Structural requirements of the short isoform of platelet derived growth factor BB (PDGF-BB) to bind dermatan sulfate (DS)/chondroitin sulfate (CS) are unknown. Meanwhile the interaction may be important for tissue repair and fibrosis which involve both high activity of PDGF-BB and matrix accumulation of DS. We examined by the solid phase assay the growth factor binding to DS chains of small proteoglycans from various fasciae as well as to standard CSs. Before the assay a structural analysis of DSs and CSs was accomplished involving the evaluation of their epimerization and/or sulfation patterns. In addition, in vivo acceptors for PDGF-BB in fibrosis affected fascia were detected. PDGF-BB binding sites on DSs/CSs are located in long chain sections with the same type of hexuronate isomer however without any apparent preference to glucuronate or iduronate residues. Alternatively, the interaction seems to involve two shorter DS chain sections assembling disaccharides with the same type of hexuronate isomer which are separated by disaccharide(s) with another hexuronate one. Moreover, DS/CS affinity to the growth factor most probably depends on an accumulation of di-2,4-O-sulfated disaccharides in binding site while the presence of 6-O-sulfated N-acetyl-galactosamine residues rather attenuates the binding. All examined fascia DSs and standard CSs showed significant PDGF-BB binding capability with the highest affinity found for normal palmar fascia decorin DS. In fibrosis affected palmar fascia DS/CS proteoglycans are able to form with PDGF-BB supramolecular complexes also including other matrix components such as type III collagen and fibronectin which bind the growth factor covalently. Our results suggest that DS chains of fascia matrix small PGs may regulate PDGF-BB availability leading to restriction of fibrosis associated with Dupuytren's disease or to control of normal fascia repair.     

Red cabbage anthocyanin extract alleviates copper-induced cytological disturbances in plant meristematic tissue and human lymphocytes

Red cabbage is a source of health beneficial substances with antioxidant and antigenotoxic properties. HPLC analysis specifying the content of the investigated extract indicated that mainly anthocyanins (ATH) were responsible for its abilities. Cytological research was conducted with two experimental models: plant tissues—meristematic cells of Vicia faba, and animal tissue elements—human lymphocytes. Positive influence of ATH extract on mitotic activity of Vicia cells exposed to Cu²⁺ stress, and inhibitory effect of ATH on cytotoxic actions of Cu²⁺ on lymphocytes were demonstrated. In all experimental series with ATH application in combinations with Cu²⁺, mitotic index (MI) were higher than those obtained for only Cu²⁺ stressed tissues. Preincubation in ATH before Cu²⁺ stress had the best effect. Similarly, after ATH applications in all tested series decrease in frequency of micronuclei (MN) appearance was noticed in comparison with only Cu²⁺ stressed material. In the case of Vicia cells ATH acted effectively even applied after Cu²⁺ stress. It suggests that this ATH mixture not only prevents and limits but also heals the cytological injury caused by Cu²⁺ stress.     

Rapid high-performance liquid chromatographic method for the determination of ketamine and its metabolite dehydronorketamine in equine serum

A simple, rapid and sensitive high-performance liquid chromatographic procedure has been developed for the determination of ketamine and dehydronorketamine in equine serum. Sample preparation consisted of mixing equal volumes of serum and acetonitrile—phosphoric acid (85%)—water (20:2:78, v/v/v), followed by ultrafiltration through a 10 000 molecular mass cut-off filter. Separation of these two analytes in the ultrafiltrate was accomplished on a reversed-phase phenyl column eluted with methanol—acetonitrile—phosphate buffer solution. Ketamine and dehydronorketamine were detected by a variable photometric UV-Vis detector set at 215 nm, and confirmed by a photodiode array detector operated in the 200–320 nm range. The limit of detection for ketamine was 5–15 ng/ml in equine serum. Additionally, the dehydronorketamine peak identity was tentatively confirmed by thermospray liquid chromatography—mass spectrometry.     

The “tomcat compound” 3-mercapto-3-methylbutanol occurs in the urine of free-ranging leopards but not in African lions or cheetahs

The felid-specific urinary odour compound 3-mercapto-3-methylbutanol and its precursors have been found in several felid species in captivity, but its presence in wild felids has not previously been investigated. We analysed the naturally deposited scent marks from three species of wild, free-ranging big cats in Northern Botswana and found 3-mercapto-3-methylbutanol in four samples of leopard urine (N = 13), but not in lion urine (N = 15) or cheetah urine (N = 6). Individual variation in the presence of the tomcat compound in samples from big cats in the wild may reconcile conflicting results from captive cats.     

Microtubular organization in protoplasts and cells of somatic embryo-regenerating and non-regenerating cultures of Larix

Protoplasts were isolated from both somatic embryo-regenerating and non-regenerating cultures of hybrid larch ( Larix x eurolepis Henry) and fractionated on a discontinuous Percoll density gradient, whereby a highly embryogenic protoplast fraction could be enriched. This fraction was cultured for 14 days, and the differentiating protoplasts, cells, proetmbryos and embryo-like cell clusters sampled at days 3, 5 and 14. Immunofluorescence studies showed that the microtubules became organized into parallel and helical arrays in protoplast-derived cells of the embryogenic tissue as early as day 3 in culture, at which time the protoplast-derived cells started to elongate. In most of the protoplasts from non-regenerating tissue the microtubules retained a random orientation for a longer period. Preprophase bands were observed in both lines. Mitotic spindles consisted mainly of kinetochore-associated microtubules and displayed broad polar regions at metaphase. The spindle poles contracted at anaphase, giving the spindles a pointed appearance. A difference between the two tissue lines was observed at telophase, when the phragmoplast in the non-regenerating tissue had a normal appearance, while a proportion of the phragmoplasts from the embryo-regenerating line were branched or Y-shaped. Y-shaped phragmoplasts resulted in two nucleated cells and one enucleated cell after fusion of the cell plate with the plasma membrane. The early rearrangement of cortical microtubules is an indication that organized growth is occurring but, as this phenomenon has been observed also in regenerating non-emhryogenic cells, it appears to be a doubtful indicator of the distinction between emhryogenic and non-embryogenic development.