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The effects of inorganic phosphate (Pi) deficiency on expression of genes encoding ADP-glucose pyrophosphorylase small and large subunits (ApS and ApL1, ApL2, ApL3 genes), UDP-glucose pyrophosphorylase (Ugp gene), sucrose synthase (Sus1), soluble and insoluble acid invertases (Inv and Invcw) and hexokinase (Hxk1 gene), all involved in carbohydrate metabolism, were investigated in Arabidopsis thaliana (L.) Heynh. We used soil-grown pho mutants affected in Pi status, as well as wild-type (wt) plants grown under Pi deficiency conditions in liquid medium, and leaves of wt plants fed with D-mannose. Generally, ApS, ApL1, Ugp and Inv genes were upregulated, although to a varied degree, under conditions of Pi-stress. The applied conditions had differential effects on expression of other genes studied. For instance, Sus1 was down regulated in pho1 (Pi-deficient) mutant, but was unaffected in wt plants grown in liquid medium under P-defi- ciency. Mannose had distinct concentration-dependent effects on expression of genes under study, possibly reflecting a dual role of mannose as a sink for Pi and as glucose analog. Feeding Pi (at up to 200 mM) to the deiached leaves of wt plants strongly affected the expression of ApL1, ApL2, Sus1 and Inv genes, possibly due to an osmotic effect exerted by Pi. The data suggest that ADP-glucose and UDP-glucose pyrophosphorylases (enzymes indirectly involved in Pi recycling) as well as invertases (sucrose hydrolysis) are transcriptionally regulated by Pi-deficiency, which may play a role in homeostatic mechanisms that acclimate the plant to the Pi-stress conditions.
This paper presents a kinetic analysis of the whole reaction course, i.e. of both the transient phase and the steady state, of open multicyclic enzyme cascade systems. Equations for fractional modifications are obtained which are valid for the whole reaction course. The steady state expressions for the fractional modifications were derived from the latter equations since they are not restricted to the condition of rapid equilibrium. Finally, the validity of our results is discussed and tested by numerical integration. Apart from the intrinsic value of knowing the kinetic behaviour of any of the species involved in any open multicyclic enzyme cascade, the kinetic analysis presented here can be the basis of future contributions concerning open multicyclic enzyme cascades which require the knowledge of their time course equations (e.g. evaluation of the time needed to reach the steady state, suggestion of kinetic data analysis, etc.), analogous to those already carried out for open bicyclic cascades.
Content available remote Stearoyl-CoA desaturase - a new player in skeletal muscle metabolism regulation
Stearoyl-CoA desaturase (SCD) is a rate-limiting enzyme catalyzing the synthesis of monounsaturated fatty acids, mainly oleate (18:1) and palmitoleate (16:1), which are a major component of tissue lipids. SCD1 deficient mice reveal increased energy expenditure and decreased body adiposity due to the upregulation of genes of fatty acid oxidation and the downregulation of genes of lipid synthesis in liver. In this review, we examine data showing that SCD is an important component in the regulation of skeletal muscle metabolism, which affects insulin sensitivity, mitochondrial fatty acid oxidation and ceramide de novo synthesis in oxidative myofibers. The lack of SCD1 gene increases the rate of fatty acid ß-oxidation through activation of the AMP-activated protein kinase (AMPK) pathway and by upregulating genes of fatty acid oxidation in soleus and red gastrocnemius muscles. Consistent with increased ß-oxidation, the contents of free fatty acids and long-chain acyl-CoAs are significantly decreased, which together with reduced mRNA level and activity of serine palmitoyltransferase led to reduced ceramide synthesis in oxidative muscles of SCD1-/- mice. Thus, reduced contents of free fatty acids, acyl-CoAs and ceramides as well as increased AMPK phosphorylation, might contribute to increased insulin sensitivity observed in muscle of SCD1-/- mice. SCD1 deficiency also results in downregulation of the expression of the protein-tyrosine phosphatase 1B, which is responsible for the sustained insulin receptor autophosphorylation despite reduced levels of plasma insulin in the SCD1-/- mice. SCD1 deficiency reduced ceramide synthesis, increased AMPK phosphorylation and carnitine palmitoyltransferase 1 activity also in soleus and red gastrocnemius muscles of leptin deficient ob/ob mice. These findings raise the possibility that SCD1 may be a downstream component of the leptin signaling pathway in skeletal muscle.
Male rats of Wistar strain (n = 48) were fed a vitamin A deficient diet for 3 days of adaptation period and then a 10 day experimental period to reduce slightly the body stores of this vitamin. Half of the animals were subjected to physical training and/or oral vitamin A supplementation. Four different doses of supplementation were used – 0, 7.5, 15 and 60 μg/d/rat, which is equivalent to 0, 25, 50 and 200 IU of vitamin A, respectively. Animals from the defined groups ran on a treadmill with a rate of 2.0 m/s for 15 minutes per day for 10 days. After overnight fasting, the rats were sacrificed, and insulin in blood serum and hepatic retinol concentrations were estimated. Daily feed intake and daily body gains were similar in groups of sedentary and physically trained rats. A moderate level of oral vitamin A supplementation (the highest supplemented dose was about 6 x the above recommended NRC level) did not cause any changes in these zootechnical parameters. Oral Vitamin A supplementation resulted in an increase in retinol concentration in the liver (F = 15.2, p < 0.001), but without significant difference between trained and untrained animals. Physical training of rats caused a statistically significant decrease of insulin concentration in blood serum (1.53 ± 0.18 vs. 1.73 ± 0.20). This difference was highly significant (F = 11.1, p < 0.001). Vitamin A supplementation was found not to influence the concentration of this hormone, which is responsible for energy metabolism regulation in the body. Based on estimated parameters, the necessity of vitamin A excessive use in physically trained subjects was not proven.
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