Regulation of pathways of glucose metabolism in kidney. The effect of experimental diabetes on the activity of the pentose phosphate pathway and the glucuronate-xylulose pathway.

Measurements have been made of the activities of enzymes of the pentose phosphate pathway, the glucuronate-xylulose pathway, hexokinase and phosphofructokinase in kidney of diabetic and normal rats. The activities of these enzymes keep pace with kidney growth, remaining constant per gram tissue but showing a marked increase on the basis of total activity per 100 g body wt. The formation of ¹⁴CO₂ from [1-¹⁴C]glucose and [6-¹⁴C]glucose by kidney slices from diabetic rats was decreased to approximately half the control value; evidence was obtained for an equivalent dilution of the glucose 6-phosphate pool. Correction of the ¹⁴CO₂ yields for the change in specific activity of glucose 6-phosphate yielded values consistent with the enzyme profile. Calculations from specific yields of ¹⁴CO₂ provided evidence for an increased flux of glucose via the pentose phosphate pathway in the kidney in diabetes. The results are discussed in relation to kidney hypertrophy in diabetes and the requirement for ribose 5-phosphate and NADPH for biosynthetic reactions and in relation to the thickening of the basement membrane in diabetes. These results are in accord with the concept of glucose overutilization by non-insulin-requiring tissues.     

Content of phenolic acids in callus culture of alfalfa (Medicago sativa): The effect of age and biochemical differentiation

Phenolic acids were separated into three fractions and determined by HPLC inMedicago sativa callus culture at the age of two, three and four weeks. The contents of free and especially of predominating ester-bound phenolic acids decreased with callus age to approx. 80 % while the content of phenolic acids nonextractable by methanol increased byca. 90 %. The proportion of benzoic acid derivatives rose from 15 to 21 % within four weeks. The determined difference in the contents of phenolic acids in the upper and lower parts of callus diminished with age. The content of bound forms was higher in the lower part regardless of the callus age. The content of free acids in two weeks old callus was half as high as in the upper part.     

Concentration of phosphoribosyl pyrophosphate in renal hypertrophy. Contrasting effects of early diabetes and unilateral nephrectomy.

Studies were made of the renal phosphoribosyl pyrophosphate (PPRibP) content and PPRibP synthetase (EC 2.7.6.1) activity in rats diabetic for 5, 14 or 20 days, or unilaterally nephrectomized (UN) for 5 days, and in doubly lesioned animals. Approximately equal degrees of renal enlargement were found after 5 days diabetes or 5 days UN. In the doubly lesioned animals the increment of growth was additive. Unilateral nephrectomy of 5 days duration, in contrast with diabetes, had no effect on the PPRibP content of the contralateral kidney, nor did it modify the renal PPRibP content when performed on animals diabetic for 5, 14 or 20 days. The activity of PPRibP synthetase was unaffected by diabetes, UN or diabetes +UN. The results pinpoint a stage of nucleotide synthesis which is differentially affected by the two stimuli, in line with evidence for differences in regulation of nucleic acid turnover in the two conditions.     

The dependence of the frequency of recessive lethal and vital mutations inArabidopsis thaliana (L.)Heynh. on the concentration of N-nitroso-N-methylurea

The dependence of the frequency of recessive lethal (two groups), chlorophyll and morphological mutations on the mutagen concentration was determined in M₂ after subjection to N-nitroso-N-methylurea applied to seeds ofArabidopsis in three concentrations (0·05, 0·10 and 0·20mm). The observed frequencies were compared with the theoretically expected ones for the linear and for two exponential types of dependence, by using the t-test, according to the formulas m=k. C, m=k. C^3/2, m=k. C². No satisfactory agreement with any expected type of dependence was found when directly observed frequencies were used. Since a considerable deficit of mutation frequency was observed in high concentrations, the correction of frequency values was done with respect to the probability of occurence of double mutations. After such a correction, a clear exponential relation was found in both types of lethals and a linear one in chlorophyll and morphological mutations. The probable occurence of multiple mutations should be, therefore, taken into account if the dependence of mutation frequency on the concentration of mutagen is discussed.     

Changes in the radioresistance of bacteria after the inhibition of proteosynthesis in the preirradiation phase

The strain ofEscherichia coli WP₂ (try_v) was irradiated with UV light, at a dosage of 240 erg/mm. Proteosynthesis was inhibited by the elimination of the essential amino acid from the cultivation medium. Changes in radioresistance were followed during 45 minutes of starvation and during the subsequent 45 minutes of restitution after the addition of the essential amino acid. The radioresistance of the cells showed a linear increase immediately after the removal of the essential amino acids, proportional to the duration of the inhibition of proteosynthesis. The increase in radioresistance was shown to be reversible. After the addition of the essential amino acid there was an immediate decrease in radioresistance which was most marked in the first 15 minutes.     

Effect of experimental diabetes on the activity of hexokinase isoenzymes in tissues of the rat

The effect of experimental diabetes on the activity of hexokinase isoenzymes was studied in a wide range of tissues of the rat. In the tissues known to require insulin for glucose phosphorylation, the activity of hexokinase was markedly decreased; the fall being mainly in the Type IV (Glucokinase) in liver and Type II in other tissues, these tissues also exhibit glucose underutilization in diabetes. In the tissues which are commonly known not to require insulin, the activity of Type I hexokinase was significantly increased, these tissues exhibit aspects of glucose overutilization in diabetes in particular kidney and lens. These changes are discussed in relation to Spiro's hypothesis of glucose under and overutilization in tissues in diabetes.     

Effect of Glutathione and N-Acetylcysteine on in Vitro and in VIVO Cardiac Toxicity of Doxorubicin

The effects of two sulfhydryl compounds, glutathione (GSH) and N-acetylcysteine (NAC), on the cardiotoxicity of doxorubicin (DXR) were tested on in vitro and in vivo models. DXR was administered to rats as 4 weekly i.v. doses of 3mg/kg. GSH (1.5 mmoles/kg), given i.v. 10 min before and 1 hr after DXR, was found to prevent the development of the delayed cardiotoxic effects of DXR, as assessed by electrocardiographic and mechanical parameters, as well as by histological examination of left ventricular preparations. In contrast, equimolar oral doses of NAC (1 hr before and 2hrs after DXR) were found to be ineffective. Both GSH and NAC prevented the negative inotropic effect produced by DXR on isolated rat atria. A good correlation exists between the cardioprotective effects of the two agents and their ability to enhance the non-protein sulfhydryl group content of the myocardium. Differences observed in vivo between GSH and NAC might be accounted for by pharmacokinetic factors.     

Study of the reversibility of the water saturation deficit as one of the methods of causal phytogeography

Jedním z kritérií pro posuzování odolnosti rostiin v??i suchu je jejich schopnost sná?et vodní deficit ani? nastanou irreversibilní změny v jejich pletivech. Tato schopnost byla zkoumána metodikou, popsanou v p?edlo?ené práci. Listy některých xerothermních trav (druhy roduStipa, Melica atd.) vysýchaly za p?esně definovaných pokusných podmínek tak, ?e dosáhly r?zně odstupňovaného vodního deficitu. Potom byly roz?ezány na segmenty a ve speciálním ráme?ku dosycovány vodou. P?vodní deficit i jeho vyronání bylo sledvváno váhově. Výsledné hodnoty byly znázorněny graficky. Zatímco ztráta vody z list? během vysýchání probíhala v některých p?ípadech lineárně, k?ivka dosycování ukazovala charakteristický zlom, který indikoval, jak dalece byla ztráta vody nahraditelná, a kdy do?lo ji? k irreversibilním změnám. Ukázalo se, ?e ze studované série rostlin druhy typicky kontinentální mají schopnost doplňovat svou zásobu vody ad integrum i p?i zna?ném vodním deficitu. Rostliny s areálem spí? oceánického charakteru tuto schopnost nemají. Je pravděpodobné, ?e i tato vlastnost bude směrodatná p?i výkladu kausální fytogeografie.