Simultaneous determination of ascorbic acid and dehydroascorbic acid in cultures of C3H/10T1/2 cells

Summary A reproducible method is described for the separation and quantification of ascorbic acid and dehydroascorbic acid by ion-pairing reverse-phase high performance liquid chromatography and detection by absorbance at 232 nm. Lowest detectable concentrations with a linear response of detection were 5 nmol for ascorbic acid and 50 nmol for dehydroascorbic acid. This method was applied to the analysis of C3H/10T1/2 cells and culture medium after influx or efflux experiments and single or multiple treatments with ascorbic acid. Subsequent measurement of the radioactivity in the eluted fractions increased the detectability of both ascorbic acid and dehydroascorbic acid to 10 to 20 pmol. This research was supported by grant CA 09320 and CA 31574 from the National Cancer Institute, Bethesda, MD, and grant BC441 from The American Cancer Society.     

A replication study of human exposure to 60-Hz fields: effects on neurobehavioral measures.

The purpose of this study was to reproduce and extend an earlier investigation of the effects of human exposure to combined, 60-Hz electric and magnetic fields. This paper presents the neurobehavioral results. Thirty men participated in one training session and four testing sessions. Subjects were randomly assigned to two groups. The 18 subjects in Group I were exposed (9 kV/m, 20 microT) and sham exposed in two counterbalanced orders. In Group II, half of 12 subjects were exposed (9 kV/m, 20 microT) every session, and the remaining half were sham exposed every session. The study was doubly blinded. Measures of cardiac interbeat interval, event-related brain potentials, and performance were obtained before, during, and after exposures. As in the earlier study, exposure to the combined field resulted in a statistically significant slowing of heart rate, in changes in late components of event-related brain potentials, and in decreased errors on a choice reaction-time task. In addition, field effects on several other measures approached statistical significance. The physiological measures obtained during exposure indicated that effects were greatest soon after the field was switched on, and again when it was switched off. The data indicate that changes in exposure level may be more important than duration of exposure for producing effects in human beings.     

Comparison of theN-glycoloylneuraminic andN-acetylneuraminic acid content of platelets and their precursors using high performance anion exchange chromatography

N-Acetylneuraminic acid (Neu5Ac) andN-glycoloylneuraminic acid (Neu5Gc) are distributed widely in nature. Using a Carbopac PA-1 anion exchange column, we have determined the ratios of Neu5Ac and Neu5Gc in hydrolysates of platelets and their precursors: a rat promegakaryoblastic (RPM) cell line and a human megakaryoblastic leukemia cell line (MEG-01). The ratio of Neu5Gc:Neu5Ac in cultured RPM cells is 16:1, whereas in platelet rich plasma and cultured MEG-01 cells it is 1:38 and 1:28, respectively. The nature of these sialic acids from RPM cells was verified using thin layer chromatography and liquid secondary ion mass spectrometry. The relevance of increased Neu5Gc levels in early stages of development is discussed.Abbreviations Neu5Ac N-acetylneuraminic acid - Neu5Gc N-glycoloylneuraminic acid - RPM rat promegakaryoblast - MEG-01 human megakaryoblastic leukaemia cell line - PAD pulsed amperometric detection - WGA wheat germ agglutinin - FCS foetal calf serum - PPEADF phosphatidylethanolamine dipalmitoyl - LSIMS liquid secondary ion mass spectrometry - HPAEC high performance anion exchange chromatography - TBA thiobarbituric acid     

Increased working capacity with hyperoxia in humans

The purpose of the study was to examine the influence of oxygen-breathing on maximal oxygen uptake (VO2max) and submaximal endurance performance. Six young women and five men rode a cycle-ergometer while breathing compressed air (normoxia, NOX) or a 55% O2 in N2 mixture (hyperoxia, HOX). The VO2max increased significantly by 12% (P less than 0.01) with HOX in the women but not in the men (+4%; nonsignificant). Maximal heart rate was also increased with HOX in the women but not in the men. Endurance time during work to exhaustion at 80% of normoxic VO2max was 41% longer in HOX than in NOX (P less than 0.025) with no significant difference between the men and the women. The variation among individuals was large. The oxygen uptake and respiratory quotient were not different in the two endurance tests, but pulmonary ventilation (VE) and blood lactate concentration were lower in HOX than in NOX, especially during the latter part of the task. Plasma base deficit (BDpl) increased initially by 3.5 mmol.l-1 during HOX and then stabilized. In NOX, a continuous increase was seen and the change was more than twice as large. Relative to BDpl, VE was higher in HOX than in NOX indicating a more efficient ventilatory compensation of the metabolic acidosis. The reduced ventilatory demand and lower metabolic acidosis in HOX in combination with lower relative exercise intensity may have contributed to the longer time to exhaustion. However, the pattern of individual variation suggested that other mechanisms were also involved.     

Oviposition choice and larval survival of Dasineura marginemtorquens (Diptera: Cecidomyiidae) on resistant and susceptible Salix viminalis

Abstract.

• 1 Substantial intraspecific variation exists in Salix viminalis resistance to the gall midge Dasineura marginemtorquens. Earlier work has found this variation to have a large genetic component. Willow clones are stable in their resistances between midge generations and different nutrient levels in both field and laboratory culture.
• 2 This study reports the results of laboratory experiments on female oviposition choice and larval survival on potted plants from clones that are very different in resistance as determined in field studies.
• 3 In choice experiments using pairs of plants, the average female midge did not prefer susceptible willow clones over resistant ones for oviposition. In about one third of the replicates, midges actually laid more eggs on the resistant clone. Further work is necessary to examine the nature of variation among midges in discrimination of these plant types.
• 4 Resistance is manifested as great differences in larval survival. Six days after oviposition survival was 92% on susceptible plants but only 6% on resistant ones. Galls developed on all of the susceptible plants, while in 73% of the resistant plants galls were not even initiated.
• 5 The plant traits causing resistance are enigmatic. Larval behaviour suggests that resistant plants interfere with feeding behaviour. On resistant plants, most larvae wander for more than 24 h without initiating any galls before dying. On susceptible plants many first instar larvae begin feeding and initiate galls within this period.

     

Isolation and characterization of porcine somatotropin containing a succinimide residue in place of aspartate129.

Aspartate129 in porcine somatotropin was converted into a cyclic imide residue (succinimide) under acidic solution conditions. Reversed-phase high performance liquid chromatography was utilized to isolate and quantitate this altered species, which accounted for approximately 30% of the total protein. The molecular mass of this modified species was determined by electrospray mass spectrometry to be 18 Da less than normal porcine somatotropin, indicative of a loss of 1 H2O molecule. Tryptic peptide mapping demonstrated that the peptide composed of residues 126-133 was altered in this modified protein. Amino acid analysis, amino acid sequencing, mass spectrometry, and capillary zone electrophoresis were used to demonstrate that aspartate129 in this peptide had been converted into a succinimide residue. Further confirmation that this peptide contained a succinimide was obtained by hydrolyzing the modified peptide at pH 9.0, which yielded both the aspartate and isoaspartate peptides.     

Effect of lowered muscle temperature on the physiological response to exercise in men

The effect of low muscle temperature on the response to dynamic exercise was studied in six healthy men who performed 42 min of exercise on a cycle ergometer at an intensity of 70% of their maximal O2 uptake. Experiments were performed under control conditions, i.e. from rest at room temperature, and following 45 min standing with legs immersed in a water bath at 12 degrees C. The water bath reduced quadriceps muscle temperature (at 3 cm depth) from 36.4 (SD 0.5) degrees C to 30.5 (SD 1.7) degrees C. Following cooling, exercise heart rate was initially lower, the mean difference ranged from 13 (SD 4) beats.min-1 after 6 min of exercise, to 4 (SD 2) beats.min-1 after 24 min of exercise. Steady-state oxygen uptake was consistently higher (0.2 l.min-1). However, no difference could be discerned in the kinetics of oxygen uptake at the onset of exercise. During exercise after cooling a significantly higher peak value was found for the blood lactate concentration compared to that under control conditions. The peak values were both reached after approximately 9 min of exercise. After 42 min of exercise the blood lactate concentrations did not differ significantly, indicating a faster rate of removal during exercise after cooling. We interpreted these observations as reflecting a relatively higher level of muscle hypoxia at the onset of exercise as a consequence of a cold-induced vasoconstriction. The elevated steady-state oxygen uptake may in part have been accounted for by the energetic costs of removal of the extra lactate released into the blood consequent upon initial tissue hypoxia.     

Cowpea aphid performance and behaviour on two resistant cowpea lines

In a series of laboratory and climate chamber tests we compared the growth and behaviour of Aphis craccivora on one susceptible (ICV-1) and two aphid-resistant (ICV-11 and ICV-12) cowpea lines. The aphids' growth rates were much lower on the resistant cowpea lines than on the susceptible one, indicating strong antibiosis. In addition, the aphids invariably settled in higher numbers on the susceptible line than on either of the resistant. Compared to ICV-1, damaged leaves of the resistant line ICV-12 were settled upon to a higher degree than undamaged leaves, and leaf discs from the same line were even less resistant.On resistant lines individual aphids waited a significantly longer time before making their first test probe. Total probing time as well as the time preceding a decision to stay or leave was also longer.These results are discussed in relation to the possible mechanisms involved, and we also consider the effects of previous leaf feeding on the expression of resistance in the field.

---

Résumé Au cours d'expériences au laboratoire et en chambres climatisées nous avons comparé la croissance et le comportement de A. craccivora sur une lignée sensible (ICV-1) et deux lignées résistantes (ICV-11 et ICV-12) de V. unguiculata. Les vitesses de croissance des pucerons ont été beaucoup plus lentes sur les lignées résistantes que sur la lignée sensible, ce qui révèle une forte antibiose. De plus, les pucerons atterrissent invariablement en plus grand nombre sur la variété sensible. Par comparaison avec ICV-1, les atterrissages sur lignée résistante ICV-12 étaient plus nombreux sur les feuilles endommagées que sur les feuilles intactes; les disques de feuilles de cette même lignée étaient encore moins résistants.Les pucerons ont séjourné individuellement un temps plus long sur les lignées résistantes avant de faire leur premier sondage. Le temps consacré aux sondages ainsi que le temps précédant de choix entre départ ou maintien sur la feuille étaient plus longs avec les lignées résistantes.Ces résultats ont été discutés en fonction des mécanismes impliqués. Nous avons aussi examiné les effets de la consommation antérieure sur les manifestations de la résistance dans la nature.

     

低温对大鼠骨骼肌能量代谢的影响——Ⅰ.高能磷酸化合物含量的变化

本实验选择对低温比较敏感的骨骼肌组织作为实验对象。将19只大鼠随机分成3组,其右后肢经冷冻处理使组织温度分别达 5℃、0℃及-5℃,于复温后不同时间测定骨骼肌中四种高能磷酸化合物AMP,ADP、ATP及CP的含量。结果表明,大鼠骨骼肌组织中高能磷酸化合物含量随冷冻时组织最低温度的下降而不同程度的减少,并且具有明显的时相过程。复温后即刻至4小时下降迅速,4～12小时三组值较为接近,其后随时间延长而明显分离。 5℃组恢复得早而且明显,0℃组恢复程度次于 5℃组,-5℃组含量下降最明显而且几乎不恢复。提示:低温使骨骼肌组织能量的产生和贮存过程受到影响。