Induction of Glutamine Synthetase by Dibutyryl Cyclic AMP in C-6 Glioma Cells

Glutamine synthetase was found to be increased in C-6 glioma cells as a result of increasing culture passage and N-6,2'-O-dibutyryl cyclic AMP (dbcAMP) treatment. At low passage dbcAMP produced a 2.5-fold increase in glutamine synthetase activity per unit of cellular protein. At high passage control glutamine synthetase was approximately double that seen at low passage, but dbcAMP produced an additional 65% increase. Lactate dehydrogenase activity was also increased by dbcAMP treatment at both low and high passage, but culture passage produced no change in the lactate dehydrogenase. With increasing culture passage, the ratio of cellular protein to DNA doubled. Therefore, expression of data per unit of protein tended to minimize the apparent changes in activity. The maximum increase in glutamine synthetase activity produced by both dbcAMP and increasing culture passage and expressed on a DNA basis was 5.6-fold. The increase in glutamine synthetase activity was generally linear during the first 20 h of drug treatment, after which enzyme activity remained nearly constant up to 72 h. Ninety percent or more of the dbcAMP remained in the medium at the end of 48-h exposure of cells to dbcAMP. 8-br-Cyclic AMP also increased glutamine synthetase activity of C-6-cels, but n-butyrate did not. Isoproterenol, which increases cyclic AMP in C-6-cells, increased glutamine synthetase activity. The effect of isoproterenol on glutamine synthetase was inhibited by the beta-adrenergic blocking agent sotalol. Cycloheximide (10 micrograms/ml) inhibited the dbcAMP effect on glutamine synthetase activity and also decreased the control enzyme activity by 60%.     

Sustainable harvesting of bog myrtle (Myrica gale L.) in Scotland

Summary

Bog myrtle communities were surveyed to determine their accessibility for harvesting and the approximate number of harvestable stems. At least 45.1 million bog myrtle stems within Scotland may be both suitable (about 55 cm long) and easily accessible (< 1 km from a tarmac road) for harvesting. In a harvesting trial in Strath Avon, plants were subjected to five different cutting treatments with and without the addition of NPK. There were significant differences between cutting treatments, but no significant effects of fertilizer. Cutting all stems to half height produced the most rapid regeneration. Stems could take eight years to regenerate to a harvestable size if this cutting method were employed. On this basis, a harvest of 5.6 million stems could be sustained annually, compared with a current harvest of about 0.6 million.     

Defect‐Engineered Ultrathin δ‐MnO2 Nanosheet Arrays as Bifunctional Electrodes for Efficient Overall Water Splitting

Recently, defect engineering has been used to intruduce half‐metallicity into selected semiconductors, thereby significantly enhancing their electrical conductivity and catalytic/electrocatalytic performance. Taking inspiration from this, we developed a novel bifunctional electrode consisting of two monolayer thick manganese dioxide (δ‐MnO₂) nanosheet arrays on a nickel foam, using a novel in‐situ method. The bifunctional electrode exposes numerous active sites for electrocatalytic rections and displays excellent electrical conductivity, resulting in strong performance for both HER and OER. Based on detailed structure analysis and density functional theory (DFT) calculations, the remarkably OER and HER activity of the bifunctional electrode can be attributed to the ultrathin δ‐MnO₂ nanosheets containing abundant oxygen vacancies lead to the formation od Mn³⁺ active sites, which give rise to half‐metallicity properties and strong H₂O adsorption. This synthetic strategy introduced here represents a new method for the development of non‐precious metal Mn‐based electrocatalysts for eddicient energy conversion.     

Hairpin RNAs derived from RNA polymerase II and polymerase III promoter-directed transgenes are processed differently in plants

RNA polymerase III (Pol III) as well as Pol II (35S) promoters are able to drive hairpin RNA (hpRNA) expression and induce target gene silencing in plants. siRNAs of 21 nt are the predominant species in a 35S Pol II line, whereas 24- and/or 22-nucleotide (nt) siRNAs are produced by a Pol III line. The 35S line accumulated the loop of the hpRNA, in contrast to full-length hpRNA in the Pol III line. These suggest that Pol II and Pol III-transcribed hpRNAs are processed by different pathways. One Pol III transgene produced only 24-nt siRNAs but silenced the target gene efficiently, indicating that the 24-nt siRNAs can direct mRNA degradation; specific cleavage was confirmed by 5' rapid amplification of cDNA ends (RACE). Both Pol II- and Pol III-directed hpRNA transgenes induced cytosine methylation in the target DNA. The extent of methylation is not correlated with the level of 21-nt siRNAs, suggesting that they are not effective inducers of DNA methylation. The promoter of a U6 transgene was significantly methylated, whereas the promoter of the endogenous U6 gene was almost free of cytosine methylation, suggesting that endogenous sequences are more resistant to de novo DNA methylation than are transgene constructs.     

Nuclear interactions in a heterokaryon: insight from the model Neurospora tetrasperma

A heterokaryon is a tissue type composed of cells containing genetically different nuclei. Although heterokaryosis is commonly found in nature, an understanding of the evolutionary implications of this phenomenon is largely lacking. Here, we use the filamentous ascomycete Neurospora tetrasperma to study the interplay between nuclei in heterokaryons across vegetative and sexual developmental stages. This fungus harbours nuclei of two opposite mating types (mat A and mat a) in the same cell and is thereby self-fertile. We used pyrosequencing of mat-linked SNPs of three heterokaryons to demonstrate that the nuclear ratio is consistently biased for mat A-nuclei during mycelial growth (mean mat A/mat a ratio 87%), but evens out during sexual development (ratio ranging from 40 to 57%). Furthermore, we investigated the association between nuclear ratio and expression of alleles of mat-linked genes and found that expression is coregulated to obtain a tissue-specific bias in expression ratio: during mycelial extension, we found a strong bias in expression for mat A-linked genes, that was independent of nuclear ratio, whereas at the sexual stage we found an expression bias for genes of the mat a nuclei. Taken together, our data indicate that nuclei cooperate to optimize the fitness of the heterokaryon, via both altering their nuclear ratios and coregulation genes expressed in the different nuclei.     

Estimates of the Daily Phase and Amplitude of the Endogenous Component of the Circadian Rhythm of Core Temperature in Sedentary Humans Living Nychthemerally

Fifteen healthy female subjects were studied for eight days while living conventionally. Subjects were free to choose the ways they spent their time within a framework of regular times of retiring and rising; in practice, much of the waking time was spent in sedentary activities. Nine of the subjects were aware of the natural light-dark cycle, this approximating to a 12:12 L:D schedule at the time of year when the study took place. Before the study, subjects were assessed for their degree of "morningness" by questionnaire; throughout the study, they wore a rectal probe, and an activity meter on their non-dominant wrist. The timing (phase) and amplitude of the circadian rectal temperature rhythm were assessed on each day by cosinor analysis as well as by a me thod based on visual inspection of the data. These two parameters were also assessed after the temperature data for each day had been "purified" by a number of methods. From these results it was possible to investigate the effect of purification upon the amplitude of the circadian rhythm of temperature. Also, the day-by-day variability of phase, and the relationship between morningness and phase, were compared using these methods of phase estimation, and using cross-correlation between data sets from adjacent days; in all cases, raw and purified temperature data were used. There was a significantly greater amount of daily variation in phase using purified rather than raw data sets, and this difference was present with all methods of purification as well as with all methods for estimating phase. Purifi cation decreased the amplitude of the circadian temperature rhythm by about 30%. Finally, there was a significant correlation between the morningness score of the subjects and the phase of the circadian temperature rhythm, the phase becoming earlier with increasing morningness; when this relationship was re-examined using purified data, it became more marked. These results reflect the masking effects exerted upon raw temperature data by lifestyle. The extent to which the purification methods enable the endogenous component of a circadian rhythm - and, by implication, the output of the endogenous circadian oscillator - to be estimated in subjects living normally is addressed.     

l-DOPA Cytotoxicity to PC12 Cells in Culture Is via Its Autoxidation

Abstract: The mechanism of cytotoxicity of l -DOPA was studied in the rat pheochromocytoma PC12 cell line. The cytotoxicity of l -DOPA to PC12 cells was time and concentration dependent. Carbidopa, which inhibited the conversion of l -DOPA to dopamine, did not protect against l -DOPA cytotoxicity in PC12 cells. Furthermore, clorgyline, a selective inhibitor of monoamine oxidase type A, and pargyline, an inhibitor of both monoamine oxidase types A and B, both did not have an effect on l -DOPA toxicity. These findings suggest that cytotoxicity was not due to dopamine formed from l -DOPA. Catalase or superoxide dismutase each partially protected against l -DOPA toxicity in PC12 cells. In combination, the effects were synergistic and provided almost total protection against cytotoxicity. 6-Cyano-7-nitroquinoxaline-2,3-dione, an antagonist of non-NMDA receptors, did not protect against l -DOPA toxicity. These data suggest that toxicity of l -DOPA is most likely due to the action of free radicals formed as a result of its autoxidation. Furthermore, these findings suggest that patients on long-term l -DOPA therapy are potentially at risk from the toxic intermediates formed as a result of its autoxidation.     

Effects of Ramadan upon Fluid and Food Intake,Fatigue, and Physical,Mental, and Social Activities: A Comparison between the UK and Libya

Two studies were performed during Ramadan, one in the UK (N=31) and the other in Libya (N=33). The aims were to assess some changes to lifestyle that are produced by fasting as well as effects due to culture. Subjects were studied on eight separate occasions: four control days (two before and two after Ramadan) and four days during the four weeks of Ramadan itself. A questionnaire was answered that asked about naps and fluid and food intake. The questions elicited if an individual had slept, drank, or eaten, plus the reasons for doing or not doing so. Also, subjects were asked to describe their physical, mental, and social activities, their fatigue, and their perceived abilities to perform physical or mental work. The questionnaire was answered five times per day: at sunrise, at 10:00 h, at 14:00 h, at sunset, and on retiring to sleep at night. Urine samples were collected at sunset and measured for osmolality. Differences between control and Ramadan days, as well as between subjects studied in UK and Libya, were assessed by analysis of variance. Correlations between fatigue and physical, mental, and social activities were also assessed, as were differences in urine osmolality. Fasting during Ramadan resulted in fewer activities and increased fatigue and frequency of napping during daytime. Changes in fluid and food intake indicated some degree of preparation for fasting before sunrise and a marked “recuperation” from fasting after sunset. The reasons given for napping in the daytime, for drinking or not drinking, and for eating or not eating, changed during Ramadan compared with control days; as a result, links between fatigue and activities, and fatigue and fluid and food intake, were all altered during Ramadan, particularly after sunset. Subjects become dehydrated during the daytime, but this was not reduced when females who were menstruating drank during this time. Several differences between the two studies were found. There was a greater frequency of napping during the daytime in the Libya study, and evidence for the conservation of energy during the daytime and reduced physical, mental, and social activities. Subjects' preparations for fasting and recovering from it—their fluid and food intakes and associated reasons for these—also differed. Possible explanations of these differences are discussed. (Author correspondence: waterhouseathome@hotmail.com)     

Daily rhythms in activity and mRNA abundance of enzymes involved in glucose and lipid metabolism in liver of rainbow trout,Oncorhynchus mykiss. Influence of light and food availability

ABSTRACT

We have investigated the magnitude of diurnal variation in back squat and bench press performance using the MuscleLab force velocity transducer. Thirty resistance-trained males (mean ± SD: age 21.7 ± 1.4 years; body mass 80.5 ± 4.5 kg; height 1.79 ± 0.06 m) underwent two sessions at different times of day: morning (M, 07:30 h) and evening (E, 17:30 h). Each session included a period when rectal temperature (T_rec) was measured at rest, a 5-min standardized 150 W warm-up on a cycle ergometer, then defined programme of bench press (at 20, 40 and 60 kg) and back squat (at 30, 50 and 70 kg) exercises. A linear encoder was attached to an Olympic bar used for the exercises and average force (AF), peak velocity (PV) and time-to-peak velocity (tPV) were measured (MuscleLab software; MuscleLab Technology, Langesund, Norway) during the concentric phase of the movements. Values for T_rec at rest were higher in the evening compared to morning values (0.48°C, P < 0.0005). Daily variations were apparent for both bench press and back squat performance for AF (1.9 and 2.5%), PV (8.3 and 12.7%) and tPV (?16.6 and ?9.8%; where a negative number indicates a decrease in the variable from morning to evening). There was a main effect for load where AF and tPV increased and PV decreased from the lightest load to the heaviest for both bench press and back squat (47.1 and 80.2%; 31.7 and 57.7%; ?42.1 and ?73.9%; P < 0.0005 where a negative number indicates a decrease in the variable with increasing load). An interaction was found only for tPV, such that the tPV occurs earlier in the evening than the morning at the highest loads (60 and 70 kg) for both bench press and back squat, respectively (mean difference of 0.32 and 0.62 s). In summary, diurnal variation in back squat and bench press was shown; and the tPV in complex multi-joint movements occurs earlier during the concentric phase of exercise when back squat or bench press is performed in the evening compared to the morning. This difference can be detected using a low cost, portable and widely available commercial instrument and enables translation of past laboratory/tightly controlled experimental research in to main-stream coaching practice.