A further extension of the method for independently measuring the amount of nitrogen fixed by a legume crop

Summary The combination of using¹⁵N for determining the amount of nitrogen fixed by a legume crop in field experiments and the labelling of only one treatment at a time in each treatment combination is shown to be conceptually and experimentally valid for determining the effect of cultural practices on the amount of nitrogen fixed by a legume crop.     

Progressive loss of mitochondrial creatine phosphokinase activity in muscular dystrophy.

Mitochondria were isolated from the pectoralis and gastrocnemius muscles of chickens with a hereditary muscular dystrophy, and age-matched controls. In the pectoralis, for dystrophic birds aged 0.12, 0.25, 0.55, and 1.55 yr, the creatine phosphokinase activity of the intact mitochondria, expressed in terms of pellet protein, was 69%, 45%, 24%, and 13% as great, respectively, as that of the controls. The corresponding figures for the gastrocnemius were 79%, 46%, 51%, and 28%. The mitochondria from dystrophic muscles exhibited satisfactory respiratory control ratios, P:0 ratios, and state 3 respiratory rates. To check whether their apparent loss of creatine phosphokinase activity was due to the presence of increasing amounts of non-mitochondrial pellet protein, the state 3 respiratory rate was used as a mitochondrial marker; the rates per mg protein were similar in mitochondria from normal and dystrophic muscles of each age group.     

Kinase GSK3β functions as a suppressor in colorectal carcinoma through the FTO-mediated MZF1/c-Myc axis

Colorectal carcinoma (CRC) poses heavy burden to human health and has an increasing incidence. Currently, the existing biomarkers for CRC bring about restrained clinical benefits. GSK3β is reported to be a novel therapeutic target for this disease but with undefined molecular mechanisms. Thus, we aimed to investigate the regulatory effect of GSK3β on CRC progression via FTO/MZF1/c-Myc axis. Firstly, the expression patterns of GSK3β, FTO, MZF1 and c-Myc were determined after sample collection. Lowly expressed GSK3β but highly expressed FTO, MZF1 and c-Myc were found in CRC. After transfection of different overexpressed and interference plasmids, the underlying mechanisms concerning GSK3β in CRC cell functions were analysed. Additionally, the effect of GSK3β on FTO protein stability was assessed followed by detection of MZF1 m6A modification and MZF1-FTO interaction. Mechanistically, GSK3β mediated ubiquitination of demethylase FTO to reduce FTO expression. Besides, GSK3β inhibited MZF1 expression by mediating FTO-regulated m6A modification of MZF1 and then decreased the proto-oncogene c-Myc expression, thus hampering CRC cell proliferation. We also carried out in vivo experiment to verify the regulatory effect of GSK3β on CRC via FTO-mediated MZF1/c-Myc axis. It was found that GSK3β inhibited CRC growth in vivo which was reversed by overexpressing c-Myc. Taken together, our findings indicate that GSK3β suppresses the progression of CRC through FTO-regulated MZF1/c-Myc axis, shedding light onto a new possible pathway by which GSK3β regulates CRC.     

The Exact Distribution of the Anderson-Kannemann Statistic

The Anderson-Kannemann test is a rank test for treatment effects in a randomized block design with K treatments and N blocks. In this paper, an algorithm for computing the exact distribution of the Anderson-Kannemann test statistic under the null hypothesis is deduced. Then, the exact distribution is compared with the asymptotic χ²-distribution, and it is shown that the exact distribution is approximated fairly well by the asymptotic distribution. Tables of the exact distribution are given for K = 3, N = 3(1)15; K = 4, N = 3(1)11; K = 5, N = 3(1)7; and K = 6, N = 3(1)5.     

Creatine kinase of heart mitochondria: Changes in its kinetic properties induced by coupling to oxidative phosphorylation

A complete kinetic analysis of the forward mitochondrial creatine kinase reaction was conducted to define the mechanism for its rate enhancement when coupled to oxidative phosphorylation. Two experimental systems were employed. In the first, ATP was produced by oxidative phosphorylation. In the second, heart mitochondria were pretreated with rotenone and oligomycin, and ATP was regenerated by a phosphoenolpyruvate-pyruvate kinase system. Product inhibition studies showed that oxidative phosphorylation did not effect the binding of creatine phosphate to the enzyme. Creatine phosphate interacted competitively with both ATP and creatine, and the E · MgATP · CrP dead-end complex was not readily detected. In a similar manner, the dissociation constants for creatine were not influenced by the source of ATP: K_ib = 29 mm; K_b = 5.3 mM, and the maximum velocity of the reaction was unchanged: V₁ = 1 μmol/ min/mg. Slight differences were noted for the dissociation constant (K_ia) of MgATP from the binary enzyme complex, E · MgATP. The values were 0.75 and 0.29 mm in the absence and presence of respiration. However, a 10-fold decrease in the steady-state dissociation constant (K_a) of MgATP from the ternary complex, E · MgATP · creatine, was documented: 0.15 mm with exogenous ATP and 0.014 mm with oxidative phosphorylation. Since K_ia × K_b does not equal K_a × K_ib under respiring conditions, the enzyme appears to be altered from its normal rapid-equilibrium random binding kinetics to some other mechanism by its coupling to oxidative phosphorylation.     

O6-Protection and Other Transformations at Guanosine and Inosine Lactam Sites with Application of Related Pyridinium Salts

Abstract

Nucleophilic displacement reactions of guanosine- and inosine-derived pyridinium salts will be discussed in a view of their preparative applications in nucleoside and oligonucleotide chemistry.     

Sleep architecture is related to the season of PSG recording in 8-month-old infants

ABSTRACT

To date, little is known about the impact of season on infant sleep. In higher latitudes, the duration of daily light time varies substantially between different seasons, and environmental light is one potential factor affecting sleep. In this cohort study, one-night polysomnography (PSG) was performed on 72 healthy 8-month-old infants in 2012 and 2013 to study the effect of season on the sleep architecture of young infants in Finland. The children were divided into four subgroups, according to the amount of light during their birth season and the amount of light during the season of the PSG recordings, corresponding to spring, summer, autumn, and winter. We found that the season of birth did not have an impact on the infants’ sleep architecture at 8 months of age, but the season of the PSG recording did have an effect on several sleep variables. In the PSGs conducted during the spring, there was less N3 sleep and more N2 sleep than in the PSGs conducted during the autumn. In addition, there was more fragmented sleep during spring than autumn. According to our data, the season has an effect on the sleep architecture of young infants and should, therefore, be considered when evaluating the PSG findings of young infants. The exact mechanisms behind this novel finding remain unclear, however. The findings imply that infants` sleep is affected by the season or light environment, as is the case in adult sleep. Since potential explanatory factors, such as direct natural or artificial light exposure and the melatonin levels of the infants, were not controlled, more research is needed in the future to better understand this phenomenon.     

pH dependence of magnesium ion binding to prothrombin fragment 1 and gamma-carboxyglutamic acid-containing peptides via 25Mg NMR.

The binding of magnesium ions to two tripeptides, $\text{L}$-Arg-$\text{D}$-Gla-$\text{D}$-Gla-OMe and Z-$\text{L}$-Arg(NO₂)-$\text{D}$-Gla-$\text{D}$-Gla-OMe, and to bovine prothrombin fragment 1 as a function of pH has been monitored by ²⁵Mg NMR spectroscopy. Binding to the tripeptide was dependent on peptide ionizations occurring at pH 4.6 – 4.8. The pH dependence of magnesium ion binding to fragment 1 reveals two inflection points 4.2 may be attributed to the deprotonation of the third side chain carboxylic acid group of the double γ-carboxyglutamic acid sequence. The origin of the increased binding of magnesium ions to fragment 1 at pH values above 7 is unknown.