Induction of leptin resistance through direct interaction of C-reactive protein with leptin

The mechanisms underlying leptin resistance are still being defined. We report here the presence in human blood of several serum leptin-interacting proteins (SLIPs), isolated by leptin-affinity chromatography and identified by mass spectrometry and immunochemical analysis. We confirmed that one of the major SLIPs is C-reactive protein (CRP). In vitro, human CRP directly inhibits the binding of leptin to its receptors and blocks its ability to signal in cultured cells. In vivo, infusion of human CRP into ob/ob mice blocked the effects of leptin upon satiety and weight reduction. In mice that express a transgene encoding human CRP, the actions of human leptin were completely blunted. We also found that physiological concentrations of leptin can stimulate expression of CRP in human primary hepatocytes. Recently, human CRP has been correlated with increased adiposity and plasma leptin. Thus, our results suggest a potential mechanism contributing to leptin resistance, by which circulating CRP binds to leptin and attenuates its physiological functions.     

Relation Between the Chlorophyll Content and Paleness of Gibberellic Acid-Treated Leaves

The rate of chlorophyll synthesis as a function of lime was examined in plants of Hordeum vitlgare L. cultivated in Knop solution containing 1 mg/1 GA₃. The action of concentration on the relations of chlorophyll components was investigated in GA₃ solutions of five different concentrations. The relations of total chlorophyll contents, including those of the a and b components, were determined from ethanol and aqueous acetone extract at various periods of treatment, at the same moment but with different concentrations of GA₃. A significant difference is noted between the chlorophyll contents of the GA₃ treated plants and those of the control. The chlorophyll contents of treated plants are decreasing. The difference increases with raising GA₃ concentration. The components were separated on thin layer with the partition method, and the spots were eluted. The density was measured with a spectrophotometer. Among the yellow components in the treated plants, the carotenes (α and β carotenes) increase as compared with the control plants. The total quantity of xanthophylls (lutein, zeaxanthine, lutein-5, 6-epoxide, violaxanthine and neoxanthine) almost equals the xanthophyll amount in the control plants; there is no significant difference. The paleness of leaves treated with GA₃ is provoked by the quantitative change of pigment components; the chlorophylls o and b decrease, the xanthophylls remain unchanged and the carotenes increase.     

In Vivo Experiments Reveal the Good,the Bad and the Ugly Faces of sFlt-1 in Pregnancy

Objective

Soluble fms-like tyrosine kinase (sFlt)-1-e15a, a primate-specific sFlt-1-isoform most abundant in the human placenta in preeclampsia, can induce preeclampsia in mice. This study compared the effects of full-length human (h)sFlt-1-e15a with those of truncated mouse (m)sFlt-1(1-3) used in previous preeclampsia studies on pregnancy outcome and clinical symptoms in preeclampsia.

Methods

Mice were injected with adenoviruses or fiber-mutant adenoviruses overexpressing hsFlt-1-e15a, msFlt-1(1-3) or control GFP under the CMV or CYP19A1 promoters on gestational day 8 (GD8) and GD11. Placentas and pups were delivered by cesarean section, and dams were monitored postpartum. Blood pressure was telemetrically recorded. Urine samples were collected with cystocentesis and examined for albumin/creatinine ratios. Tissue specimens were evaluated for transgene as well as endogenous mFlt-1 and msFlt-1-i13 expression. H&E-, Jones- and PAS-stained kidney sections were histopathologically examined. Placental GFP expression and aortic ring assays were investigated with confocal microscopy.

Results

Mean arterial blood pressure (MAP) was elevated before delivery in hsFlt-1-e15a-treated mice compared to controls (GD18: ΔMAP = 7.8 mmHg, p = 0.009), while ΔMAP was 12.8 mmHg (GD18, p = 0.005) in msFlt-1(1-3)-treated mice. Urine albumin/creatinine ratio was higher in hsFlt-1-e15a-treated mice than in controls (GD18, p = 0.04; PPD8, p = 0.03), and msFlt-1(1-3)-treated mice had marked proteinuria postpartum (PPD8, p = 4×10⁻⁵). Focal glomerular changes were detected in hsFlt-1-e15a and msFlt-1(1-3)-treated mice. Aortic ring microvessel outgrowth was decreased in hsFlt-1-e15a (p = 0.007) and msFlt-1(1-3)-treated (p = 0.02) mice. Full-length msFlt-1-i13 expression was unique for the placenta. In hsFlt-1-e15a-treated mice, the number of pups (p = 0.046), total weight of living pups (p = 0.04) and maternal weights (p = 0.04) were higher than in controls. These differences were not observed in truncated msFlt-1(1-3)-treated mice.

Conclusions

Truncated msFlt-1(1-3) simulated the preeclampsia-promoting effects of full-length hsFlt-1. MsFlt-1(1-3) had strong effect on maternal endothelium but not on placentas and embryos. In contrast, hsFlt-1-e15a induced preeclampsia-like symptoms; however, it also increased litter size. In accord with the predominant placental expression of hsFlt-1-e15a and msFlt-1-i13, full-length sFlt-1 may have a role in the regulation of embryonic development. These observations point to the difference in the biological effects of full-length and truncated sFlt-1 and the changes in the effect of full-length sFlt-1 during pregnancy, and may have important implications in the management of preeclampsia.     

Glutathione as an Antioxidant and Regulatory Molecule in Plants Under Abiotic Stress Conditions

The glutathione (GSH)/glutathione disulfide (GSSG) redox couple is involved in several physiologic processes in plants under both optimal and stress conditions. It participates in the maintenance of redox homeostasis in the cells. The redox state of the GSH/GSSG couple is defined by its reducing capacity and the half-cell reduction potential, and differs in the various organs, tissues, cells, and compartments, changing during the growth and development of the plants. When characterizing this redox couple, the synthesis, degradation, oxidation, and transport of GSH and its conjugation with the sulfhydryl groups of other compounds should be considered. Under optimal growth conditions, the high GSH/GSSG ratio results in a reducing environment in the cells which maintains the appropriate structure and activity of protein molecules because of the inhibition of the formation of intermolecular disulfide bridges. In response to abiotic stresses, the GSH/GSSG ratio decreases due to the oxidation of GSH during the detoxification of reactive oxygen species (ROS) and changes in its metabolism. The lower GSH/GSSG ratio activates various defense mechanisms through a redox signalling pathway, which includes several oxidants, antioxidants, and stress hormones. In addition, GSH may control gene expression and the activity of proteins through glutathionylation and thiol-disulfide conversion. This review discusses the size and redox state of the GSH pool, including their regulation, their role in redox signalling and defense processes, and the changes caused by abiotic stress.     

Effects of tail-like substituents on the binding of competitive inhibitors to the Q(B) site of photosystem II

The QB quinone-binding site of photosystem II is an important target for herbicides. Two major classes of herbicides are based on s-triazine and phenylurea moieties. A small library of triazine and phenylurea compounds has been synthesized which have tail-like substituents in order to test the effects of charge, hydrophobicity and size of the tail on binding properties. It is found that a tail can be attached to one of the alkylamino groups of triazine-type herbicides or to the para position of phenylurea-type herbicides without loss of binding, provided that the tail is hydrophobic. This indicates that the herbicides must be oriented in the QB site such that these positions point toward the natural isoprenyl tail-binding pocket that extends out of the Q(B) site. In turn, the requirement that the tail must extend out of the QB site constrains the size of the other herbicide substituents in the pocket. This is in agreement with the presumed orientation and fit of ligands in the QB site. When longer hydrophobic tails are used, the binding penalty that occurs upon adding a charged substituent at the distal end is reduced. This allows the use of a series of tail substituents possessing a distal charge as an approximate molecular ruler to measure the distance from the QB site to the aqueous phase. Even a 10-carbon alkyl chain still shows a 4-fold effect from the presence or absence of a distal charge. Such a chain does not appear to be long enough to extend from the bulk aqueous phase to the QB site because binding is completely lost when a large hydrophilic domain (PEG(4000)) is attached to the distal end. Longer tails are effective only if they are sufficiently hydrophobic. An effort was made to use tailed herbicides for affinity binding of photosystem II. It was found that hydrophobic linkers promote nonspecific binding, but careful choice of solvent conditions, such as the use of excess nonionic detergent well above its critical micelle concentration, might obviate this problem during affinity-binding applications.     

Effect of osmotic stress on glutathione and hydroxymethylglutathione accumulation in wheat

The effect of osmotic stress on glutathione and hydroxymethylglutathione levels was compared in three wheat genotypes and two 5A chromosome substitution lines. Freezing-tolerant genotypes seemed also to be tolerant to osmotic stress induced by polyethylene glycol (PEG), since their fresh weight was not affected by the treatment. However, the growth of freezing-sensitive genotypes was reduced by 7-day PEG treatment and they had greater injuries after osmotic stress. The reduced forms of the two glutathione precursors, cysteine and gamma-glutamylcysteine, and of hydroxymethylglutathione (hmGSH) and glutathione (GSH) were present in greater quantities after PEG treatment in the two tolerant genotypes than in the sensitive ones. Similarly, osmotic stress resulted in a higher ratio of the reduced to the oxidised form of these thiols and in greater activity of gamma-glutamylcysteine synthetase and glutathione reductase in the tolerant genotypes compared to the sensitive ones. Following in vivo glutathione synthesis, a greater incorporation of radioactivity from [35S]sulphate into the four thiols was found in the tolerant genotypes than in the sensitive ones during osmotic stress. The present results indicate that hmGSH and GSH may contribute to the improvement of tolerance against osmotic stress in wheat and that the 5A chromosome influences the stress-induced changes in GSH and hmGSH levels.     

Parallel-stranded guanine quadruplex interactions with a copper cationic porphyrin

G-quadruplexes are formed by association of DNA strands containing multiple contiguous guanines. The capability of drugs to induce formation of or stabilize G-quadruplexes is an active area of investigation. We report the interactions of CuTMpyP4, the Cu(2+) derivative of 5,10,15,20-tetrakis(1-methyl-4-pyridyl)-21H,23H-porphine, with the parallel-stranded G-quadruplexes formed by d(T(4)G(4)T(4)) (1) and d(T(4)G(8)T(4)) (3). Absorption titrations of CuTMpyP4 with (1)(4) or (3)(4) cause both bathochromicity and hypochromicity of the porphyrin Soret band, with larger changes observed for the longer oligonucleotide. An approximate binding constant for (1)(4) and CuTMpyP4 according to the Scatchard model is 5.6 x 10(6) M(-)(1) in terms of quadruplexes and according to the McGhee-von Hippel model is 1.3 x 10(6) M(-)(1) in terms of potential binding sites. An approximate binding constant for (3)(4) and CuTMpyP4 according to the Scatchard model is 5.2 x 10(7) M(-)(1) in terms of quadruplexes and in terms of the McGhee-von Hippel model is 2.4 x 10(6) M(-)(1) in terms of potential binding sites. The site size for CuTMpyP4 and (1)(4) is four using the McGhee-von Hippel model. We find a 2:1 binding stoichiometry for CuTMpyP4 and (1)(4) and a 3:1 binding stoichiometry for CuTMpyP4 and (3)(4) using the method of continuous variation analysis. Induced emission spectra of CuTMpyP4 with (1)(4) or (3)(4) indicate a mode of binding in which the ligand is protected from the solvent. Electron paramagnetic resonance spectra of CuTMpyP4 with added oligonucleotide show an increase in the Cu-N superhyperfine coupling constant as the length of the oligonucleotide increases. On the basis of these data, we propose that for both (1)(4) and (3)(4), CuTMpyP4 molecules externally stack at each end of the run of guanines, similar to other planar G-quadruplex ligands. For (3)(4), our data are consistent with intercalation of a CuTMpyP4 molecule into the G-quadruplex.     

Deception Undermines the Stability of Cooperation in Games of Indirect Reciprocity

Indirect reciprocity is often claimed as one of the key mechanisms of human cooperation. It works only if there is a reputational score keeping and each individual can inform with high probability which other individuals were good or bad in the previous round. Gossip is often proposed as a mechanism that can maintain such coherence of reputations in the face of errors of transmission. Random errors, however, are not the only source of uncertainty in such situations. The possibility of deceptive communication, where the signallers aim to misinform the receiver cannot be excluded. While there is plenty of evidence for deceptive communication in humans the possibility of deception is not yet incorporated into models of indirect reciprocity. Here we show that when deceptive strategies are allowed in the population it will cause the collapse of the coherence of reputations and thus in turn it results the collapse of cooperation. This collapse is independent of the norms and the cost and benefit values. It is due to the fact that there is no selection for honest communication in the framework of indirect reciprocity. It follows that indirect reciprocity can be only proposed plausibly as a mechanism of human cooperation if additional mechanisms are specified in the model that maintains honesty.     

Mice expressing the variant rs1143679 allele of ITGAM (CD11b) show impaired DC-mediated T cell proliferation

Mammalian Genome - Genome-wide association studies (GWAS) and functional genomic analyses have implicated several ITGAM (CD11b) single-nucleotide polymorphisms (SNPs) in the development of SLE and...