Prenatal stress modulates HPA axis homeostasis of offspring through dentate TERT independently of glucocorticoids receptor.

In response to stressful events, the hypothalamic-pituitary-adrenal (HPA) axis is activated, and consequently glucocorticoids are released by the adrenal gland into the blood circulation. A large body of research has illustrated that excessive glucocorticoids in the hippocampus exerts negative feedback regulation of the HPA axis through glucocorticoid receptor (GR), which is critical for the homeostasis of the HPA axis. Maternal prenatal stress causes dysfunction of the HPA axis feedback mechanism in their offspring in adulthood. Here we report that telomerase reverse transcriptase (TERT) gene knockout causes hyperactivity of the HPA axis without hippocampal GR deficiency. We found that the level of TERT in the dentate gyrus (DG) of the hippocampus during the developmental stage determines the responses of the HPA axis to stressful events in adulthood through modulating the excitability of the dentate granular cells (DGCs) rather than the expression of GR. Our study also suggests that the prenatal high level of glucocorticoids exposure-induced hypomethylation at Chr13:73764526 in the first exon of mouse Tert gene accounted for TERT deficiency in the DG and HPA axis abnormality in the adult offspring. This study reveals a novel GR-independent mechanism underlying prenatal stress-associated HPA axis impairment, providing a new angle for understanding the mechanisms for maintaining HPA axis homeostasis.

Some new sharp bounds for the spectral radius of a nonnegative matrix and its application.

In this paper, we give some new sharp upper and lower bounds for the spectral radius of a nonnegative irreducible matrix. Using these bounds, we obtain some new and improved bounds for the signless Laplacian spectral radius of a graph or a digraph.

The synthesis of TiO2 and TiO2-Pt and their application in the removal of Cr (VI).

The deposition of noble metal on titanium dioxide (TiO2) has been considered as an effective strategy to improve the activity of TiO2. In this paper, TiO2 nanoparticles were prepared via a sol-gel route, followed by heat treatment at elevated temperatures. TiO2-Pt catalyst was prepared by deposition of platinum (Pt) on the surface of as-prepared TiO2 nanoparticles. TiO2 and TiO2-Pt were characterized by X-ray diffraction, transmission electron microscopy, Raman spectroscopy, ultraviolet-visible differential reflectance spectra and infrared spectroscopy. TiO2 and TiO2-Pt were used as heterogeneous catalysts for the removal of Cr (VI) under ultraviolet-light illumination. TiO2 prepared at low temperature had smaller particle size and higher specific surface area, and consequently had higher activity on the removal of Cr (VI). The Pt deposited on the surface of TiO2 favoured the separation of photo-produced electrons (e-) and holes (h+), inhibited the recombination of e- and h+, and enhanced Cr (VI) removal. However, its blocking of active sites also inhibited the removal of Cr (VI). The deposition of 1% (wt.%) Pt to TiO2 produced the optimum activity for the removal of Cr (VI). A lower pH favoured the adsorption of Cr (VI) on the surface of TiO2, and correspondingly enhanced the removal of Cr (VI).

Non-covalent binding of azo compound to peptide chain: interactions of biebrich scarlet and naphthochrome green with four model proteins.

We studied the non-specific interactions of two azo compounds: biebrich scarlet (BS) and naphthochrome green (NG), with four model proteins: bovine serum albumin, ovalbumin, poly-L-lysine and hemoglobin by UV-VIS spectrometry, fluorophotometry and circular dichroism melting technique. The optimal acidities of NG and BS for binding to proteins correspond to the physiological pHs of skin and gastro tissues. The saturation binding numbers of BS and NG on peptide chains were determined and the effects of electrolytes and temperature were investigated. These interactions were fitted by the Temkin absorption model and their thermodynamic parameters were calculated. The different bindings of BS and NG to proteins were compared from their molecular structures. We inferred that an ion-pair electrostatic interaction first fixes azo compounds to basic amino acid residues and subsequent binding involves the collective action of other non-covalent bonds: hydrogen bond, van der Waals force, and hydrophobic interaction. This combination of bonds caused a change of secondary conformation of protein from beta-sheet to helix and the possible process was illustrated. The potential protein toxicity resulting from such a non-specific binding was analyzed. Besides, the interaction of BS with peptide chains was applied to protein assay.

Interaction of brilliant red X-3B with bovine serum albumin and application to protein assay.

The interaction of brilliant red X-3B (BRX) with bovine serum albumin (BSA) in three pH media has been characterized by the spectral correction technique. The binding number maximum of BRX was determined to be 102 at pH 2.03, 82 at pH 3.25 and 38 at pH 4.35 and the binding mechanism was analyzed in detail. The effects of ionic strength from 0 to 1 mol L(-1) and temperature from 20 to 70 degrees C on the binding were investigated. The results showed that the interaction of BRX with BSA responded to the Langmuir adsorption isothermal model and the binding constant was determined. From the correlation between the binding number and the number of basic amino acid residues, the ion-pair attraction induced the union of non-covalent bonds including H-bond, van der Waals force and hydrophobic bond and the binding model was illustrated. The binding of BRX to BSA has resulted in change of the BSA conformation confirmed by means of circular dichroism. Using this interaction at pH 2.03, a sensitive method named the absorbance ratio difference spectrometry was established and applied to the protein assay and the limit of detection of protein was only 6 microg L(-1). Two samples were determined and the results were in agreement with those obtained by the classical coomassie brilliant blue colorimetry.

Non-covalent interaction of 2', 4', 5', 7'-tetrabromo-4, 5, 6, 7-tetrachlorofluorescein with proteins and its application.

The interactions of 2', 4', 5', 7'-tetrabromo-4, 5, 6, 7-tetrachlorofluorescein (TBTCF) with BSA, ovalbumin (OVA) and poly-L-lysine (PLYS) at pH 3.70 have been investigated by combination of the spectral correction technique and the Langmuir isothermal adsorption. The active connection actions such as ion pairs, van der Waals' force, hydrogen bond, hydrophobic bond were proposed to explain the non-covalent interaction between TBTCF and BSA, OVA and PLYS. Effects of the electrolyte and high temperature indicated that union of the active connections between TBTCF and BSA and OVA was too firm to be destroyed. The relationship between the binding number of TBTCF and variety fraction of the amino acid residues was analyzed. The binding number of TBTCF depended on the number of positively charged amino acid residues. The other amino acid residues surrounded and seized TBTCF by hydrogen bonds and hydrophobic bonds when the electrostatic attraction pulled TBTCF to link protein. In addition, a novel method named the absorbance ratio difference was established for determination of protein in trace level and was applied with much higher sensitivity than the ordinary method.

Effects of rare earth ions on the conformational stability of anticoagulation factor II from Agkistrodon acutus venom probed by fluorescent spectroscopy.

Anticoagulation factor II (ACF II) isolated from the venom of Agkistrodon acutus is an activated coagulation factor X-binding protein in a Ca(2+)-dependent fashion with marked anticoagulant activity. The equilibrium unfolding of rare earth ions (RE(3+))-reconstituted ACF II in guanidine hydrochloride (GdnHCl) solution was studied by fluorescence. The GdnHCl-induced unfolding of RE(3+) (Nd(3+), Sm(3+), Eu(3+), Gd(3+))-reconstituted ACF II follows a three-state transition with a stable intermediate state. Substitutions of the RE(3+) ions for Ca(2+) in ACF II decrease the conformational stability of its native state but markedly increase the conformational stability of its intermediate state. The free energy change of RE(3+)-ACF II from the intermediate state to denatured state linearly increases with the increase of ionic potentials of bound metal ions (Ca(2+), Nd(3+), Sm(3+), Eu(3+), and Gd(3+)). The refolding of ACF II from the unfolded state to the intermediate state can be induced merely by adding 10 microM RE(3+) ions without changing the concentration of the denaturant. The kinetic results of the RE(3+)-induced refolding provide evidence indicating that the intermediate state of RE(3+)-ACF II consists of at least two refolding phases and that the refolding rate constant values of the faster phase decrease with the increase of the difference between the radii of Ca(2+) and RE(3+), but the refolding rate constant values of the slower phase are similar to each other. The results of this study indicate that the size of metal ion is the major factor responsible for the metal ion-induced conformational stabilization of the native ACF II, while the metal ionic potential plays a predominant role in stabilizing the conformation for the intermediate state.

Metal-ion- and pH-induced conformational changes of acutolysin D from Agkistrodon acutus venom probed by fluorescent spectroscopy.

Acutolysin D isolated from the venom of Agkistrodon acutus is a protein of 44 kDa with marked hemorrhagic and proteolytic activities. The metal-ion- and pH-induced conformational changes of acutolysin D have been studied by following fluorescence and activity measurements. Here we provide evidence for the fact that native holo-acutolysin D adopts two different conformations, native state a, stable in the weak acidic pH range from 5.5 to 7.0 with low activity, and native state b, stable in the weak alkaline pH range from 8.0 to 9.0 with high activity. Holo-acutolysin D has an optimum pH of 9.0 for caseinolytic activity and a maximum fluorescence at pH 9.0. The protein adopts the most stable conformation at pH 9.0. The addition of 1 mM Zn(2+) shifts both the alkali-induced unfolding transition curve and the alkali-induced inactivation curve toward higher pH value but has little effect on the acid-induced unfolding transition curve. No obvious effects on the pH-induced unfolding transition curve and the pH-dependent activity curve have been observed after the addition of 1 mM Ca(2+) to holo-acutolysin D. The results indicate that Zn(2+) is essential for its CA, while Ca(2+) is not essential for its CA. Removal of Ca(2+) and Zn(2+) from the protein enhances its sensitivity to pH and significantly reduces its overall stability during acid-induced denaturation. The kinetic results of the demetalization of holo-acutolysin D show that the demetalization rate constant K(1) for a slower reaction linearly decreases with the pH increase from 5.0 to 9.0, while K(2) for the faster reaction linearly increases with the pH change from 5.0 to 7.0. It is also evident from the present work that the free Zn(2+)-induced inactivation in the pH range from 8.0 to 9.0 should be attributed to the effect of Zn(OH)(2) precipitation on the protein.