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An evaluation of beta-turn prediction methods.

MOTIVATION: beta-turn is an important element of protein structure. In the past three decades, numerous beta-turn prediction methods have been developed based on various strategies. For a detailed discussion about the importance of beta-turns and a systematic introduction of the existing prediction algorithms for beta-turns and their types, please see a recent review (Chou, Analytical Biochemistry, 286, 1-16, 2000). However at present, it is still difficult to say which method is better than the other. This is because of the fact that these methods were developed on different sets of data. Thus, it is important to evaluate the performance of beta-turn prediction methods. RESULTS: We have evaluated the performance of six methods of beta-turn prediction. All the methods have been tested on a set of 426 non-homologous protein chains. It has been observed that the performance of the neural network based method, BTPRED, is significantly better than the statistical methods. One of the reasons for its better performance is that it utilizes the predicted secondary structure information. We have also trained, tested and evaluated the performance of all methods except BTPRED and GORBTURN, on new data set using a 7-fold cross-validation technique. There is a significant improvement in performance of all the methods when secondary structure information is incorporated. Moreover, after incorporating secondary structure information, the Sequence Coupled Model has yielded better results in predicting beta-turns as compared with other methods. In this study, both threshold dependent and independent (ROC) measures have been used for evaluation.

Algorithms↗

Neoplastic transformation and DNA-binding of 4,4'-methylenebis(2-chloroaniline) in SV40-immortalized human uroepithelial cell lines.

The tumorigenic transformation of certain occupationally significant chemicals, such as N-hydroxy-4-4'-methylenebis[2-chloroaniline] (N-OH-MOCA), N-hydroxy-ortho-toluidine (N-OH-OT), 2-phenyl-1,4-benzoquinone (PBQ) and N-hydroxy-4-aminobiphenyl (N-OH-ABP) were tested in vitro using the well established SV40-immortalized human uroepithelial cell line SV-HUC.PC. SV-HUC cells were exposed in vitro to varying concentrations of N-OH-MOCA, N-OH-OT, N-OH-ABP and PBQ that caused approximately 25% and 75% cytotoxicity. The carcinogen treated cells were propagated in culture for about six weeks and subsequently injected subcutaneously into athymic nude mice. Two of the fourteen different groups of SV-HUC.PC treated with different concentrations of N-OH-MOCA, and one of the three groups exposed to N-OH-ABP, formed carcinomas in athymic nude mice. 32P-postlabeling analyses of DNA isolated from SV-HUC.PC after exposure to N-OH-MOCA revealed one major and one minor adduct. The major adduct has been identified as the N-(deoxyadenosin-3',5'-bisphospho-8-yl)-4-amino-3-chlorob enz yl alcohol (pdAp-ACBA) and the minor adduct as N-(deoxyadenosin-3',5'-bisphospho-8-yl)-4-amino-3-chlorot oluene (pdApACT). Furthermore, SV-HUC.PC cytosols catalyzed the binding of N-OH-MOCA to DNA, in the presence of acetyl-CoA, to yield similar adducts. The same adducts were also formed by chemical interaction of N-OH-MOCA with calf thymus DNA, suggesting that the aryl nitrenium ion may be the ultimate reactive species responsible for DNA binding. The tumorigenic activity of N-OH-MOCA in this highly relevant in vitro transformation model, coupled with the findings that SV-HUC.PC cells formed DNA-adducts in vitro and contained enzyme systems that activated N-OH-MOCA to reactive electrophilic species that bound to DNA, strongly suggest that MOCA could be a human bladder carcinogen. These findings are consistent with the International Agency for Research on Cancer's classification of MOCA as a probable human carcinogen.

Animals↗

Coupling with packaging explains apparent nonreciprocality of Chi-stimulated recombination of bacteriophage lambda by RecA and RecBC functions.

Chi (chi, 5'-GCTGGTGG) is a recombinator in RecA- and RecBC-mediated recombination in Escherichia coli. In vegetative recombination between two bacteriophage lambda strains, one with and the other without Chi (a+ chi +b- X a- chi 0b+), the chi-containing recombinant (a- chi +b-) is less abundant than the non-chi-containing recombinant (a+ chi 0b+). Previously this was taken was evidence for nonreciprocality of chi-stimulated exchange. This inequality, however, is now seen to result from an event at cos (lambda's packaging origin) that both activates Chi and initiates DNA packaging. An event at rightward cos leads to activation of leftward chi on the same chromosome for an exchange to its left. From the resulting circulating dimer (--cos-a+- chi 0-b+-cos-a-- chi +-b- --), the cos that activated chi is more likely to be used for rightward packaging initiation than is the cos from the other parent. Consistent with this coupling model is "biased packaging" in lambda carrying two cos sites per monomer genome. When their maturation is dependent on dimerization by chi-stimulated exchange, the phage particles result more often from packaging from the cos that activates chi than from packaging from the other cos. Since Chi activation and packaging can be uncoupled, we infer that some early and reversible step in packaging activates chi. A strong candidate for this step is a double-strand break at cos that provides an oriented entry site for a recombinase.

Bacterial Proteins↗

Promoter-associated displacement of hypermutations.

The distribution of somatic hypermutations around the rearranged V(D)J in antigen-selected B cells is asymmetrical. At the 5' end of the gene a high frequency of mutations does not occur until approximately 200 bp downstream of the V gene promoter in the leader intron. This finding seems inconsistent with recently proposed, transcription-coupled models of hypermutation. Here we describe studies on extensively mutated copies of a kappa light chain transgene which appear to exist as passenger genes for a significant portion of their mutational history. These transgenes contain between one and four in-frame stop codons, and have a ratio of replacement to silent mutations in framework regions that is near random; the ratio in their functional counterparts is clearly non-random. When non-functional passenger and functional transgenes are compared, the patterns of mutation in the leader intron are not significantly different; the frequency 3' is greater in the passenger transgenes. This result indicates that the low level mutational activity immediately 3' of the promoter followed by rapid rise in activity is an intrinsic feature of the mutational process. One inference from this finding is that there is a structural feature in V region DNA or one induced during transcription which is critical to a functioning mutator.

Animals↗

Azidodeoxythymidine and didehydrodeoxythymidine as inhibitors and substrates of the human herpesvirus 8 thymidine kinase.

Human herpesvirus 8 (HHV-8), the aetiological agent of Kaposi's sarcoma (KS), encodes many core genes that have been maintained during evolution of the Herpesviridae. Among these is a thymidine kinase (TK) homologue (ORF21), which has 12% homology to the related TK encoded by herpes simplex virus. We show that the HHV-8 TK is a functional deoxythymidine (dT) kinase, with Michaelis constants (K(m)) for dT and ATP of 18.5 and 6.6 microM, respectively. Using homology modelling coupled with site-directed mutagenesis, we identify Gly265, Asp362 and Phe372 as key amino acid residues involved in the catalytic process. The HHV-8 TK is competitively inhibited by azidodeoxythymidine (zidovudine) and didehydrodeoxythymidine (stavudine) and can also accept these anti-retroviral compounds as substrates. These data have implications for our understanding of changes in AIDS-KS incidence following the clinical licensing of these compounds and in the development of new therapies for KS.

Amino Acid Sequence↗

Gene positional changes relative to the nuclear substructure correlate with the proliferating status of hepatocytes during liver regeneration.

In the interphase nucleus the DNA of higher eukaryotes is organised in loops anchored to a proteinaceous substructure variously named but commonly known as the nuclear matrix. Important processes of nuclear physiology, such as replication, transcription and processing of primary transcripts, occur at macromolecular complexes located at discrete sites upon the nuclear substructure. The topological relationships between gene sequences located in the DNA loops and the nuclear substructure appear to be non-random, thus posing the question of whether such relationships remain invariant or change after the critical nuclear transitions associated with cell proliferation and tissue regeneration in vivo. The hepatocytes are cells that preserve a proliferating capacity that is readily displayed after partial ablation of the liver, leading to liver regeneration in experimental animals such as the rat. Using this animal model coupled to a recently developed PCR-based method for mapping the position of specific DNA sequences relative to the nuclear substructure, we provide evidence that transient changes in the topological relationships between specific genes and the nuclear substructure occur during liver regeneration and that such changes correlate with the actual proliferating status of the cells, thus suggesting that specific transitions in the higher-order DNA structure are characteristic of the quiescent (G0) and replicating (S) phases of the cell cycle in vivo.

Active Transport, Cell Nucleus↗

Medium chain triglyceride in the therapy of gastroesophageal reflux.

Carbohydrate solutions that empty rapidly from the stomach have been shown to produce less gastroesophageal reflux (GER) during the postcibal period than more slowly emptying formulas. These observations have not been extended to complete infant formulas containing fat, carbohydrate, and protein constituents. Medium chain triglyceride has been shown to significantly accelerate gastric emptying compared to long chain triglyceride for preterm infants when investigated in commercial infant formulas. Using a previously reported acidified formula model coupled with esophageal pH probe testing, we have documented in a group of 28 infants that modification of formula by enrichment with medium chain triglyceride produced no improvement in rates of GER during the first 2 h after feeding.

Gastroesophageal Reflux↗

Acute microvascular action of vascular endothelial growth factor in skeletal muscle ischemia/reperfusion injury.

BACKGROUND: The purpose of this study was to investigate the acute action of vascular endothelial growth factor (VEGF) in the microcirculation of skeletal muscle subject to ischemia/reperfusion in vivo and to determine the role of nitric oxide synthase in VEGF-induced microvascular protection. METHODS: A vascular pedicle isolated rat cremaster muscle model coupled with local intraarterial infusion technique was used. Each muscle underwent 4 hours of zero-flow warm ischemia followed by 2 hours of reperfusion. Femoral artery cannulation was performed before reperfusion. The infusate was administered by continuous infusion into the arterial tree of the muscle beginning at 1 minute before reperfusion and at the rate of 0.1 ml/hour throughout the entire reperfusion period. Three groups were designed: (1) the ischemia/reperfusion group, with infusion normal saline; (2) the VEGF plus ische-mia/reperfusion group, with infusion of recombinant human VEGF165 protein; and (3) the L-NA plus VEGF plus ischemia/reperfusion group, with infusion of N-nitro-L-arginine (L-NA; a nonselective nitric oxide synthase antagonist) mixed with VEGF165 protein. After 2 hours of reperfusion, microcirculation measurements including arteriole diameter, capillary perfusion, and endothelium-dependent and endothelium-independent vasodilatation were performed. The muscle was harvested and processed for reverse-transcriptase polymerase chain reaction for measuring eNOS and endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) gene expression. RESULTS: Reperfusion caused significant microvascular alterations including vasoconstriction, poor capillary perfusion, and endothelial dysfunction in the skeletal muscle. These alterations were significantly attenuated by intraarterial infusion of VEGF during reperfusion, but the beneficial effect of VEGF was reduced significantly by coadministration of L-NA. Reverse-transcriptase polymerase chain reaction study revealed that ischemia/reperfusion depressed eNOS mRNA expression but enhanced iNOS mRNA expression. Intraarterial infusion of VEGF during reperfusion amplified mRNA expression of eNOS but not of iNOS. CONCLUSIONS: Local intraarterial infusion of VEGF produced significant microvascular protection from skeletal muscle ischemia/reperfusion injury. The VEGF-induced enhancement of eNOS may play an important mechanistic role.

Animals↗

Equilibrium and stationary nonequilibrium states in a chain of colliding harmonic oscillators

Equilibrium and nonequilibrium properties of a chain of colliding harmonic oscillators (ding-dong model) are investigated. Our chain is modeled as harmonically bounded particles that can only interact with neighboring particles by hard-core interaction. Between the collisions, particles are just independent harmonic oscillators. We are especially interested in the stationary nonequilibrium state of the ding-dong model coupled with two stochastic heat reservoirs (not thermostated) at the ends, whose temperature is different. We check the Gallavotti-Cohen fluctuation theorem [G. Gallavoti and E. G. D. Cohen, Phys. Rev. Lett. 74, 2694 (1995)] and also the Evans-Searles identity [D. Evans and D. Searles, Phys. Rev. E. 50, 1994 (1994)] numerically. It is verified that the former theorem is satisfied for this system, although the system is not a thermostated system.

Journal Article↗

Convective structures in a two-layer gel-liquid excitable medium

The onset of convection due to wave propagation is investigated in the framework of the Belousov-Zhabotinsky reaction. Numerical calculations are based on a three variable Oregonator model coupled with the Navier-Stokes hydrodynamic equations under the Boussineq approximation for a system consisting of two layers, a liquid and a gel, both in close contact through an interface where chemical concentration exchange is allowed. The influence on the formation of convective rolls associated to wave front propagation is studied in terms of the exchange rate through the interface, the liquid layer width, and the coupling strength between the fluid flow and chemical dynamics. Waves are initiated on the surface of the gel and this perturbation is allowed to propagate into the liquid initiating either two counterrotating convective cells (at both sides of the front) or a disordered pattern.

Journal Article↗

Microscopic dynamics of molecular liquids and glasses: role of orientations and translation-rotation coupling.

We investigate the dynamics of a fluid of dipolar hard spheres in its liquid and glassy phases, with emphasis on the microscopic time or frequency regime. This system shows rather different glass transition scenarios related to its rich equilibrium behavior, which ranges from a simple hard sphere fluid to long range ferroelectric orientational order. In the liquid phase close to the ideal glass transition line and in the glassy regime a medium range orientational order occurs leading to a softening of an orientational mode. To investigate the role of this mode we use the molecular mode-coupling equations to calculate the spectra straight phi"lm(q,omega) and chi"lm(q,omega). In the center of mass spectra straight phi"00(q,omega) and chi"00(q,omega) we found, besides a high frequency peak at omega(hf), a peak at omega(op), about one decade below omega(hf) x omega(op) has almost no q dependence and exhibits an "isotope" effect omega(op) proportional to I(-1/2), with I the moment of inertia. We give evidence that the existence of this peak is related to the occurrence of medium range orientational order. It is shown that some of these features also exist for schematic mode coupling models.

Journal Article↗

Molecular dynamics study of cage decay, near constant loss, and crossover to cooperative ion hopping in lithium metasilicate.

Molecular dynamics (MD) simulations of lithium metasilicate (Li2SiO3) in the glassy and supercooled liquid states have been performed to illustrate the decay with time of the cages that confine individual Li+ ions before they hop out to diffuse cooperatively with each other. The self-part of the van Hove function of Li+ ions, G(s)(r,t), is used as an indicator of the cage decay. At 700 K, in the early time regime t of Li+ ions also increases very slowly with time approximately as t(0.1) and has weak temperature dependence. Such can be identified with the near constant loss (NCL) observed in the dielectric response of ionic conductors. At longer times, when the cage decays more rapidly as indicated by the increasing buildup of the intensity of G(s)(r,t) at the distance between Li+ ion sites, broadly crosses over from the NCL regime to another power law t(beta) with beta approximately 0.64 and eventually it becomes t(1.0), corresponding to long-range diffusion. Both t(beta) and t(1.0) terms have strong temperature dependence and they are the analogs of the ac conductivity [sigma(omega) proportional, variant omega(1-beta)] and dc conductivity of hopping ions. The MD results in conjunction with the coupling model support the following proposed interpretation for conductivity relaxation of ionic conductors: (1) the NCL originates from very slow initial decay of the cage with time caused by few independent hops of the ions because t(x1)< and are about the same for t of the fast ions increases much more rapidly for t>t(x2). The self-part of the van Hove function of Li+ reveals that first jumps for some Li+ ions, which are apparently independent free jumps, have taken place before t(x2). While after t(x2) the angle between the first jump and the next is affected by the other ions, again indicating cooperative jump motion. The dynamic properties are analogous to those found in supercooled colloidal particle suspension by confocal microscopy.

Journal Article↗

Long-ranged correlations in sheared fluids.

The presence of long-ranged correlations in a fluid undergoing uniform shear flow is investigated. An exact relation between the density autocorrelation function and the density-momentum correlation function implies that the former must decay more rapidly than 1/r, in contrast to predictions of simple mode-coupling theory. Analytic and numerical evaluation of a nonperturbative mode-coupling model confirms a crossover from 1/r behavior at "small" r to a stronger asymptotic power-law decay. The characteristic length scale is l approximately sqrt[lambda(0)/a], where lambda(0) is the sound damping constant and a is the shear rate.

Journal Article↗

Changes in dynamic crossover with temperature and pressure in glass-forming diethyl phthalate.

Dielectric relaxation measurements have been used to study the crossover in dynamics with temperature and pressure, onset of breakdown of the Debye-Stokes-Einstein law, and the relation between the alpha and the beta relaxations in diethyl phthalate. The measurements made over 10 decades in frequency and a broad range of temperature and pressure enable the dc conductivity and the alpha- and the beta-relaxations to be studied altogether. The isobaric data show that the alpha-relaxation time tau(alpha) has temperature dependence that crosses over from one Vogel-Fulcher-Tammann-Hesse form to another at T(B) approximately 227 K and tau(alpha) approximately 10(-2) s. The dc conductivity sigma exhibits similar crossover at the same T(B). At temperatures above T(B), tau(alpha) and sigma have the same temperature dependence, but below T(B) they become different and the Debye-Stokes-Einstein law breaks down. The breadth of the alpha relaxation is nearly constant for T T(B). The time dependence of tau(beta) is Arrhenius, which when extrapolated to higher temperatures intersects tau(alpha) at T(beta) nearly coincident with T(B). Isothermal measurements at various applied pressures when compared with isobaric data show that the shape of the alpha-relaxation depends only on tau(alpha), and not on the T and P combinations. At a constant temperature, while tau(alpha) increases rapidly with pressure, the beta-relaxation time tau(beta) is insensitive to applied pressure. This behavior is exactly the same as found in 1,1(')-bis (p-methoxyphenyl) cyclohexane. The findings are discussed in the framework of the coupling model.

Journal Article↗

Relation between the activation energy of the Johari-Goldstein beta relaxation and T(g) of glass formers.

For glass-forming substances, we show that the ratio E(beta)/RT(g) can be predicted quantitatively from the coupling model. Here E(beta) is the glassy state activation enthalpy of the Johari-Goldstein beta relaxation, T(g) is the glass transition temperature of the alpha relaxation, and R is the gas constant. The calculated value is in good agreement with the experimental value in many glass formers. The results locate the origin of this cross correlation between E(beta) of the Johari-Goldstein beta relaxation and T(g) of the alpha relaxation, although there are some notable exceptions to this cross correlation.

Journal Article↗

Theory of one-dimensional swelling dynamics of polymer gels under mechanical constraint.

Recently, the swelling kinetics of thin-plate gels with rectangular surfaces under mechanical constraint was experimentally investigated [J. Chem. Phys. 114, 5012 (2001)]]. In this system, the top and bottom surfaces of gels were chemically clamped on the glass plates, and the gels could swell and shrink only along the thickness direction when the osmotic pressure of the solvent is changed. Here, we analyze this process using the linearized stress-diffusion coupling model of gels based on the two fluids model. The result is somewhat unusual in that the time evolution of the thickness is described by a single exponential even though the swelling is governed by the diffusion of solvent. This result and that the characteristic relaxation time depends on the lengths of the rectangular surfaces and not on the thickness of gels agree well with the experiment.

Journal Article↗

Swelling dynamics of constrained thin-plate gels under an external force.

We analyze the swelling kinetics of constrained thin-plate gels using the linearized stress-diffusion coupling model proposed in the previous paper [Phys. Rev. E 69, 041402 (2004)]]. The gel is chemically clamped on the disk-like glass plates at the top and the bottom surfaces and can swell and shrink only along the thickness direction. We analyze how the top plate moves when a force is applied at a certain point on the top plate while the bottom plate is fixed. We predict that (i) the translation and the rotation of the top plate are described by a single exponential relaxation process, that (ii) the rotational relaxation process is three times faster than the translational one, and that (iii) when the force is applied to the edge, the displacement by the rotation is four times larger than that by the translation at the edge point where the force is applied. We also analyze how the gel deforms when it is clamped on the flexible film on which external load is applied.

Journal Article↗

Effects of water on the primary and secondary relaxation of xylitol and sorbitol: implication on the origin of the Johari-Goldstein relaxation.

Dielectric spectroscopy was employed to study the effects of water on the primary alpha -relaxation and the secondary beta -relaxation of xylitol. The measurements were made on anhydrous xylitol and mixtures of xylitol with water with three different water concentrations over a temperature range from 173 K to 293 K. The alpha -relaxation speeds up with increasing concentration of water in xylitol, whereas the rate of the beta -relaxation is essentially unchanged. Some systematic differences in the behavior of alpha -relaxation for anhydrous xylitol and the mixtures were observed. Our findings confirm all the observations of Nozaki et al. [J. Non-Cryst. Solids 307, 349 (2002)]] in sorbitol/water mixtures. Effects of water on both the alpha - and beta -relaxation dynamics in xylitol and sorbitol are explained by using the coupling model.

Journal Article↗