Interaction of MgATP2− with DNA: Assessment of metal binding sites and DNA conformations by spectroscopic and thermal denaturation studies

Spectroscopic (IR, UV, CD and fluorescence) and thermal denaturation studies of native calf thymus DNA, DNAMgATP²⁻ and DNAMg²⁺ have been carried out in aqueous KBr medium (introduced by the present authors as a very effective solvent for DNA). The IR data recorded for the systems indicate that MgATP²⁻ binds to the N₇ and C₆O of the guanine residue of DNA forming a five-membered chelate ring. The data also suggest that despite binding to the guanine bases, Mg²⁺ binds more strongly to the phosphate moiety of DNA. Solution CD spectra of DNA, DNAMgATP²⁻ and DNAMg²⁺ indicate that in each case DNA exists in the B conformation. Thin-film CD studies reveal that irrespective of the relative humidity conditions, pure DNA as well as that after interaction with Mg²⁺ show a structural transition B → C, conformationally, although belonging to the B family. A similar study shows that DNA on interaction with MgATP²⁻ assumes a more packed conformation (B)_n giving rise to a ψ⁻ spectrum. Steady-state as well as dynamic fluorimetric studies clearly indicate that MgATP²⁻ does not intercalate between CGGC base pairs. The thermal denaturation studies support the IR data with respect to the metal binding sites and the mode of binding in both cases.     

Identification of spawning sites and natural nurseries of fishes in the upper Paraná River,Brazil

Synopsis We studied the timing of migratory fish spawning in the last dam-free stretch of the upper Paraná River and in Itaipu Reservoir. Eggs were more common in the Amambai and Ivaï Rivers, while larvae predominated in the Paraná River and in Itaipu Reservoir. Both eggs and larvae were more abundant at night. The highest abundance of eggs was in October and that of larvae in November. Migratory species predominated in the Amambai and Paraná Rivers, and non-migratory species in the Ivaï River and Itaipu Reservoir. The predominance of eggs in the upper and middle portions, and larvae in the lower, infer that there are spawning sites in the former and nurseries in the latter. The high nocturnal abundance of eggs is associated with spawning at sunset and that of larvae with feeding, avoidance of predators and nocturnal disorientation. The presence of tributaries such as the Amambai and Ivaí Rivers in the last dam-free stretch of the Paraná River is extremely important to the maintenance of regional fish diversity and fish stocks in both the Paraná River and Itaipu Reservoir.     

Somatosensory impairments in patients with multiple sclerosis: association with dynamic postural control and upper extremity motor function

Abstract

Purpose: We planned this study to bring attention to the somatosensory impairments in patients with multiple sclerosis (PwMS) and to investigate relationship of somatosensory impairments with dynamic postural stability and upper extremity motor function.

Methods: Seven males and 23 females, 30 patients with mean EDSS 2.9 (SD = 1.4), aged between 18 and 65 years (mean = 41.43?±?14.90 years) were included in this clinical study. Light touch sensorial assessment was made with Semmes Weinstein monofilament test and proprioception by distal proprioception test. Hand strength was measured by the Jamar dynamometer, fine motor skill was examined with nine-hole peg test, functional reach test in sitting and standing position was applied. Nottingham Extended Activities of Daily Living Scale (NEADLS) was used to measure everyday activities.

Results: We found a negative and moderate correlation between FRT in standing and light touch of the middle of the heel (right: –0.515), metatarsal bone (right r: 0.453, left r: –0.426), and medial of the foot (right r: –0.462). There was a negative and moderate correlation between NEADLS and light touch of the metatarsal bone (right r: –0.564, left r: –0.472), medial of the foot (right r: –0.531, left r: –0.479), and lateral of the foot (right r: –0.526). We found a positive and moderate correlation between proprioception of the ankle (right r: 0.421 left r: 0.588) and NEADLS.

Conclusions: We found impairment in light touch and proprioception and, associations between sensorial functions and dynamic postural stability in PwMS. Also impaired sensorial functions cause dependent patients in daily living activities. In the assessment of balance and falling risk, independency in daily living activities; foot light touch and proprioception sense should be taken into account, hence it may provide guidance in planning rehabilitation programmes.

Abbreviations: MS: multiple sclerosis; PwMS: patients with multiple sclerosis; VAS: visual analogue scale; FRT: functional reach test; 9-HPT: Nine-hole peg test; EDSS: The Expanded Disability Status Scale; NEADLS: Nottingham Extended Activities of Daily Living Scale     

Does the examiner's experience matter in evaluation of the kinematics of the upper limb?

This study aimed to evaluate test and retest reliability according to examiner experience with the three-dimensional kinematics of the trunk, scapula, and arm segments during flexion and unilateral abduction of the arm. Ten men and 10 women (mean age, 25.1 [1.1] years) participated in this study. Each volunteer participated in six test sessions, four on the first day (two for each examiner) and two on the second day (one for each examiner). A 48-h interval was given between test days. The assessments were made by one examiner with movement analysis experience and a second examiner without experience. For each session (intra-day), the volunteers performed five repetitions of unilateral arm flexions and abductions using their dominant arms. After 1 h, the data were re-collected and all markers were replaced. Data from the trunk, scapula, and arm were analysed at 30°, 60°, 90°, and 120° of arm flexion and abduction using intraclass coefficient correlation, standard error of the measurement, and analysis of variance. The results did not differ between the experienced and inexperienced examiners except for trunk axial rotation at all studied angles and for arm rotation at 120° of abduction. The examiner previously trained in movement analysis marker placement demonstrated the same intra-tester reliability as the inexperienced tester when marker placement accuracy was the variable of interest.     

The association−dissociation behavior of the ApoE proteins: kinetic and equilibrium studies

The apolipoprotein E family consists of three major protein isoforms: apolipoprotein E4 (ApoE4), ApoE3, and ApoE2. The isoforms, which contain 299 residues, differ only by single-amino acid changes, but of the three, only ApoE4 is a risk factor for Alzheimer’s disease. At micromolar concentrations, lipid-free ApoE exists predominantly as tetramers. In more dilute solutions, lower-molecular mass species predominate. Using fluorescence correlation spectroscopy (FCS), intermolecular fluorescence resonance energy transfer (FRET), and sedimentation methods, we found that the association?dissociation reaction of ApoE can be modeled with a monomer?dimer?tetramer process. Equilibrium constants have been determined from the sedimentation data, while the individual rate constants for association and dissociation were determined by measurement of the kinetics of dissociation of ApoE and are in agreement with the equilibrium constants. Dissociation kinetics as measured by intermolecular FRET show two phases reflecting the dissociation of tetramer to dimer and of dimer to monomer, with dissociation from tetramer to dimer being more rapid than the dissociation from dimer to monomer. The rate constants differ for the different ApoE isoforms, showing that the association?dissociation process is isoform specific. Strikingly, the association rate constants are almost 2 orders of magnitude slower than expected for a diffusion-controlled process. Dissociation kinetics were also monitored by tryptophan fluorescence in the presence of acrylamide and the data found to be consistent with the monomer?dimer?tetramer model. The approach combining multiple methods establishes the reaction scheme of ApoE self-association.     

Locating the binding sites of retinol and retinoic acid with milk β-lactoglobulin

β-lactoglobulin (β-LG) is a member of lipocalin superfamily of transporters for small hydrophobic molecules such as retinoids. We located the binding sites of retinol and retinoic acid on β-LG in aqueous solution at physiological conditions, using FTIR, CD, fluorescence spectroscopic methods, and molecular modeling. The retinoid-binding sites and the binding constants as well as the effect of retinol and retinoic acid complexation on protein stability and secondary structure were determined. Structural analysis showed that retinoids bind strongly to β-LG via both hydrophilic and hydrophobic contacts with overall binding constants of K (retinol-) (β) (-LG?)=?6.4 (±?.6)?×?10(6)?M(-1) and K (retinoic acid-) (β) (-LG?)=?3.3 (±?.5)?×?10(6)?M(-1). The number of retinoid molecules bound per protein (n) is 1.1 (±?.2) for retinol and 1.5 (±?.3) for retinoic acid. Molecular modeling showed the participation of several amino acids in the retinoid-protein complexes with the free binding energy of -8.11?kcal/mol for retinol and -7.62?kcal/mol for retinoic acid. Protein conformation was altered with reduction of β-sheet from 59 (free protein) to 52-51% and a major increase in turn structure from 13 (free protein) to 24-22%, in the retinoid-β-LG complexes, indicating a partial protein destabilization.     

Locating the binding sites of retinol and retinoic acid with milk β-lactoglobulin

β-lactoglobulin (β-LG) is a member of lipocalin superfamily of transporters for small hydrophobic molecules such as retinoids. We located the binding sites of retinol and retinoic acid on β-LG in aqueous solution at physiological conditions, using FTIR, CD, fluorescence spectroscopic methods, and molecular modeling. The retinoid-binding sites and the binding constants as well as the effect of retinol and retinoic acid complexation on protein stability and secondary structure were determined. Structural analysis showed that retinoids bind strongly to β-LG via both hydrophilic and hydrophobic contacts with overall binding constants of K _{retinol- β -LG?}=?6.4 (±?.6)?×?10⁶?M^?1 and K _{retinoic acid- β -LG?}=?3.3 (±?.5)?×?10⁶?M^?1. The number of retinoid molecules bound per protein (n) is 1.1 (±?.2) for retinol and 1.5 (±?.3) for retinoic acid. Molecular modeling showed the participation of several amino acids in the retinoid–protein complexes with the free binding energy of ?8.11?kcal/mol for retinol and ?7.62?kcal/mol for retinoic acid. Protein conformation was altered with reduction of β-sheet from 59 (free protein) to 52–51% and a major increase in turn structure from 13 (free protein) to 24–22%, in the retinoid–β-LG complexes, indicating a partial protein destabilization.     

Novel parameters of surface EMG in patients with Parkinson’s disease and healthy young and old controls

The aim of this study was to evaluate a variety of traditional and novel surface electromyography (SEMG) characteristics of biceps brachii muscle in patients with Parkinson’s disease (PD) and compare the results with the healthy old and young control subjects. Furthermore, the aim was to define the optimal biceps brachii loading level that would most likely differentiate patients from controls. The results indicated that such nonlinear SEMG parameters as %Recurrence, %Determinism and SEMG distribution kurtosis, correlation dimension and sample entropy were significantly different between the PD patients and healthy controls. These novel nonlinear parameters, unlike traditional spectral or amplitude parameters, correlated with the Unified Parkinson’s Disease Rating Scale (UPDRS) and finger tapping scores. The most significant between group differences were found in the loading condition where no additional weights were applied in isometric elbow flexion. No major difference of SEMG characteristics was detected between old and young control subjects. In conclusion, the novel SEMG parameters can differentiate the patients with PD from healthy control subjects and these parameters may have potential in the assessment of the severity of PD.     

Measurement of scapular kinematics with the moiré fringe projection technique

The aim of this study was to investigate the applicability of the moiré fringe projection technique (MFPT) to quantify the scapular motions relative to the thorax. This system was composed of a LCD projector, a digital photographic camera, and a microcomputer. To automatically obtain the scapular profiles, the phase shifting method was combined with the MFPT. Four fringes were projected on the scapula and four on the reference planes. By the simple subtraction of the reference values from the scapular phase maps, the map due to the moiré fringes could be digitally obtained. After the phase decoding, the tridimensional (3D) profiles were obtained without prior information about the samples and the calculations of the scapular kinematics were carried out using dedicated software. On average, the movements of lateral rotation ranged from ?1.8±6.1° to ?26.5±3.5°; the protraction from 28.4±4.7° to 27.7±6.8°, and the posterior tilt from ?6.4±7.8° to ?21.7±6.1°, during the arm elevation in the scapular plane performed by six healthy subjects. For the test–retest reliability, the intra-class correlation coefficients ranged from 0.92 to 0.997 and the maximum estimated error was 0.8%. The MFPT allowed the scapular 3D measurements to be obtained in a digital and non-invasive manner. The main advantages compared with other existing systems were its ease in implementation, the use of standard optical components, and its possible clinical applications.     

Assessment of association of estrogen receptor-α gene polymorphism with physical activity and bone metabolism

Genetic and environmental factors have long been suspected to influence on athletic performance and bone metabolism. We have therefore investigated a population-based association study of Korean elite athletes relative to the general population with PvuII polymorphism of the ER-α gene. In addition, interactions between the ER-a genotype distribution and variation of biochemical parameters for bone metabolism during short-duration aerobic performance development were analyzed in a Korean middle-aged women population. The ER-a gene locus was found to be deviation from Hardy-Weinberg equilibrium (p<0.05), but not in the female athlete group. The most common ER-α genotype wasPp in elite athletes (44/72) and controls (74/109), with higher than expected number of heterozygotes (p<0.05). No statistically significant difference in the distribution of ER-a genotype frequency was observed between the elite athletes and control groups. In contrast, almost all biochemical markers studied here appeared to be significantly higher levels after 12-week exercise training compared with baseline in both groups of Korean women, although there was no genotype effect on the change of levels of the bone metabolic parameters. Thus, our data imply that regularly exercise training may affect bone metabolism, but the genetic marker of ER-a gene polymorphism may not be valuable indicator for the development of elite athletes and physical therapy educational program.     

Meta-analysis demonstrates lack of association of the GSK3B −50C/T polymorphism with risk of bipolar disorder

Published data on the association between GSK3B ?50C/T (rs334558) and bipolar disorder (BD) are inconclusive. We performed this meta-analysis to evaluate the relationship of this single-nucleotide polymorphism with the susceptibility, and with the age at onset of BD. A literature search was conducted though PubMed, EMBASE, Web of Science and China National Knowledge Infrastructure databases to identify relevant studies up to February 14, 2014. We identified a total of 6 publications including 1,251 cases and 1,804 controls to investigate the effect of GSK3B ?50C/T on BD risk, and found no significant association in any genetic models (C vs. T: OR = 1.03, 95 % CI: 0.92–1.15; CC vs. TT+TC: OR = 1.04, 95 % CI: 0.84–1.28; TC+CC vs. TT: OR = 1.16, 95 % CI: 0.97–1.39; and CC vs. TC vs. TT: OR = 1.08, 95 % CI: 0.96–1.22). Subgroup analysis by ethnicity did not change the results. The association between GSK3B ?50C/T and age at onset of BD was explored by 6 identified studies with a total of 659 BD type I patients. Similarly, we did not observe significant results in any genetic models (TC+CC vs. TT: SMD = 0.20, 95 % CI: ?0.07 to 0.47; CC vs. TT+TC: SMD = 0.11, 95 % CI: ?0.10 to 0.32; CC vs. TT: SMD = 0.32, 95 % CI: ?0.13 to 0.77). The power analysis and tests for publication bias ensured the reliability of our results. In summary, this meta-analysis suggests that the functional polymorphism ?50C/T within the GSK3B gene promoter is unlikely to relate with BD risk. However, more larger and well-designed studies are still needed to yield a conclusive result on the topic.     

Probing the binding sites of resveratrol,genistein, and curcumin with milk β-lactoglobulin

We determined the binding sites of curcumin (cur), resveratrol (res), and genistein (gen) with milk β-lactoglobulin (β-LG) at physiological conditions. Fourier transform infrared spectroscopy, circular dichroism, and fluorescence spectroscopic methods as well as molecular modeling were used to determine the binding of polyphenol–protein complexes. Structural analysis showed that polyphenols bind β-LG via both hydrophilic and hydrophobic contacts with overall binding constants of K_{curcumin–β-LG}?=?4.4 (±?.4)?×?10⁴ M^?1, K_{resveratrol–β-LG}?=?4.2 (±?.2)?×?10⁴ M^?1, and K_{genistein–β-LG}?=?1.2 (±?.2)?×?10⁴?M^?1. The number of polyphenol molecules bound per protein (n) was 1 (cur), 1.1 (res), and 1 (gen). Molecular modeling showed the participation of several amino acid residues in polyphenol–protein complexation with the free binding energy of ?12.67 (curcumin–β-LG), ?12.60 (resveratrol–β-LG), and ?10.68?kcal/mol (genistein–β-LG). The order of binding was cur?>?res?>?gen. Alteration of the protein conformation was observed in the presence of polyphenol with a major reduction of β-sheet and an increase in turn structure, causing a partial protein structural destabilization. β-LG might act as a carrier to transport polyphenol in vitro.     

GSM 900 MHz radiation inhibits ants' association between food sites and encountered cues

The kinetics of the acquisition and loss of the use of olfactory and visual cues were previously obtained in six experimental colonies of the ant Myrmica sabuleti meinert 1861, under normal conditions. In the present work, the same experiments were conducted on six other naive identical colonies of M. sabuleti, under electromagnetic radiation similar to those surrounding GSM and communication masts. In this situation, no association between food and either olfactory or visual cues occurred. After a recovery period, the ants were able to make such an association but never reached the expected score. Such ants having acquired a weaker olfactory or visual score and still undergoing olfactory or visual training were again submitted to electromagnetic waves. Not only did they lose all that they had memorized, but also they lost it in a few hours instead of in a few days (as under normal conditions when no longer trained). They kept no visual memory at all (instead of keeping 10% of it as they normally do). The impact of GSM 900?MHz radiation was greater on the visual memory than on the olfactory one. These communication waves may have such a disastrous impact on a wide range of insects using olfactory and/or visual memory, i.e., on bees.     

The association of prostaglandins E2 and F2α with erythropoiesis regulatory factors

Prostaglandins E₂ and F_2α have regulatory roles, respectively, in the potentiation and inhibition of erythropoietin. Antisera to PGF_2α and EIF, an erythropoiesis inhibitory factor, can be used to resolve the reason for improvement in erythropoiesis following renal dialysis. Concurrent loss of PGE₂ with Ep during treatment with neuraminidase suggests that PGE₂ is bound to Ep by way of sialic acid.     

Matrix metalloproteinases’s association of polymorphisms of MMP3 and MMP9 with development of genital endometriosis

We present a comparative analysis of the allelic polymorphisms of the matrix metalloproteinase (MMP) gene family, including MMP3 (rs3025058), MMP7 (rs11568818), MMP9 (rs17576, rs2250889), MMP12 (rs2276109), and MMP13 (rs2252070), in patients with external genital endometriosis (EGE) and in a control group of healthy women proven to be free of disease by laparoscopic inspection. We found significant differences in the incidence of particular MMP3 and MMP9 alleles, which substantiate the role of matrix metalloproteinases in EGE pathogenesis. We used the Multifactor Dimensionality Reduction (MDR) analysis to show that 14 allelic combinations of the MMP containing MMP3 (rs3025058) × MMP7 (rs11568818) × MMP9 (rs17576) alleles showed a statistically significant association with an increased risk of EGE, while 10 other combinations correlated with a reduced risk of the disease. MDR analysis produced two statistically significant models for MMP allelic combinations involved in EGE progression, both with 100% penetrance and 83 and 78% accuracy.