Non-empirical study of the phosphorylation reaction catalyzed by 4-methyl-5-β-hydroxyethylthiazole kinase: relevance of the theory of intermolecular interactions

The subject of this study was an analysis of the role of active site residues in the phosphoryl transfer reaction catalyzed by 4-methyl-5-β-hydroxyethylthiazole kinase (ThiK). The ThiK-catalyzed reaction is of special interest due to the lack of a highly conserved aspartate residue serving as a catalytic base. ONIOM(B3LYP:PM3) models of stationary points along the reaction pathway consisted of reactants, two magnesium ions and several highly conserved ThiK active site residues. The results indicate that an S_N2-like mechanism of ThiK, with γ-phosphate acting as an alcohol-activating base is reasonable. Geometries of substrates, transition state and products were utilized in the non-empirical analysis of the physical nature of catalytic interactions taking place in the ThiK active site. The role of particular residues was investigated in terms of their ability to preferentially stabilize the transition state relative to substrates (differential transition state stabilization, DTSS) or products (differential product stabilization, DPS). It seems that Mg2, Glu126 and Cys198 play a major catalytic role, whereas Mg1 and the same Cys198 are responsible for product release. It is remarkable that no dominant role of an electrostatic term in the interactions involved in catalytic activity is observed for product release. Determination of catalytic fields expressing differential electrostatic potential of the transition state with respect to substrates revealed the optimal electrostatic features of an ideal catalyst for the studied reaction. The predicted catalytic environment is in agreement with experimental data showing increased catalytic activity of ThiK upon mutation of Cys198 to aspartate. Figure Catalytic fields for ThiK-catalyzed reaction juxtaposed with the positions of active site residues of a model system. Magnesium ions are considered part of the transition state/reactants. The surface of constant electronic density is colored according to differential electrostatic potential of transition state with respect to reactants. The sign of the differential potential reflects the electrostatic properties of a complementary molecular environment. Red (green) color denotes regions where a negative (positive) charge would be optimal for catalytic activity     

Pulsed electromagnetic fields regulate osteocyte apoptosis,RANKL/OPG expression,and its control of osteoclastogenesis depending on the presence of primary cilia

Growing evidence has shown that pulsed electromagnetic fields (PEMF) can modulate bone metabolism in vivo and regulate the activities of osteoblasts and osteoclasts in vitro. Osteocytes, accounting for 95% of bone cells, act as the major mechanosensors in bone for transducing external mechanical signals and producing cytokines to regulate osteoblastic and osteoclastic activities. Targeting osteocytic signaling pathways is becoming an emerging therapeutic strategy for bone diseases. We herein systematically investigated the changes of osteocyte behaviors, functions, and its regulation on osteoclastogenesis in response to PEMF. The osteocyte-like MLO-Y4 cells were exposed to 15 Hz PEMF stimulation with different intensities (0, 5, and 30 Gauss [G]) for 2 hr. We found that the cell apoptosis and cytoskeleton organization of osteocytes were regulated by PEMF with an intensity-dependent manner. Moreover, PEMF exposure with 5 G significantly inhibited apoptosis-related gene expression and also suppressed the gene and protein expression of the receptor activator of nuclear factor κB ligand/osteoprotegerin (RANKL/OPG) ratio in MLO-Y4 cells. The formation, maturation, and osteoclastic bone-resorption capability of in vitro osteoclasts were significantly suppressed after treated with the conditioned medium from PEMF-exposed (5 G) osteocytes. Our results also revealed that the inhibition of osteoclastic formation, maturation, and bone-resorption capability induced by the conditioned medium from 5 G PEMF-exposed osteocytes was significantly attenuated after abrogating primary cilia in osteocytes using the polaris siRNA transfection. Together, our findings highlight that PEMF with 5 G can inhibit cellular apoptosis, modulate cytoskeletal distribution, and decrease RANKL/OPG expression in osteocytes, and also inhibit osteocyte-mediated osteoclastogenesis, which requires the existence of primary cilia in osteocytes. This study enriches our basic knowledge for further understanding the biological behaviors of osteocytes and is also helpful for providing a more comprehensive mechanistic understanding of the effect of electromagnetic stimulation on bone and relevant skeletal diseases (e.g., bone fracture and osteoporosis).     

Evaluation of the neuroprotective effects of electromagnetic fields and coenzyme Q10 on hippocampal injury in mouse

Electromagnetic fields (EMFs) are reported to interfere with chemical reactions involving free radical production. Coenzyme Q₁₀ (CoQ10) is a strong antioxidant with some neuroprotective activities. The purpose of this study was to examine and compare the neuroprotective effects of EMF and CoQ10 in a mouse model of hippocampal injury. Hippocampal injury was induced in mature female mice (25–30 g), using an intraperitoneal injection of trimethyltin hydroxide (TMT; 2.5 mg/kg). The experimental groups were exposed to EMF at a frequency of 50 Hz and intensity of 5.9 mT for 7 hr daily over 1 week or treated with CoQ10 (10 mg/kg) for 2 weeks following TMT injection. A Morris water maze apparatus was used to assess learning and spatial memory. Nissl staining and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) tests were also performed for the histopathological analysis of the hippocampus. Antiapoptotic genes were studied, using the Western blot technique. The water maze test showed memory improvement following treatment with CoQ10 and coadministration of CoQ10 + EMF. The Nissl staining and TUNEL tests indicated a decline in necrotic and apoptotic cell count following treatment with CoQ10 and coadministration of CoQ10 + EMF. The Western blot study indicated the upregulation of antiapoptotic genes in treatment with CoQ10, as well as coadministration. Also, treatment with EMF had no significant effects on reducing damage induced by TMT in the hippocampus. According to the results, EMF had no significant neuroprotective effects in comparison with CoQ10 on hippocampal injury in mice. Nevertheless, coadministration of EMF and CoQ10 could improve the neuroprotective effects of CoQ10.     

鄱阳湖越冬白鹤在农业用地的食物组成

白鹤(Grus leucogeranus)是IUCN极危物种,全球仅存3 500～4 000只,其中约98%的个体在鄱阳湖越冬。以前白鹤在鄱阳湖主要在浅水生境挖掘苦草(Vallisneria spp.)的冬芽为食。然而,近年来农业用地已成为白鹤的重要觅食地。目前对于白鹤在农业用地的食物组成尚不清楚。本研究采用粪便显微镜检法对2017年11月至2018年4月采集的70份白鹤粪便样品进行分析。结果表明,白鹤的食物来源于10科15种植物,其中,水稻(Oryza sativa)、莲藕(Nelumbo nucifera)和紫云英(Astragalus sinicus)为最主要的食物,这3种植物在检测到的食物中的相对密度分别为34.34%、22.99%和10.61%,而白鹤的传统食物苦草冬芽所占的比例极低(2.05%)。由此可见,白鹤的食物组成已经发生变化,农作物已成为白鹤的重要食物来源。白鹤在农业用地中觅食使该物种的保护面临一系列问题,尤其是高强度的人为干扰,对此,提出了相应的保护措施,希望为白鹤提供安全、稳定的觅食环境。     

Early‐Life Exposure to Pulsed LTE Radiofrequency Fields Causes Persistent Changes in Activity and Behavior in C57BL/6 J Mice

Despite much research, gaps remain in knowledge about the potential health effects of exposure to radiofrequency (RF) fields. This study investigated the effects of early‐life exposure to pulsed long term evolution (LTE) 1,846 MHz downlink signals on innate mouse behavior. Animals were exposed for 30 min/day, 5 days/week at a whole‐body average specific energy absorption rate (SAR) of 0.5 or 1 W/kg from late pregnancy (gestation day 13.5) to weaning (postnatal day 21). A behavioral tracking system measured locomotor, drinking, and feeding behavior in the home cage from 12 to 28 weeks of age. The exposure caused significant effects on both appetitive behaviors and activity of offspring that depended on the SAR. Compared with sham‐exposed controls, exposure at 0.5 W/kg significantly decreased drinking frequency (P ≤ 0.000) and significantly decreased distance moved (P ≤ 0.001). In contrast, exposure at 1 W/kg significantly increased drinking frequency (P ≤ 0.001) and significantly increased moving duration (P ≤ 0.005). In the absence of other plausible explanations, it is concluded that repeated exposure to low‐level RF fields in early life may have a persistent and long‐term effect on adult behavior. Bioelectromagnetics. 2019;40:498–511. © 2019 The Authors. Bioelectromagnetics Published by Wiley Periodicals, Inc.     

Complications of nonionizing radiofrequency on divided attention

Exposure to electromagnetic fields is considered as a potential hazard for biological systems. The objective of our investigation is the study of probable consequences of radiofrequency electromagnetic fields from Wi-Fi router devices on the short-term memory, and attention's levels. A population consisting of 312 female college students (14 to 17 years old) was elected by cluster random sampling. Teenagers were divided into two groups of control group (Wi-Fi nonusers; n = 138), and experiment group (Wi-Fi users; n = 174). Both groups have been examined using short-term memory tests; selective attention, and also divided attention tests. According to the results, there was no significant difference between using Wi-Fi router devices on levels of selective attentions and short-term memory of the sample students with the control group. However, analyses revealed that there is a significant correlation between the use of Wi-Fi routers and declining levels of divided attentions. Our investigation has demonstrated the adverse consequences of 2.4-2.48 GHz radiofrequency electromagnetic fields of Wi-Fi router devices on divided attention levels of female university students that should be mentioned as a technological risk factor and taken into account by healthcare organizations.     

Evidence for a slow time-scale of interaction for magnetic fields inhibiting tamoxifen’s antiproliferative action in human breast cancer cells

One critical biophysical feature of environmental-level magnetic field (MF) interactions with biological systems is the time-scale of interaction. A recently proposed fast/slow hypothesis states that a fast mechanism can only sense the instantaneous absolute value of the MF, and that a slow mechanism is potentially capable of sensing features such as frequency and relative orientation and magnitude of the field components. Here we applied the fast/slow hypothesis to a breast cancer model system: A 1.2 μT(rms), 60-Hz field inhibits tamoxifen’s (TAM’s) cytostatic action in MCF-7 cells via a MF interaction. We measured the growth of MCF-7 cells treated with TAM over 7 d, within different MFs: a sinusoidal, 60-Hz, 0.2-μT(rms) field; a sinusoidal, 60-Hz, 1.2-μT(rms) field; and a full-wave rectified version of the 1.2-μT(rms) sinusoidal field. A fast mechanism should not be able to distinguish between the latter two exposures. We observe that the rectified 1.2-μT field does not inhibit TAM’s action, but that the 1.2-μT sinusoidal field does. Therefore, the 1.2-μT MF inhibition of TAM’s cytostatic action operates via a relatively slow mechanism, and we predict that there exists a biologically dynamic complex capable of sensing a 1.2-μT, 60-Hz sinusoidal MF with an intrinsic time-scale of 17 ms or longer, the period of the 60-Hz applied field.     

Influence of non-bonded parameters on the quality of NMR structures: A new force field for NMR structure calculation

The effects of different non-bonded parameters of force fields for NMR structure calculation on the quality of the resulting NMR solution structures were investigated using Interleukin 4 as a model system. NMR structure ensembles were calculated with an ab initio protocol using torsion angle dynamics. The calculations were repeated with five different non-bonded energy functions and parameters. The resulting ensembles were compared with the available X-ray structures, and their quality was assessed with common structure validation programs. In addition, the impact of torsion angle restraints and dihedral energy terms for the sidechains and the backbone was studied. The further improvement of the quality by refinement in explicit solvent was demonstrated. The optimal parameters, including those necessary for water refinement, are available in the new version of the PARALLHDG force field.     

Separation of Spatio-Temporal Receptive Fields into Sums of Gaussian Components

Visual cortical simple cells have been experimentally shown to reveal non-trivial spatio-temporal orientation tuning functions comprising different phases of specifically tuned enhanced and suppressed activity. A recently developed analytical method based on nonlinear neural field models suggests that such space-time responses should be approximately separable into a sum of temporally amplitude modulated Gaussian spatial components. In the present work, we investigate this possibility by means of numerical fits of sums of Gaussians to response functions observed in experiments and computer simulations. Because the theory relates each single component to a particular connectivity kernel between the underlying cell classes shaping the response, the relative contribution of feedforward and cortex-intrinsical excitatory and inhibitory feedback mechanisms to single cell tuning can be approached and quantified in experimental data.