Extraction of mandibular third molars: relationship of preoperative anxiety with body mass index, serum high-sensitivity C-reactive protein levels, and visual analog scale scores and predictors of postoperative complications.

Objectives: Patients undergoing oral surgery exhibit high anxiety, which may elevate their cortisol levels and affect postoperative recovery. Overweight patients are often encountered in the dental clinic due to the increasing prevalence of overweight. We aimed to investigate the relationships between preoperatively assessed body mass index (BMI), serum cortisol and high-sensitivity C-reactive protein (hs-CRP) levels, and visual analog scale (VAS) scores and preoperative anxiety in patients undergoing mandibular third molar (MM3) extraction and to identify predictors of postoperative complications. Patients and Methods: We analyzed 43 patients (age, 20-42 years) undergoing MM3 extraction. At the first visit, patients completed the Modified Dental Anxiety Scale (MDAS) and Amsterdam Preoperative Anxiety and Information Scale (APAIS) questionnaires. Their BMI and VAS scores were also calculated. The participants underwent blood tests 1 hour before MM3 extraction. On the first postoperative day, the participants' VAS scores and serum hs-CRP levels were reevaluated. Results: We found that BMI was significantly correlated with preoperative VAS scores. Further, BMI and preoperative hs-CRP levels were significantly correlated among women and patients undergoing extractions of fully impacted MM3s. No correlations were found between serum cortisol and other variables. The preoperative MDAS and VAS scores were significantly positively correlated, especially among patients undergoing extractions of fully impacted MM3s. Multiple linear regression showed that BMI and the eruption status of the MM3 were significant predictors of postoperative hs- CRP levels and VAS scores, respectively. Conclusion: In MM3 removals, patients with higher BMI showed elevated hs-CRP and higher VAS scores before surgery. Patients with higher anxiety among those undergoing extractions of fully impacted MM3s showed higher preoperative VAS scores. The two main predictors of postoperative complications were BMI and MM3 eruption status.

[Spatiotemporal Evolution Characteristics of PM2.5-O3 Compound Pollution in Chinese Cities from 2015 to 2020].

Based on the monitoring data of PM2.5 and O3 concentrations in 333 cities in China from 2015 to 2020, using spatial clustering, trend analysis, and the geographical gravity model, this study quantitatively analyzed the characteristics of PM2.5-O3 compound pollution concentrations and its spatiotemporal dynamic evolution pattern in major cities in China. The results showed that:① there was a synergistic change in PM2.5 and O3 concentrations. When ρ(PM2.5_mean) ≤ 85 µg·m-3, for every 10 µg·m-3 increase in ρ(PM2.5_mean), the peak of the mean value of ρ(O3_perc90) increased by 9.98 µg·m-3. When ρ(PM2.5_mean) exceeded the national Grade II standards of (35±10) µg·m-3, the peak of the mean value of ρ(O3_perc90) increased the fastest, with an average growth rate of 11.81%. In the past six years, on average, 74.97% of Chinese cities with compound pollution had a ρ(PM2.5_mean) in the range of 45 to 85 µg·m-3. When ρ(PM2.5_mean)>85 µg·m-3, the mean value of ρ(O3_perc90) showed a significant decreased trend. ② The spatial clustering pattern of PM2.5 and O3 concentrations in Chinese cities was similar, and hot spots of the six-year mean values of ρ(PM2.5_mean) and ρ(O3_perc90) were distributed in the Beijing-Tianjin-Hebei urban agglomeration and other cities in the Shanxi, Henan, and Anhui provinces. ③ The number of cities with PM2.5-O3 compound pollution showed an interannual variation trend of increasing first (2015-2018) and then decreasing (2018-2020) and a seasonal trend of gradually decreasing from spring to winter. Further, the compound pollution phenomenon mainly occurred in the warm season (April to October). ④ The spatial distribution of PM2.5-O3 compound polluted cities was changing from dispersion to aggregation. From 2015 to 2017, the compound polluted areas spread from the eastern coastal areas to the central and western regions of China, and by 2017, a large-scale polluted area centered on the Beijing-Tianjin-Hebei urban agglomeration, the Central Plains urban agglomeration, and surrounding areas was formed. ⑤ The migration directions of PM2.5 and O3 concentration centers were similar, and there were obvious trends of moving westward and northward. The problem of high-concentration compound pollution was concentrated and highlighted in cities in central and northern China. In addition, since 2017, the distance between the centers of gravity of PM2.5 and O3 concentrations in the compound polluted areas had been significantly reduced, with a reduction of nearly 50%.

Effect of aromatherapy on quality of life in maintenance hemodialysis patients: a systematic review and meta-analysis.

Objective: Aromatherapy has been used for patients on maintenance hemodialysis (MHD), but the outcomes are still controversial. Thus, we conducted this study to systematically evaluate the effect of aromatherapy on the quality of life of patients on MHD.Methods: We searched the PubMed, Embays, Scopus, Web of Science, and CNKI databases for randomized controlled trials that evaluated the use of aromatherapy in dialysis patients and reported at least one outcome of interest.Results: Twenty-two relevant studies were included in the meta-analysis. The meta-analysis revealed that aromatherapy significantly increased subjective sleep quality (a lower score indicates better sleep quality) [standardized mean difference (SMD) = -1.52, 95% CI (-2.38, -0.67), p < 0.01] and reduced fatigue [SMD = -1.14, 95% CI (-1.95, -0.33), p = 0.01], anxiety [SMD = -1.38, 95% CI (-2.09, -0.67), p < 0.01], symptoms of restless legs syndrome [RLS; SMD = -1.71, 95% CI (-2.09, -1.33), p < 0.01], and arteriovenous fistula puncture pain [SMD= -1.56, 95% CI (-2.60, -0.52), p < 0.01].Conclusions: Aromatherapy may be used as a novel complementary and alternative therapy to improve sleep quality and reduce fatigue, anxiety, symptoms of RLS, and arteriovenous fistula puncture pain in patients on MHD.

Ozone exposure and health risks of different age structures in major urban agglomerations in People's Republic of China from 2013 to 2018.

High concentration of surface ozone (O3) will cause health risks to people. In order to analyze the spatiotemporal characteristics of O3 and assess O3 exposure and health risks for different age groups in China, we applied multiple methods including standard deviation ellipse, spatial autocorrelation, and exposure-response functions. Results show that O3 concentrations increased in 64.5% of areas in China from 2013 to 2018. The central plain urban agglomeration (CPU), Beijing-Tianjin-Hebei (BTH), and Yangtze River Delta (YRD) witnessed the greatest incremental rates of O3 by 16.7%, 14.3%, and 13.1%. Spatially, the trend of O3 shows a significant positive autocorrelation, and high trend values primarily in central and east China. The proportion of the total population exposed to high O3 (above 160 µg/m3) increased annually. Compared to 2013, the proportion of the young, adult, and old populations exposed to high O3 increased to different extents in 2018 by 26.8%, 29.6%, and 27.2%, respectively. The extent of population exposure risk areas in China expanded in size, particularly in north and east China. The total premature respiratory mortalities attributable to long-term O3 exposure in six urban agglomerations were about 177,000 in 2018 which has increased by 16.4% compared to that in 2013. Among different age groups, old people are more vulnerable to O3 pollution, so we need to strengthen their relevant health protection of them.

Characterization of a novel Hoxa5eGFP mouse line.

BACKGROUND: Hox genes encode transcription factors that are important for establishing the body plan. Hoxa5 is a member of the mammalian Hox5 paralogous group that regulates the patterning and morphology of the cervical-thoracic region of the axial skeleton. Hoxa5 also plays crucial functions in lung morphogenesis. RESULTS: We generated a Hoxa5eGFP reporter mouse line using CRISPR technology, allowing real-time visualization of Hoxa5 expression. Hoxa5eGFP recapitulates reported embryonic Hoxa5 mRNA expression patterns. Specifically, Hoxa5eGFP can be visualized in the developing mouse neural tube, somites, lung, diaphragm, foregut, and midgut, among other organs. In the stomach, posteriorly biased Hoxa5eGFP expression correlates with a drastic morphological reduction of the corpus in Hox5 paralogous mutants. Expression of Hoxa5eGFP in the lung continues in all lung fibroblast populations through postnatal and adult stages. CONCLUSIONS: We identified cell types that express Hoxa5 in postnatal and adult mouse lungs, including various fibroblasts and vascular endothelial cells. This reporter line will be a powerful tool for studies of the function of Hoxa5 during mouse development, homeostasis, and disease processes.

Extraction of mandibular third molars: relationship of preoperative anxiety with body mass index, serum high-sensitivity C-reactive protein levels, and visual analog scale scores and predictors of postoperative complications

Objectives@#Patients undergoing oral surgery exhibit high anxiety, which may elevate their cortisol levels and affect postoperative recovery. Overweight patients are often encountered in the dental clinic due to the increasing prevalence of overweight. We aimed to investigate the relationships between preoperatively assessed body mass index (BMI), serum cortisol and high-sensitivity C-reactive protein (hs-CRP) levels, and visual analog scale (VAS) scores and preoperative anxiety in patients undergoing mandibular third molar (MM3) extraction and to identify predictors of postoperative complications. @*Patients and Methods@#We analyzed 43 patients (age, 20-42 years) undergoing MM3 extraction. At the first visit, patients completed the Modified Dental Anxiety Scale (MDAS) and Amsterdam Preoperative Anxiety and Information Scale (APAIS) questionnaires. Their BMI and VAS scores were also calculated. The participants underwent blood tests 1 hour before MM3 extraction. On the first postoperative day, the participants’ VAS scores and serum hs-CRP levels were reevaluated. @*Results@#We found that BMI was significantly correlated with preoperative VAS scores. Further, BMI and preoperative hs-CRP levels were significantly correlated among women and patients undergoing extractions of fully impacted MM3s. No correlations were found between serum cortisol and other variables. The preoperative MDAS and VAS scores were significantly positively correlated, especially among patients undergoing extractions of fully impacted MM3s. Multiple linear regression showed that BMI and the eruption status of the MM3 were significant predictors of postoperative hsCRP levels and VAS scores, respectively. @*Conclusion@#In MM3 removals, patients with higher BMI showed elevated hs-CRP and higher VAS scores before surgery. Patients with higher anxiety among those undergoing extractions of fully impacted MM3s showed higher preoperative VAS scores. The two main predictors of postoperative complications were BMI and MM3 eruption status.

The Lung Elastin Matrix Undergoes Rapid Degradation Upon Adult Loss of Hox5 Function.

Hox genes encode transcription factors that are critical for embryonic skeletal patterning and organogenesis. The Hoxa5, Hoxb5, and Hoxc5 paralogs are expressed in the lung mesenchyme and function redundantly during embryonic lung development. Conditional loss-of-function of these genes during postnatal stages leads to severe defects in alveologenesis, specifically in the generation of the elastin network, and animals display bronchopulmonary dysplasia (BPD) or BPD-like phenotype. Here we show the surprising results that mesenchyme-specific loss of Hox5 function at adult stages leads to rapid disruption of the mature elastin matrix, alveolar enlargement, and an emphysema-like phenotype. As the elastin matrix of the lung is considered highly stable, adult disruption of the matrix was not predicted. Just 2 weeks after deletion, adult Hox5 mutant animals show significant increases in alveolar space and changes in pulmonary function, including reduced elastance and increased compliance. Examination of the extracellular matrix (ECM) of adult Tbx4rtTA; TetOCre; Hox5a f a f bbcc lungs demonstrates a disruption of the elastin network although the underlying fibronectin, interstitial collagen and basement membrane appear unaffected. An influx of macrophages and increased matrix metalloproteinase 12 (MMP12) are observed in the distal lung 3 days after Hox5 deletion. In culture, fibroblasts from Hox5 mutant lungs exhibit reduced adhesion. These findings establish a novel role for Hox5 transcription factors as critical regulators of lung fibroblasts at adult homeostasis.

[Spatial Variation of Surface Ozone Concentration During the Warm Season and Its Meteorological Driving Factors in China].

The concentration of surface ozone (O3) in China increased consistently from 2015 to 2018, and became an important air pollutant, followed by particulate matter. This study uses real-time O3 and meteorological data, obtained in 337 cities in China during the warm seasons (April to September) of 2015 to 2018, to determine the spatial variation of surface O3 and its meteorological driving factors in major cities in China, via trend analysis, spatial autocorrelation, hotspot analysis, and multi-scale geographically weighted regression (MGWR) modeling. The results show that: ① during the warm season, O3 concentrations showed a significant growth trend (P<0.05), with an average growth rate of 0.28 µg·(m3·a)-1, while more than 55% of urban O3 concentrations increased by 0.50 µg·m-3 annually. ② There were significant regional differences in O3 concentration. High values (>60 µg·m-3) were distributed over east China, north China, central China, and northwest China, while low values (<20 µg·m-3) were distributed over south China and southwest China. ③ The spatial agglomeration of O3 concentration has been enhanced year by year, with hotspots mainly distributed over east China and central China. In contrast, there are cold spots in northeast China, southwest China, and southern China. ④Analysis of the MGWR model indicated that temperature, wind speed, cloud coverage, and precipitation all have a significant effect on the distribution of O3, although there are also discrepancies in driving factor priorities between the different regions. Temperature was the main meteorological driving factor of O3 variation during the warm season in China, and its impact on O3 concentration was significantly higher in north China, northwest China, and northeast China than in other regions; overall, there was a significant positive correlation between O3 concentration and temperature, except in Guangxi, Yunnan, and Jiangxi. O3 concentration was negatively correlated with wind speed in most regions of south China, east China, and central China, and positively correlated with wind speed in north China and northeast China. O3 concentration was significantly negatively correlated with cloud cover, except in Liaoning, Shandong, Hebei, Gansu, Guangdong, and some areas in southwest China. O3 concentration was significantly negatively correlated with precipitation, except in the northwest and southwest regions.

Global, continental, and national variation in PM2.5, O3, and NO2 concentrations during the early 2020 COVID-19 lockdown.

Lockdowns implemented in response to COVID-19 have caused an unprecedented reduction in global economic and transport activity. In this study, variation in the concentration of health-threatening air pollutants (PM2.5, NO2, and O3) pre- and post-lockdown was investigated at global, continental, and national scales. We analyzed ground-based data from >10,000 monitoring stations in 380 cities across the globe. Global-scale results during lockdown (March to May 2020) showed that concentrations of PM2.5 and NO2 decreased by 16.1% and 45.8%, respectively, compared to the baseline period (2015-2019). However, O3 concentration increased by 5.4%. At the continental scale, concentrations of PM2.5 and NO2 substantially dropped in 2020 across all continents during lockdown compared to the baseline, with a maximum reduction of 20.4% for PM2.5 in East Asia and 42.5% for NO2 in Europe. The maximum reduction in O3 was observed in North America (7.8%), followed by Asia (0.7%), while small increases were found in other continents. At the national scale, PM2.5 and NO2 concentrations decreased significantly during lockdown, but O3 concentration showed varying patterns among countries. We found maximum reductions of 50.8% for PM2.5 in India and 103.5% for NO2 in Spain. The maximum reduction in O3 (22.5%) was found in India. Improvements in air quality were temporary as pollution levels increased in cities since lockdowns were lifted. We posit that these unprecedented changes in air pollutants were mainly attributable to reductions in traffic and industrial activities. Column reductions could also be explained by meteorological variability and a decline in emissions caused by environmental policy regulations. Our results have implications for the continued implementation of strict air quality policies and emission control strategies to improve environmental and human health.

Segmentation Method for Ship-Radiated Noise Using the Generalized Likelihood Ratio Test on an Ordinal Pattern Distribution.

Due to the diversity of ship-radiated noise (SRN), audio segmentation is an essential procedure in the ship statuses/categories identification. However, the existing segmentation methods are not suitable for the SRN because of the lack of prior knowledge. In this paper, by a generalized likelihood ratio (GLR) test on the ordinal pattern distribution (OPD), we proposed a segmentation criterion and introduce it into single change-point detection (SCPD) and multiple change-points detection (MCPD) for SRN. The proposed method is free from the acoustic feature extraction and the corresponding probability distribution estimation. In addition, according to the sequential structure of ordinal patterns, the OPD is efficiently estimated on a series of analysis windows. By comparison with the Bayesian Information Criterion (BIC) based segmentation method, we evaluate the performance of the proposed method on both synthetic signals and real-world SRN. The segmentation results on synthetic signals show that the proposed method estimates the number and location of the change-points more accurately. The classification results on real-world SRN show that our method obtains more distinguishable segments, which verifies its effectiveness in SRN segmentation.

Inhibition kinetics of acetosyringone on xylanase in hydrolysis of hemicellulose.

Many phenolic compounds, derived from lignin during the pretreatment of lignocellulosic biomass, could obviously inhibit the activity of cellulolytic and hemicellulolytic enzymes. Acetosyringone (AS) is one of the phenolic compounds produced from lignin degradation. In this study, we investigated the inhibitory effects of AS on xylanase activity through kinetic experiments. The results showed that AS could obviously inhibit the activity of xylanase in a reversible and noncompetitive binding manner (up to 50% activity loss). Inhibitory kinetics and constants of xylanase on AS were conducted by the HCH-1 model (ß = 0.0090 ± 0.0009 mM-1). Furthermore, intrinsic and 8-anilino-1-naphthalenesulfonic (ANS)-binding fluorescence results showed that the tertiary structure of AS-mediated xylanase was altered. These findings provide new insights into the role of AS in xylanase activity. Our results also suggest that AS was an inhibitor of xylanase and targeting AS was a potential strategy to increase xylose production.

Co-targeting CK2α and YBX1 suppresses tumor progression by coordinated inhibition of the PI3K/AKT signaling pathway.

Protein kinase CK2 alpha (CK2α) is involved in the development of multiple malignancies. Overexpression of Y-box binding protein 1 (YBX1) is related to tumor proliferation, drug resistance, and poor prognosis. Studies have demonstrated that both CK2 and YBX1 could regulate the PI3K/AKT pathway. In addition, we predicted that CK2 might be the upstream kinase of YBX1 through the Human Protein Reference Database (HPRD). Herein, we hypothesize that CK2 may interact with YBX1 and they regulate the PI3K/AKT signaling pathway together. Expressions of CK2α and YBX1 in cancer cell lines were evaluated by immunoblotting. The results showed that CK2α could regulate the expression of YBX1 at the transcriptional level, which is dependent on its enzymatic activity. Synergistic effects of PI3K/AKT pathway inactivation could be observed through combined inhibition of CK2α and YBX1, and YBX1 was required for CK2α-induced PI3K/AKT pathway activation. Further results demonstrated that CK2α could interact with YBX1 and PI3K/AKT antagonist decreased cell resistance to doxorubicin induced by co-activation of CK2α and YBX1. These results indicated that combined inhibition of CK2α and YBX1 showed synergistic effects in inactivating the PI3K/AKT signaling pathway and may be one of the mechanisms involved in tumor growth and migration.

Baicalin Regulates Proliferation, Apoptosis, Migration, and Invasion in Mesothelioma.

BACKGROUND Baicalin, one of the main bioactive components extracted from the traditional Chinese medicine baical Skullcap root, has an anti-tumor activity which had been studied in several cancers. However, its role in human mesothelioma remains unknown. In this study, we investigated the anti-tumor mechanisms of baicalin in the mesothelioma cell line MESO924. MATERIAL AND METHODS Effects of baicalin on mesothelioma were assessed by measuring cell viability, apoptosis, migration, invasion, inactivation of signaling intermediates, and cell-cycle alterations. RESULTS Baicalin inhibited the proliferation, migration, and invasion of human mesothelioma cells and increased their apoptosis, all in a dose-dependent manner. Specifically, baicalin decreased the expression of p-EGFR, p-AKT, p-MAPK, p-S6, Bcl-2, and VEGF and increased the expression of Bax in mesothelioma cells. The suppressed mesothelioma cellular proliferation is due to the arrest of the S cell cycle by baicalin. Inhibition of the PI3K/AKT/mTOR signaling pathway by a PI3K/AKT/mTOR inhibitor augmented the anti-proliferation effects induced by baicalin. In addition, baicalin increased the sensitivity of MESO924 to the chemotherapeutic drugs doxorubicin, cisplatin, and pemetrexed. CONCLUSIONS These results highlight the roles of baicalin in inhibiting cell growth, migration, and invasion of mesothelioma cells while increasing apoptosis and sensitizing cells to chemotherapeutic agents through the PI3K/AKT/mTOR signaling pathway, which indicates that baicalin could be a useful drug for mesothelioma therapy.

Application of Differential Entropy in Characterizing the Deformation Inhomogeneity and Life Prediction of Low-Cycle Fatigue of Metals.

The relation between deformation inhomogeneity and low-cycle-fatigue failure of T2 pure copper and the nickel-based superalloy GH4169 under symmetric tension-compression cyclic strain loading is investigated by using a polycrystal representative volume element (RVE) as the material model. The anisotropic behavior of grains and the strain fields are calculated by crystal plasticity, taking the Bauschinger effect into account to track the process of strain cycles of metals, and the Shannon's differential entropies of both distributions of the strain in the loading direction and the first principal strain are employed at the tension peak of the cycles as measuring parameters of strain inhomogeneity. Both parameters are found to increase in value with increments in the number of cycles and they have critical values for predicting the material's fatigue failure. Compared to the fatigue test data, it is verified that both parameters measured by Shannon's differential entropies can be used as fatigue indicating parameters (FIPs) to predict the low cycle fatigue life of metal.

Effect of Cadmium Ion on alpha-Glucosidase: An Inhibition Kinetics and Molecular Dynamics Simulation Integration Study.

α-Glucosidase is a critical metabolic enzyme that produces glucose molecules by catalyzing carbohydrates. The aim of this study is to elucidate biological toxicity of Cd(2+) based on α-glucosidase activity and conformational changes. We studied Cd(2+)-mediated inactivation as well as conformational modulation of α-glucosidase by using kinetics coupled with simulation of molecular dynamics. The enzyme was significantly inactivated by Cd(2+) in a reversibly binding behavior, and Cd(2+) binding induced a non-competitive type of inhibition reaction (the K i was calculated as 0.3863 ± 0.033 mM). Cd(2+) also modulated regional denaturation of the active site pocket as well as overall partial tertiary structural change. In computational simulations using molecular dynamics, simulated introduction of Cd(2+) induced in a depletion of secondary structure by docking Cd(2+) near the saccharides degradation at the active site, suggesting that Cd(2+) modulating enzyme denaturation. The present study elucidated that the binding of Cd(2+) triggers conformational changes of α-glucosidase as well as inactivates catalytic function, and thus suggests an explanation of the deleterious effects of Cd(2+) on α-glucosidase.

Integration of Inhibition Kinetics and Molecular Dynamics Simulations: A Urea-Mediated Folding Study on Acetaldehyde Dehydrogenase 1.

Understanding the mechanism of acetaldehyde dehydrogenase 1 (ALDH1) folding is important because this enzyme is directly involved in several types of cancers and other diseases. We investigated the urea-mediated unfolding of ALDH1 by integrating kinetic inhibition studies with computational molecular dynamics (MD) simulations. Conformational changes in the enzyme structure were also analyzed using intrinsic and 1-anilinonaphthalene-8-sulfonate (ANS)-binding fluorescence measurements. Kinetic studies revealed that the direct binding of urea to ALDH1 induces inactivation of ALDH1 in a manner of mixed-type inhibition. Tertiary structural changes associated with regional hydrophobic exposure of the active site were observed. The urea binding regions on ALDH1 were predicted by docking simulations and were partly shared with active site residues of ALDH1 and with interface residues of the oligomerization domain for tetramer formation. The docking results suggest that urea prevents formation of the ALDH1 normal shape for the tetramer state as well as entrance of the substrate into the active site. Our study provides insight into the structural changes that accompany urea-mediated unfolding of ALDH1 and the catalytic role associated with conformational changes.

Adenoviral delivery of the EMX2 gene suppresses growth in human gastric cancer.

BACKGROUND: EMX2 is a human orthologue of the Drosophila empty spiracles homeobox gene that has been implicated in embryogenesis. Recent studies suggest possible involvement of EMX2 in human cancers; however, the role of EMX2 in carcinogenesis needs further exploration. RESULTS: In this study, we reported that down-regulation of EMX2 expression was significantly correlated with EMX2 promoter hypermethylation in gastric cancer. Restoring EMX2 expression using an adenovirus delivery system in gastric cancer cell lines lacking endogenous EMX2 expression led to inhibition of cell proliferation and Wnt signaling pathway both in vitro and in a gastric cancer xenograft model in vivo. In addition, we observed that animals treated with the adenoviral EMX2 expression vector had significantly better survival than those treated with empty adenoviral vector. CONCLUSION: Our study suggests that EMX2 is a putative tumor suppressor in human gastric cancer. The adenoviral-EMX2 may have potential as a novel gene therapy for the treatment of patients with gastric cancer.

Effect of SNPs on creatine kinase structure and function: identifying potential molecular mechanisms for possible creatine kinase deficiency diseases.

Single-nucleotide polymorphisms (SNPs) are common genetic material changes that often occur naturally. SNPs can cause amino acid replacements that may lead to severe diseases, such as the well-known sickle-cell anemia. We constructed eight SNP mutants of human brain-type creatine kinase (CKB) based on bioinformatics predictions. The biochemical and biophysical characteristics of these SNP mutants were determined and compared to those of the wild-type creatine kinase to explore the potential molecular mechanisms of possible creatine kinase SNP-induced diseases. While the reactivation of six SNP mutants after heat shock dropped more than 45%, only three of them showed notable increases in ANS fluorescence intensity and decreases in catalytic efficiency. Among them, H26Y and P36T bind substrates as well as the wild-type form does, but the melting temperatures (T(m)) dropped below body temperature, while the T59I mutant exhibited decreased catalytic activity that was most likely due to the much reduced binding affinity of this mutant for substrates. These findings indicate that SNPs such as H26Y, P36T and T59I have the potential to induce genetic diseases by different mechanisms.

Metallothioneins protect cytosolic creatine kinases against stress induced by nitrogen-based oxidants.

The formation of intracellular nitrogen-based oxidants has important physiological and pathological consequences. CK (creatine kinase), which plays a key role in intracellular energy metabolism, is a main target of low concentrations of oxidative and nitrative stresses. In the present study, the interaction between cytosolic CKs [MM-CK (muscle-type CK) and BB-CK (brain-type CK)] and MTs [metallothioneins; hMT2A (human MT-IIA) and hMT3 (human MT-III)] were characterized by both in vitro and intact-cell assays. MTs could successfully protect the cytosolic CKs against inactivation induced by low concentrations of PN (peroxynitrite) and NO both in vitro and in hMT2A-overexpressing H9c2 cells and hMT3-knockdown U-87 MG cells. Under high PN concentrations, CK formed granule-like structures, and MTs were well co-localized in these aggregated granules. Further analysis indicated that the number of cells containing the CK aggregates negatively correlated with the expression levels of MTs. In vitro experiments indicated that MTs could effectively protect CKs against aggregation during refolding, suggesting that MT might function as a chaperone to assist CK re-activation. The findings of the present study provide direct evidence of the connection between the two well-characterized intracellular systems: the precisely balanced energy homoeostasis by CKs and the oxidative-stress response system using MTs.

Trehalose has a protective effect on human brain-type creatine kinase during thermal denaturation.

We investigated the effects of trehalose on thermal inactivation and aggregation of human brain-type creatine kinase (hBBCK) in this study. In the presence of 1.0 M trehalose, the midpoint temperature of thermal inactivation (T (m)) of hBBCK increased by 4.6 °C, and the activation energy (E (a)) for thermal inactivation increased from 29.7 to 41.1 kJ mol(-1). Intrinsic fluorescence spectra also showed an increase in the apparent transition temperature (T (1/2)) of hBBCK from 43.0 °C to 46.5 °C, 47.7 °C, and 49.9 °C in 0, 0.6, 0.8, and 1.2 M trehalose, respectively. In addition, trehalose significantly blocked the aggregation of hBBCK during thermal denaturation. Our results indicate that trehalose has potential applications as a thermal stabilizer and may aid in the folding of other enzymes in addition to hBBCK.