ABSTRACT
Physical exercise plays a crucial role in the development of cognition and brain functions in children. Inhibitory control is an advanced cognition that affects children's life and learning. In the current study, the relationship between physical training and inhibitory control was explored. In total, 80 children were randomly but equally assigned to the experimental and control groups. The experimental group underwent physical training (volleyball) for 60â¯min, thrice a week for 12 weeks. In contrast, the control group did not undergo any training and continued with their daily routines. The flanker task and functional near-infrared spectroscopy (fNIRS) were employed to investigate the effects of 12-week physical training on inhibitory control and changes in the oxy-Hb concentration in the prefrontal cortex (PFC) during the task. The behavioral results revealed that the experimental group performed better on the flanker task (e.g., shorter response time [F(1,74) = 18.420, p < 0.001, ηp2 = 0.199] and higher accuracy [F(1,74) = 15.00, p < 0.001, ηp2 = 0.169] than the control group. The oxy-Hb concentration in the right dorsolateral PFC (R-DLPFC) was higher and the activation level of this region was higher during the flanker task [F(1,74) = 6.216, p < 0.05, ηp2 = 0.077]. Moreover, the McNemar test revealed improved cognitive performance in response time or accuracy and R-DLPFC activation induced by physical exercise coincided significantly (χ2 = 5.49, p < 0.05; χ2 = 6.081, p < 0.05). These findings suggest that the R-DLPFC is likely the neural substrate for improved cognitive performance elicited by 12-week physical training.
Subject(s)
Cognition , Spectroscopy, Near-Infrared , Child , Humans , Spectroscopy, Near-Infrared/methods , Cognition/physiology , Prefrontal Cortex/physiology , Exercise , Reaction Time/physiologyABSTRACT
Koumiss, a traditional fermented dairy product made from fresh mare milk, is a sour beverage that contains an abundance of microbial communities, including lactic acid bacteria, yeast and others. Firstly, probiotics such as Lacticaseibacillus in koumiss can induce the secretion of immunoglobulin G in serum and interleukin-2 in the spleen while beneficial Saccharomyces can secrete antibacterial compounds such as citric acid and ascorbic acid for specific immunopotentiation. Additionally, more isoflavone in koumiss can regulate estrogen levels by binding to its receptors to prevent breast cancer directly. Bile salts can be converted into bile acids such as taurine or glycine by lactic acid bacteria to lower cholesterol levels in vivo. Butyric acid secretion would be increased to improve chronic gastrotis by regulating intestinal flora with lactic acid bacteria. Finally, SCFA and lCFA produced by Lacticaseibacillus inhibit the reproduction of pathogenic microorganisms for diarrhea prevention. Therefore, exploring the mechanisms underlying multiple physiological functions through utilizing microbial resources in koumiss represents promising avenues for ameliorating chronic diseases.
ABSTRACT
The search for topological superconductivity (TSC) is currently an exciting pursuit, since non-trivial topological superconducting phases could host exotic Majorana modes. However, the difficulty in fabricating proximity-induced TSC heterostructures, the sensitivity to disorder and stringent topological restrictions of intrinsic TSC place serious limitations and formidable challenges on the materials and related applications. Here, we report a new type of intrinsic TSC, namely intrinsic surface topological superconductivity (IS-TSC) and demonstrate it in layered AuSn4 with Tc of 2.4 K. Different in-plane and out-of-plane upper critical fields reflect a two-dimensional (2D) character of superconductivity. The two-fold symmetric angular dependences of both magneto-transport and the zero-bias conductance peak (ZBCP) in point-contact spectroscopy (PCS) in the superconducting regime indicate an unconventional pairing symmetry of AuSn4. The superconducting gap and surface multi-bands with Rashba splitting at the Fermi level (EF), in conjunction with first-principle calculations, strongly suggest that 2D unconventional SC in AuSn4 originates from the mixture of p-wave surface and s-wave bulk contributions, which leads to a two-fold symmetric superconductivity. Our results provide an exciting paradigm to realize TSC via Rashba effect on surface superconducting bands in layered materials.
ABSTRACT
Age-associated cognitive and motor decline is related to central nervous system injury in older adults. Motor imagery training (MIT), as an emerging rehabilitative intervention, can activate neural basis similar to that in actual exercise, so as to promote motor function in older adults. The complex motor skills rely on the functional integration of the cerebral cortex. Understanding the neural mechanisms underlying motor imagery in older adults would support its application in motor rehabilitation and slowing cognitive decline. Based on this, the present study used functional near infrared spectroscopy (fNIRS) to record the changes in oxygen saturation in older adults (20 participants; mean age, 64.8 ± 4.5 years) during Baduanjin motor execution (ME) and motor imagery (MI). ME significantly activated the left postcentral gyrus, while the oxy-hemoglobin concentration in the right middle temporal gyrus increased significantly during motor imagery. These results indicate that advanced ME activates brain regions related to sensorimotor function, and MI increases the activation of the frontal-parietal cortex related to vision. In older adults, MI overactivated the temporo-parietal region associated with vision, and tend to be activated in the right brain.
Subject(s)
Imagination , Movement , Humans , Aged , Middle Aged , Movement/physiology , Imagination/physiology , Brain/physiology , Brain Mapping , Somatosensory Cortex , Magnetic Resonance ImagingABSTRACT
Shengxian decoction (SXT) is clinically used in chronic obstructive pulmonary disease (COPD) treatment. This study aimed to explore the mechanism and target genes of SXT acting on COPD. Differentially expressed genes (DEGs) between COPD and controls were identified and then performed enrichment analysis. The effective active compounds and corresponding target genes were obtained from the traditional Chinese medicine systems pharmacology database. We also compiled COPD related genes from the GeneCards database. Through the protein-protein interaction (PPI) network and least absolute shrinkage and selection operator (LASSO) regression was performed to identify key genes. Molecular docking was used for docking of key genes and compounds. The expression of key genes was detected by quantitative real-time PCR in COPD patients and bronchial epithelial cells stimulated with cigarette stroke extract (CSE). We identified 1,458 intersected DEGs from GSE47460 and GSE57148 datasets. Compared with intersected DEGs, we obtained 33 SXT target COPD-related genes. PI3K-Akt signaling pathway, MAPK signaling pathway, and focal adhesion were enriched by these 33 genes, as well as intersected DEGs. According to LASSO regression, there were 12 genes considered as signature genes. Then we constructed active compounds and corresponding six target genes. Finally, HIF1A and IL1B were selected as key genes by combining PPI network. HIF1A and IL1B were all upregulated expression in COPD and CSE stimulated cells and recovered in SXT treated CSE stimulated cells. This study provides a scientific basis for the identification of active compounds and target genes of SXT in the treatment of COPD.
Subject(s)
Network Pharmacology , Pulmonary Disease, Chronic Obstructive , Humans , Molecular Docking Simulation , Phosphatidylinositol 3-Kinases , Databases, Factual , Pulmonary Disease, Chronic Obstructive/drug therapyABSTRACT
This paper studies the active force characteristics of the neck muscles under the condition of rapid braking, which can provide theoretical support for reducing the neck injury of pilots when carrier-based aircraft blocks the landing. We carried out static loading and real vehicle braking experiments under rapid braking conditions, collected the active contraction force and electromyography (EMG) signals of neck muscles, and analyzed the response characteristics of neck muscle active force response. The results showed that the head and neck forward tilt time was delayed and the amplitude decreased during neck muscle pre-tightening. The duration of the neck in the extreme position decreased, and the recovery towards the seat direction was faster. The EMG signals of trapezius muscle was higher than sternocleidomastoid muscle. This suggests that pilots can reduce neck injury by pre-tightening the neck muscles during actual braking flight. In addition, we can consider the design of relevant fittings for pre-tightening the neck muscles.
Subject(s)
Neck Muscles , Neck , Electromyography , HeadABSTRACT
Long non-coding RNAs (lncRNA) have an extensive role in the progression and chemoresistance of gastric cancer (GC). Deeply study the regulatory role of lncRNAs could provide potential therapeutic targets. The aim of this study is to explore the regulatory role of HOTAIR in the progression and oxaliplatin resistance of GC. The expression of HOTAIR in GC and cell lines were detected by using qRT-PCR. Cell proliferation and apoptosis were analysed by CCK-8, EdU incorporation and flow cytometry. Luciferase reporter assay was used to identify the interaction between HOTAIR and ABCG2 (ATP-binding cassette (ABC) superfamily G member 2, ABCG2) via miR-195-5p. The regulatory functions were verified by using molecular biology experiments. HOTAIR was significantly overexpressed in GC and associated with poor prognosis. Knock-down of HOTAIR inhibited the GC cells proliferation and oxaliplatin resistance, while overexpression of HOTAIR showed opposite functions. Further studies found that HOTAIR acted as a competing endogenous RNA (ceRNA) to absorb miR-195-5p and elevated the expression of ABCG2, which leads to resistance of GC cells to oxaliplatin. Taken together, our findings demonstrated that HOTAIR regulates ABCG2 induced resistance of GC to oxaliplatin through miR-195-5p signalling and illustrate the great potential of developing new therapeutic targets for GC patients.
Subject(s)
MicroRNAs , RNA, Long Noncoding , Stomach Neoplasms , Humans , Stomach Neoplasms/drug therapy , Stomach Neoplasms/genetics , Stomach Neoplasms/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Oxaliplatin/pharmacology , Cell Line, Tumor , Cell Proliferation/genetics , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Gene Expression Regulation, Neoplastic , ATP Binding Cassette Transporter, Subfamily G, Member 2/genetics , ATP Binding Cassette Transporter, Subfamily G, Member 2/metabolism , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolismABSTRACT
Objective: The study aimed to investigate the effects of a 12-week Tai Chi exercise intervention on working memory in older adults using ERP-fNIRS. Method: Fifty older adults were randomly assigned to either an experimental group receiving a 12-week Tai Chi exercise intervention or a control group receiving regular daily activities. Working memory was assessed using the n-back task before and after the intervention, and spatial and temporal components of neural function underlying the n-back task were measured using ERP-fNIRS. Results: The experimental group demonstrated significant improvements in reaction time and accuracy on the 2-back task and showed higher activation levels in the R-DLPFC. Additionally, the Tai Chi group displayed significant increases in P3 amplitude in the overall n-back task. Conclusion: These findings suggest that Tai Chi interventions can enhance working memory in older adults, as evidenced by increasing neural activity and improving HbO in the R-DLPFC during the 2-back task.
ABSTRACT
ABSTRACT: Transcranial direct current stimulation (tDCS) is used in neuromodulation to regulate the excitability of the cerebral cortex and induce neural plasticity. It was initially used to rehabilitate patients with neurological diseases. However, with the increasing number of studies involving healthy individuals, this technology is currently used in the field of sports as well. The administration of tDCS to the cerebral cortex, especially over the primary motor cortex (M1), has been found to improve muscle strength, enhance endurance, and promote motor skills in humans. This study mainly summarizes the effects of tDCS on motor function, mainly involving motor promotion of tDCS in healthy athletes and nonathletes, and in patients diagnosed with neurological diseases. The tDCS is a promising and effective tool used to promote motor function by regulating cortical excitability. However, no consensus is available regarding individually appropriate models of tDCS.
Subject(s)
Electroconvulsive Therapy , Motor Cortex , Nervous System Diseases , Transcranial Direct Current Stimulation , Humans , Motor Cortex/physiology , Evoked Potentials, Motor/physiology , Transcranial Magnetic StimulationABSTRACT
BACKGROUND: To identify a novel marker for gastric cancer, we examined the usefulness of phosphoglycerate mutase 1 (PGAM1) as a potential diagnostic marker using isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics and evaluated its clinical significance. METHODS: Proteins from a discovery group of four paired gastric cancer tissues and adjacent gastric tissues were labeled with iTRAQ reagents and then identified and quantified using LC-MS/MS. The expression of PGAM1 was further validated in 139 gastric cancer patients using immunohistochemistry. Furthermore, the correlation of PGAM1 expression with clinical parameters was analyzed. Gene set enrichment analysis (GSEA) was performed to identify gene sets that were activated in PGAM1-overexpressing patients with gastric cancer. RESULTS: PGAM1 was significantly overexpressed in most cancers but particularly so in gastric cancer, with a sensitivity of 82.01% (95% confidence interval [CI]: 75.5%-88.5%) and specificity of 79.13% (95% CI: 72.3%-86%). Its expression was significantly associated with histological grade II and III tumors (p = 0.033), lymph node metastasis (p = 0.031), and TNM III-IV staging (p = 0.025). The area under the receiver operating characteristic (ROC) curve for the detection of PGAM1 overexpression in gastric cancer was 0.718 (p < 0.01). Furthermore, GSEA revealed that several important pathways such as glycolysis pathway and immune pathways were significantly enriched in patients with gastric cancer with PGAM1 overexpression. CONCLUSIONS: This study provided a sensitive method for detecting PGAM1, which may serve as a novel indicator for poor prognosis of gastric cancer, as well as a potent drug target for gastric cancer.
Subject(s)
Phosphoglycerate Mutase , Stomach Neoplasms , Humans , Phosphoglycerate Mutase/genetics , Phosphoglycerate Mutase/metabolism , Stomach Neoplasms/diagnosis , Stomach Neoplasms/metabolism , Chromatography, Liquid , Tandem Mass Spectrometry , Glycolysis , Neoplasm StagingABSTRACT
OBJECTIVE: The sensitivity and specificity of current biomarkers for gastric cancer were insufficient. The aim of the present study was to screen novel biomarkers and determine the diagnostic values of ornithine aminotransferase (OAT) and carbamoyl phosphate synthetase 1 (CPS1) for detecting gastric cancer. METHODS: With stable isotope tags, we labelled an initial discovery group of four paired gastric cancer tissue samples and identified with LC-ESI-MS/MS. A validation group of 159 gastric cancer samples and 30 healthy controls were used to validate the candidate targets. GSEA was used to explore the pathways activated in gastric cancer. RESULTS: Four hundred and thirty one proteins were found differentially expressed in gastric cancer tissues. Of these proteins, OAT and CPS1 were found over-expressed in gastric cancer patients, with sensitivity of 70.4% (95% CI: 63.3%-77.6%) and specificity of 80.5% (95% CI: 74.3%-86.7%) for ornithine aminotransferase, and with sensitivity of 68.6% (95% CI: 61.3%-75.8%) and specificity of 73% (95% CI: 66%-79.9%) for carbamoyl phosphate synthetase 1. The co-expression of OAT and CPS1 in gastric cancer tissues has a sensitivity of 81% (95% CI: 73.2%-88.8%) and specificity of 89% (95% CI: 83%-95%). Furthermore, both OAT and CPS1 were overexpressed in patients with local invasion T3 and T4 stages than those in patients with T1 and T2 stages. The co-expression of OAT and CPS1 was strongly correlated with histological grade I 68% (95% CI: 58.7%-77.3%) and TNM stage I/II 52% (95% CI: 42%-62%). The areas under ROC curves were up to 0.758 for the co-expression of OAT and CPS1 in gastric cancer. GSEA results showed that two gene sets and 30 gene sets were activated in OAT high- and CPS1 high-expression patients with gastric cancer, respectively. CONCLUSIONS: The present findings indicated a tight correlation between the co-expression of OAT and CPS1 and the histological grade, local invasion, and TNM stages of gastric cancer. Therefore, OAT and CPS1 might be predictors for gastric cancer invasion and potential targets for anticancer drug design for gastric cancer.
Subject(s)
Antineoplastic Agents , Stomach Neoplasms , Ammonia , Biomarkers , Carbamoyl-Phosphate Synthase (Ammonia)/genetics , Carbamoyl-Phosphate Synthase (Ammonia)/metabolism , Carbamyl Phosphate/metabolism , Humans , Ornithine-Oxo-Acid Transaminase/genetics , Stomach Neoplasms/pathology , Tandem Mass SpectrometryABSTRACT
OBJECTIVE: To verify the reliability and validity of the simplified Chinese version of the Yale Physical Activity Survey (YPAS) scale in the measurement of physical activity of elderly women in mainland China to provide a new standardized scale for evaluating the level of physical activity. METHODS: Twenty-one healthy elderly women completed all the research procedures. The participants completed the questions on the YPAS and wore an Omegawave Sport Technology®System diagnostic system that recorded maximum oxygen uptake and the adaptation index of the energy metabolism system. The reliability of the YPAS was evaluated based on the consistency of the two measurements (pre-test and post-test), and its validity was verified based on the relevant indicators in the Omegawave diagnostic system. Descriptive statistics, intraclass correlation coefficient and Pearson correlation analysis were used. The significance level was set at p < 0.05. RESULTS: In terms of reliability, the Pearson correlation coefficient and ICC of the total physical activity time of YPAS (r = 0.917, ICC = 0.897) was high. In terms of total calories, the Pearson correlation coefficient and ICC of the two test results was high (r = 0.958, ICC = 0.934). On the YPAS total index, the Pearson correlation coefficient and ICC of the two test was high (r = 0.930, ICC = 0.920). In terms of validity, there was a moderate correlation between the energy metabolism system adaptation index and the YPAS-total time (r = 0.472) and a moderate correlation with total calories (r = 0.472). There was a high correlation between the YPAS-total index and the maximum oxygen uptake (r = 0.782). CONCLUSION: The simplified Chinese version of the YPAS can measure the level of physical activity and energy metabolism of elderly women in mainland China. It is a reliable tool for measuring the physical activity of the elderly.
Subject(s)
Oxygen Consumption , Oxygen , Aged , China , Exercise , Female , Humans , Psychometrics , Reproducibility of Results , Surveys and QuestionnairesABSTRACT
Background: Tai Chi Chuan (TCC) is an effective method for delaying cognitive decline in older adults. However, in older adults, the association between long-term TCC practice and working memory updating has not been extensively studied. Objective: This cross-sectional study investigated how updating function operationalized via Reaction Times (RTs) and Accuracy Rates (ARs) of N-Back tasks being measured in a laboratory setting is related to long term practice of TCC. Methods: Twenty-six healthy elderly people participated in this experiment. According to the duration of time TCC was practiced, 13 subjects in the TCC group had more than 5 years of experience with TCC exercise, and 13 elderly subjects who had not been systematically exposed to mind-body exercise were assigned to the control group. The N-back task was administered to every participant to evaluate the updating function. Results: The TCC group had faster RTs than the control group (p < 0.05). For the 1-back task, the TCC group showed faster RTs than the control group; for the 2-back task, the TCC group exhibited faster RTs than the control group. The TCC group had higher ARs than the control group (p < 0.05). For the 1-back task, the TCC group showed higher ARs than the control group; for the 2-back task, the TCC group exhibited higher ARs than the control group. Conclusions: Long-term TCC practitioners exhibit a better updating function as compared to controls who did not practice TCC. Thus, our findings suggest that long-term TCC positively influences the updating function of older adults, making it, in turn, an effective mind-body exercise to maintain specific aspects of cognitive functioning.
Subject(s)
Tai Ji , Aged , Cognition , Cross-Sectional Studies , Exercise , Exercise Therapy , Humans , Tai Ji/methodsABSTRACT
BACKGROUND: Gastric cancer is the third leading cause of cancer-related death in the world. The purpose of the present study is to investigate the expression and prognostic significance of 6-phosphogluconolactonase (PGLS) in gastric cancer. METHODS: The protein extracted from a panel of four pairs of gastric cancer tissues and adjacent tissues, labeled with iTRAQ (8-plex) reagents, and followed by LC-ESI-MS/MS. The expressions of proteins were further validated by immunohistochemistry analysis. The expression levels of mRNA were analyzed and validated in the Oncomine database. The correlations of PGLS with prognostic outcomes were evaluated with Kaplan-Meier plotter database. RESULTS: The present study found that PGLS was significantly up-regulated in gastric cancer by using iTRAQ-based proteomics and immunohistochemistry analysis. The sensitivity of PGLS in gastric cancer was 72.9%. The high expression of PGLS was significantly correlated with TNM staging in gastric cancer (p = 0.02). The overexpression of PGLS predicts worse overall survival (OS) and post-progression survival (PPS) for gastric cancer (OS, HR = 1.48, p = 2.1e-05; PPS, HR = 1.35, p = 0.015). Specifically, the high PGLS expression predicts poor OS, PPS in male gastric cancer patients, in patients with lymph node metastasis and in patients with Her-2 (-). CONCLUSIONS: These findings suggested that PGLS was aberrantly expressed in gastric cancer and predicts poor overall survival, post-progression survival for gastric cancer patients. The present study collectively supported that PGLS is an important target for early determining and follow-up monitoring for gastric cancer.
Subject(s)
Biomarkers, Tumor/analysis , Carboxylic Ester Hydrolases/analysis , Early Detection of Cancer/methods , Proteome/analysis , Stomach Neoplasms/diagnosis , Stomach/chemistry , Biomarkers, Tumor/metabolism , Carboxylic Ester Hydrolases/metabolism , Female , Humans , Immunohistochemistry , Kaplan-Meier Estimate , Male , Middle Aged , Neoplasm Staging , Prognosis , Proteomics , Stomach/pathology , Stomach Neoplasms/metabolism , Stomach Neoplasms/mortality , Stomach Neoplasms/pathologyABSTRACT
We used long-term data on the variation in harem size in Yunnan snub-nosed monkeys to research the effects of harem size on reproductive success and the ratio of grooming received to given (RGRG). The results suggest that harem holders derive reproductive benefits commensurate with harem size, whereas the females' reproductive success is unaffected by harem size. Males of larger harems groomed less and had higher RGRG than males of smaller harems. In the case of females, grooming given increased, and RGRG decreased with an increase in harem size. The males' reproductive success seems to be a driver of harem size maximization. From the females' perspective, dwindling social benefits appear to set the upper limit for harem enlargement. We also showed that males of monogamous units ('single-female harems') invested more into grooming their female, presumably to prevent unit disintegration and loss of mating privileges.
ABSTRACT
Repetitive transcranial magnetic stimulation (rTMS) is a noninvasive brain-stimulation technique that transiently modulates cerebral cortex excitability, achieving overall positive results in poststroke motor-function recovery. Excessive inhibition of the ipsilesional-affected hemisphere by the contralesional-unaffected hemisphere has seriously hindered poststroke motor-function recovery. Hence, intracortical disinhibition can be used as an approach to managing poststroke brain injury. This technique promotes neural plasticity for faster motor-function recovery. rTMS relieves unilateral inhibition of the brain function by regulatinga interhemispheric-imbalanced inhibition. This paper summarized 12 studies from 2016 to date, focusing on rTMS on motor function after acute and chronic stroke by regulating the interhemispheric imbalance of inhibitory inputs. Although rTMS studies have shown promising outcomes on recovery of motor functions in stroke patients, different intervention methods may lead to discrepancies in results. A uniform optimal stimulus model cannot routinely be used, mainly due to the stimulus schemes, stroke types and outcome-measuring differences among studies. Thus, the effect of rTMS on poststroke motor-function recovery should be investigated further to standardize the rTMS program for optimal poststroke motor-function recovery. More randomized, placebo-controlled clinical trials with standardized rTMS protocols are needed to ensure the effectiveness of the treatment.
ABSTRACT
The acceleration characteristics of fragments generated from explosively-driven cylindrical shells are important issues in warhead design. However, there is as yet no reasonable theory for predicting the acceleration process of a specific metallic shell; existing approaches either ignore the effects of shell disintegration and the subsequent gas leakage on fragment acceleration or treat them in a simplified manner. In this paper, a theoretical model was established to study the acceleration of discrete fragments under the combined effect of shell disintegration and gas leakage. Firstly, an equation of motion was developed, where the acceleration of a cylindrical shell and the internal detonation gas was determined by the motive force impacting the inner surface of the metallic cylinder. To account for the force decrease induced by both the change in fragment area after the shell disintegrates and the subsequent drop in gas pressure due to gas leakage, the equation of motion was then associated with an equation for the locally isentropic expansion of the detonation gas and a modified gas-leakage equation. Finally, theoretical analysis was conducted by solving the associated differential equations. The proposed model showed good agreement with experimental data and numerical simulations, indicating that it was suitable for predicting the acceleration of discrete fragments generated from a disintegrated warhead shell. In addition, this study facilitated a better understanding of the complicated interaction between fragment acceleration and gas outflow.
ABSTRACT
BACKGROUND: Cryptochrome is a flavin-binding blue-light photoreceptor that functions in growth and development in plants, the circadian clock in animals and navigation in birds. However, a lack of purified cryptochrome has hindered studies of the structure and function of this protein. In this study, we obtained a substantial amount of the Columbia livia Cryptochrome1 (ClCry1) protein by using a prokaryotic expression system. In addition, we performed comprehensive experiments to assess the influence of several factors on the purification and yield of ClCry1, such as the inducer that was used, temperature, duration of expression and type of growth medium. These assays clearly indicated that such factors influenced the purification and yield of ClCry1. Moreover, Flavin Adenine Dinucleotide (FAD) was added during expression and purification of ClCry1, which resulted in production of large amounts of ClCry1 protein with the FAD cofactor from the Escherichia coli (E. coli) heterologous expression system. We believe that this study provides a novel avenue to acquire large amounts of ClCry1 that contains FAD and lays the foundation for studies of the geomagnetic navigation mechanism of Aves. OBJECTIVE: In this article, our motivation is to sufficiently acquire functional ClCry1 protein. METHOD: In this article, we performed series of experiments to optimize the yields of ClCry1 protein expression by conducting with expression-vectors, variable inducers, temperatures, medias and durations of induction, which also identified the most appropriate conditions for obtaining functional ClCry1. Moreover, we identified a solution for the FAD abscission of ClCry1 by adding additional FAD into the dialysis buffer during the purification. RESULTS: Following our performed series of experiments, we assessed several crucial parameters, such as inducer, temperature, duration of induction, culture medium and recombinant expression vector. The highest yields of ClCry1 were observed with 0.01 mM IPTG and expressing for 8 h with pET21a-ClCry1 as recombinant expression vectors. CONCLUSION: We demonstrated the feasibility of heterologous expression of ClCry1 in E. coli. In addition, we identified a solution for the low yield and FAD abscission of ClCry1 by conducting several experiments with variable inducers, temperatures, medias and durations of induction, which also identified the most appropriate conditions for obtaining functional ClCry1. Moreover, the typical yield was approximately 6 mg of ClCry1 from 2-liter culture, and 50% of the final protein contained the FAD cofactor. These results strongly suggest that our expression strategy is useful.
Subject(s)
Columbidae/genetics , Cryptochromes/genetics , Cryptochromes/isolation & purification , Escherichia coli/genetics , Animals , Gene Expression , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purificationABSTRACT
Metabolomics is the study of metabolic changes in biological systems and provides the small molecule fingerprints related to the disease. Extracting biomedical information from large metabolomics data sets by multivariate data analysis is of considerable complexity. Therefore, more efficient and optimizing metabolomics data processing technologies are needed to improve mass spectrometry applications in biomarker discovery. Here, we report the findings of urine metabolomic investigation of hepatitis C virus (HCV) patients by high-throughput ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) coupled with pattern recognition methods (principal component analysis, partial least-squares, and OPLS-DA) and network pharmacology. A total of 20 urinary differential metabolites (13 upregulated and 7 downregulated) were identified and contributed to HCV progress, involve several key metabolic pathways such as taurine and hypotaurine metabolism, glycine, serine and threonine metabolism, histidine metabolism, arginine and proline metabolism, and so forth. Metabolites identified through metabolic profiling may facilitate the development of more accurate marker algorithms to better monitor disease progression. Network analysis validated close contact between these metabolites and implied the importance of the metabolic pathways. Mapping altered metabolites to KEGG pathways identified alterations in a variety of biological processes mediated through complex networks. These findings may be promising to yield a valuable and noninvasive tool that insights into the pathophysiology of HCV and to advance the early diagnosis and monitor the progression of disease. Overall, this investigation illustrates the power of the UPLC-MS platform combined with the pattern recognition and network analysis methods that can engender new insights into HCV pathobiology.
Subject(s)
Mass Spectrometry , Metabolic Networks and Pathways , Metabolomics/methods , Pattern Recognition, Automated , Biomarkers/urine , Chromatography, High Pressure Liquid , Humans , Multivariate Analysis , UrinalysisABSTRACT
Metabolomics represents an emerging and powerful discipline concerned with the comprehensive analysis of small molecules and provides a powerful approach to discover biomarkers in biological systems. Recent development of biomarkers for diagnosis and therapeutic monitoring of liver-stagnation and spleen-deficiency syndrome (LSS)-type disease remains challenging. This study was undertaken to discover novel potential biomarkers for the non-invasive early diagnosis of human LSS. Urine samples which are potentially a rich source of metabolites were collected from patients with LSS, together with healthy control samples. Metabolite profiling was performed by ultra-performance liquid-chromatography/electrospray-ionization synapt high-definition mass spectrometry (UPLC-Q-TOF-HDMS) in conjunction with multivariate data analysis and ingenuity pathway analysis that were used to select the metabolites to be used for the non-invasive diagnosis of LSS. Twelve urinary differential metabolites contributing to the complete separation of LSS patients from matched healthy controls were identified involving several key metabolic pathways such as pentose and glucuronate interconversions, ascorbate, aldarate, cysteine, methionine, tyrosine, tryptophan, amino sugar and nucleotide sugar metabolism. More importantly, of the 12 differential metabolites, 4 metabolite markers, prolylhydroxyproline, L-homocystine, 2-octenoylcarnitine and α-N-phenylacetyl-L-glutamine, were effective for the diagnosis of human LSS, with an achieved sensitivity of 93.0%. These results demonstrate that robust metabolomics has the potential as a non-invasive strategy and promising screening tool to evaluate the potential of these metabolites in the early diagnosis of LSS patients and provides new insight into pathophysiological mechanisms.