Replacement of Loops at the Entrance of the Active Pocket of Streptococcus thermophilus 4,6-α-Glucanotransferase Changes Its Catalytic Activity and Product Specificity.

To explore the roles of loops around active pocket in the reuteran type 4,6-α-glucanotransferase (StGtfB) from S. thermophilus, they were individually or simultaneously replaced with those of an isomalto/maltopolysaccharides type 4,6-α-glucanotransferase from L. reuteri. StGtfB with the replaced loops A1, A2 (A1A2) and A1, A2, B (A1A2B), respectively, showed 1.41- and 0.83-fold activities of StGtfB. Two mutants reduced crystallinity and increased starch disorder at 2, 4, and 8 U/g more than StGtfB and increased DP ≤ 5 short branches of starch by 38.01% at 2 U/g, much more than StGtfB by 4.24%. A1A2B modified starches had the lowest retrogradation over 14 days. A1A2 modified starches had the highest percentage of slowly digestible fractions, ranging from 40.32% to 43.34%. StGtfB and its mutants bind substrates by hydrogen bonding and van der Waals forces at their nonidentical amino acid residues, suggesting that loop replacement leads to a different conformation and changes activity and product structure.

A mutation in CsGME encoding GDP-mannose 3,5-epimerase results in little and wrinkled leaf in cucumber.

KEY MESSAGE: Map-based cloning revealed that a mutation in a highly conserved amino acid of the CsGME gene encoding GDP-mannose 3,5-epimerase, causes the phenotype of little and wrinkled leaves in cucumbers. Leaf size is a critical determinant of plant architecture in cucumbers, yet only a few genes associated with this trait have been mapped or cloned. Here, we identified and characterized a mutant with little and wrinkled leaves, named lwl-1. Genetic analysis revealed that the phenotype of the lwl-1 was controlled by a single recessive gene. Through map-based cloning, the lwl-1 locus was narrowed down to a 12.22-kb region exclusively containing one fully annotated gene CsGME (CsaV3_2G004170). CsGME encodes GDP-mannose 3,5-epimerase, which is involved in the synthesis of ascorbic acid (ASA) and one of the components of pectin, RG-II. Whole-length sequencing of the 12.22 kb DNA fragment revealed the presence of only a non-synonymous mutation located in the sixth exon of CsGME in lwl-1, resulting in an amino acid alteration from Pro363 to Leu363. This mutation was unique among 118 inbred lines from cucumber natural populations. CsGME expression significantly reduced in various organs of lwl-1, accompanied by a significant decrease in ASA and pectin content in leaves. Both CsGME and Csgme proteins were localized to the cytoplasm. The mutant phenotype exhibited partial recovery after the application of exogenous boric acid. Silencing CsGME in cucumber through VIGS confirmed its role as the causal gene for lwl-1. Transcriptome profiling revealed that CsGME greatly affected the expression of genes related to the cell division process and cell plate formation. This study represents the first report to characterize and clone the CsGME in cucumber, indicating its crucial role in regulating leaf size and development.

A mutation in CsDWF7 gene encoding a delta7 sterol C-5(6) desaturase leads to the phenotype of super compact in cucumber (Cucumis sativus L.).

KEY MESSAGE: A novel super compact mutant, scp-3, was identified using map-based cloning in cucumber. The CsDWF7 gene encoding a delta7 sterol C-5(6) desaturase was the candidate gene of scp-3. Mining dwarf genes is important in understanding stem growth in crops. However, only a small number of dwarf genes have been cloned or characterized. Here, we characterized a cucumber (Cucumis sativus L.) dwarf mutant, super compact 3 (scp-3), which displays shortened internodes and dark green leaves with a wrinkled appearance. The photosynthetic rate of scp-3 is significantly lower than that of the wild type. The dwarf phenotype of scp-3 mutant can be partially rescued by the exogenous brassinolide (BL) application, and the endogenous brassinosteroids (BRs) levels in the scp-3 mutant were significantly lower compared to the wild type. Microscopic examination revealed that the reduced internode length in scp-3 resulted from a decrease in cell size. Genetic analysis showed that the dwarf phenotype of scp-3 was controlled by a single recessive gene. Combined with bulked segregant analysis and map-based cloning strategy, we delimited scp-3 locus into an 82.5 kb region harboring five putative genes, but only one non-synonymous mutation (A to T) was discovered between the mutant and its wild type in this region. This mutation occurred within the second exon of the CsGy4G017510 gene, leading to an amino acid alteration from Leu156 to His156. This gene encodes the CsDWF7 protein, an analog of the Arabidopsis DWF7 protein, which is known to be involved in the biosynthesis of BRs. The CsDWF7 protein was targeted to the cell membrane. In comparison to the wild type, scp-3 exhibited reduced CsDWF7 expression in different tissues. These findings imply that CsDWF7 is essential for both BR biosynthesis as well as growth and development of cucumber plants.

Effect of physical activity on risk of Alzheimer's disease: A systematic review and meta-analysis of twenty-nine prospective cohort studies.

OBJECTIVE: Physical activity (PA) is beneficial in reductions of all-cause mortality and dementia. However, whether Alzheimer's disease (AD) risk is modified by PA remains disputable. This meta-analysis aims to disclose the underlying relationship between PA and incident AD. METHODS: Pubmed, Embase, Cochrane Library, and Web of Science were retrieved from inception to June 2023. Random-effects models were employed to derive the effect size, represented by hazard ratio (HR) and 95% confidence interval (CI). RESULTS: Twenty-nine prospective cohort studies involving 2068,519 participants were included. The pooled estimate showed a favorable effect of PA on AD risk decline (HR 0.72, 95% CI 0.65-0.80). This association remained robust after adjusting for maximum confounders (HR 0.85, 95% CI 0.79-0.91). Subgroup analysis of PA intensity demonstrated an inverse dose-response relationship between PA and AD, effect sizes of which were significant in moderate (HR 0.85, 95% CI 0.80-0.93) and high PA (HR 0.56, 95% CI 0.45-0.68), but not in low PA (HR 0.94, 95% CI 0.77-1.15). Regardless of all participants or the mid-life cohort, the protection of PA against AD appeared to be valid in shorter follow-up (<15 years) rather than longer follow-up (≥15 years). In addition to follow-up, the robustness of the estimates persisted in supplementary meta-analyses, meta-regression analyses, and sensitivity analyses. CONCLUSION: PA intervention reduces the incidence of AD, but merely in moderate to vigorous PA with follow-up of less than 15 years, thus conditionally recommending the popularization of PA as a modifiable lifestyle factor to prevent AD.

Paraformaldehyde as C1 Synthon: Electrochemical Three-Component Synthesis of Tetrahydroimidazo[1,5-a]quinoxalin-4(5H)-ones in Aqueous Ethanol.

A green and practical method for the electrochemical synthesis of tetrahydroimidazo[1,5-a]quinoxalin-4(5H)-ones through the three-component reaction of quinoxalin-2(1H)-ones, N-arylglycines and paraformaldehyde was reported. In this strategy, EtOH played dual roles (eco-friendly solvent and waste-free pre-catalyst) and the in situ generated ethoxide promoted triple sequential deprotonations.

New innovations and educational process in undergraduate neurology education in blended learning.

Many undergraduate students suffer from 'neurophobia,' which refers to a lack of knowledge or confidence in neurology, and this can influence their career choices. Various measures have been taken to address this issue, including the implementation of new technologies and methodologies. Significant advancements have been made in the development of blended learning, and the integration of student-centered learning modules, multimedia, and web-based devices has become a common teaching approach. Nonetheless, the optimal delivery form, as well as assessment for the selected learning format and teaching quality in both theory and clinical practice, are being investigated. The purpose of this review is to provide a summary of the current understanding of blended learning as well as innovative methods, technologies, and assessments of undergraduate neurology education. It aims to highlight opportunities for implementing a novel, comprehensive learning model with a suitable blended learning method within a framework of customized technology-assessment processes for future neurology classes, encompassing both theoretical and clinical training.

Specific and efficient hydrolysis of all outer glucosyls in protopanaxadiol type and protopanaxatriol type ginsenosides by a ß-glucosidase from Thermoclostridium stercorarium.

To develop a new method for enzymatic preparation of minor ginsenosides, T. stercorarium ß-glucosidase (Tsbgl) was characterized and its activities of deglycosylation towards natural ginsenosides were examined. The substrates of 1 mmol l-1 were incubated with the enzyme of 38.3 U ml-1 at 65 â„ƒ and pH 5.0. The Km values of Tsbgl for ginsenosides Rb1, Rg1 and pNPG were 0.37 ± 0.03, 3.26 ± 0.19, and 1.24 ± 0.03 mmol l-1, and the Vmax values were 183.63 ± 7.15, 85.03 ± 4.90, and 117.66 ± 1.96 µmol mg-1 min-1, respectively. The molar conversion of ginsenosides Rb1, Rb2, Rb3, Rc, Re, Rg1, and Rf by Tsbgl within 6 h was 100%, 50.1%, 42.7%, 92.0%, 57.3%, 67.9%, and 76.8%, respectively. The yield of aglycone protopanaxadiol was 35.5 µmol l-1 h-1 for Rb1, while the yields of aglycone protopanaxatriol were 64.2 and 70.4 µmol l-1 h-1 for Rg1 and Rf. Tsbgl with good organic solvent tolerance, mild reaction conditions and broad substrate specificity, could completely remove all outer glucosyls at the C-3 and C-20 hydroxyls of protopanaxadiol-type ginsenosides, and the C-6 and C-20 hydroxyls of protopanaxatriol-type ginsenosides through various pathways, providing a specific and efficient way to produce minor ginsenosides.

Electrocatalytic three-component synthesis of 4-halopyrazoles with sodium halide as the halogen source.

The first example of the electrocatalytic multicomponent synthesis of 4-chloro/bromo/iodopyrazoles from hydrazines, acetylacetones and sodium halides under chemical oxidant- and external electrolyte-free conditions has been developed. Sodium halides played a dual role as a halogenation reagent and a supporting electrolyte. Mechanism studies revealed that the bromination reaction proceeded via an ionic pathway, whereas both chlorination and iodination proceeded via a radical pathway.

Potato starch modified by Streptococcus thermophilus GtfB enzyme has low viscoelastic and slowly digestible properties.

Potato starch with high viscosity and digestibility cannot be added into some foods. To address this issue, a novel starch-acting enzyme 4,6-α-glucosyltransferase from Streptococcus thermophilus (StGtfB) was used. StGtfB decreased the iodine affinity and the molecular weight, but increased the degree of branching of starch at a mode quite different from glycogen 1,4-α-glucan branching enzyme (GBE). StGtfB at 5 U/g substrate mainly introduced DP 1-7 into amylose (AMY) or DP 1-12 branches into amylopectin (AMP), and increased the ratio of short- to long-branches from 0.32 to 2.22 or from 0.41 to 2.50. The DP 3 branch chain was the most abundant in both StGtfB-modified AMY and StGtfB-modified AMP. The DP < 6 branch chain contents in StGtfB-modified AMY were 42.68%, much higher than those of GBE-modified AMY. StGtfB significantly decreased viscoelasticity but still kept pseudoplasticity of starch. The modifications also slowed down the glucose generation rate of products at the mammalian mucosal α-glucosidase level. The slowly digestible fraction in potato starch increased from 34.29% to 53.22% using StGtfB of 5 U/g starch. This low viscoelastic and slowly digestible potato starch had great potential with respect to low and stable postprandial blood glucose.

Severe Fatigue is an Important Factor in the Prognosis of Patients with Advanced Hepatocellular Carcinoma Treated with Sorafenib.

OBJECTIVE: To investigate the effects of fatigue on the survival of patients with advanced hepatocellular carcinoma treated with sorafenib. PATIENTS AND METHODS: A retrospective analysis of 182 cases of patients with advanced hepatocellular carcinoma treated with sorafenib in our hospital from October 1, 2008, to October 31, 2017, showed clinical and pathological data and follow-up results. The clinical and pathological data as well as follow-up results of 182 patients with advanced hepatocellular carcinoma treated with sorafenib in our hospital from October 1, 2008, to October 31, 2018, were retrospectively analyzed. All patients were treated for at least 3 months. Patients were divided into three groups: fatigue grade I (n=74), fatigue grade II (n=62), and fatigue grade III (n=46), according to National Cancer Institute common terminology criteria for adverse events (NCI CTCAE) version 5.0. Survival analysis between groups was performed by the Kaplan-Meier method (Log rank test), continuous variables were analyzed by t-test, and categorical variables were analyzed by chi-square test. RESULTS: The overall survival (OS) of patients who were relieved of fatigue was 33.0±9.3 months, whereas the OS of patients who were not relieved of fatigue was 15.0±1.8 months (P<0.000). Furthermore, the time to progress (TTP) of patients who were relieved of fatigue by resting was 20.3 ± 10.9 months compared to a TTP of 7.7 ± 1.0 months in patients who were not relieved of fatigue (P<0.000). CONCLUSION: Patients, especially the elderly and infirm, were more susceptible to toxicity.

Highly selective and sensitive chemosensor for Al(III) based on isoquinoline Schiff base.

As a colorimetric and fluorescent turn-on sensor to Al3+, N'-(2-hydroxybenzylidene)isoquinoline-3-carbohydrazide (HL) has been easily synthesized. The fluorescence intensity increases by 273 times in the presence of Al3+ at 458 nm. Meanwhile, the experiment data indicate that the limit of detection for Al3+ is 1.11 × 10-9 M. Remarkably, the blue fluorescence signal of HL-Al3+ could be specially observed by the naked eye under UV light and is significantly different from those of other metal ions. Fluorescence switch based on the control of Al3+ and EDTA proved HL could act as a reversible chemosensor. According to ESI-MS result and the Job's plots, the 2:1 coordination complex formed by HL and Al3+ could be produced. Density functional theory calculations were performed to illustrate the structures of HL and complex. The cell imaging experiment indicates that HL can be applied for monitoring intracellular Al3+ levels in cells.

Belief in and use of traditional Chinese medicine in Shanghai older adults: a cross-sectional study.

BACKGROUND: Traditional medicine is broadly used across Asian societies for various medical conditions and health concerns. However, there remains a wide disparity between users and non-use, which makes it imperative to understand the factors affecting the lay perception and utilization of traditional medicine. This study systematically examined the demographic, medical, and socioeconomic factors affecting belief in and use of traditional medicine among older adults of Shanghai, China. METHODS: We used the data from Survey of Life and Opinion on Shanghai Older Adults in 2013 with a sample of 3418 older adults aged 50 years or older. The multilevel logistic models were applied to examine the associations between faith and utilization of traditional medicine and a set of factors of the respondents, including demographics (gender, age, rural/urban residence), socioeconomic status (educational attainment, income, primary occupation), social support (marital status, social network), and disease/conditions. The associations between individual use of traditional medicine and the profile of socioeconomic development and the medical services conditions of local communities were also modelled. RESULTS: We found that cardiovascular diseases, lung diseases, cancer, prostatitis, arthritis, and nervous system diseases were positive correlates for using traditional medicine. Older adults who had a cancer, a prostatitis, or a fracture had more faith in traditional treatment. Rural living, higher educational attainment, and white-collar occupation promoted the use of traditional medicine. A higher number of strong social ties and a tie connected with medical staff were positive factors of use as well. CONCLUSION: The belief in and use of traditional medicine were prevalent among older adults in Shanghai, China. Though not conclusive, our study suggested that traditional medicine in China appears to serve two distinct functions, namely complementary medicine for those socioeconomically advantaged whereas alternative medicine for those socioeconomically disadvantaged.