Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

A Neural-network-based Alternative Scheme to Include Nonhydrostatic Processes in an Atmospheric Dynamical Core

Here,a nonhydrostatic alternative scheme(NAS)is proposed for the grey zone where the nonhydrostatic impact on the atmosphere is evident but not large enough to justify the necessity to include an implicit nonhydrostatic solver in an atmospheric dynamical core.The NAS is designed to replace this solver,which can be incorporated into any hydrostatic models so that existing well-developed hydrostatic models can effectively serve for a longer time.Recent advances in machine learning(ML)provide a potential tool for capturing the main complicated nonlinear-nonhydrostatic relationship.In this study,an ML approach called a neural network(NN)was adopted to select leading input features and develop the NAS.The NNs were trained and evaluated with 12-day simulation results of dry baroclinic-wave tests by the Weather Research and Forecasting(WRF)model.The forward time difference of the nonhydrostatic tendency was used as the target variable,and the five selected features were the nonhydrostatic tendency at the last time step,and four hydrostatic variables at the current step including geopotential height,pressure in two different forms,and potential temperature,respectively.Finally,a practical NAS was developed with these features and trained layer by layer at a 20-km horizontal resolution,which can accurately reproduce the temporal variation and vertical distribution of the nonhydrostatic tendency.Corrected by the NN-based NAS,the improved hydrostatic solver at different horizontal resolutions can run stably for at least one month and effectively reduce most of the nonhydrostatic errors in terms of system bias,anomaly root-mean-square error,and the error of the wave spatial pattern,which proves the feasibility and superiority of this scheme.

Improved genome annotation of Brassica oleracea highlights the importance of alternative splicing

Brassica oleracea has been developed into many important crops,including cabbage,kale,cauliflower,broccoli and so on.The genome and gene annotation of cabbage(cultivar JZS),a representative morphotype of B.oleracea,has been widely used as a common reference in biological research.Although its genome assembly has been updated twice,the current gene annotation still lacks information on untranslated regions(UTRs)and alternative splicing(AS).Here,we constructed a high-quality gene annotation(JZSv3)using a full-length transcriptome acquired by nanopore sequencing,yielding a total of 59452 genes and 75684 transcripts.Additionally,we re-analyzed the previously reported transcriptome data related to the development of different tissues and cold response using JZSv3 as a reference,and found that 3843 out of 11908 differentially expressed genes(DEGs)underwent AS during the development of different tissues and 309 out of 903 cold-related genes underwent AS in response to cold stress.Meanwhile,we also identified many AS genes,including BolLHCB5 and BolHSP70,that displayed distinct expression patterns within variant transcripts of the same gene,highlighting the importance of JZSv3 as a pivotal reference for AS analysis.Overall,JZSv3 provides a valuable resource for exploring gene function,especially for obtaining a deeper understanding of AS regulation mechanisms.

2,3,5,4’-Tetrahydroxystilbene-2-O-b-D-Glucoside modulates CHEK2 and CCND1 alternative splicing to inhibit MCF-7 cells proliferation

Background:In our previous study,we observed a synergistic effect of 2,3,5,4’-Tetrahydroxystilbene-2-O-b-D-glucoside combined with adriamycin to induce apoptosis in MCF-7 breast cancer cells.However,the underlying mechanisms of epigenetic modifications,such as alternative splicing,have not been explored.In this study,we aimed to investigate the mechanism by which THSG inhibits MCF-7 cell proliferation using full-length transcriptome sequencing.Methods:First,cell viability was examined using the methyl thiazolyl tetrazolium method and full-length transcriptome sequencing was performed to identify genes and pathways.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes were used to identify the principal pathways and targets of THSG.Flow cytometry analysis of cell cycle distribution was performed.Meanwhile,the analysis of alternative splicing and domains of the key proteins was conducted.Quantitative polymerase chain reaction and western blotting were performed for verification.Results:THSG showed significant cytotoxic activity in MCF-7 cells.Full-length transcriptome sequencing revealed differential alternative splicing with 173 upregulated and 263 downregulated genes.Further analysis identified distinct differential expression of genes(CHEK2-211 and CCND1-201)involved in the cell cycle in the THSG-treated group.Subsequently,alternative splicing types of CHEK2(mutually exclusive exon)and CCND1(intron retention).We found that THSG downregulated mRNA expression,as confirmed by quantitative polymerase chain reaction analysis.Interestingly,protein structural analysis revealed that THSG treatment led to the generation of CHK2-211,which was the result of a mutation in the amino acid residues(GLU-150,ASN-151)of the CHEK2 domain(VAL-150,GLY-151).and CyclinD1-201 were obtained when an amino acid(ASP-267)in the domain was lost in CyclinD1.Moreover,molecular docking analysis demonstrated that the domains of key proteins could bind THSG more effectively,with no difference in affinity.Western blotting confirmed that THSG inhibited the expression of CHK2 and CyclinD1.Conclusion:THSG modulated the alternative splicing of CHEK2 and CCND1 by inducing G0/G1 cell cycle arrest,consequently suppressing MCF-7 cell proliferation.

Molecular targets and mechanisms of different aberrant alternative splicing in metastatic liver cancer

Metastasis remains a major challenge in the successful management of malignant diseases.The liver is a major site of metastatic disease and a leading cause of death from gastrointestinal malignancies such as colon,stomach,and pancreatic cancers,as well as melanoma,breast cancer,and sarcoma.As an important factor that influences the development of metastatic liver cancer,alternative splicing drives the diversity of RNA transcripts and protein subtypes,which may provide potential to broaden the target space.In particular,the dysfunction of splicing factors and abnormal expression of splicing variants are associated with the occurrence,progression,aggressiveness,and drug resistance of cancers caused by the selective splicing of specific genes.This review is the first to provide a detailed summary of the normal splicing process and alterations that occur during metastatic liver cancer.It will cover the role of alternative splicing in the mechanisms of metastatic liver cancer by examining splicing factor changes,abnormal splicing,and the contribution of hypoxia to these changes during metastasis.

Workshop “Navigating the Transition: Standards Powering the Journey of Alternative Fuel Infrastructure”

April 18, Brussels, Belgium In light of recent significant regulatory changes within the European Union impacting the transport sector, such as the Alternative Fuels Infrastructure Regulation(AFIR), Re Fuel EU and Fuel EU Maritime, the role of European standardization becomes increasingly critical in facilitating the transition towards the deployment of alternative fuels.

Use of Topical Complementary and Alternative Medicines (CAM) in Diabetic Septic Foot Disease in Makkah Region, Western of Saudi Arabia, an Exploratory Study

Diabetes is a chronic condition that is significantly impacted daily by environment. At least one in five Saudis, suffer from diabetes. “Diabetes is a pandemic of unprecedented magnitude spiraling out of control” (Boulton). Saudi Arabia ranks the second highest in the Middle East, and is the seventh in the world for the prevalence of diabetes (WHO). Diabetes mellitus (DM) has been found to be related to high mortality, morbidity accompanied by poor general health and lower quality of life. Current Saudi diabetes patients’ behaviors, practices, and beliefs regarding foot health problems such as ulcers, are not well investigated. Diabetic patients frequently utilize natural remedies and Herbs for self-medication as a part of complementary and alternative Medicine. Aim of study: To explore the widespread use of the alternative remedies in the local communities of the Western Saudi Arabia, the drives behind using and to focus on the potential adverse effects following their usage. Method: A descriptive, cross-sectional study of 386 diabetic patients with feet problems in Makkah Region, West of Saudi Arabia during the period April - June 2023. Results: Out of the 531 participants, only 386 participants reported diabetic foot disorder (DFD). Most of the participant were above 65 years of age group (24.61 %). Male (52.08%) more than female (47.92%) in the research group. All of the participants had diabetic foot disorders at least once. The prevalence of complementary and alternative medicines (CAM) use among them was 67.88%, as solitary or in combination with hospital medicines. Honey headed the list followed by myrrh and black seeds. Acquaintances advice (59.54%) was the most reason for using the alternative medicine. 60.30% of Diabetic foot patient complained of adverse effects during CAM remedies use. Conclusion: Consumption of CAM remedies among diabetic patients for Diabetic Foot Disorder (DFD) is common. Honey headed the list followed by myrrh and black seeds. Additional wide-scale research is required to establish their actual efficacy, safety and potential adverse out-comes and to generalize the outcomes.

A Comparative Study on Alternative Education in China and Korea

Alternative educational practice has something in common that has been raised as an alternative to the crisis of traditional public education,but the causes,like background,and development patterns of alternative education may be different.As an alternative to restore education,various progressive schools or programs have been operated in Korea and China.The purpose of this study was to shed light on the alternative education in Korea and China,and seek to research its development process and practice.

Influence of electric parameters on MAO of AZ91D magnesium alloy using alternative square-wave power source

Micro-arc oxidation （MAO） process was carried out on AZ91D alloy in alkaline borate solution using an alternative square-wave power source with different parameters. The effects of voltage, frequency and duty cycle on the coatings were investigated by orthogonal experiment. It is found that the thickness of coatings increases with the increase of voltage and duty cycle, but decreases with the increase of frequency. The structure and morphology of the coatings also depend on voltage, frequency and duty cycle. The coatings become more porous and crack with increasing voltage and duty cycle. The coating is thin and transparent when the voltage is lower than 120 V. The corrosion resistances of different coatings were evaluated by polarization curves and electrochemical impedance spectroscopy （EIS） in 3.5% NaCl （mass fraction） solution. When the optimized values of voltage, frequency and duty cycle are 140 V, 2 000 Hz and 0.4, respectively, the anodic coating shows the best corrosion resistance.

A Comparative Study on Detection of the Expression of Alternative Oxidase in Tobacco Callus with the Monoclonal Antibody Against Alternative Oxidase and Antibody Against-Synthetic Polypeptide Antibody

从经过不同温度处理的烟草 (NicotianarusticaL .)愈伤组织中提取并纯化线粒体蛋白 ,分别与交替氧化酶的单克隆抗体和抗合成多肽抗体进行免疫杂交。结果表明 :交替氧化酶的含量随温度的下降而显著上升 ;单克隆抗体的特异性较高于抗合成多肽抗体 ,但后者与交替氧化酶同样有良好的亲和性。因此 ,用合成多肽方法制备的抗体可以用于交替氧化酶的研究中。

弱Hopf代数上的弱Alternative Doi-Hopf模

该文首先引入了弱Hopf代数上的弱Alternative Doi—Hopf模,然后构造了从弱Alter—native Doi-Hopf模范畴到模范畴(余模范畴)忘却函子的伴随函子.

Comparison of the Effects of H_2O_2 and Salicylic Acid on Alternative Respiratory Pathway in Aged Potato Tuber Slices

The influences of exogenous H 2O 2 and salicylic acid (SA) treatments on the alternative respiratory pathway (ARP) in aged potato (Solanum tuberosum L.) tuber slices were compared. The results showed that both H 2O 2(5.0 mmol/L) and SA (0.1 mmol/L) treatments had a significant inducing effect on ARP capacity (V alt ) and its ratio to total respiration (V alt /V t) in potato slices aged for 24 h. With a monoclonal antibody against the alternative oxidase (AOX), Western blotting results showed that both H 2O 2 and SA treatments increased the AOX expression levels in aged potato tuber slices. However, the results of oxygen isotope discrimination experiments showed that H 2O 2 had no influence on the in vivo ARP activity (ρV alt ) and its contribution to V t(expressed as ρV alt /V t) in potato slices aged for 24 h, but SA had a significant influence on the ρV alt and ρV alt /V t values of the aged potato tuber slices. These results indicate that there are differences between the effects of H 2O 2 and SA on ARP in plant tissues. Both of them possess the ability to induce ARP capacity through inducing AOX expression. However, SA can simultaneously stimulate the operation of ARP, but H 2O 2 can not.

Alternative Assessment in an EFL Project-based Learning Context: Perceptions and Correlations

This study aims to explore Chinese university EFL learners'perceptions toward alternative assessment in a context of a project-based learning digital storytelling presentation in Speaking Course.It also seeks to compare the relationship between alternative assessment and teacher assessment.The findings showed that a strong correlation between alternative assessment and teacher assessment occurred.Alternative assessment activities are viewed by students as"authentic"assessments,as they mimic how the student will be using their knowledge in the future.Alternative assessment as a form of formative assessment can be a powerful day-to-day tool for teachers and students.Alternative assessment is an enabler of process of learning.The study suggests that alternative assessment can encourage learners to become more fully responsible for their learning and can result in more and better learning.Alternative assessment can thus be used as a golden key to the"deaf and dumb"phenomenon for Chinese university EFL learners.

Dynamic regulation of alternative splicing and chromatin structure in Drosophila gonads revealed by RNA-seq

Both transcription and post-transcriptional processes, such as alternative splicing, play crucial roles in controlling developmental programs in metazoans. Recently emerged RNA-seq method has brought our understanding of eukaryotic transcriptomes to a new level, because it can resolve both gene expression level and alternative splicing events simultaneously. To gain a better understanding of cellular differentiation in gonads, we analyzed mRNA profiles from Drosophila testes and ovaries using RNA-seq. We identified a set of genes that have sex-specific isoforms in wild-type （WT） gonads, including several transcription factors. We found that differentiation of sperms from undifferentiated germ cells induced a dramatic downregulation of RNA splicing factors. Our data confirmed that RNA splicing events are significantly more frequent in the undifferentiated cell-enriched bag of marbles （barn） mutant testis, but downregulated upon differentiation in WT testis. Consistent with this, we showed that genes required for meiosis and terminal differentiation in WT testis were mainly regulated at the transcriptional level, but not by alternative splicing. Unexpectedly, we observed an increase in expression of all families of chromatin remodeling factors and histone modifying enzymes in the undifferentiated cell-enriched bam testis. More interestingly, chromatin regulators and histone modifying enzymes with opposite enzymatic activities are coenriched in undifferentiated cells in testis, suggesting that these cells may possess dynamic chromatin architecture. Finally, our data revealed many new features of the Drosophila gonadal transcriptomes, and will lead to a more comprehensive understanding of how differential gene expression and splicing regulate gametogenesis in Drosophila. Our data provided a foundation for the systematic study of gene expression and alternative splicing in many interesting areas of germ cell biology in Droso- phila, such as the molecular basis for sexual dimorphism and the regulation of the proliferation vs terminal differentiation programs in germline stem cell lineages. The GEO accession number for the raw and analyzed RNA-seq data is GSE16960.

Complementary and alternative medicines in irritable bowel syndrome: An integrative view

Irritable bowel syndrome(IBS)is a common gastrointestinal disorder with a high incidence in the general population.The diagnosis of IBS is mainly based on exclusion of other intestinal conditions through the absence of inflammatory markers and specific antigens.The current pharmacological treatment approaches available focus on reducing symptom severity while often limiting quality of life because of significant side effects.This has led to an effectiveness gap for IBS patients that seek further relief to increase their quality of life.Complementary and alternative medicines(CAM)have been associated with a higher degree of symptom management and quality of life in IBS patients.Over the past decade,a number of important clinical trials have shown that specific herbal therapies(peppermint oil and Iberogast?),hypnotherapy,cognitive behavior therapy,acupuncture,and yoga present with improved treatment outcomes in IBS patients.We propose an integrative approach to treating the diverse symptoms of IBS by combining the benefits of and need for pharmacotherapy with known CAM therapies to provide IBS patients with the best treatment outcome achievable.Initial steps in this direction are already being considered with an increasing number of practitioners recommending CAM therapies to their patients if pharmacotherapy alone does not alleviate symptoms sufficiently.

New Hybrid Parallel Algorithm for Variable-sized Batch Splitting Scheduling with Alternative Machines in Job Shops

The batch splitting scheduling problem has recently become a major target in manufacturing systems, and the researchers have obtained great achievements, whereas most of existing related researches focus on equal-sized and consistent-sized batch splitting scheduling problem, and solve the problem by fixing the number of sub-batches, or the sub-batch sizes, or both. Under such circumstance and to provide a practical method for production scheduling in batch production mode, a study was made on the batch splitting scheduling problem on alternative machines, based on the objective to minimize the makespan. A scheduling approach was presented to address the variable-sized batch splitting scheduling problem in job shops trying to optimize both the number of sub-bathes and the sub-batch sizes, based on differential evolution（DE）, making full use of the finding that the sum of values of genes in one chromosome remains the same before and after mutation in DE. Considering before-arrival set-up time and processing time separately, a variable-sized batch splitting scheduling model was established and a new hybrid algorithm was brought forward to solve both the batch splitting problem and the batch scheduling problem. A new parallel chromosome representation was adopted, and the batch scheduling chromosome and the batch splitting chromosome were treated separately during the global search procedure, based on self-adaptive DE and genetic crossover operator, respectively. A new local search method was further designed to gain a better performance. A solution consists of the optimum number of sub-bathes for each operation per job, the optimum batch size for each sub-batch and the optimum sequence of sub-batches. Computational experiments of four test instances and a realistic problem in a speaker workshop were performed to testify the effectiveness of the proposed scheduling method. The study takes advantage of DE＇s distinctive feature, and employs the algorithm as a solution approach, and thereby deepens and enriches the content of batch splitting scheduling.

Diets, functional foods, and nutraceuticals as alternative therapies for inflammatory bowel disease: Present status and future trends

Inflammatory bowel disease(IBD) is a serious health concern among western societies. The disease is also on the rise in some East Asian countries and in Australia. Health professionals and dietitians around the world are facing an unprecedented challenge to prevent and control the increasing prevalence of IBD. The current therapeutic strategy that includes drugs and biological treatments is inefficient and are associated with adverse health consequences. In this context, the use of natural products is gaining worldwide attention. In vivo studies and clinical evidence suggest that wellplanned dietary regimens with specific nutrients can alleviate gastrointestinal inflammation by modulating inflammatory cytokines, such as tumor necrosis factor α(TNF-α), interleukin 1(IL-1), IL-6, IL-1β, and IL-10. Alternatively, the avoidance of high-fat and highcarbohydrate diets is regarded as an effective tool to eliminate the causes of IBD. Many functional foods and bioactive components have received attention for showing strong therapeutic effects against IBD. Both animal and human studies suggest that bioactive functional foods can ameliorate IBD by downregulating the pro-inflammatory signaling pathways, such as nuclear factor κB, STAT1, STAT6, and pro-inflammatory cytokines, including IL-1β, IL-4, IL-6, COX-2, TNF-α, and interferon γ. Therefore, functional foods and diets have the potential to alleviate IBD by modulating the underlying pathogenic mechanisms. Future comprehensive studies are needed to corroborate the potential roles of functional foods and diets in the prevention and control of IBD.

Allele-specific expression and alternative splicing in horse×donkey and cattle×yak hybrids

Divergence of gene expression and alter native splicing is a crucial driving force in the evolution of species;to date, however the molecular mechanism remains unclear. Hybrids of closely related species provide a suitable model to analyze allele-specific expressi on (ASE) and allele-specific alter native splicing (ASS). Analysis of ASE and ASS can uncover the differences in cis-regulatory elements between closely related species, while eliminating interferenee of trans-regulatory elements. Here, we provide a detailed characterization of ASE and ASS from 19 and 10 transcriptome datasets across five tissues from reciprocal-cross hybrids of horsex don key (mule/hi nny) and cattlexyak (dzo), respectively. Results showed that 4.8%-8.7% and 10.8%-16.7% of genes exhibited ASE and ASS, respectively. Notably, IncRNAs and pseudogenes were more likely to show ASE than protein-coding genes. In addition, genes showing ASE and ASS in mule/hinny were found to be involved in the regulation of muscle strength, whereas those of dzo were involved in high-altitude adaptati on. In con clusi on, our study dem on strated that explorati on of genes showing ASE and ASS in hybrids of closely related species is feasible for species evolution research.

Job shop scheduling problem with alternative machines using genetic algorithms

The classical job shop scheduling problem(JSP) is the most popular machine scheduling model in practice and is known as NP-hard.The formulation of the JSP is based on the assumption that for each part type or job there is only one process plan that prescribes the sequence of operations and the machine on which each operation has to be performed.However,JSP with alternative machines for various operations is an extension of the classical JSP,which allows an operation to be processed by any machine from a given set of machines.Since this problem requires an additional decision of machine allocation during scheduling,it is much more complex than JSP.We present a domain independent genetic algorithm(GA) approach for the job shop scheduling problem with alternative machines.The GA is implemented in a spreadsheet environment.The performance of the proposed GA is analyzed by comparing with various problem instances taken from the literatures.The result shows that the proposed GA is competitive with the existing approaches.A simplified approach that would be beneficial to both practitioners and researchers is presented for solving scheduling problems with alternative machines.

Magnesium alloys as extremely promising alternatives for temporary orthopedic implants-A review

Mg alloys are emerging as potential and very promising alternatives for replacing permanent metallic implant materials such as steels and titanium in applications where the implants need to be removed following healing through revision surgery.Use of Mg alloys for implant application is seen as a game changer and Mg alloys are almost perfect materials for the future in both engineering and biomedical applications.Present review therefore focuses on highlighting significance of Mg alloys in biomedical field and risks of using permanent metallic implants particularly when the implants are no longer required after the injury is healed.In this review,importance of orthopedic implants in present scenario,serious concern related to accidents that are causing permanent disabilities,demand in orthopedic implant market worldwide,potential applications of Mg based materials and their compatibility in biological environment is presented and discussed.In addition,degradation rate,major reactions associated with Mg based materials and effect of alloying elements on implant performance are also discussed based on in-vivo results.Recent advances in development of Mg alloys through various techniques and their performance in in-vitro conditions are also outlined.Possible ways to eliminate the limitations of Mg alloys include alloying,melt purification,surface alterations,surface modifications,chemical treatment,secondary processing etc.are discussed.Challenges and opportunities for Mg alloys to become ideal implant material is also addressed.