Category: 3. Business

  • The 6th Round of Bilateral Political Consultations between Pakistan and Finland

    The 6th round of Bilateral Political Consultations between Pakistan and Finland was held today in Islamabad. 

    The Pakistan side was led by Additional Secretary Europe Muhammad Khalid Jamali and the Finnish side was led by Under-Secretary of State for Foreign and Security Policy Outi Holopainen. 

    The two sides discussed the entire spectrum of bilateral relations including cooperation in trade, investment, education, science and technology and climate change. Both sides also held a detailed exchange of views on global and regional issues of mutual interest.

    The Additional Secretary (Europe) appreciated the positive momentum in bilateral ties and reaffirmed Pakistan’s commitment to further strengthen cooperation with Finland. 

    Furthermore, the Additional Secretary thanked the Finnish side for its support on Pakistan’s GSP Plus status and invited Finnish companies to participate in the upcoming Pakistan-EU Business Forum scheduled for April 2026. 

    It was agreed to hold the next round of consultations in Finland.

     

    Islamabad
    November 18, 2025

    Continue Reading

  • Press release: Climate Change Performance Index 2026

    Press release: Climate Change Performance Index 2026

    Climate ranking shows: The world is making progress, but the US and other Petrostates are resisting change

    Germanwatch, NewClimate Institute, and CAN International publish the 21st edition of the Climate Change Performance Index (CCPI 2026): Denmark remains at the top, followed by UK and Morocco. Petrostates rank at the bottom: Saudi Arabia, Iran and USA. Dynamics in many G20 countries not yet strong enough.

    Belém (18 Nov 2025). Ten years after the Paris Agreement, progress is clearly visible: Global per capita emissions are falling, renewable energies are growing massively, and more than 100 countries now have their own climate targets for net-zero emissions. However, the pace is still too slow to meet the Paris climate targets. The Climate Change Performance Index (CCPI) published today by Germanwatch and the NewClimate Institute also paints this ambivalent picture.

    As in previous years, the top three ranks remain vacant. Countries must accelerate climate action to align with the Paris Agreement’s temperature limit. Denmark remains the top-ranked country (4th). It ranks first in climate policy and is among the only three countries that achieve a very high rating in the field of renewable energies – the country is a leader in offshore energy. Denmark is followed by the United Kingdom (5th), which has climbed one place compared to last year. Overall, the country’s years of climate policy efforts are paying off – for example the UK already had the coal phase-out last year. However, the country still has some catching up to do, especially when it comes to renewable energy (ranked ‘low‘). Morocco (6th) scores ‘good’ in all categories except renewable energies, where the trend is nevertheless ‘good’. The country still has very low per capita emissions and impresses with major investments in public transport and a respectable new climate target for 2035.

    Niklas Höhne (NewClimate Institute), author of the CCPI: “While we cannot yet attest to any country having very good climate mitigation performance overall, there are pioneers in some categories who are demonstrating ambitious performance. Pakistan, for example, is a surprise contender in terms of emissions and energy consumption thanks to its very low per capita figures. As in previous years, Norway, Denmark and Sweden are setting the standards in renewable energies.”

    In contrast, there are the countries that perform worst in the ranking. The three last-placed countries in the CCPI are Saudi Arabia (67th), Iran (66th), and the USA (65th). Thea Uhlich (Germanwatch), author of the CCPI: “The USA has suffered a particularly remarkable decline – ranking third to last in the overall standings just behind Russia. The largest oil- and gas-producing countries are virtually among themselves and show no sign of departing from fossil fuels as a business model. This means they are missing out on an opportunity to embrace the future.”

    G20 countries: Only one good – ten very weak

    “There is positive momentum for renewable energies and electrification worldwide. However, we also see a worrying picture among the major emitters – the G20 countries – with only one country in the ‘high’ category and ten in the ‘very low’ category,” says Uhlich. Although the G20 countries are responsible for more than 75 percent of global greenhouse gas (GHG) emissions and have a special responsibility, only one G20 country, the UK, achieves a ‘high‘ rating in the ranking. It is particularly worrying that ten G20 countries are still classified as ‘very low‘ (Turkey, China, Australia, Japan, Argentina, Canada, Korea, Russia, USA, Saudi Arabia), followed by three more in the low category: South Africa, Indonesia, and Italy.

    The largest carbon emitter, China (54th), has improved by one place but – despite the dynamics on the way to the first electro state – still has a very low rating. Only in the area of climate policy is China achieving a ‘high‘.

    In the first quarter of 2025, China’s emissions declined – this could be an indication that emissions in China have peaked. Although China is a pioneer in green technologies – there is an ongoing boom in electric cars, batteries and renewable energies – and has set a relatively ambitious climate target, it is simultaneously expanding its fossil fuel production. It is important for China that the expansion of renewable energy and e-mobility goes hand in hand with the phase-out of fossil fuels, not only to prevent emissions from rising further, but also to bring them down quickly.

    India (23th), one of the largest emitters, is one of the biggest faller in this year’s ranking, landing in the ‘medium‘ category. Jan Burck (Germanwatch), author of the CCPI: “The decline in the ranking is due to a combination of factors. India ranks last in terms of emissions trends, as emissions have been rising steadily in recent years. At the same time, energy consumption is increasing. India has also lost many places in climate policy rankings mainly due to its lack of a plan to phase out coal or even a concrete phase-out date. If India will reduce the building of new coal power plants and continues the promising trend for renewables, the country can achieve a much better ranking again next year.”

    Continue Reading

  • BAE Systems, Boeing and Saab to Collaborate on Next-Gen Pilot Training – BAE Systems

    1. BAE Systems, Boeing and Saab to Collaborate on Next-Gen Pilot Training  BAE Systems
    2. Boeing says partnership for T-7 trainer export bid imminent  Reuters
    3. Boeing, Saab, and BAE Plot Trainer Jet Power Move–Hawk Successor Could Shake Up Market  TradingView
    4. BAE Systems ties up with Boeing, Saab for UK jet trainer bid  MarketScreener
    5. Boeing hints at new alliance as U.K. eyes Hawk replacement (BA:NYSE)  Seeking Alpha

    Continue Reading

  • Genprex Announces U.S. Patent for Reqorsa® Gene Therapy in Combination with PD-L1 Antibodies to Treat Cancers

    Genprex Announces U.S. Patent for Reqorsa® Gene Therapy in Combination with PD-L1 Antibodies to Treat Cancers

    Strengthens Intellectual Property Portfolio and Provides Protection for Therapeutic Combination in Acclaim-3 Clinical Trial

    AUSTIN, Texas, Nov. 18, 2025 /PRNewswire/ — Genprex, Inc. (“Genprex” or the “Company”) (NASDAQ: GNPX), a clinical-stage gene therapy company focused on developing life-changing therapies for patients with cancer and diabetes, today announced that the United States Patent and Trademark Office (USPTO) has granted Genprex a patent that covers the use of the Company’s lead drug candidate, Reqorsa® Gene Therapy, in combination with PD-L1 antibodies, such as Tecentriq®, through 2037.

    “We continue to build protection around our lead drug candidate, REQORSA, and this new patent provides the necessary technology protection applicable to our Acclaim-3 clinical trial in small cell lung cancer,” said Thomas Gallagher, Senior Vice President of Intellectual Property and Licensing at Genprex. “In the Acclaim-3 clinical trial, we are combining REQORSA with Tecentriq, a PD-L1 antibody, and this patent secures exclusivity for this drug combination for the treatment of cancer, preventing would-be competitors from making, using or selling this drug combination.”

    Genprex has been granted patents for the use of REQORSA in combination with PD-L1 antibodies in the U.S. and Korea. Genprex is pursuing additional patent applications in Europe, Canada, Brazil, China and Israel. Should these applications grant, they would also be applicable to Genprex’s Acclaim-3 clinical trial.

    PD-L1 antibodies are a type of targeted immunotherapy that block the activity of PD-L1 immune checkpoint proteins present on the surface of cells.

    About Acclaim-3
    Acclaim-3 is a Phase 1/2 clinical trial evaluating the combination of REQORSA and Genentech’s Tecentriq® (atezolizumab) as maintenance therapy in patients with extensive stage small cell lung cancer (ES-SCLC) who are candidates for maintenance therapy after receiving Tecentriq and chemotherapy as standard of care initial treatment. In this study, patients will be treated with REQORSA and Tecentriq until disease progression or unacceptable toxicity is experienced.

    The Phase 2 expansion study follows the successful completion of the Phase 1 dose escalation portion of the study, which showed REQORSA was generally well tolerated. There were no dose limiting toxicities, and in Acclaim-3, the Phase 2 patients are receiving the same dose of REQORSA as patients in the Phase 2 portion of Acclaim-1.

    The Phase 2 expansion portion is expected to enroll approximately 50 patients. The primary endpoint of the Phase 2 portion is to determine the 18-week progression-free survival rate from the time of the start of maintenance therapy with REQORSA and Tecentriq in patients with ES-SCLC. Patients will also be followed for survival. Genprex’s team plans to conduct an interim analysis after the 25th patient enrolled and treated reaches 18 weeks of follow up. The Company expects to complete enrollment of the first 25 patients for interim analysis in the Phase 2 expansion portion of the study in the first half of 2026. The Acclaim-3 clinical trial is supported by U.S. Food and Drug Administration (FDA) Fast Track Designation and Orphan Drug Designation.

    About Genprex, Inc.
    Genprex, Inc. is a clinical-stage gene therapy company focused on developing life-changing therapies for patients with cancer and diabetes. Genprex’s technologies are designed to administer disease-fighting genes to provide new therapies for large patient populations with cancer and diabetes who currently have limited treatment options. Genprex works with world-class institutions and collaborators to develop drug candidates to further its pipeline of gene therapies in order to provide novel treatment approaches. Genprex’s oncology program utilizes its systemic, non-viral Oncoprex® Delivery System which encapsulates the gene-expressing plasmids using lipid-based nanoparticles in a lipoplex form. The resultant product is administered intravenously, where it is taken up by tumor cells that then express tumor suppressor proteins that were deficient in the tumor. The Company’s lead product candidate, Reqorsa® Gene Therapy (quaratusugene ozeplasmid), is being evaluated in two clinical trials as a treatment for NSCLC and SCLC. Each of Genprex’s lung cancer clinical programs has received a Fast Track Designation from the FDA for the treatment of that patient population, and Genprex’s SCLC program has received an FDA Orphan Drug Designation. Genprex’s diabetes gene therapy approach is comprised of a novel infusion process that uses an AAV vector to deliver Pdx1 and MafA genes directly to the pancreas. In models of Type 1 diabetes, GPX-002 transforms alpha cells in the pancreas into functional beta-like cells, which can produce insulin but may be distinct enough from beta cells to evade the body’s immune system. In a similar approach for Type 2 diabetes, where autoimmunity is not at play, GPX-002 is believed to rejuvenate and replenish exhausted beta cells.

    Interested investors and shareholders are encouraged to sign up for press releases and industry updates by visiting the Company Website, registering for Email Alerts and by following Genprex on Twitter, Facebook and LinkedIn.

    Cautionary Language Concerning Forward-Looking Statements
    Statements contained in this press release regarding matters that are not historical facts are “forward-looking statements” within the meaning of the Private Securities Litigation Reform Act of 1995. These forward-looking statements are made on the basis of the current beliefs, expectations and assumptions of management, are not guarantees of performance and are subject to significant risks and uncertainty. These forward-looking statements should, therefore, be considered in light of various important factors, including those set forth in Genprex’s reports that it files from time to time with the Securities and Exchange Commission and which you should review, including those statements under “Item 1A – Risk Factors” in Genprex’s Annual Report on Form 10-K for the year ended December 31, 2024.

    Because forward-looking statements are subject to risks and uncertainties, actual results may differ materially from those expressed or implied by such forward-looking statements. Such statements include, but are not limited to, statements regarding: Genprex’s ability to advance the clinical development, manufacturing and commercialization of its product candidates in accordance with projected timelines and specifications; the timing and success of Genprex’s clinical trials, its intended regulatory submissions and any resulting regulatory approvals; the effect of Genprex’s product candidates, alone and in combination with other therapies, on cancer and diabetes; Genprex’s future growth and financial status, including Genprex’s ability to maintain compliance with the continued listing requirements of The Nasdaq Capital Market and to continue as a going concern and to obtain capital to meet its long-term liquidity needs on acceptable terms, or at all; Genprex’s commercial and strategic partnerships, including those with its third party vendors, suppliers and manufacturers and their ability to successfully perform and scale up the manufacture of its product candidates; Genprex’s intellectual property and licenses, including the potential for future grants of patent applications globally; and Genprex’s current expectations, estimates, forecasts and projections about the industry and markets in which it operates.

    These forward-looking statements should not be relied upon as predictions of future events and Genprex cannot assure you that the events or circumstances discussed or reflected in these statements will be achieved or will occur. If such forward-looking statements prove to be inaccurate, the inaccuracy may be material. You should not regard these statements as a representation or warranty by Genprex or any other person that Genprex will achieve its objectives and plans in any specified timeframe, or at all. You are cautioned not to place undue reliance on these forward-looking statements, which speak only as of the date of this press release. Genprex disclaims any obligation to publicly update or release any revisions to these forward-looking statements, whether as a result of new information, future events or otherwise, after the date of this press release or to reflect the occurrence of unanticipated events, except as required by law.

    Genprex, Inc.
    (877) 774-GNPX (4679)

    GNPX Investor Relations
    [email protected]

    GNPX Media Contact
    Kalyn Dabbs
    [email protected]

    SOURCE Genprex, Inc.


    Continue Reading

  • Vertiv and Caterpillar Announce Energy Optimization Collaboration to Expand End-to-End Power and Cooling Offerings for AI Data Centers

    Vertiv and Caterpillar Announce Energy Optimization Collaboration to Expand End-to-End Power and Cooling Offerings for AI Data Centers

    New agreement aims to enhance data center efficiency, resiliency and deployment timelines through integrated energy solutions.

    COLUMBUS, Ohio, Nov. 18, 2025 /PRNewswire/ — Vertiv (NYSE: VRT), a global leader in critical digital infrastructure, and Caterpillar Inc. (NYSE: CAT), a global leader in power systems, today announced the signing of a strategic undertaking to collaborate on advanced energy optimization solutions for data centers. This initiative will integrate Vertiv’s power distribution and cooling portfolio with Caterpillar’s, and its subsidiary Solar Turbines’, product and expertise in power generation and CCHP (Combined Cooling, Heat and Power) to deliver pre-designed architectures that simplify deployment, accelerate time-to-power and optimize performance for data center operations.

    A Powerful Collaboration:
    This collaboration directly addresses the growing demand for on-site energy solutions that deliver reliable power and cooling. Together, the companies are able to offer a fully integrated solution with validated interfaces and performance, enabling customers to accelerate design, installation and deployment.

    • Caterpillar and Solar Turbines will supply power generation solutions, such as natural gas turbines and reciprocating engines, to deliver dependable, scalable electric power and thermal energy for CCHP.
    • Vertiv will provide a complete portfolio of power and cooling solutions and services, packaged as modular, pre-designed blocks, to shorten design cycles and standardize deployment.

    The Customer Advantages:

    • Accelerates Time-to-Power – by utilizing predesigned, modular reference architectures to speed up deployment time.
    • Lowers PUE (Power Usage Effectiveness) – enables improved energy efficiency and carbon footprint because the system is optimized end-to-end: power, cooling, distribution and dynamic load management, compared to traditional design.
    • Global lifecycle support – the offering is backed by the trusted, global service and support networks of both Vertiv and Caterpillar.

    “This collaboration with Caterpillar and Solar Turbines is a cornerstone of our Bring Your Own Power & Cooling (BYOP&C) strategy and aligns seamlessly with our grid-to-chip framework by offering resilient, on-site power generation solutions. This is optimal for customers looking to reduce or eliminate grid dependence,” said Gio Albertazzi, CEO, at Vertiv. “By combining our complementary technologies, portfolios and expertise, we are enabling coordinated integration. Our pre-engineered, interoperability-tested building blocks let customers execute design, build and deploy concurrently, with predictable system performance.”

    “As AI-driven workloads continue to accelerate, the demand for robust and scalable power infrastructure and cooling is becoming increasingly critical,” said Jason Kaiser, group president of Caterpillar Power & Energy. “Our collaboration with Vertiv will enable us to deliver integrated, on-site energy solutions that lower PUE and meet customers’ evolving needs.”

    This initiative directly addresses the growing demand for on-site energy solutions and offers a coordinated, customer-first approach to solution design and implementation. The Vertiv and Caterpillar Memorandum of Understanding (MOU) represents a pivotal step in further refining this ecosystem, enabling customers to overcome energy constraints and deploy optimized AI centers.

    To learn more about Vertiv’s end-to-end power and thermal management solutions, visit Vertiv.com.
    To learn more about the Caterpillar capability, visit Caterpillar.com / SolarTurbines.com.

    About Vertiv
    Vertiv (NYSE: VRT) brings together hardware, software, analytics and ongoing services to enable its customers’ vital applications to run continuously, perform optimally and grow with their business needs. Vertiv solves the most important challenges facing today’s data centers, communication networks and commercial and industrial facilities with a portfolio of power, cooling and IT infrastructure solutions and services that extends from the cloud to the edge of the network. Headquartered in Westerville, Ohio, USA, Vertiv does business in more than 130 countries. For more information, and for the latest news and content from Vertiv, visit Vertiv.com.

    About Caterpillar
    With 2024 sales and revenues of $64.8 billion, Caterpillar Inc. is the world’s leading manufacturer of construction and mining equipment, off-highway diesel and natural gas engines, industrial gas turbines and diesel-electric locomotives. For 100 years, we’ve been helping customers build a better, more sustainable world and are committed and contributing to a reduced-carbon future. Our innovative products and services, backed by our global dealer network, provide exceptional value that helps customers succeed. Caterpillar does business on every continent, principally operating through three primary segments – Construction Industries, Resource Industries and Energy & Transportation – and providing financing and related services through our Financial Products segment. Visit us at caterpillar.com or join the conversation on our social media channels.

    About Solar Turbines
    Solar Turbines Incorporated, headquartered in San Diego, is a wholly owned subsidiary of Caterpillar Inc. Solar manufactures the world’s most widely used family of mid-sized industrial gas turbines from the 1 – 39 MW range. More than 15,000 Solar units are operating in 100 countries around the world. Primary applications include electric power generation, oil and natural gas production and natural gas transmission.

    Forward-looking statements
    This release contains forward-looking statements within the meaning of the Private Securities Litigation Reform Act of 1995, Section 27 of the Securities Act, and Section 21E of the Securities Exchange Act. These statements are only a prediction. Actual events or results may differ materially from those in the forward-looking statements set forth herein. Readers are referred to Vertiv’s filings with the Securities and Exchange Commission, including its most recent Annual Report on Form 10-K and any subsequent Quarterly Reports on Form 10-Q for a discussion of these and other important risk factors concerning Vertiv and its operations. Vertiv is under no obligation to, and expressly disclaims any obligation to, update or alter its forward-looking statements, whether as a result of new information, future events or otherwise.

    VERTIV CONTACT
    [email protected]

    CATERPILLAR CONTACT
    [email protected]

    SOURCE Vertiv Holdings Co

    Continue Reading

  • Jia H, Li M, Li W, Liu L, Jian Y, Yang Z, et al. A serine/threonine protein kinase encoding gene KERNEL NUMBER PER ROW6 regulates maize grain yield. Nat Commun. 2020;11(1):988. https://doi.org/10.1038/s41467-020-14746-7.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Guo W, Guo X, Xu L, Shao L, Zhu B, Liu H, et al. Effect of whole-plant corn silage treated with lignocellulose-degrading bacteria on growth performance, rumen fermentation, and rumen microflora in sheep. Animal. 2022;16(7):100576. https://doi.org/10.1016/j.animal.2022.100576.

    Article 
    PubMed 

    Google Scholar 

  • Zhang H, Wu J, Zhao X, Yan P, Yang R, Yan J, et al. Improving aerobic stability and methane production of maize stover silage with lactic acid bacteria inoculants: focus on pentose-fermentation. IND CROP PROD. 2023;201:116861. https://doi.org/10.1016/j.indcrop.2023.116861.

    Article 

    Google Scholar 

  • Tahir M, Wang T, Zhang J, Xia T, Deng X, Cao X, Zhong J. Compound lactic acid bacteria enhance the aerobic stability of Sesbania Cannabina and maize mixed silage. BMC Microbiol. 2025;25(1):68. https://doi.org/10.1186/s12866-025-03781-3.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Xin Y, Chen C, Zhong Y, Bu X, Huang S, Tahir M, Du Z, Liu W, Yang W, Li J, Wu Y, Zhang Z, Lian J, Xiao Q, Yan Y. Effect of storage time on the silage quality and microbial community of mixed maize and Faba bean in the Qinghai-Tibet plateau. Front Microbiol. 2023;13:1090401. https://doi.org/10.3389/fmicb.2023.1161337.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Liao C, Na B, Tang X, Zhao M, Zhang C, Chen S, et al. Contribution of the bacterial community of poorly fermented oat silage to biogas emissions on the Qinghai Tibetan plateau. Sci Total Environ. 2023;897:165336. https://doi.org/10.1016/j.scitotenv.2023.165336.

    Article 
    PubMed 

    Google Scholar 

  • Li F, Jia M, Chen H, Chen M, Su R, Usman S, Ding Z, Hao L, Franco M, Guo X. Responses of microbial community composition and cazymes encoding gene enrichment in ensiled Elymus nutans to altitudinal gradients in alpine region. Appl Environ Microbiol. 2024;90(10):e0098624. https://doi.org/10.1128/aem.00986-24.

    Article 
    PubMed 

    Google Scholar 

  • Jatkauskas J, Vrotniakiene V, Eisner I, Witt KL, do Amaral RC. Comparison of the chemical and microbial composition and aerobic stability of high-moisture barley grain ensiled with either chemical or viable lactic acid bacteria application. Fermentation. 2024;10:62. https://doi.org/10.3390/fermentation10010062.

    Article 

    Google Scholar 

  • Chen L, Bai S, You M, Xiao B, Li P, Cai Y. Effect of a low temperature tolerant lactic acid bacteria inoculant on the fermentation quality and bacterial community of oat round bale silage. Anim Feed Sci Technol. 2020;269:114669. https://doi.org/10.1016/j.anifeedsci.2020.114669.

    Article 

    Google Scholar 

  • Wang X, Han X, Wang H, Jing X, Liu C, Zhou Y. Studying progress of Lactobacillus’s responses in a variety of stress. Sci Technol Food Ind. 2015;6:365–9.

    Google Scholar 

  • Wang S, Dong Z, Li J, Chen L, Shao T. Pediococcus acidilactici strains as silage inoculants for improving the fermentation quality, nutritive value and in vitro ruminal digestibility in different forages. J Appl Microbiol. 2018;126:1–10. https://doi.org/10.1111/jam.14146.

    Article 

    Google Scholar 

  • Liu H, Zeng T, Zhang Y, Wen X, Liu H, Zhang L, Xiao O, Li X, Yan Y. Screening and identification of low temperature resistant lactic acid bacteria and its effect on fermentation quality of oat silage. Pratacultural Sci. 2025;42(3):669–78. https://doi.org/10.11829/j.issn.1001-0629.2023-0688.

    Article 

    Google Scholar 

  • Chen C, Xin Y, Li X, Ni H, Zeng T, Du Z, et al. Effects of acremonium cellulase and heat-resistant lactic acid bacteria on lignocellulose degradation, fermentation quality, and microbial community structure of hybrid elephant grass silage in humid and hot areas. Front Microbiol. 2022;13:1066753. https://doi.org/10.3389/fmicb.2022.1066753.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Broderick GA, Kang JH. Automated simultaneous determination of ammonia and total amino acids in ruminal fluid and in vitro media. J Dairy Sci. 1980;63:64–75. https://doi.org/10.3168/jds.s0022-0302(80)82888-8.

    Article 
    PubMed 

    Google Scholar 

  • Murphy RP. A method for the extraction of plant samples and the determination of total soluble carbohydrates. J Sci Food Agric. 1958;9:714–7. https://doi.org/10.1029/2001JB000884.

    Article 

    Google Scholar 

  • Van Soest PJ, Robertson JB, Lewis BA. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J Dairy Sci. 1991;74:3583–97. https://doi.org/10.3168/jds.s0022-0302(91)78551-2.

    Article 
    PubMed 

    Google Scholar 

  • AOAC. Official methods of analysis. J Pharm Sci. 1975;60:414–5. https://doi.org/10.1002/jps.2600600253.

    Article 

    Google Scholar 

  • Zhu L, Zhao M, Yan Y, Sun P, Yan X, Liu M, Na R, Jia Y, Cha S, Ge G. Characteristics of isolated lactic acid bacteria at low temperature and their effects on the silage quality. Microbiol Spectr. 2025;6:e0319424. https://doi.org/10.1128/spectrum.03194-24.

    Article 

    Google Scholar 

  • Ren H, Shi R, Yang D, Tian H, Wang L, Ling Z, et al. Innovative strategy to enhance bioconversion of sweet sorghum Bagasse (SSB) by the combination of bio-fortified ensiling and dilute alkali pretreatment. Ind Crop Prod. 2024;211:118208. https://doi.org/10.1016/j.indcrop.2024.118208.

    Article 

    Google Scholar 

  • Gaspar P, Neves AR, Shearman CA, Gasson MJ, Baptista AM, Turner DL, et al. The lactate dehydrogenases encoded by the Ldh and LdhB genes in Lactococcus lactis exhibit distinct regulation and catalytic properties – comparative modeling to probe the molecular basis. FEBS J. 2007;274(22):5924–36. https://doi.org/10.1111/j.1742-4658.2007.06115.x.

    Article 
    PubMed 

    Google Scholar 

  • Yang Y, Bao C, Li K, Pan Y, Wang X, Fang G, Chen H, Zhang S, Chen G, Gang W. Identification, low temperature growth characteristics and tolerance mechanism of low temperature-resistant strain. China Brew. 2022;41(5):47–51. https://doi.org/10.11882/j.issn.0254-5071.2022.05.009.

    Article 

    Google Scholar 

  • Bernardes T, Daniel J, Adesogan T, McAllister A, Nussio D. Silage review: unique challenges of silages made in hot and cold regions. J Dairy Sci. 2018;101:4001–19. https://doi.org/10.3168/jds.2017-13703.

    Article 
    PubMed 

    Google Scholar 

  • Liu X, Yu X, He J, Guo J, Wang P, Wang Z. Interactions between nine probiotics and mechanisms of cooperative symbiosis. Food Ferment Ind. 2019;45(13):65–70. https://doi.org/10.13995/j.cnki.11-1802/ts.019868.

    Article 

    Google Scholar 

  • Kleinschmit DH Jr. A meta-analysis of the effects of Lactobacillus buchneri on the fermentation and aerobic stability of corn and grass and small-grain silages. J Dairy Sci. 2006;89(10):4005–13. https://doi.org/10.3168/jds.S0022-0302(06)72444-4.

    Article 
    PubMed 

    Google Scholar 

  • Yin X, Zhao J, Wang S, Dong Z, Li J, Shao T. The effects of epiphytic microbiota and chemical composition of Italian ryegrass harvested at different growth stages on silage fermentation. J Sci Food Agric. 2023;103(3):1385–93. https://doi.org/10.1002/jsfa.12232.

    Article 
    PubMed 

    Google Scholar 

  • You L, Bao W, Yao C, Zhao F, Jin H, Huang W, Li B, Kwok L, Liu W. Changes in chemical composition, structural and functional Microbiome during alfalfa (Medicago sativa) ensilage with Lactobacillus plantarum PS-8. Anim Nutr. 2022;24:100–9. https://doi.org/10.1016/j.aninu.2021.12.004.

    Article 

    Google Scholar 

  • Yang S, Xing Y, Gao J, Jin R, Lin R, Weng W, et al. Lacticaseibacillus paracasei fermentation broth identified peptide, Y2Fr, and its antibacterial activity on vibrio parahaemolyticus. Microb Pathog. 2023;182:106260. https://doi.org/10.1016/j.micpath.2023.106260.

    Article 
    PubMed 

    Google Scholar 

  • Zhao J, Dong Z, Li J, Chen L, Bai Y, Jia Y, et al. Ensiling as pretreatment of rice straw: the effect of hemicellulase and Lactobacillus plantarum on hemicellulose degradation and cellulose conversion. Bioresour Technol. 2018;266:158–65. https://doi.org/10.1016/j.biortech.2018.06.058.

    Article 
    PubMed 

    Google Scholar 

  • Cheng X, Huang L, Li K. Antioxidant activity changes of exopolysaccharides with different carbon sources from Lactobacillus plantarum LPC-1 and its metabolomic analysis. World J Microbiol Biotechnol. 2019;35(5):68. https://doi.org/10.1007/s11274-019-2645-6.

    Article 
    PubMed 

    Google Scholar 

  • Li F, Ding Z, Ke W, Xu D, Zhang P, Bai J, et al. Ferulic acid esterase-producing lactic acid bacteria and cellulase pretreatments of maize stalk silage at two different temperatures: ensiling characteristics, carbohydrates composition, and enzymatic saccharification. Bioresour Technol. 2019;282:211–21. https://doi.org/10.1016/j.biortech.2019.03.022.

    Article 
    PubMed 

    Google Scholar 

  • Gong M, Wang Y, Bao D, Jiang S, Chen H, Shang J, Wang X, Hnin Y, Zou G. Improving cold-adaptability of mesophilic cellulase complex with a novel mushroom cellobiohydrolase for efficient low-temperature ensiling. Bioresour Technol. 2023;376:128888. https://doi.org/10.1016/j.biortech.2023.128888.

    Article 
    PubMed 

    Google Scholar 

  • Santos MAB, Morais F, Mandelli EA, Lima RY, Miyamoto PMR, Higasl EAA, Araujo DAA, Paixao JMJ, Motta ML, Streit RSA, Morao LG, Silva CBC, Wolf CRF, Terrasan NR, Bulka JA, Diogo FJ, Fuzita FM, Colombar CR, Santos PT, Rodrigues DB, Silva SG, Bernardes N, Terrapon V, Lombard AJC, Henrissat B, Bissaro MJGB, Persinoti GF, Berrin MT. A metagenomic ‘dark matter’ enzyme catalyses oxidative cellulose conversion. Nature. 2025;639:1076–83. https://doi.org/10.1038/s41586-024-08553-z.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Weissbach F, Kuhla S, Schmidt L. Estimation of the metabolizable energy in forages by a cellulase-method. Proc Soc Nutr Physiol. 1996;5:11–25. https://doi.org/10.2134/1994.foragequality.c16.

    Article 

    Google Scholar 

  • Zong C, Xiao Y, Shao T, Chiou AJ, Wu A, Huang Z, Chen C, Jiang W, Zhu J, Dong Z, Liu Q, Li M. Alfalfa as a vegetable source of β-carotene: the change mechanism of β-carotene during fermentation. Food Res Int. 2023;172:113104. https://doi.org/10.1016/j.foodres.2023.113104.

    Article 
    PubMed 

    Google Scholar 

  • Ferrero F, Tabacco E, Piano S, Casale M, Borreani G. Temperature during conservation in laboratory silos affects fermentation profile and aerobic stability of corn silage treated with Lactobacillus buchneri, Lactobacillus hilgardii, and their combination. J Dairy Sci. 2021;104(2):1696–713. https://doi.org/10.3168/jds.2020-18733.

    Article 
    PubMed 

    Google Scholar 

  • Bai J, Ding Z, Su R, Wang M, Cheng M, Xie D, Guo X. Storage temperature is more effective than lactic acid bacteria inoculations in manipulating fermentation and bacterial community diversity, co-occurrence and functionality of the whole-plant maize silage. Microbiol Spectr. 2022;10:e00101–22. https://doi.org/10.1128/spectrum.00101-22.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Li M, Fan X, Cheng Q, Chen Y, Long J, Lei Y, et al. Effect of Amomum villosum essential oil as an additive on the chemical composition, fermentation quality, and bacterial community of paper mulberry silage. Front Microbiol. 2022;13:951958. https://doi.org/10.3389/fmicb.2022.951958.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Ogunade IM, Jiang Y, Cervantes AAP, Kim DH, Oliveira AS, Vyas D, Weinberg ZG, Jeong KC, Adesogan AT. Bacterial diversity and composition of alfalfa silage as analyzed by illumina miseq sequencing: effects of Escherichia coli O157:H7 and silage additives. J Dairy Sci. 2017;101:2048–59. https://doi.org/10.3168/jds.2017-12876.

    Article 
    PubMed 

    Google Scholar 

  • Zong C, Wu Q, Wu A, Chen S, Dong D, Zhao J, et al. Exploring the diversity mechanism of fatty acids and the loss mechanisms of polyunsaturated fatty acids and fat-soluble vitamins in alfalfa silage using different additives. Anim Feed Sci Technol. 2021;280:115044. https://doi.org/10.1016/j.anifeedsci.2021.115044.

    Article 

    Google Scholar 

  • Li L, Zhang H, Meng D, Yin H. Transcriptomics of Lactobacillus paracasei: metabolism patterns and cellular responses under high-density culture conditions. Front Bioeng Biotechnol. 2023;11:1274020. https://doi.org/10.3389/fbioe.2023.1274020.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • Shen S, Choi O, Park SH, Kim CG, Park CS. Root colonizing and biocontrol competency of Serratia plymuthica A21-4 against phytophthora blight of pepper. Plant Pathol J. 2005;21:64–7. https://doi.org/10.5423/PPJ.2005.21.1.064.

    Article 

    Google Scholar 

  • Dai J, Han R, Xu Y, Li N, Wang J, Dan W. Recent progress of antibacterial natural products: future antibiotics candidates. Bioorg Chem. 2020;101:103922. https://doi.org/10.3390/fermentation8040158.

    Article 
    PubMed 

    Google Scholar 

  • Bansal K, Kumar S, Kaur A, Singh A, Patil PB. Deep phylo-taxono genomics reveals xylella as a variant lineage of plant-associated Xanthomonas and supports their taxonomic reunification along with Stenotrophomonas and Pseudoxanthomonas. Genomics. 2021;113:3989–4003. https://doi.org/10.1016/j.ygeno.2021.09.021.

    Article 
    PubMed 

    Google Scholar 

  • Zeng T, Li X, Guan H, Yang W, Yan Y. Dynamic microbial diversity and fermentation quality of the mixed silage of corn and soybean grown in strip intercropping system. Bioresour Technol. 2020;123655. https://doi.org/10.1016/j.biortech.2020.123655.

    Article 
    PubMed 

    Google Scholar 

  • Du Z, Lin Y, Sun L, Yang F, Cai Y. Microbial community structure, co-occurrence network and fermentation characteristics of Woody plant silage. J Sci Food Agric. 2021;102:193–1204. https://doi.org/10.1002/jsfa.11457.

    Article 

    Google Scholar 

  • Kleerebezem M, Bachmann H, van PeltKleinJan E, Douwenga S, Smid EJ, Teusink B, van Mastrigt O. Lifestyle, metabolism and environmental adaptation in Lactococcus lactis. FEMS Microbiol Rev. 2020;24:804–20. https://doi.org/10.1093/femsre/fuaa033.

    Article 

    Google Scholar 

  • Zhou J, Huo T, Sun J, Feng Y, Pan J, Zhao Y, et al. Response of amino acid metabolism to decreased temperatures in anammox consortia: strong, efficient and flexible. Bioresour Technol. 2022;352:127099. https://doi.org/10.1016/j.biortech.2022.127099.

    Article 
    PubMed 

    Google Scholar 

  • Rabinowitz JD, Enerbäck S. Lactate: the ugly duckling of energy metabolism. Nat Metab. 2020;2(7):566–71. https://doi.org/10.1038/s42255-020-0243-4.

    Article 
    PubMed 
    PubMed Central 

    Google Scholar 

  • McDonald P, Henderson AR, Heron SJJW. The biochemistry of silage. 2nd ed. Chalcombe; 1992. p. 145–50. https://doi.org/10.1017/S0014479700023115.

    Book 

    Google Scholar 

Continue Reading

  • Eviden and AMD to Power Europe’s New Exascale Supercomputer, the First Based in France

    Eviden and AMD to Power Europe’s New Exascale Supercomputer, the First Based in France

    More than a single supercomputer, Alice Recoque will expand Europe’s AI and research capabilities while ensuring energy efficiency and sovereignty

    Paris, France – November 18, 2025

    Eviden, the Atos Group product brand leading in advanced computing, and AMD (NASDAQ: AMD) announced their selection to build Alice Recoque, a next-generation supercomputer to support the need for scientific computing (HPC) and artificial intelligence (AI), serving as an AI Factory. Alice Recoque will be France’s first and Europe’s second Exascale supercomputer, a machine to expand Europe’s AI and research capabilities while ensuring energy efficiency and sovereignty.

    For more information, please click here.

    Continue Reading

  • Stock market today: Live updates

    Stock market today: Live updates

    Traders work on the floor of the New York Stock Exchange.

    NYSE

    Stock futures fell on Tuesday, as continued weakness in tech ahead of Nvidia’s major earnings report this week put pressure on the broader market.

    Futures tied to the Dow Jones Industrial Average lost 185 points, or 0.4%. S&P 500 futures shed 0.2%, while Nasdaq-100 futures were dipped 0.2%.

    Nvidia fell about 0.9% in the premarket, while Palantir Technologies dipped 1.2%. Amazon and Microsoft were also down more than 1%.

    Also putting pressure on futures was a 1.5% decline in Home Depot shares. The home improvement pulled back after reporting an earnings miss and cutting its full-year outlook.

    The three major U.S. indexes closed in the red in the previous trading session. The 30-stock Dow Jones Industrial Average plunged more than 550 points, or 1.2%, while the S&P 500 and Nasdaq Composite each lost around 0.9%.

    Nvidia notably declined about 2% ahead of the chipmaker’s third-quarter results due after Wednesday’s close. The company, which is reporting toward the end of a strong earnings season, has been at the center of a debate about the strength of the artificial intelligence-powered market rally this year. Concerns have grown about weak market breadth, pricey tech valuations and the soundness of AI fundamentals due to a boom in Big Tech debt offerings and the pace of AI chip depreciation.

    The tech-heavy Nasdaq is on pace to snap its seven-month win streak, while the S&P 500 is down 2.5% in November after rallying for six months in a row.

    “The market narrative has certainly shifted dramatically over the past few weeks, as the market’s reaction function with respect to AI has taken a sharp left turn from rewarding ever-growing capex spend to rapidly growing skepticism of further investment and future returns,” said Garrett Melson, portfolio strategist at Natixis Investment Managers Solutions. “Pair that with crowded positioning across real money and systematic accounts and you’ve got all the ingredients for a sharp de-risking and an accompanying narrative reset.”

    To be sure, Melson remains bullish that a cooling labor market and an overall improving inflation picture will power a year-end rally. “Despite the fears, the AI cycle remains alive and well, something we expect NVDA will confirm on Wednesday. That certainly isn’t a bearish backdrop,” he said.

    Aside from Nvidia’s report, investors this week will monitor data points that can inform the trajectory of upcoming interest rate decisions, which have scaled back in recent weeks. Fed funds futures traders are pricing in roughly 40% chance of a cut, significantly lower than the more than 90% chance priced in a month ago, according to the CME FedWatch tool. The Federal Reserve’s October meeting minutes and September nonfarm payrolls release, which will be the first piece of economic data released following the U.S. government shutdown, are on deck for Wednesday and Thursday releases, respectively.

    Continue Reading

  • tRF-His-GTG-1 promotes Salmonella survival through modulation of lipid metabolism and immune signaling | Cell Communication and Signaling

    tRF-His-GTG-1 promotes Salmonella survival through modulation of lipid metabolism and immune signaling | Cell Communication and Signaling

    Subjects

    In total, 25 patients with SLE who fulfilled the 1997 revised criteria of the American College of Rheumatology [17] and the 2012 Systemic Lupus International Collaborating Clinics classification criteria [18] were included. Among them, five had NTS bacteremia. Twenty healthy volunteers without rheumatic disease served as non-SLE controls, and an additional five non-SLE patients with NTS bacteremia were enrolled. The demographic and clinical characteristics of patients with SLE and control subjects are summarized in Table 1. Sex was not considered a biological variable. The Institutional Review Board of Taichung Veterans General Hospital approved this study (CF21176A), and written informed consent was obtained from all participants in accordance with the Declaration of Helsinki.

    Table 1 Demographics and clinical characteristics of systemic lupus erythematosus (SLE) patients and non-SLE control subjects with or without non-typhoidal Salmonella (NTS) bacteremia

    Human PBMC isolation

    PBMCs were isolated immediately after venous blood collection using Ficoll-Paque PLUS (GE Healthcare Biosciences AB, Sweden) density gradient centrifugation.

    Salmonella infection

    Salmonella enterica serovar Typhimurium strain ATCC14028 was cultured on LB agar plates at 37 °C in an incubator with 5% CO₂. Human PBMCs were infected with strain ATCC14028 at a multiplicity of infection (MOI) of 10. At 3 h post-infection, the cells were incubated with medium containing 50 µg/mL gentamicin for 1 h to eliminate extracellular bacteria, washed three times with phosphate-buffered saline (PBS), and then maintained in antibiotic-free medium. Bacterial growth was assessed by colony-forming unit (CFU) assays. At the indicated time points post-infection, cells were lysed in sterile PBS containing 0.1% Triton X-100 to release intracellular bacteria. The lysates were serially diluted, plated on LB agar plates, and incubated at 37 °C for 18–24 h to determine CFU counts.

    EV isolation

    Samples were centrifuged at 350 × g for 10 min at 4 °C to remove cell debris. For EV characterization and functional assays, 2.5 mL of each sample was diluted with 7.5 mL of PBS and concentrated using Amicon Ultra-0.5 centrifugal filter units (Millipore, 100 K cutoff) at 3000 × g for 30 min at 4 °C. The retentate (100 µL) was diluted with 1.4 mL of PBS and centrifuged at 10,000 × g for 30 min at 4 °C. The resulting pellets were resuspended in 1.5 mL of PBS and ultracentrifuged at 120,000 × g for 90 min at 4 °C. Finally, the pellets were resuspended in 50 µL of PBS and stored at − 80 °C [16].

    IC isolation

    ICs were isolated from the sera of patients with clinically active SLE, defined as those with an SLE Disease Activity Index of >6, by polyethylene glycol (PEG) precipitation as previously described [19]. Briefly, serum samples were mixed with an equal volume of ice-cold 6% (wt/vol) PEG 6000 (Sigma-Aldrich, USA) to achieve a final concentration of 3%, incubated for 60 min at 4 °C, and centrifuged at 2,000 × g for 20 min. The resulting precipitates were washed three times with sterile PBS and resuspended to the original serum volume in PBS. Anti-DNA enzyme-linked immunosorbent assay (ELISA) (CUSABIO, USA) was performed on both the PEG pellets and corresponding supernatants to confirm that the precipitated material contained DNA-binding ICs. More than 75% of total anti-DNA reactivity was recovered in the PEG pellet fraction of SLE sera, whereas pellets from healthy controls showed undetectable signal. The concentrations of PEG-precipitated ICs were determined using a circulating IC C1q ELISA (BioVendor, Czech Republic) and expressed as aggregated human IgG equivalents (µg Eq/mL).

    ICs from three different patients with active SLE were quantified and diluted in PBS to a final concentration of 50 µg Eq/mL. For stimulation assays, 40 µL of the IC suspension was added to PBMC cultures and incubated for 24 h, unless otherwise indicated. Each experiment used ICs derived from one patient with SLE and was performed in triplicate; the full set of experiments was independently repeated with ICs from three different patients to confirm reproducibility and minimize donor-specific bias.

    IC-primed pEV Preparation

    Human platelets were isolated from peripheral blood by centrifugation at 230 × g for 15 min at 25 °C, followed by centrifugation at 1,000 × g for 10 min. The platelet pellets were resuspended in Tyrode’s buffer (Sigma-Aldrich, USA) containing one-sixth volume of acid citrate dextrose (Sigma-Aldrich, USA) and 1 µM prostaglandin I₂ (Sigma-Aldrich, USA), then centrifuged again at 1,000 × g for 10 min at 25 °C. The pellets were resuspended in Tyrode’s buffer containing 1 µM prostaglandin I₂ and 0.04 U/mL apyrase (Sigma-Aldrich, USA) and gently agitated on a shaker. Before stimulation, the platelets (4 × 10⁷ per tube) were washed once by centrifugation at 1,000 × g for 10 min and resuspended in fresh Tyrode’s buffer. SLE ICs (2 µg aggregated human IgG equivalents) were added to the platelet suspension and incubated for 2 h at 37 °C. The supernatants were then collected and sequentially centrifuged at 2,000 × g for 20 min, 10,000 × g for 30 min, and 100,000 × g for 70 min to isolate pEVs. The resulting pellets were washed in PBS and resuspended in 40 µL of sterile PBS for subsequent assays. The pEVs were quantified by nanoparticle tracking analysis and characterized by immunoblotting for CD41, CD63, CD9, and CD81. For stimulation experiments, approximately 1 × 10⁸ pEV particles were added to 2 × 10⁶ PBMCs and incubated for 24 h, unless otherwise indicated.

    Transient transfection

    Human PBMCs (1 × 10⁶ cells) were transiently transfected with 30 nM Toll-like receptor (TLR)7/8 siRNA (Dharmacon, Horizon, USA) or control siRNA using Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions and incubated at 37 °C for 24 h. Knockdown efficiency of TLR7/8 was confirmed by immunoblotting.

    Quantitative polymerase chain reaction (PCR) for TsRNAs

    tsRNAs were extracted from PBMCs using the rtStar tRF&tiRNA Pretreatment Kit (Arraystar, USA) to remove RNA modifications. Twenty-five femtomoles of synthetic Caenorhabditis elegans miRNA (cel-miR-39; Thermo Fisher Scientific, USA) were added to each sample as an internal control. For cDNA synthesis, 250 ng of total tsRNA was reverse-transcribed using the rtStar cDNA Synthesis Kit (Arraystar, USA). Real-time PCR quantification of specific tsRNAs was performed with the LightCycler 480 SYBR Green I Master (Roche, Germany) and specific primers (Supplementary Table 1) and analyzed on the LightCycler 96 real-time PCR System (Roche, Germany) using the standard thermoprofile recommended by the manufacturer. Relative expression was calculated using the comparative threshold cycle (Ct) method and expressed as 2ΔCt, where ΔCt = [mean of control subjects (CttsRNA − Ctcel−miR−39)] − [patient (CttsRNA − Ctcel−miR−39)].

    Immunofluorescence assay

    Human PBMCs were fixed with 4% paraformaldehyde (Sigma-Aldrich, USA) for 10 min at room temperature, washed with PBS (Thermo Fisher Scientific, USA), permeabilized in PBS containing 1% bovine serum albumin (Thermo Fisher Scientific, USA) and 0.2% saponin (Sigma-Aldrich, USA), and then blocked in PBS containing 2% bovine serum albumin for 1 h. LDs were stained with 2 µM BODIPY 493/503 (Thermo Fisher Scientific, USA) for 30 min. Coverslips were mounted using SlowFade mounting medium (Thermo Fisher Scientific, USA), and images were acquired with an Olympus FV1000 confocal microscope. Image analysis was performed using FV10-ASW software version 4.2 (Olympus).

    Flow cytometry

    For LD analysis, PBMCs were stained with 2 µM BODIPY 493/503 (Thermo Fisher Scientific, USA) for 30 min at 37 °C in the dark, washed twice with PBS, and resuspended in PBS for acquisition. LD content was quantified by the mean fluorescence intensity (MFI) of the BODIPY 493/503 signal in the FITC channel. The gating strategy included: (i) exclusion of debris based on forward scatter (FSC) and side scatter (SSC) profiles, (ii) elimination of doublets using FSC-A versus FSC-H plots, and (iii) selection of viable PBMCs for analysis of BODIPY 493/503 fluorescence. Samples were acquired on a FACSCanto II flow cytometer (BD Biosciences, USA), and data were analyzed using CellQuest (version 6.0, BD Biosciences) or FlowJo (version 10.10.0, BD Biosciences).

    Immunoblotting

    Cells were lysed in RIPA lysis and extraction buffer (Thermo Fisher Scientific, USA) supplemented with protease inhibitors (Roche, Germany). Equal amounts (40 µg) of total protein were separated by SDS–polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes (Millipore, USA), and incubated with primary antibodies followed by horseradish peroxidase–conjugated secondary antibodies (listed in Supplementary Materials). Signals were detected using enhanced chemiluminescence (Millipore, USA) and visualized with a CCD imaging system (GE Healthcare, USA). Band intensities were quantified using ImageJ software, with β-actin (Santa Cruz Biotechnology, USA) serving as the loading control. All experiments were performed in triplicate. Data are presented as mean ± standard deviation (SD). Statistical comparisons were made using a two-tailed unpaired Student’s t-test (GraphPad Prism version 8). Densitometric quantification is provided in Supplementary File 2.

    RNA-seq analysis

    Total RNA (1 µg) from neutrophils was used for library preparation with the TruSeq Stranded mRNA Library Prep Kit (Illumina, RS-122–2001/2002) according to the manufacturer’s protocol. mRNA was enriched using oligo(dT) beads, fragmented, and reverse-transcribed into cDNA. After adaptor ligation and PCR amplification, libraries were purified with the AMPure XP system (Beckman Coulter, USA), quality-checked with the Qsep400 System (Bioptic Inc., Taiwan), and quantified using a Qubit 2.0 Fluorometer (Thermo Fisher Scientific, USA). Paired-end sequencing (150 bp) was performed on an Illumina NovaSeq platform. Raw reads were quality-checked with FastQC and trimmed using Trimmomatic. Clean reads were aligned to the human reference genome (GRCh38) using HISAT2, and gene counts were generated with featureCounts. Differential gene expression analysis was performed with DESeq2, and significantly altered genes were defined as those with an adjusted P value < 0.05 and |log₂ fold change| ≥ 1.

    RNA-protein pull-down assay

    A single biotinylated nucleotide was attached to the 3′ terminus of the tRF-His-GTG-1 mimic or control using the Pierce RNA 3′ End Desthiobiotinylation Kit (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions. Human neutrophils (2 × 10⁶ cells) were transiently transfected with 30 nM biotin-labeled tRF-His-GTG-1 mimic or control using Lipofectamine RNAiMAX Transfection Reagent (Thermo Fisher Scientific, USA) and incubated for 24 h at 37 °C. After incubation, cells were washed with PBS and lysed with lysis buffer. Following centrifugation at 12,000 × g for 15 min at 4 °C, the supernatant was collected and quantified for protein concentration. RNA-binding proteins associated with the tRF-His-GTG-1 mimic were isolated using the Pierce Magnetic RNA–Protein Pull-Down Kit (Thermo Fisher Scientific, USA) according to the manufacturer’s instructions. The immunoprecipitated proteins were analyzed by immunoblotting with the indicated antibodies.

    Statistics

    Results are presented as mean ± SD. Unpaired two-tailed Student’s t-tests and Mann–Whitney U tests were used for between-group comparisons. One-way analysis of variance with Bonferroni post hoc correction was applied for multiple comparisons. Correlations were assessed using Spearman’s correlation coefficient. P values of < 0.05 were considered statistically significant. All statistical analyses were performed using GraphPad Prism version 8.

    Continue Reading

  • Perceived fatigue progression tracking during manual handling tasks using sEMG recordings | Journal of NeuroEngineering and Rehabilitation

    Perceived fatigue progression tracking during manual handling tasks using sEMG recordings | Journal of NeuroEngineering and Rehabilitation

    Fig. 1

    The flowchart of the procedure in this study includes: A Preprocessing the sEMG signals; B Manual handling task segmentation using the joint angles obtained by IMUs; C Segmenting sEMG signals based on IMU-derived time windows; D Identifying and extracting linear and complexity-based MMF indicators; E Analyzing the correlation between MMF indicators and RPE scales; and F Classifying performance fatigue via a long short-term memory (LSTM) model and a convolutional neural network combined with LSTM (CNN-LSTM) using MMF indicators

    Figure 1 provides an overview of the experimental process followed in this study, with subsequent sections offering detailed explanations of each step .

    Experimental procedure

    The study recruited eight able-bodied male participants (average age: 24 ± 2 years, average body mass: 73 ± 11 kg, average body height: 179 ± 4 cm). The participants were instructed to perform repetitive cycles of a manual handling task including lifting a 16-lbs load from a 15-cm high table, turning, placing it on a 75-cm high table, and going to the rest position; lifting the load again from the 75-cm high table, turning, and placing it back on the 15-cm high table (Fig. 2). The participant performed these movements repeatedly and reported their perceived fatigue level using the Borg RPE scale every 2 min. The experiment concluded when participants reported a fatigue level of 9 out of 10 on the RPE scale. Participants performed an average of 146 ± 43 cycles, and each participant completed two recording sessions on different days, each starting in a rested state to minimize potential carryover effects such as residual fatigue or motor adaptation. The Research Ethics Board of the University of Alberta approved the experimental protocol (Approval no. Pro00089234), and the participants provided written consent prior to testing. We used the data used in our previous publications [4, 8].

    Fig. 2
    figure 2

    Manual handling task cycle including lifting (a), carrying, (bd), and lowering (e) activities, followed by a resting position (f)

    Ten sEMG sensors (Trigno, Delsys, USA) were placed on the following muscles on the right side of the participants’ bodies, as all participants were right-handed: Biceps Brachii (Biceps), Flexor Carpi Radialis, Trapezius, Deltoideus Posterior, Erector Spinae Longissimus (L), Erector Spinae Iliocostalis (I), Rectus Femoris, Tibialis Anterior, Biceps Femoris, and Lateral Gastrocnemius with a sampling frequency of 1200 Hz. The sEMG data were first mean-subtracted and then filtered using a zero-lag, 8th-order Butterworth bandpass filter set to 10–500 Hz. Following this, the data were smoothed with a 50 ms window size. The sEMG time series was then normalized to the maximum amplitude during the first five cycles (non-fatigue states) of the experiment to help reduce inter-session variability.

    Additionally, seven IMUs (MTws, Xsens Technologies, NL) were positioned on their sternum, sacrum, right upper arm, forearm, thigh, shank, and foot to monitor joint angles’ kinematics, with a sampling frequency of 100 Hz. Similar to the EMG setup, IMUs were placed only on the right side, as all participants were right-handed. This setup helped reduce the number of sensors required, minimizing participant discomfort and motion constraints during the prolonged trials. Prior to the main experiment, a functional calibration was performed according to the protocol outlined in [23]. This calibration aimed to synchronize the inertial frames of the IMUs with the body’s anatomical frames for measuring joint angles. Participants were instructed to maintain stillness for 5 s, followed by performing ten flexions and extensions of both legs and arms with locked knee and elbow joints. Subsequently, the IMU readings were utilized to calculate body joint angles based on the orientation derived from the Xsens sensor fusion algorithm. Data was preprocessed using MATLAB R2022a (The MathWorks Inc., Natick, MA, USA).

    Data segmentation based on activities

    Our experiment involved a manual handling task with different activities of lifting, carrying, and lowering, and each of them included different body movements and muscle engagements; therefore, the sEMG recordings were first segmented based on these three distinct activities. This segmentation was performed using joint angle data processed in OpenSim, which simulated the motion and identified activity boundaries according to predefined activity definitions. Additionally, each muscle primarily influences the motion of one or more joints and the positions of adjacent joints can also affect the level of muscle engagement due to changes in torque demands, moment arms, and muscle–tendon length. Specifically: (1) Biceps and Flexor Carpi Radialis were associated with the elbow & shoulder flexion/extension, (2) Trapezius and Deltoideus Posterior influenced the shoulder flexion/extension, (3) Erector Spinae L and I influenced the trunk flexion/extension, (4) Rectus Femoris and Biceps Femoris influenced the hip and knee flexion/extension, (5) Tibialis Anterior and Lateral Gastrocnemius influenced the ankle and knee flexion/extension. The sEMG signals were further segmented based on the relevant joints’ ROM, dividing each into two equal segments (0–50% and 50–100% of the ROM).

    sEMG data processing

    Amplitude analysis

    Root Mean Square (RMS) values were used to characterize sEMG signal amplitude for monitoring muscle fatigue [24]. For a discrete signal with N samples, the RMS can be computed using the following formula:

    $$:RMS=:sqrt{frac{1}{N}{int:}_{i=1}^{N}x{left[iright]}^{2}}$$

    (1)

    Where (:xleft[iright]) represents each sample within the analyzed sEMG signal segment.

    As muscle fatigue progresses, the sEMG amplitude generally rises due to the recruitment of additional motor units, and potentially, their excitation at higher firing frequencies to compensate for the declining force-generating capacity of individual muscle fibers. Consequently, higher RMS values are expected in the later stages of RPE scales.

    Spectral analysis

    This approach identifies muscle fatigue by observing a shift in the power spectrum of sEMG signals toward lower frequencies, as indicated by the median frequency. The median frequency is calculated using the following equation:

    $$:{int:}_{0}^{MDF}PSDleft(fright)df=:{int:}_{MDF}^{infty:}PSDleft(fright)df=frac{1}{2}{int:}_{0}^{infty:}PSDleft(fright)df$$

    (2)

    Where (:PSDleft(fright)) is the power spectral density at a given frequency (:f), calculated by the spectrogram function, which performs the Short-Time Fourier Transform of the preprocessed signal [25]. This process involved segmenting the signal with a 128-sample window, 64-sample overlap, and 256 Fast Fourier Transform points, with a sampling frequency of 1200 Hz. The spectrogram function provided the PSD matrix, reflecting the power at various frequencies and times, which was used for further analysis.

    Mobility

    Mobility is quantified as the square root of the ratio of the variance of the signal’s first derivative to the variance of the original signal [22] calculated as follows:

    $$:Mobility=:sqrt{frac{{{sigma:}_{1}}_{x1}^{x2}}{{{sigma:}_{0}}_{x1}^{x2}}}:$$

    (3)

    where (:{{sigma:}_{0}}_{x1}^{x2}) and (:{{sigma:}_{1}}_{x1}^{x2}) are the variance of the sEMG signal and the variance of the first derivative of the sEMG signal, respectively. Both were computed over the full duration of each activity segment (i.e., lifting, carrying, or lowering), with (:{x}_{1}) and (:{x}_{2}) denoting the start and end time points of each segment.

    As fatigue sets in, the mobility of the sEMG signal is expected to decrease due to a reduction in conduction velocity [22].

    Entropy

    Fuzzy entropy, a measure of complexity and regularity in time-series data, was investigated in this study due to its demonstrated robustness in assessing MMF. Fuzzy entropy was computed using the following procedure [16]:

    1. 1.

      The sEMG time-series were divided into overlapping intervals of length (:m).

    2. 2.

      For two sEMG intervals (:{S}_{i}=[{x}_{1},{x}_{2},dots:,:{x}_{m}]) and (:{S}_{j}=[{y}_{1},{y}_{2},dots:,:{y}_{m}]), the distance was calculated as:

    $$:{d}_{i,j}^{m}=text{m}text{a}text{x}left(right|{x}_{1}-{y}_{1}|,|{x}_{2}-{y}_{2}|,dots:,|{x}_{m}-{y}_{m}left|right)$$

    (4)

    1. 3.

      For each pair of intervals, the similarity function was calculated as:

    $$:{Omega:}left({d}_{i,j}^{m},text{n},text{r}right)=text{e}text{x}text{p}(-frac{{left({d}_{ij}^{m}:right)}^{n}}{r})$$

    (5)

    Where (:r) is the similarity threshold and (:n) is the power of the similarity function.

    1. 4.

      (:{C}_{i}^{m}left(rright)), the similarity for interval (:i), was calculated as the average similarity across all other intervals (:j) as:

    $$:{C}_{i}^{m}left(rright)={left(N-m+1right)}^{-1}{sum:}_{j=1,jne:i}^{N-m+1}{Omega:}({d}_{i,j}^{m},text{r})$$

    (6)

    Where N is the total number of sEMG time-serious data points.

    1. 5.

      (:{C}^{m}left(rright)) was calculated as the average of (:{C}_{i}^{m}left(rright)) for all the intervals.

    2. 6.

      Previous steps were repeated for intervals of length (:m+1) to calculate (:{C}^{m+1}left(rright)).

    3. 7.

      Finally, the fuzzy entropy was calculated as:

    $$:FuzzyEnleft(m,n,r,Nright)=-text{l}text{n}left(frac{{C}^{m+1}left(rright)}{{C}^{m}left(rright)}right)$$

    (7)

    The fuzzy entropy was calculated using the FuzzyEn MATLAB function [26] with an embedding dimension of (:m=2), a power factor of (:n=2) for the similarity function, and a similarity threshold of (:r=0.25) [27]. In higher fatigue stages, lower entropy values are anticipated due to the decreased complexity of sEMG signals, which is caused by a reduction in the number of active motor units [16].

    Dimitrov’s index

    To address the low sensitivity of traditional spectral parameters like mean frequency and median frequency for muscle fatigue monitoring, previous studies investigated the relationship between the sEMG power content in low and high frequencies [28]. However, a challenge is defining the boundaries of the high- and low-frequency bands, as their selection can significantly impact the interpretation of muscle fatigue. To overcome this issue, the Dimitrov’s index, a highly sensitive spectral metric, has been proposed. It effectively defines these boundaries using frequency weighting factors, as shown in Eq. 8. The Dimitrov’s index is derived from sEMG spectral characteristics using FFT. Here, the Dimitrov’s index [28] is calculated in the band frequency from 10 Hz to 500 Hz:

    $$:FI=frac{underset{f=10}{overset{500}{int:}}{f}^{-1}.PSleft(fright).df}{underset{f=10}{overset{500}{int:}}{f}^{5}.PSleft(fright).df}$$

    (8)

    Where (:f) denotes frequency (which is the variable of integration), (:PSleft(fright)) is the sEMG power-frequency spectrum as a function of frequency (:f). (:{f}^{-1}) is a frequency weighting factor that gives more emphasis to lower frequencies, increasing their contribution and highlighting the power in the lower frequency range of the sEMG signal. Conversely, (:{f}^{5}) in the denominator is a frequency weighting factor that prioritizes higher frequencies by assigning more weight to larger frequency values, thereby focusing on the power in the higher frequency range.

    As fatigue progresses, the Dimitrov’s index is expected to increase due to the higher power spectral density in the low and ultra-low frequencies compared to the higher frequencies in the higher fatigue stages [28].

    Statistical analysis

    Our approach aimed to initially investigate the correlation between the RPE scales, known as a perceived fatigue evaluator, and the MMF indicator calculated in this study. Thus, Spearman’s correlation coefficients and their corresponding p-values were calculated between each of the MMF indicators and fatigue levels (RPE scales of 1 to 10), utilizing the Fisher’s Z transformation and the generalized Bonferroni-Holm procedure, respectively [29, 30]. Spearman’s correlation was chosen because the analysis focused on monotonic, rather than linear, relationships between the MMF indicators and fatigue levels. For each RPE level, 10 segments were randomly sampled for each activity type (lifting, carrying, and lowering) to reduce autocorrelation and maintain consistency across participants, and MMF indicators were calculated over the full duration of each segment without additional windowing.

    Spearman’s correlation coefficients were computed separately for each session between each MMF indicator and RPE scale. For analysis, correlations from the two sessions of each participant were first averaged (after Fisher Z-transformation) to obtain a single value per participant. The resulting correlation coefficients were transformed using Fisher’s Z transformation and averaged to obtain a group-level correlation value, which was then back-transformed to obtain the final reported Spearman’s ρ. Corresponding session-level p-values were adjusted for multiple comparisons using the generalized Bonferroni-Holm procedure. All statistical analyses were conducted in MATLAB using built-in and custom-written functions.

    Multi-class fatigue detection

    Then, we developed deep learning algorithms to classify multiple stages of perceived fatigue using the MMF indicators as inputs. For this purpose, the extracted MMF indicators from 5 consecutive repetitions were grouped together into a window and labeled into five discrete fatigue level bins using the reported RPE scales: 1–2, 3–4, 5–6, 7–8, and 9–10. The five fatigue levels were chosen because they provided reasonable accuracy while enabling multi-level classification without overcomplicating the process. We first used a long short-term memory (LSTM) network to capture temporal dependencies and complex sequential patterns in time-series data [31]. The model was developed using TensorFlow, incorporating regularization techniques and the Adam optimizer to enhance training efficiency and prevent overfitting. The neural architecture consisted of three LSTM layers with progressively smaller units: 128, 64, and 32, respectively. Each LSTM layer utilized L2 regularization with a strength of 0.01 to mitigate overfitting, and dropout was applied with a rate of 0.5 following each LSTM layer to further address overfitting. The model concluded with three dense layers with 32, 16, and 8 neurons, respectively, each employing the Rectified Linear Unit (ReLU) activation function. The output layer was comprised of five neurons with a SoftMax activation function designed to produce class probabilities corresponding to the five bins of RPE ratings. Leave-one-out cross-validation (LOOCV) was performed to ensure the model’s generalizability across different subjects. This method involved training the model on data from all but one participant and evaluating it on the excluded participant, ensuring a solid assessment of the model’s performance across diverse data subsets.

    To improve predictive accuracy by capturing spatial dependencies of MMF indicators, a CNN-LSTM architecture was implemented. The CNN part of the model included two convolutional layers with 64 and 32 filters, respectively, using a kernel size of 3 and ‘tanh’ activation. Max-pooling layers with a pool size of 2 followed each convolutional layer to reduce spatial dimensions. These convolutional layers were followed by LSTM layers to capture temporal dynamics, allowing the model to learn both spatial and temporal dependencies in the data.

    Continue Reading