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Optimized protoplast isolation and transfection with a breakpoint:accelerating Cas9/sgRNA cleavage efficiency validation in monocot and dicot

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aBIOTECH 2024年第2期5卷 151-168页

作者： Debasmita Panda Subhasis Karmakar Manaswini Dash Swagat Kumar Tripathy Priya Das sagar banerjee Yiping Qi Sanghamitra Samantaray Pradipta Kumar Mohapatra Mirza J.Baig Kutubuddin A.Molla ICAR National Rice Research Institute CuttackOdisha 753006India Department of Botany Ravenshaw UniversityCuttackOdisha 753003India Department of Plant Science and Landscape Architecture University of MarylandCollege ParkMD 20742USA

The CRISPR-Cas genome editing tools are revolutionizing agriculture and basic biology with their simplicity and precision ability to modify target genomic ***-predicted guide RNAs(gRNAs)often fail to induce efficient cleavage at target *** target loci are inaccessible due to complex chromatin ***,there is no suitable tool available to predict the architecture of genomic target sites and their ***,significant time and resources are spent on performing editing experiments with inefficient *** in vitro-cleavage assay could provide a rough assessment of gRNA efficiency,it largely excludes the interference of native genomic *** in-vivo testing gives a proper assessment of the cleavage ability of editing reagents in a native genomic ***,we developed a modified protocol that offers highly efficient protoplast isolation from rice,Arabidopsis,and chickpea,using a sucrose gradient,transfection using PEG(polyethylene glycol),and validation of single guide RNAs(sgRNAs)cleavage efficiency of *** have optimized various parameters for PEG-mediated protoplast transfection and achieved high transfection efficiency using our protocol in both monocots and *** introduced plasmid vectors containing Cas9 and sgRNAs targeting genes in rice,Arabidopsis,and chickpea *** dual sgRNAs,our CRISPRdeletion strategy offers straightforward detection of genome editing success by simple agarose gel *** sequencing of PCR products confirmed the editing efficiency of specific ***,we demonstrated that isolated protoplasts can be stored for up to 24/48 h with little loss of viability,allowing a pause between isolation and *** high-efficiency protocol for protoplast isolation and transfection enables rapid(less than 7 days)validation of sgRNA cleavage efficiency before proceeding with stable *** isolation and transfection method can als

关键词： Protoplast Genome editing Transgene-free edited plants Rice Chickpea

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Nanozyme‑Engineered Hydrogels for Anti‑Inflammation and Skin Regeneration

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Nano-Micro Letters 2024年第6期16卷 127-179页

作者： Amal George Kurian Rajendra K.Singh Varsha sagar Jung‑Hwan Lee Hae‑Won Kim Institute of Tissue Regeneration Engineering(ITREN) Dankook UniversityCheonan 31116Republic of Korea Department of Nanobiomedical Science&BK21 NBM Global Research Center for Regenerative Medicine Dankook UniversityCheonan 31116Republic of Korea Department of Biomaterials Science School of DentistryDankook UniversityCheonan 31116Republic of Korea UCL Eastman‑Korea Dental Medicine Innovation Centre Dankook UniversityCheonan 31116Republic of Korea Cell and Matter Institute Dankook UniversityCheonan 31116Republic of Korea Mechanobiology Dental Medicine Research Center Dankook UniversityCheonan 31116Republic of Korea

Inflammatory skin disorders can cause chronic scarring and functional impairments,posing a significant burden on patients and the healthcare *** therapies,such as corticosteroids and nonsteroidal anti-inflammatory drugs,are limited in efficacy and associated with adverse ***,nanozyme(NZ)-based hydrogels have shown great promise in addressing these ***-based hydrogels possess unique therapeutic abilities by combining the therapeutic benefits of redox nanomaterials with enzymatic activity and the water-retaining capacity of *** multifaceted therapeutic effects of these hydrogels include scavenging reactive oxygen species and other inflammatory mediators modulating immune responses toward a pro-regenerative environment and enhancing regenerative potential by triggering cell migration and *** review highlights the current state of the art in NZ-engineered hydrogels(NZ@hydrogels)for anti-inflammatory and skin regeneration *** also discusses the underlying chemo-mechano-biological mechanisms behind their ***,the challenges and future directions in this ground,particularly their clinical translation,are *** insights provided in this review can aid in the design and engineering of novel NZ-based hydrogels,offering new possibilities for targeted and personalized skin-care therapies.

关键词： Nanozymes Hydrogels ROS scavenging Anti-inflammation Skin regeneration

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A case report on a rare case of primary tuberculous otitis media

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Journal of Otology 2024年第1期19卷 40-45页

作者： Dr Yadav sagar Shyamlal Dr Shikha Gianchand Dr Rahul Kurkure Dept of ENT and Head Neck Surgery 5 Air Force HospitalJorhatAssam785005India Dept of ENT and Head Neck Surgery Armed Forces Medical CollegePuneMaharashtra411040India

Although tuberculosis has become more common in recent years, it still accounts for just a small percentage of cases of chronic otitis media. Common symptoms of tuberculous otitis media(TOM) include otorrhoea, hearing loss, and multiple tympanic membrane perforations. Due to its rarity, the illness is frequently misdiagnosed. When a patient does not improve with standard antibacterial and antifungal treatment, this may be a possible diagnosis to explore. Complete healing is possible if the illness is diagnosed and treated timely to prevent consequences.

关键词： otitis healing diagnosis

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An arrow in the quiver: evaluating the performance of Hyptis suaveolens(L.) Poit. in different light levels

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Ecological Processes 2024年第2期 130-147页

作者： Kanhaiya Shah Gyan Prakash Sharma R.sagar Department of Environmental Studies University of Delhi Department of Botany Banaras Hindu University

Background Hyptis suaveolens（L.） Poit.,native to the American tropics,is a pantropical annual plant and a major invasive species throughout *** was anticipated that the availability of sunlight,coupled with its superior reproductive potential,persistent propagule bank,and dispersal ability,could lead to an increase in the growth and spread of this invader,thus potentially impeding herbaceous growth and diversity in non-native *** its ecological fitness and competitive performance will be useful to manage the spread of *** in natural ecosystems that are facing a wide range of anthropogenic *** The present study is a three-tier *** the first tier,a field study was conducted to assess the patterns of *** abundance and herbaceous species diversity in response to light availability(sun,842-1072μmol m-2s-1and shade 253-341 μmol m-2S-1) in the tropical dry deciduous ecosystems in the Vindhyan highlands,***,the impact of *** abundance on the resident native and non-native species abundance and diversity was also *** the second tier,a randomized common garden experiment was conducted to understand the trait fitness of *** in sun(940 μmol m-2s-1) and shade(300 μmol m-2s-1) *** the third tier,a plant growth chamber experiment with high-light(940 μmol m-2s-1) and low-light(300 μmol m-2s-1)treatments was done to learn how *** partitions its biomass between aboveground and belowground plant *** The field study indicated that the sunlit areas had a higher abundance of *** and a lower diversity of resident herbaceous species than the shaded *** common garden experiment showed that sun-dwelling *** individuals performed better in germinative,vegetative,eco-physiological,and reproductive traits than the shade-dwelling *** growth chamber experiment exhibited that plants grown in high-light environment had greater seed germi

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A Harmonic Approach to Handwriting Style Synthesis Using Deep Learning

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Computers, Materials & Continua 2024年第6期79卷 4063-4080页

作者： Mahatir Ahmed Tusher Saket Choudary Kongara sagar Dhanraj Pande Seong Ki Kim Salil Bharany School of Computer Science and Engineering VIT-AP UniversityAmaravatiAndhra Pradesh522237India School of Engineering&Technology Pimpri Chinchwad UniversityPuneMaharashtra412106India Department of Computer Engineering Chosun UniversityGwangju61452Republic of Korea Independent Researcher AmritsarPunjab143001India

The challenging task of handwriting style synthesis requires capturing the individuality and diversity of human *** majority of currently available methods use either a generative adversarial network(GAN)or a recurrent neural network(RNN)to generate new handwriting *** is why these techniques frequently fall short of producing diverse and realistic text pictures,particularly for terms that are not commonly *** resolve that,this research proposes a novel deep learning model that consists of a style encoder and a text generator to synthesize different handwriting *** network excels in generating conditional text by extracting style vectors from a series of style *** model performs admirably on a range of handwriting synthesis tasks,including the production of text that is *** works more effectively than previous approaches by displaying lower values on key Generative Adversarial Network evaluation metrics,such Geometric Score(GS)(3.21×10^(-5))and Fréchet Inception Distance(FID)(8.75),as well as text recognition metrics,like Character Error Rate(CER)and Word Error Rate(WER).A thorough component analysis revealed the steady improvement in image production quality,highlighting the importance of specific handwriting *** fields include digital forensics,creative writing,and document security.

关键词： Recurrent neural network generative adversarial network style encoder fréchet inception distance geometric score character error rate mixture density network word error rate

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尼泊尔巴迪亚国家公园缓冲区的人象冲突

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Journal of Resources and Ecology 2024年第2期15卷 412-421页

作者： PAHARI sagar JOSHI Rajeev PAUDEL Umesh 南十字星大学澳大利亚利斯莫尔2480 尼泊尔农林大学自然资源管理学院林学系尼泊尔卡塔里56310 尼泊尔巴迪亚自然保护国家信托基金会尼泊尔巴迪亚21800

亚洲象(Elephasmaximus)是尼泊尔特莱(Terai)低地保护区重要的大型动物之一,它们经常在保护区森林边界附近遇到人类及其生计支持活动。人象冲突一直是保护区附近人类居住区的主要问题之一,每年都会造成经济损失,甚至威胁人类的生命安全... 详细信息

亚洲象(Elephasmaximus)是尼泊尔特莱(Terai)低地保护区重要的大型动物之一,它们经常在保护区森林边界附近遇到人类及其生计支持活动。人象冲突一直是保护区附近人类居住区的主要问题之一,每年都会造成经济损失,甚至威胁人类的生命安全,这一问题阻碍了保护区的可持续管理。本研究的目的是分析巴迪亚国家公园缓冲区人象冲突的原因,并评估人们对这种大型动物的感知。为此,在巴迪亚国家公园缓冲区内的三个市镇进行了结构化问卷调查。此外,作为问卷调查的补充,还对关键知情人士进行了访谈。结果显示,93%的受访者在过去三年中曾遭遇大象的袭击。由于大象袭击,去年每个家庭平均损失了大约9690尼泊尔卢比。大象的攻击行为大多数发生在7月至9月,其次是10月至12月。这表明大象对作物的偏好是研究区大象发起攻击/袭击行为的主要原因。大象袭击的第二个重要原因是由于农田向森林的扩张而导致大象食物不足。90%的受访者对大象攻击的反应是驱赶它们(使用火或噪音);51%的受访者接受人象共存,他们认为大象具有生物学和经济价值;然而,还有41%的人拒绝与大象共存,因为大象对当地居民及其生计构成了威胁。人象冲突已对保护区管理活动产生了负面影响,因此,必须通过保护区、当地居民和行政利益相关者的合作来解决相关问题。建议以后开展更多的研究,以了解大象在森林边界和邻近定居点附近人们的居住和迁徙模式。

关键词：攻击补偿作物的损害感知偏好

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Minimally Invasive Total Arterial Coronary Artery Bypass Grafting in Left Main Stem Disease

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World Journal of Cardiovascular Surgery 2024年第7期14卷 107-114页

作者： Pradeep Nambiar Radwan Husseini Prashant sagar Dr. Sulaiman Al Habib Hospital Dubai UAE Aster Hospital Dubai UAE

Objective: The aim was to show that Minimally Invasive total arterial revascularization for left main stem coronary artery disease, via a left anterior Mini thoracotomy using bilateral internal thoracic arteries is feasible, reproducible and safe. Further, there has been no exclusive data or experience with minimally invasive coronary artery bypass grafting in left main stem disease. Methods: From April 2019 to March 2024, 41 patients with left main stem stenosis, left main equivalent disease and unprotected left main with triple vessel disease underwent off pump minimally invasive multivessel coronary artery bypass grafting using either in situ pedicled Bilateral Internal Thoracic arteries or Left and Right Internal Thoracic artery Y composite conduits at three centers. Bilateral Internal Thoracic arteries were harvested under direct vision. All patients had an Intra-Aortic Balloon Pump inserted via the femoral artery prior to induction of anesthesia, to prevent any hemodynamic instability, arrhythmias, and was removed following completion of the procedure in the operating room without any complications. Efficacy and outcomes were evaluated by i) Primary (MACCE)-Major Adverse Cardiac and Cardiovascular events and ii) Secondary outcome measures including total length of stay, return to full physical activity and quality of life. Mean follow-up was 1.4 years (Maximum was 2.5 years). Results: 41 patients with left main stem coronary artery stenosis, underwent total arterial revascularization using bilateral internal thoracic arteries. Left main stem stenosis was present in 29 patients, Unprotected left main stem stenosis with triple vessel disease in 7 and left main equivalence in 5 patients. In this cohort, 29 patients with only left main stem stenosis had 2 grafts each, 7 patients with left main and triple vessel disease had 3 grafts and 5 patients with left main equivalent disease had 2 grafts respectively. The average number of grafts was 2.2. One patient was conv

关键词： Left Main Stem Minimally Invasive CABG Bilateral Internal Mammary Arteries

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Physico-chemical properties of wild edible fruit Meyna laxifora Robyns(Alu):production of Alu fruit powder by using drying aid

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Journal of Future Foods 2024年第4期4卷 353-360页

作者： Rohini Vijay Dhenge Arjun Mohanakumar Suhel Bhendawade Prashant Chavhan sagar Deshpande Pooja Lade Abhijeet Gatade Akshaya kumar Sahoo Deparment of Food and Dnug.University of Parma Parma 43124.Italy Deparoment of Food Science and Technolog Shivaji UniversityKolhapurMaharashtra 416004.India

The study focuses on the physico-chemical characteritics of the wild fruit Meyna laxiflora Robyns(Alu)and fruit powder,which was gathered from the Konkan region of Maharashtra Alu fruit powder is made by using a drying aid called maltodextrin(MD).Powder samples(MP1,MP2,MP3,MP4 and MP5)were made using different amounts of MD(05,10,15,2.0 and 2.5 g).Preliminary phytochemical screening of fnuit pulp and powder showed the presence of some phytoconstituents such as reducing sugar,saponin,flavonoids,and terpenoids,whereas akaloids,tannins,steroids and protein were absent in *** parameters of the powder are durectly infuenced by the amount of MD,as evidenced by the variations in bulk density,total soluble solids,pH,specific gravity and the proximate among the powder *** solubility was found in the sample with a medium of *** highest of particle distribution was found in MD-added samples MP2(1 g),MP3(1.5 g),MP4(2 g)whereas the raw(MPO)showed the *** analysis was done using 9-point hedonic scale,which revealed that MP2,MP3,and MP4 were highly *** study assumed that fresh Alu fuit,palp and in the form of powder can be useful for the development of new products.

关键词： Meyna laxiflora Robyns Maltodextrin Solubility Particle size distribution Sensory analysis

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Metabolic collaboration between cells in the tumor microenvironment has a negligible effect on tumor growth

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The Innovation 2024年第2期5卷 107-114页

作者： Johan Gustafsson Fariba Roshanzamir Anders Hagnestål sagar M.Patel Oseeyi I.Daudu Donald F.Becker Jonathan L.Robinson Jens Nielsen Department of Life Sciences Chalmers University of TechnologySE-41296 GothenburgSweden Hagnesia AB SE-43854 HindåsSweden Department of Biochemistry and Redox Biology Center University of Nebraska-LincolnLincolnNE 68588USA BioInnovation Institute DK2200 CopenhagenDenmark

The tumor microenvironment is composed of a complex mixture of different cell types interacting under conditions of nutrient deprivation,but the metabolism therein is not fully understood due to difficulties in measuring metabolic fluxes and exchange of metabolites between different cell types in ***-scale metabolic modeling enables estimation of such exchange fluxes as well as an opportunity to gain insight into the metabolic behavior of individual cell ***,we estimated the availability of nutrients and oxygen within the tumor microenvironment using concentration measurements from blood together with a metabolite diffusion *** addition,we developed an approach to efficiently apply enzyme usage constraints in a comprehensive metabolic model of human *** combined modeling reproduced severe hypoxic conditions and the Warburg effect,and we found that limitations in enzymatic capacity contribute to cancer cells’preferential use of glutamine as a substrate to the citric acid ***,we investigated the common hypothesis that some stromal cells are exploited by cancer cells to produce metabolites useful for the cancer *** identified over 200 potential metabolites that could support collaboration between cancer cells and cancer-associated fibroblasts,but when limiting to metabolites previously identified to participate in such collaboration,no growth advantage was *** work highlights the importance of enzymatic capacity limitations for cell behaviors and exemplifies the utility of enzyme-constrained models for accurate prediction of metabolism in cells and tumor microenvironments.

关键词： metabolism effect limitations

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Effect of ankle versus thigh tourniquets on post-operative pain in foot and ankle surgery

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World Journal of Orthopedics 2024年第2期15卷 163-169页

作者： Ashish Mishra Ahmed Barakat Jitendra Mangwani Jakub Kazda sagar Tiwatane Sana Mohammed Aamir Shaikh Linzy Houchen-Wolloff Vipul Kaushik Department of Trauma and Orthopedics University Hospitals LeicesterLeicester LE15WWUnited Kingdom Department of Trauma and Orthopedics Leicester University Hospitals-NHS TrustLeicester LE15WWLeicestershireUnited Kingdom Academic Team of Musculoskeletal Surgery University Hospitals of Leicester NHS TrustLeicester LE39QPUnited Kingdom Department of Anaesthesia York&Scarborough Teaching Hospitals NHS Foundation TrustNorth Yorkshire YO318HEUnited Kingdom Department of Anaesthesia Royal Free London NHS TrustLondon NW32QGUnited Kingdom Department of Trauma&Orthopaedics Breach Candy Hospital TrustMumbai 400026India Department of Physiotherapy University Hospitals of Leicester NHS TrustLeicester LE39QPUnited Kingdom Department of Anasthesia University Hospitals of Leicester NHS TrustLeicester LE15WWUnited Kingdom

BACKGROUND Tourniquets are commonly used in elective extremity orthopaedic surgery to reduce blood loss,improve visualization in the surgical field,and to potentially reduce surgical *** is a lack of consensus in existing guidelines regarding the optimal tourniquet pressure,placement site,and duration of *** is a paucity of data on the relationship between the site of a tourniquet and postoperative pain in foot and ankle *** To explore the relationship between tourniquet site and intensity of post-operative pain scores in patients undergoing elective foot and ankle *** Retrospective analysis of prospectively collected data on 201 patients who underwent foot and ankle surgery in a single institution was *** tourniquet duration,tourniquet pressure and site,and postoperative pain scores using Visual Analogue Score were collected in immediate recovery,at six hours and at 24 h *** plots were used to analyse the data and to assess for the statistical correlation between tourniquet pressure,duration,site,and pain scores using Pearson correlation *** All patients who underwent foot and ankle surgery had tourniquet pressure of 250 mmHg for ankle tourniquet and 300 mmHg for *** was no correlation between the site of the tourniquet and pain scores in recovery,at six hours and after 24 *** was a weak correlation between tourniquet time and Visual Analogue Score immediately post-op(r=0.14,P=0.04)but not at six or 24 h *** This study shows that there was no statistically significant correlation between tourniquet pressure,site and postop pain in patients undergoing foot and ankle *** choice of using a tourniquet is based on the surgeon's preference,with the goal of minimizing the duration of its application at the operative site.

关键词： Lower limb surgery Tourniquet time Tourniquet pressure Tourniquet site Post-operative pain Pain scores

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