bluuebunny/crossref_metadata_2025 huggingface dataset:
Application of Blockchain in case of Engineering Data Processes and Product Lifecycle Management System
Authors: Salehi Vahid , Vilsmeier Josef
Published: February 2025

In the following paper, the possibilities of the latest industry 4.0 and digitization technologies, such as the use of Blockchain (BC) in Model-Based Systems Engineering with the associated product data (PDM) and product life cycle (PLM) management information, will be considered holistically. A Blockchain Engineering approach called EngineeringBlock will be researched in Systems engineering for the management of software data and engineering data within product lifecycle management. At this point, data input refers, for example, to the uploading of software versions to a control unit in the Engineering Data like Requirements and Functions. The two most important directions of the research paper will show the use of Blockchain technology with the related PLM System. This paper will present a Framework which enables the presentation of all product-relevant requirements, functions and logical aspects of the product up to the usage phase in a holistic and consistent way.
Application of Systems Engineering in context of building 3D Environment for Autonomous Vehicle Systems in an industrial context for Central Car Computing
Authors: Salehi Vahid
Published: February 2025

This paper outlines a robust framework for the creation of intelligent 3D traffic environments driven by advanced data insights. The primary objective is to enhance traffic process efficiency using virtualization and comprehensive data integration. Smart 3D environments offer a sophisticated means to replicate real-life traffic scenarios digitally, thereby supporting well-informed decision-making, optimizing traffic flows, and enabling predictive maintenance strategies. By incorporating real-time sensor data and leveraging innovative AI models, these environments build a dynamic and precise digital representation of traffic systems, enabling continuous monitoring and actionable data insights. The presented framework incorporates essential elements such as sensor networks, IoT-based systems, and AI-driven algorithms to construct an all-encompassing digital counterpart of the actual traffic environment. This digital replica facilitates the simulation of various traffic conditions, allowing for the identification and mitigation of inefficiencies before they impact real-world systems. Additionally, the implementation of these intelligent environments enables the seamless integration of novel technologies, including autonomous vehicles and connected infrastructure, into current traffic systems. Moreover, this paper emphasizes the importance of the Systems Modelling Language (SysML) for effectively organizing and managing the complex data sets required for the development and ongoing maintenance of these environments. The study’s findings illustrate that intelligent 3D traffic environments contribute significantly to improved operational efficiency, resource optimization, and adaptability within dynamic traffic ecosystems. This research provides valuable insights for engineers and scholars aiming to apply intelligent 3D modelling solutions in the domain of traffic management and urban planning.
SYNERGISTIC EFFECTS OF BORON AND ZINC ON THE MORPHO-PHYSIOLOGICAL AND NUTRITIONAL TRAITS OF CUCUMBER (CUCUMIS SATIVUS L.) AS INFLUENCED BY DIFFERENT APPLICATION METHODS
Authors: Zahid Hussain  , Khalid Khan  , Sajjad Ali  , Wajid Ali Shah  , Manzoor Ahmad  , Roohul Amin  , Rahamdad Khan
Published: February 2025

A pot experiment was conducted to investigate the effects of various concentrations of boron (B) and zinc (Zn) applied through different methods on cucumber. The results showed significant increases in morphological attributes, biochemical parameters (chlorophyll, carotenoids, sugar, protein, proline, phenols, and flavonoids), and nutrient contents (iron, Zn, manganese, B, magnesium, nitrogen, phosphorus, potassium, and calcium) with the application of B and Zn. The combined root and leaf application method (R+L) was more effective than separate applications. Optimal levels of B (3.8 mg/L) and Zn (3.8 mg/L) with R+L application method produced the best results. This study concludes that B and Zn application at optimal levels with R+L method can improve cucumber growth, biochemical properties, and nutrient uptake.
GENDER-BASED VIOLENCE IN DIGITAL SPACES: EXPLORING CHALLENGES, AND POTENTIAL SOLUTIONS FOR UNIVERSITY STUDENTS IN PAKISTAN
Authors: Jalal Khan  , Dr Tazeen Saeed Ali  , Ambreen Ahmed  , Qiamuddin  , Sofia Mansoor  , Anny Ashiq Ali  , Mehdi Muhammad
Published: February 2025

Digital gender-based violence (GBV) poses a serious challenge for young adults, especially for university students in Pakistan. The speedy rise in online engagement has increased risks of harassment, abuse, mistreatment and exploitation within digital sitting room. This study explores key definitions, societal dynamics, and fundamental factors driving digital GBV, together with potential strategies to reduce its impact. Proposed solutions include policy development, awareness campaigns, and the implementation of technological protections to foster safer online environments. By analyzing the interplay between societal norms and evolving digital interactions, this study aims to guide future research and inform comprehensive interventions. The findings are expected to support the creation of evidence-based strategies that enhance positive digital engagement while addressing the vital need for protection against online abuse. Through a multidisciplinary perspective, this effort highlights the importance of collaboration to combat digital GBV and promote a safer digital culture for students.
IMPACT OF NITROGEN FERTILIZER MANAGEMENT ON DISTRIBUTION OF NITROGEN CONTENT IN DIFFERENT ORGANS OF WINTER WHEAT
Authors: Muhammad Saleem Kubar  , Asif Ali Kaleri  , Danish Manzoor  , Urooj Rehmani  , Shafiq Ur Rehman  , Wahid Baksh  , Hafsa Munir  , Rabia Laghari  , Vik Ram Meghwar  , Muhammad Luqman Khan Bazai  , Yang Wude
Published: February 2025

However, increasing the rates and adjusting the time of nitrogen treatment might worsen nitrogen loss and lower grain yields, which will have a detrimental effect on the quality of the environment. The relationship between nitrogen nutrition and grain production has been extensively studied, but much has been discovered about how nitrogen concentration in winter wheat, higher up in components like leaves, stems, and grains, reacts to nitrogen nutrition. At Shanxi Agricultural University's Taigu Experimental Agricultural Centre in Shanxi Province, China, experiments in the field were carried out. Three repetitions of these split-plot tests were conducted. In the main plots, four nitrogen application levels—75, 150, 225, and 300 kg ha-1 were tested. Different nitrogen levels were used in the subplots, including 5:5 (50%, 50%) and 6:4 (60%, 40%). Nitrogen fertilizer was applied at pre-sowing, jointing, flowering, and grain filling stages, respectively. The 12 m² (3 m × 4 m) experimental plots were used. As nitrogen fertilizer rates increased, there was a considerable increase in the total nitrogen content of the dry matter. In comparison to other growing phases and nitrogen rates, plants got nitrogen nutrition at 60% during sowing and 40% during the jointing stage, increasing the average total nitrogen content from 40.00 to 60.00% under 225 kg N ha-1. Less than 225 kg N ha-1 at the jointing stage, when nitrogen was given in 6:4 ratios, the maximum nitrogen content in leaves was recorded at 32–34 g kg-1. Similarly, at 225 kg N ha-1, the maximum nitrogen contents in the stem were measured at 13–15 g kg-1, meaning that plants absorbed 60% of their nitrogen nutrition during sowing and 40% during the jointing stage. 225 kg ha-1 of nitrogen, applied in the following proportions: 60%, 40%, 0%, and 0% (amount applied at the sowing, jointing, blooming, and grain filling stages) successfully increased the nitrogen content of winter wheat's leaves, stems, and grains above ground.
ANALYSIS OF MICROBIOLOGICAL AND PHYSICOCHEMICAL PARAMETERS OF WASTEWATER FROM TAMBUWAL ABATTOIR, SOKOTO STATE, NIGERIA
Authors: Atiku Yari Dogon Daji  , Sa’adu Abdullahi Kaffe  , Ahmad Bello  , Nura Maikawai Salah  , Zayyanu Danbaba kware
Published: February 2025

Pollution is threatening water quality worldwide. Abattoirs are examples of sources of polluted waste water that can affect biota. This study examined the microbiological and physicochemical parameters of wastewater from the Sokoto Modern Abattoir in Nigeria. The key findings are: Microbiological analysis results include, 1. Bacterial isolates (12 bacterial isolates were identified, including Staphylococcus chromogene, Proteus Vulgaris, Enterobacter Aerogene, and others); 2. Fungal isolates (Fungal isolates were also identified, with varying macroscopic characteristics). 3. Coliform count: The coliform count was high, indicating potential contamination. Physicochemical Analysis; Temperature, Dissolved oxygen, Biochemical oxygen demand (BOD), and Chemical oxygen demand (COD) were lower, but pH was high, indicating potential pollution. Conclusively, the study highlights the need for quality analysis at abattoirs to safeguard public health and protect the interests of animal rearers.
ASSESSING CYBERSECURITY CHALLENGES AND RESPONSE READINESS IN PAKISTAN: A COMPREHENSIVE ANALYSIS
Authors: Muhammad Saad Qasim  , Zeeshan Ahmad  , Saima Maqsood  , Shahroz Zafar  , Dr. Muhammad Azam
Published: February 2025

The rapid transformation of the globe into a global village that has been brought about by the enormous rise of information technology. As a result, the distances between places have become more manageable, and information can now travel quickly throughout the world as events unfold. On the other hand, at the same time, it has also provided a boost to vulnerabilities, dangers, frauds, and criminals in the online. The ease of access, hacking tools that are favourable to users, and complexity of cyber assaults have all contributed to an invasion of privacy on the part of people, organisations, and nations. In this day and age, Pakistan is confronted with a myriad of different kinds of cyber threats. In order to determine the nature and dynamics of the danger posed to Pakistan's information and communications technology (ICT) infrastructure and resources, this study conducts an analysis of recent cyber assaults that have been launched against the government, corporate, and private sectors. The research sheds light on a broad spectrum and diversity of cyber risks, including anything from simple website defacement to complex and enduring cyber threat. In addition, the current reaction capacity at the organisational and government levels has been examined, and the deficiencies have been brought to light. It is thought that Pakistan's lack of any regulations pertaining to cyber security, the absence of any reaction mechanisms, and the absence of any organisational structure in the nation may make the country's internet a paradise for criminals and operators and users who intentionally do harm. It should be underlined that the government of Pakistan not only needs to grasp the dangers posed by the internet and the repercussions of its uncontrolled use, but it also needs an effective reaction system in order to protect itself from these dangers. This paper proposes a high level organisational structure for the development of critical cyber security organisations at multiple tiers. These bodies will be responsible for safeguarding the nation's cyber space by enacting appropriate laws and designing the reaction mechanism at different levels of government.
HARNESSING COCONUT HUSK: SOLVENT-BASED EXTRACTION AND ITS ANTIMICROBIAL ACTIVITY AGAINST FOODBORNE PATHOGENS
Authors: Sana Khan  , Syeda Bushra Zafar  , Marium Shaikh  , Mehwish Faiz  , Syeda Hafiza Afsheen Zafar  , Shahzad Nasim
Published: February 2025

Natural sources of antimicrobial agents have gained attention as antibiotic resistance continues to threaten global health. Coconut husk, a renewable agro-industrial byproduct rich in phenolics, flavonoids, and tannins, demonstrates antimicrobial properties. This study evaluates coconut husk extracts prepared with ethanol, methanol, and water solvents for antimicrobial activity against four food borne pathogens including Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Salmonella typhi. The ethanol extract showed significant antimicrobial activity, particularly at 48 hours, producing the largest inhibition zones (up to 29 mm for P. aeruginosa), due to its ability to extract a broader range of bioactive compounds. Methanol and water extracts showed moderate activity but were less effective compared to ethanol. Structural changes observed in microbial cell membranes after coconut husk extraction were confirmed by SEM, which indicated disruption of cellular integrity as mode of action of the antimicrobial activity. Active compounds present in phytochemical analysis confirmed to be alkaloids, tannins, flavonoids, and phenolics. The antimicrobial activity of coconut husk extracts indicating their potential to use in food preservation reducing pathogen load and extending shelf life.
ASSESSMENT OF KNOWLEDGE AND AWARENESS OF ANEMIA IN DISTRICT SIALKOT, PUNJAB, PAKISTAN
Authors: Fatima Afzal  , Sherjeel Adnan  , Jannat Asif  , Laiba Zia  , Maila Iftikhar  , Manal Akmal  , Mariam Amir  , Maryam Asif  , Umme Kalsoom
Published: February 2025

Background: Anemia is a major global health issue that impacts public health by reducing quality of life and productivity. This study explores how much people in Sialkot, Punjab, Pakistan knows about anemia and identifies gaps in their awareness.
Objective: To assess knowledge, identify risk factors, and evaluate perceptions about anemia and its management.
Methods: A descriptive cross-sectional study surveyed 424 participants from urban and rural areas using a structured questionnaire. Data were analyzed using Microsoft Excel 2019, with descriptive statistics highlighting trends.
Results: Of 424 respondents, 78.8% had general awareness of anemia, with healthcare professionals being the most common source of information (23.5%). About 71.9% recognized risk factors, while 67.1% were familiar with symptoms. Challenges remain in understanding diagnostic methods and available treatment options.
Conclusion: The study highlights a moderate level of awareness with notable differences between urban and rural populations. Targeted interventions are essential to address these gaps and enhance public health outcomes.
A DEEP LEARNING APPROACH TO OPTIMIZING COVID-19 MORTALITY PREDICTIONS WITH RECURRENT NEURAL NETWORKS (RNNs)
Authors: Fazal Malik Fazal Malik , Hamid Ullah  , Ashraf Ullah  , Said Khalid Shah  , Rahmat Hussain  , Muhammad Javed
Published: February 2025

The COVID-19 pandemic, caused by the SARS-CoV-2 virus, has posed significant challenges to global health since its onset in December 2019. While numerous models have been developed to predict COVID-19 outcomes, many fall short in accurately forecasting mortality rates, focusing instead on clinical severity. This study addresses these gaps by proposing an advanced approach using Recurrent Neural Networks (RNNs) to improve mortality prediction. Although existing models have employed various machine learning (ML) and deep learning (DL) techniques, they often rely on limited data sources and lack a dedicated focus on global mortality forecasting. Our research introduces RNNs as a primary tool for addressing this limitation, enhancing predictive accuracy in the context of COVID-19 deaths. The methodology involves four key phases: (1) Data Acquisition from the "COVID-19 Symptoms Dataset," which includes variables such as symptoms, confirmed cases, and death rates; (2) Data Processing, which encompasses data cleaning, visualization, and feature selection; (3) Supervised Learning using the RNN model for prediction; and (4) Performance Evaluation, where the RNN's performance is assessed in terms of accuracy, precision, recall, and F1-score. Results show that the RNN model significantly outperforms previous approaches, achieving an accuracy of 94%, precision of 92%, recall of 93%, and F1-score of 92%. This study demonstrates the effectiveness of RNNs in enhancing COVID-19 mortality predictions, providing critical insights for better global forecasting and aiding public health efforts in resource allocation and response planning.
BREAKING BOUNDARIES: THE REPRESENTATION OF PAKISTANI WOMEN IN BAPSI SIDHWA’S PAKISTANI BRIDE
Authors: Dr. Syeda Sughra Naqvi
Published: February 2025

Bapsi Sidhwa’s Pakistani Bride (1983) presents a profound exploration of the lives of Pakistani women navigating the complexities of tradition, culture, and gender in a patriarchal society. This article examines how Sidhwa portrays the limitations imposed on women and how they challenge these boundaries, whether through defiance or passive resistance. Drawing from feminist literary theory, this paper analyzes the representation of women’s agency, social roles, and personal identity within the socio-cultural framework of Pakistan. It argues that through her nuanced characters, Sidhwa critiques the structures of patriarchy while also highlighting the potential for transformation and change in Pakistani women’s lives. The study focuses on the central character, Zaitoon, and her journey from submission to empowerment, emphasizing the significance of education, love, and self-discovery as pivotal in challenging the societal constraints placed on women.
ناول بستی میں تہذیبی بکھراؤ اور ناسٹلجیائی بیانیہ
Authors: Meharin Saba  , Saeeda Irum
Published: February 2025

Intezar Hussain, who suffered from the pain of migration, writes about the memories of the past. Migration, the feeling of exil, the reflection of the glorious past and happy memories are prominent in his writings. There is a lot of nostalgia in his novels.which is a psychological phenomenon. The environment, living conditions, and cultural customs constantly remind the diasporas of their past. This research article explores how Intzar Hussain has described nostalgic feelings of Diaspora in his novel Basti.
COST EFFECTIVE ROUTE OPTIMIZATION FOR DAIRY PRODUCT DELIVERY
Authors: Nasir Khan  , Shahbaz Ahmad  , Salman Raza  , Dr. Ahmad Khan  , Muhammad Younas
Published: February 2025

Milk is an essential component of human nutrition, particularly for the growth and cognitive development of children in undernourished populations. In Pakistan, milk production has more than doubled, with the country now the third-largest milk producer in the world. Most milk in Pakistan, however, is consumed in the form of unpackaged loose milk not only making hygiene an issue but logistical as well. An optimized route planner caters to the cost-effective and environmentally friendly delivery of dairy products. Vehicle routing has only played a minor role in large-scale, dimension-driven, and technology-defined approaches in developing traditional solutions. To address these limitations, sophisticated optimization methods like metaheuristics, machine learning, and deep reinforcement learning (DRL), are studied. In this paper, we proposed a DRL-based delivery model that optimizes routes of dairy products in both capacity and time windows constraint using Proximal Policy Optimization (PPO). By integrating vehicle, customer, and global states, the framework makes informed decisions while satisfying constraints including vehicle capacity and delivery deadlines. We analyze performance measurements such as total route length, inter-customer gaps, time constraints, and capacity utilization to assess efficiency. Experimental results demonstrate that the DRL-PPO model significantly outperforms traditional benchmarks such as Deep Q-Network (DQN) and Advantage Actor-Critic (A2C). Specifically, DRL-PPO yielded advantages in several key areas: it achieved reduced route lengths, shorter delivery times, minimized inter-customer gaps, increased vehicle capacity utilization, and higher success rates across varying levels of task complexity. These findings indicate the potential of Deep Reinforcement Learning (DRL)-based optimization methods to effectively address logistical inefficiencies prevalent in Pakistan's dairy industry. The implications of adopting DRL-PPO are substantial, offering cost-effective solutions that can enhance profitability while concurrently reducing operational expenses. By leveraging advanced machine learning techniques, businesses can pave the way toward sustainable logistics systems for perishable goods. This not only boosts overall supply chain efficiency but also ensures better access to high-quality dairy products for consumers. As such, the use of DRL-PPO represents a strategic opportunity to transform logistics practices within the dairy sector, leading to improved outcomes for both producers and consumers alike.
CHEMICAL COPRECIPITATION SYNTHESIS AND ENHANCED PHOTOCATALYTIC CHARACTERISTICS OF NICKEL-DOPED ZINC SULFIDE NANOPARTICLES
Authors: Muhammad Iqbal  , Zeeshan Mahmood  , Waseem Sajjad Tahir  , Kanwal Akhtar  , Muhammad Salman  , Aurangzaib  , Aneela Din Muhammad  , Yasir Javeed
Published: February 2025

Metal sulfide semiconductors having wide bandgap like ZnS experience a significant decrease in band gap when transition metal ions like Ni2⁺ are doped. This alteration creates new composite materials with various uses in photonics, solar cells, optoelectronic devices, and spintronic devices. These modifications have led to improvements in ZnS nanoparticle production techniques. Nickel-doped zinc sulfide nanoparticles are synthesized by using a simple chemical coprecipitation method. Ni-doped ZnS nanoparticles have been characterized using UV-visible spectra and Fourier-transform infrared spectroscopy. Zn–S vibrational bands were detected by Fourier-transform infrared (FTIR) spectroscopy; changes in these bands suggested that Ni had been successfully incorporated into the ZnS lattice. UV–visible spectroscopy revealed a decrease in the optical band gap from 3.70 eV for 3% Ni-doped ZnS to 3.46 eV for 1% Ni-doped ZnS, demonstrating tunable optical properties due to Ni doping, which could enhance photocatalytic performance under visible light.
Design and Development of Self Microemulsifying Drug Delivery System of Clopidogrel Bisulphatea
Authors: Patel Priyank B.
Published: February 2025

The aim of this investigation was to design and develop SMEDDS formulation of clopidogrel for enhancing its solubility. Oral bioavailabilityof clopidogrel is very low (less than 50%), due to its poor water solubility. For this purpose, initially, solubility of clopidogrel was determined in various vehicles. SMEDDS was prepared with Capmul MCM NF as oil, Acrysol K140 as surfactant and Propylene Glycol as co-surfactant after preliminary screening of excipients. The system was optimized by ternary phase diagram. The prepared formulations were characterized for Drug content, % Transmittance, Visual assessment of self emulsification, particle size and Invitro drug release. The optimized system contains mean particle size of 16.13 nm and zeta potential (ζ) -8.25 mV. In-Vitro drug release of optimized formulation carried out which gives 98.48% within 40 minute. Hence, by formulating into SMEDDS, the solubility ofClopidogrel was found to be significantly enhanced.
Using social media in the business of small and medium-sized enterprises in Serbia
Authors: Jakšić Krsto , Trajković Slaviša , Đurović Mihajlo
Published: February 2025

Today's era of digitalization and rapid technological change presents an opportunity for small and medium-sized enterprises (SMEs) to significantly improve their competitiveness by following and applying these achievements. One such aspect is the use of social media, which can significantly increase their presence in new markets and thus increase sales volume. Social media technologies enable new ways of communicating with consumers and other partners, thus overcoming resource constraints. This paper aims to analyze the use of social media by small and medium-sized enterprises in the Republic of Serbia and provides a comparative analysis of the use of web sites and social networks in enterprises from neighboring countries, as well as in the European Union. The results indicate lower use of social media by small and medium-sized enterprises compared to large ones in the Republic of Serbia. At the same time, the results show higher use of social media by enterprises in the Republic of Serbia compared to neighboring countries.
La révolte des sans-travail montréalais et la révolte ouvrière au Québec (1919-1925)
Authors: Marsan Benoit
Published: February 2025

À travers l’analyse de la contestation des personnes sans travail qui se produit à Montréal entre 1919 et 1925, cet article vise deux objectifs. Le premier est de faire ressortir l’importance du chômage et des mobilisations des sans-travail lors de la révolte ouvrière au Canada (1917-1925). Le second est d’apporter une nouvelle contribution à l’historiographie québécoise de la révolte ouvrière en montrant que ce mouvement de protestation doit-être apprécié plus largement qu’à la seule lumière de l’histoire des syndicats internationaux et de la vague de grèves de 1919-1921.
ТЕЛА ЗА КРИВИЧНУ И ПОЛИЦИЈСКУ САРАДЊУ У ЕВРОПСКОЈ УНИЈИ, ЕВРОПСКИ ЈАВНИ ТУЖИЛАЦ И ЦОРПУС ИУРИС
Authors: Симовић Миодраг Н. , Хрустић Амна
Published: February 2025

Европска унија већ дуго улаже одређене напоре да развије кривично право Европске уније, чији би главни циљ био заштита финансијских интереса Уније. У ту сврху уведени су одређени нови кривичноправни институти типични за ову грану права. Међутим, да би се постигла стварна имплементација и примена механизама заштите интереса Уније, постојала је потреба да се унутар Уније успоставе тела полиције и криминалне сарадње, која ће бити независна од кривичних закона држава чланица, али на истовремено остварују висок степен сарадње између себе и са националним органима држава чланица, а све у циљу брже, ефикасније и једноставније сарадње и заштите вредности Уније. У том контексту уводе се и Европски јавни тужилац, те академска студија Цорпус Иурис, за коју многи сматрају да је претеча кривичног закона Европске уније. У раду су детаљније представљени органи за правосудну сарадњу између држава чланица – Евројуст, Европска правосудна мрежа, магистрати за везу, Еуропол, Европска канцеларија за борбу против превара – ОЛАФ, Европски јавни тужилац и Цорпус иурис. Поред тога, предмет пажње су и тенденције у заштити људских права.
Characterization of direct Purkinje cell outputs to the brainstem
Authors: Chen Christopher H , Yao Zhiyi , Wu Shuting , Regehr Wade G
Published: February 2025

Purkinje cells (PCs) primarily project to cerebellar nuclei but also directly innervate the brainstem. Some PC-brainstem projections have been described previously, but most have not been thoroughly characterized. Here we use a PC-specific cre line to anatomically and electrophysiologically characterize PC projections to the brainstem. PC synapses are surprisingly widespread, with the highest densities found in the vestibular and parabrachial nuclei. However, there are pronounced regional differences in synaptic densities within both the vestibular and parabrachial nuclei. Large optogenetically-evoked PC-IPSCs are preferentially observed in subregions with the highest densities of PC synapses, suggesting that PCs selectively influence these areas and the behaviors they regulate. Unexpectedly, the pontine central gray and nearby subnuclei also contained a low density of PC synapses, and large PC-IPSCs are observed in a small fraction of cells. We combined electrophysiological recordings with immunohistochemistry to assess the molecular identities of two putative PC targets: PC synapses onto mesencephalic trigeminal neurons were not observed even though these cells are in close proximity to PC boutons. PC synapses onto locus coeruleus neurons are exceedingly rare or absent, even though previous studies concluded that PCs are a major input to these neurons. The availability of a highly selective cre line for PCs allowed us to study functional synapses, while avoiding complications that can accompany the use of viral approaches. We conclude that PCs directly innervate numerous brainstem nuclei, but only inhibit a small fraction of cells in many nuclei. This suggests that PCs target cell types with specific behavioral roles in brainstem regions.
Flexibility in PAM recognition expands DNA targeting in xCas9
Authors: Hossain Kazi A , Nierzwicki Lukasz , Orozco Modesto , Czub Jacek , Palermo Giulia
Published: February 2025

xCas9 is an evolved variant of the CRISPR-Cas9 genome editing system, engineered to improve specificity and reduce undesired off-target effects. How xCas9 expands the DNA targeting capability of Cas9 by recognising a series of alternative protospacer adjacent motif (PAM) sequences while ignoring others is unknown. Here, we elucidate the molecular mechanism underlying xCas9’s expanded PAM recognition and provide critical insights for expanding DNA targeting. We demonstrate that while wild-type Cas9 enforces stringent guanine selection through the rigidity of its interacting arginine dyad, xCas9 introduces flexibility in R1335, enabling selective recognition of specific PAM sequences. This increased flexibility confers a pronounced entropic preference, which also improves recognition of the canonical TGG PAM. Furthermore, xCas9 enhances DNA binding to alternative PAM sequences during the early evolution cycles, while favouring binding to the canonical PAM in the final evolution cycle. This dual functionality highlights how xCas9 broadens PAM recognition and underscores the importance of fine-tuning the flexibility of the PAM-interacting cleft as a key strategy for expanding the DNA targeting potential of CRISPR-Cas systems. These findings deepen our understanding of DNA recognition in xCas9 and may apply to other CRISPR-Cas systems with similar PAM recognition requirements.
Megakaryocytes assemble a three-dimensional cage of extracellular matrix that controls their maturation and anchoring to the vascular niche
Authors: Masson Claire , Scandola Cyril , Rinckel Jean-Yves , Proamer Fabienne , Janus-Bell Emily , Batool Fareeha , Osmani Naël , Goetz Jacky G , Mallo Léa , Léon Catherine , Bornert Alicia , Poincloux Renaud , Destaing Olivier , Mansson Alma , Qian Hong , Lehmann Maxime , Eckly Anita
Published: February 2025

Megakaryocytes, the progenitor cells of blood platelets, play a crucial role in hemostasis by residing in the bone marrow and ensuring continuous platelet production. Unlike other hematopoietic cells, megakaryocytes do not enter the blood circulation intact. They remain anchored within the bone marrow while extending cytoplasmic protrusions called proplatelets through the sinusoidal endothelial barrier. These proplatelets subsequently fragment into functional platelets. This unique process of intravasation facilitates efficient platelet production while maintaining the megakaryocyte cell body within the bone marrow niche, thus preventing potential thrombotic complications. How the extracellular matrix (ECM) influences the delicate balance between megakaryocyte retention and proplatelet extension remains largely unknown. Here, we investigate the spatial organization and functional role of ECM components in the megakaryocyte vascular niche. Our findings reveal that laminin and collagen IV form three-dimensional (3D) ECM cages encompassing megakaryocytes and anchor them to the sinusoidal basement membrane. Gene deletion shows the existence of laminin α4 in the ECM cage that is necessary to maintain megakaryocyte-sinusoid interactions. Notably, megakaryocytes actively contribute to the ECM cage assembly; β1/β3 integrin knockout weakens these structures, increasing intravasation and entire megakaryocyte entry into circulation. The retention of megakaryocytes by these 3D ECM cages depends on dynamic remodeling processes. Inhibition of ECM proteolysis results in denser cage formation, increasing the frequence of immature megakaryocytes with impaired demarcation membrane system (DMS) development. Thus, the ECM cage represents a novel concept of an active and dynamic 3D microenvironment that is continuously remodeled and essential for maintaining megakaryocyte perivascular positioning. This specific microarchitecture guides megakaryocyte maturation and intravasation, underscoring the critical role of ECM microarchitecture and dynamics in megakaryocyte function.
iC3 Research Data Management Plan
Authors: Sarti Fabio , Wadham Jemma , Winsborrow Monica , Assmy Philipp , Langebroek Petra , Souster Terri
Published: February 2025

The iC3 DMP's purpose is to provide clear guidance to iC3 members, to ensure that data is proficiently managed at all stages of the project and that it remains accessible, secure and sustainable in the long term. Furthermore, it will ensure better project implementation by providing a clear overview of all data management activities. To achieve these goals the DMP includes measures to comply with FAIR principles and OpenScience by Sikt.
Romalılar Gıdalarını Nasıl Muhafaza Ettiler: Tahıl, Üzüm ve Zeytin
Authors: Kapka Emine
Published: February 2025

Kendi ürününü yetiştirmeyi ve işlemeyi, medeniyet göstergesi ve erdemlilik olarak kabul eden Romalılar için gıdayı muhafaza etmek önemli olmuş olmalıdır. Bu varsayımla yola çıktığımız bu çalışma, “Romalılar gıdalarını nasıl muhafaza ettiler?”, sorusunu yanıtlamayı amaçlamaktadır. Lakin yapmış olduğumuz araştırmalar konunun, birkaç müstakil çalışma kapsamında olduğunu göstermiştir. Dolayısıyla, konu sınırlandırılması zorunluluğu hasıl olmuştur. Bu anlamda önceliğimiz Roma beslenme kültürünün temelini oluşturan tahıl, üzüm ve zeytin ürünleri yönünde olmuş, çalışmamızın içeriği de bu yönde şekillenmiştir. Çalışmamızın tarihi sınırları olarak, MÖ 753-MS 281 yılları belirlenirken, bugünkü İtalya, coğrafi sınır olarak saptanmıştır. Bu çerçeve içerisinde yer alan Latince eserler, çalışmamızın ana kaynaklarını oluşturmaktadır. Ek olarak, modern literatür taranmıştır. Nitekim ilgili alanda yapılan çalışmaların, Roma özelinde olmayıp konuyu dağınık bir şekilde ele aldıkları görülür. Bu sebepten yapmış olduğumuz çalışmanın, literatürdeki boşluğun doldurulmasına katkı sağlayarak, okuyucuyu yeni araştırma sorularına yönlendireceği kanaatindeyiz. Yapılan çalışmalar, Romalıların, günümüz gıda muhafazası metotlarının bilgisine sahip olduklarını, tek farkın teknolojik donanın olduğunu göstermiştir. Romalılar için gıda muhafazası, en ilkel ihtiyaçlardan biri olan beslenebilmeyi sağlamakla kalmamış siyaset, edebiyat, mimari gibi hayatlarının pek çok alanına nüfuz ederek onları şekillendirmiştir.
Impact of proton PBS machine operating parameters on the effectiveness of layer rescanning for interplay effect mitigation in lung SBRT treatment
Authors: Liang Xiaoying , Liu Chunbo , Shen Jiajian , Flampouri Stella , Park Justin C. , Lu Bo , Yaddanapudi Sridhar , Tan Jun , Furutani Keith M. , Beltran Chris J.
Published: April 2024

Background

Rescanning is a common technique used in proton pencil beam scanning to mitigate the interplay effect. Advances in machine operating parameters across different generations of particle therapy systems have led to improvements in beam delivery time (BDT). However, the potential impact of these improvements on the effectiveness of rescanning remains an underexplored area in the existing research.

Methods

We systematically investigated the impact of proton machine operating parameters on the effectiveness of layer rescanning in mitigating interplay effect during lung SBRT treatment, using the CIRS phantom. Focused on the Hitachi synchrotron particle therapy system, we explored machine operating parameters from our institution's current (2015) and upcoming systems (2025A and 2025B). Accumulated dynamic 4D dose were reconstructed to assess the interplay effect and layer rescanning effectiveness.

Results

Achieving target coverage and dose homogeneity within 2% deviation required 6, 6, and 20 times layer rescanning for the 2015, 2025A, and 2025B machine parameters, respectively. Beyond this point, further increasing the number of layer rescanning did not further improve the dose distribution. BDTs without rescanning were 50.4, 24.4, and 11.4 s for 2015, 2025A, and 2025B, respectively. However, after incorporating proper number of layer rescanning (six for 2015 and 2025A, 20 for 2025B), BDTs increased to 67.0, 39.6, and 42.3 s for 2015, 2025A, and 2025B machine parameters. Our data also demonstrated the potential problem of false negative and false positive if the randomness of the respiratory phase at which the beam is initiated is not considered in the evaluation of interplay effect.

Conclusion

The effectiveness of layer rescanning for mitigating interplay effect is affected by machine operating parameters. Therefore, past clinical experiences may not be applicable to modern machines.
Patient‐Led Urate Self‐Monitoring to Improve Clinical Outcomes in People With Gout: A Feasibility Study
Authors: Michael Toni J. F. , Wright Daniel F. B. , Chan Jian S. , Coleshill Matthew J. , Aslani Parisa , Hughes Dyfrig A. , Day Richard O. , Stocker Sophie L.
Published: April 2024

Objective

Self‐monitored point‐of‐care urate‐measuring devices are an underexplored strategy to improve adherence to urate‐lowering therapy and clinical outcomes in gout. This study observed patient‐led urate self‐monitoring practice and assessed its influence on allopurinol adherence, urate control, and health‐related quality of life.

Methods

People with gout (n = 31) and prescribed allopurinol self‐monitored their urate concentrations (HumaSens2.0plus) at baseline and thereafter monthly for 12 months (3 months per quarter). Adherence to allopurinol was measured using medication event monitoring technology (Medication Event Monitoring System cap). Time spent below the target urate concentration (<0.36 mmol/L) was determined. Health‐related quality of life was measured using a survey (EuroQoL EQ‐5D‐5L). Gout flares were recorded. Two‐tailed Spearman correlation and the Wilcoxon matched‐pairs signed‐rank test (P < 0.05) were used for statistical comparisons.

Results

Most participants were male (94%) and had urate concentrations below the target (74%) at baseline. Overall, seven participants demonstrated repeated periods of “missed doses” (two or fewer allopurinol doses missed consecutively) and “drug holidays” (three or more missed doses). Most participants (94%) persisted with allopurinol. Time spent within the target urate concentration increased 1.3‐fold (from 79% to 100%; P = 0.346), and the incidence of gout flares decreased 1.6‐fold (from 8 to 5; P = 0.25) in the final quarter compared to that in the first quarter of the study. Health‐related quality of life was reduced for participants reporting at least one gout flare (median utility values 0.9309 vs 0.9563, P = 0.04).

Conclusion

Patient‐led urate self‐monitoring may support the maintenance of allopurinol adherence and improve urate control, thus reducing the incidence of gout flares. Further research on patient‐led urate self‐monitoring in a randomized controlled study is warranted.
Programmable Bacteria‐Based Biohybrids as Living Biotherapeutics for Enhanced Cancer Sonodynamic‐Immunotherapy
Authors: Wang Cheng , Chen Linfu , Zhu Jiafei , Wang Chunjie , Li Maoyi , Miao Yu , Liu Nanhui , Ji Zhaoxin , Pan Feng , Liu Yi , Zhu Junjie , Yang Yang , Chen Qian
Published: April 2024

Synthetic biology is propelling medicine into a new era through its capacity to genetically program living cells. One of the particular interests is engineering bacteria as a live and targeted therapeutic delivery system. Herein, the bacterial biohybrid (E. coli‐pE@PCN) is developed by genetically engineering Escherichia coli BL21 to overexpress catalase (E. coli‐pE) and electrostatically adsorbing nano‐sonosensitizers (PCN NPs) for enhanced and targeted sonodynamic therapy (SDT). Leveraging the ability to colonize and penetrate deep in tumors, engineered bacteria can not only sustainably express catalase to relieve tumor hypoxia, but also facilitate the enriched and expanded distribution of the carried sonosensitizer at the tumor site, so as to trigger effective SDT. More interestingly, it is found that E. coli‐pE@PCN‐based SDT can successfully inhibit the growth of subcutaneous and orthotopic colorectal tumors by inducing potent antitumor immune responses due to the released tumor‐associated antigens and native immunogenicity of bacterial pathogen‐associated molecular patterns. Furthermore, E. coli‐pE@PCN‐based SDT can not only prime a strong immune memory response to prevent tumor recurrence but also elicit a potent abscopal effect to inhibit tumor metastasis. Therefore, the programmable bacteria‐based biohybrids developed here pave an avenue to prepare next‐generation sonodynamic‐immunotherapeutics to eliminate cancer and prevent its relapse and metastasis.
A Simple and Sensitive Approach for Real‐Time Sensing of Enzymatically Catalyzed Hydrogelation
Authors: Putz Sebastian , Kassar Meriem , Oelschlaeger Claude , Franzreb Matthias , Kabay Gözde
Published: April 2024

Despite the critical role of hydrogels in material science and biotechnology, current methods for analyzing their formation lack real‐time monitoring and require complex sample preparation and instrumentation. In this work, an innovative methodology is introduced for the real‐time analysis of enzymatically catalyzed hydrogelation. Electrochemical impedance spectroscopy (EIS) coupled with interdigitated electrodes (IDEs) to sense and transduce the gelation reaction of model precursor carboxymethyl cellulose‐tyramine (CMC‐TA) conjugates that undergoes enzymatic cross‐linking by horseradish peroxidase (HRP) and hydrogen peroxide (H2O2). Real‐time monitoring involves single‐frequency analyses at 3 × 105 Hz, where the measured impedance consists solely of a resistive component, and the admittance equates to solution conductance. The gelation trajectories for all tested enzymatically cross‐linked hydrogel component combinations are determined by substituting the conductance data in the modified Michaelis–Menten kinetic model. Specifically, for CMC‐TA cross‐linked by HRP, the authors calculate apparent KM and kcat values of 82.1 µM and 95.5 s−1, respectively. These findings are further validated through rheological characterization, including oscillatory shear measurements and microrheology. Overall, this research paves the way for a streamlined, accurate, and cost‐effective approach to controllable enzymatically initiated hydrogel synthesis, enhancing their successful application in various fields ranging from material science to biotechnology.
Tailoring Photoweldable Shape Memory Polyurethane with Intrinsic Photothermal / Fluorescence Via Engineering Metal‐Phenolic Systems
Authors: Ping Zhongxin , Xie Fang , Gong Xiaobo , Zhang Fenghua , Zheng Jie , Liu Yanju , Leng Jinsong
Published: April 2024

Multifunctional shape memory polymers are significant for next‐generation intelligent systems as smart materials. However, their practical applications are limited by toughness, manufacturability, and functional integration strategies. Besides, while 4D printing is used to construct deformable devices, the limited accessible shapes and difficulty of integrating multiple materials hindered the development of multifunctional devices. Herein, metal‐phenolic coordination is used as crosslinks to tailor tough shape memory polyurethanes with intrinsic photothermal effect and/or tunable fluorescence. The reversible dissociation of metal‐phenolic complexes upon loading provides an efficient energy dissipation for the polymer network. The photothermal/fluorescent properties of target polyurethane can be effortlessly programmed by changing the coordinated metals and functional ligands. The obtained fluorescent polyurethane displays tunable fluorescence through the reversible dissociation‐association of dynamic bonds upon thermal stimulus. Moreover, relying on light‐triggered hybrid exchange reactions among various metal‐phenolic coordination bonds, heterogeneous interfaces can be merged to manufacture exquisite 3D shape‐shifting devices and integrate more advanced functions. As demonstrations, intrinsic photothermal‐responsive shape memory stent, and 3D fluorescent encryption device are produced based on photo‐welding instead of 4D printing. This study not only provides a convenient supramolecular strategy for constructing multifunctional smart materials but also demonstrates a feasible method for making personalized integrated architectures.
2D Ca2Nb3O10 Optoelectronic Neuromorphic Device for Ultrasensitive UV‐C Vision and Encrypted Communication
Authors: Chen Jiaxin , Liu Xinya , Zhu Qingshan , Fang Xiaosheng , Xu Wentao
Published: April 2024

Neuromorphic device that percepts ultraviolet (UV) radiations is essential for supplementing human visual perception. However, UV‐responsive synapses with high sensitivity and wavelength selectivity have not yet been developed so far. Herein, 2D optoelectronic synapses that are able to distinguish between UV‐A, UV‐,B and UV‐C radiations using a single material are demonstrated. The synapses use Ca2Nb3O10 nanosheets with wide bandgap and persistent photoconductivity effect (PPC) as UV‐responsive layer. The devices can detect UV‐C rays at ultralow intensity of 70 nW cm−2, which is the lowest record among 2D UV synaptic devices. Moreover, such a device demonstrates UV image vision, recognition, and memorization functions based on its diverse synaptic behaviors. The atomically thin UV synapses provide implications for enhancing human visual perception capabilities with spectral selectivity, ultrahigh sensitivity, and low‐energy consumption.
Hydrogen‐Induced Disproportionation of Samarium‐Cobalt Intermetallics Enabling Promoted Hydrogen Evolution Reaction Activity and Durability in Alkaline Media
Authors: Chen Ziliang , Mebs Stefan , Mondal Indranil , Yang Hongyuan , Dau Holger , Kang Zhenhui , Haumann Michael , Ghosh Suptish , Cen Wanglai , Driess Matthias , Menezes Prashanth W.
Published: April 2024

Transition‐metal nanoparticles hold great promise as electrocatalysts for alkaline hydrogen evolution reaction (HER), however, addressing the simultaneous challenges of ensuring sufficient active sites, promoting favorable water dissociation, and optimizing binding energy toward hydrogen intermediates remains a formidable task. To overcome these hurdles, a novel gaseous hydrogen engineering strategy is proposed by in situ embedding cobalt nanoparticles within a samarium hydride matrix (Co/SmH2) via hydrogen‐induced disproportionation of SmCo5 particles for efficient alkaline HER. The as‐designed Co/SmH2 delivered an overpotential as low as 252 mV at 100 mA cm−2, surpassing the performance of pristine Co by 100 mV. Notably, this catalyst lasts remarkably long maintaining a durability at ≈500 mA cm−2 for 120 h. A combination of in situ Raman spectroscopy, in situ X‐ray absorption spectroscopy, density functional theory calculation and post‐HER characterizations unambiguously unveiled that the surface SmH2 transforms into samarium (hydr)oxide during electrocatalysis. This transformation not only inhibits the aggregation of the ultrafine cobalt nanoparticles but also significantly enhances the water dissociation and optimizes the binding energy of active cobalt species toward hydrogen intermediate, resulting in concurrent improvement of kinetics, thermodynamics, and stability of the HER process.