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Abstract Denosumab is a monoclonal anti-RANKL antibody that inhibits bone resorption, increases bone mass, and reduces fracture risk. Denosumab discontinuation causes an extensive wave of rebound resorption, but the cellular mechanisms remain poorly characterized. We utilized in situ hybridization (ISH) as a direct approach to identify the cells that activate osteoclastogenesis through the RANKL/OPG pathway. ISH was performed across species, skeletal sites, and following recombinant OPG (OPG:Fc) and parathyroid hormone 1–34 (PTH) treatment of mice. OPG:Fc treatment in mice induced an increased expression of RANKL mRNA mainly in trabecular, but not endocortical bone surface cells. Additionally, a decreased expression of OPG mRNA was detected in bone surface cells and osteocytes of both compartments. A similar but more pronounced effect on RANKL and OPG expression was seen one hour after PTH treatment. These findings suggest that bone surface cells and osteocytes conjointly regulate the activation of osteoclastogenesis, and that OPG:Fc treatment induces a local accumulation of osteoclastogenic activation sites, ready to recruit and activate osteoclasts upon treatment discontinuation. Analysis of publicly available single-cell RNA sequencing (scRNAseq) data from murine bone marrow stromal cells revealed that Tnfsf11 + cells expressed high levels of Mmp13, Limch1, and Wif1, confirming their osteoprogenitor status. ISH confirmed co-expression of Mmp13 and Tnfsf11 in bone surface cells of both vehicle- and OPG:Fc-treated mice. Under physiological conditions of human/mouse bone, RANKL is expressed mainly by osteoprogenitors proximate to the osteoclasts, while OPG is expressed mainly by osteocytes and bone-forming osteoblasts.

Mnaza Noreen,1 Muhammad Bilal,1,2 Muhammad Usman Qamar,3,4 Nasir Rasool,1 Abid Mahmood,5 Sobia Umar Din,1 Tawaf Ali Shah,6 Yousef A Bin Jardan,7 Mohammed Bourhia,8 Lahcen Ouahmane9 1Department of Chemistry, Government College University Faisalabad, Faisalabad, 38000, Pakistan; 2School of Chemistry and Chemical Engineering, Shandong University, Jinan, 250100, People’s Republic of China; 3Institute of Microbiology, Faculty of Life Sciences, Government College, University Faisalabad, Faisalabad, 38000, Pakistan; 4Division of Infectious Disease and Department of Medicine, University of Geneva, Geneva, Switzerland; 5Department of Pharmaceutical Chemistry, Government College University Faisalabad, Faisalabad, 38000, Pakistan; 6College of Agriculture Engineering and Food Science, Shandong University of Technology, Zibo, 25500, People’s Republic of China; 7Department of Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia; 8Department of Chemistry and Biochemistry, Faculty of Medicine and Pharmacy, Ibn Zohr University, Laayoune, 70000, Morocco; 9Laboratory of Microbial Biotechnologies, Agrosciences and Environment (Biomage), Labeled Research Unit-CNRSTN° 4, Cadi Ayyad University, Marrakesh, 40000, MoroccoCorrespondence: Nasir Rasool; Mohammed Bourhia, Email nasirrasool@gcuf.edu.pk; m.bourhia@uiz.ac.maIntroduction: New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae (NDM-1-KP) sequence type (ST) 147 poses a significant threat in clinical settings due to its evolution into two distinct directions: hypervirulence and carbapenem resistance. Hypervirulence results from a range of virulence factors, while carbapenem resistance stems from complex biological mechanisms. The NDM-1-KP ST147 clone has emerged as a recent addition to the family of successful clones within the species.Methods: In this study, we successfully synthesized 5-bromo-N-alkylthiophene-2-sulfonamides ( 3a-c) by reacting 5-bromothiophene-2-sulfonamide ( 1) with various alkyl bromides ( 2) using LiH. We also synthesized a series of compounds ( 4a-g) from compound ( 3b) using the Suzuki-Miyaura cross-coupling reaction with fair to good yields (56– 72%). Further, we screened the synthesized molecules against clinically isolated New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. Subsequently, we conducted in-silico tests on compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I.Results: The compound ( 3b) with favourable drug candidate status, MIC of 0.39 μg/mL, and MBC of 0.78 μg/mL. This low molecular weight compound exhibited the highest potency against the resistant bacterial strains. The in-silico tests revealed that the compound 3b against a protein extracted from NDM-KP ST147 with PDB ID: 5N5I demonstrated H-bond and hydrophobic interactions.Conclusion: The 5-bromo-N-alkylthiophene-2-sulfonamides displayed antibacterial efficacy against New Delhi Metallo-β-lactamase producing Klebsiella pneumoniae ST147. After the in-vivo trial, this substance might offer an alternative therapeutic option. Keywords: NDM-1, antibacterial, drug resistance, Klebsiella pneumoniae, ST, sulfonamides

The aggregation of the protein α-synuclein is closely associated with several neurodegenerative disorders and as such the structures of the amyloid fibril aggregates have high scientific and medical significance. However, there are dozens of unique atomic-resolution structures of these aggregates, and such a highly polymorphic nature of the α-synuclein fibrils hampers efforts in disease-relevant in vitro studies on α-synuclein amyloid aggregation. In order to better understand the factors that affect polymorph selection, we studied the structures of α-synuclein fibrils in vitro as a function of pH and buffer using cryo-EM helical reconstruction. We find that in the physiological range of pH 5.8–7.4, a pH-dependent selection between Type 1, 2, and 3 polymorphs occurs. Our results indicate that even in the presence of seeds, the polymorph selection during aggregation is highly dependent on the buffer conditions, attributed to the non-polymorph-specific nature of secondary nucleation. We also uncovered two new polymorphs that occur at pH 7.0 in phosphate-buffered saline. The first is a monofilament Type 1 fibril that highly resembles the structure of the juvenile-onset synucleinopathy polymorph found in patient-derived material. The second is a new Type 5 polymorph that resembles a polymorph that has been recently reported in a study that used diseased tissues to seed aggregation. Taken together, our results highlight the shallow amyloid energy hypersurface that can be altered by subtle changes in the environment, including the pH which is shown to play a major role in polymorph selection and in many cases appears to be the determining factor in seeded aggregation. The results also suggest the possibility of producing disease-relevant structure in vitro.

The rising problem of plastic pollution is becoming one of the major environmental issues for the world. In the ocean, plastics undergo degradation into smaller microplastics (MPs) and nanoplastics (NPs). Wild fish and farmed salmon would likely be exposed to these NPs and MPs both through skin and through skin wounds. Keratocyte cells, located in the skin epithelial layer, are scavenger cells which may remove foreign materials and maintain the salmon’s health. They are therefore first in line to handle and to suffer from MP and NP exposure. While the impacts of MPs have been well studied in many different organisms, much less is known about the effects of NP exposure, particularly at the subcellular level. Here, we have used holotomographic and fluorescence microscopy to show that both skin and corneal salmon keratocyte cells fully internalize 500–1000 nm polystyrene particles, as well as inorganic 500 nm silica particles. The fact that corneal epithelial cells also take up particles is novel. Furthermore, some of these particles likely end up in lysosomal compartments within 2 hours of exposure. Here, we show that both conventional and new modalities of microscopy have a role to play to understand how micro- and nano particles affect epithelial cells.

In the last decade, the detection and attribution science that links climate change to extreme weather and climate events has emerged as a growing field of research with an increasing body of literature. This paper overviews the methods for extreme event attribution (EEA) and discusses the new insights that EEA provides for infrastructure adaptation. We found that EEA can inform stakeholders about current climate risk, support vulnerability-based and hazard-based adaptations, assist in the development of cost-effective adaptation strategies, and enhance justice and equity in the allocation of adaptation resources. As engineering practice shifts from a retrospective approach to a proactive, forward-looking risk management strategy, EEA can be used together with climate projections to enhance the comprehensiveness of decision making, including planning and preparing for unprecedented extreme events. Additionally, attribution assessment can be more useful for adaptation planning when the exposure and vulnerability of communities to past events are analyzed, and future changes in the probability of extreme events are evaluated. Given large uncertainties inherent in event attribution and climate projections, future research should examine the sensitivity of engineering design to climate model uncertainties, and adapt engineering practice, including building codes, to uncertain future conditions. While this study focuses on adaptation planning, EEA can also be a useful tool for informing and enhancing decisions related to climate mitigation.

Shazia Naheed,1 Irum Umar Din,1 Muhammad Usman Qamar,2 Nasir Rasool,1 Matloob Ahmad,1 Muhammad Bilal,1 Aqsa Khalid,3 Gulraiz Ahmad,1 Sami A Al-Hussain,4 Magdi EA Zaki4 1Department of Chemistry, Government College University Faisalabad, Faisalabad, 38000, Pakistan; 2Institute of Microbiology, Faculty of Life Sciences, Government College University, Faisalabad, 38000, Pakistan; 3School of Interdisciplinary Engineering & Science (SINES), National University of Sciences and Technology (NUST), Islamabad, 44000, Pakistan; 4Department of Chemistry, Faculty of Science, Imam Mohammad Ibn Saud Islamic University, Riyad, 11623, Saudi ArabiaCorrespondence: Nasir Rasool; Magdi E A Zaki, Email nasirrasool@gcuf.edu.pk; mezaki@imamu.edu.saIntroduction: Global public health concerns include the emergence and spread of methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum beta-lactamase Escherichia coli (ESBL-E. coli). These pathogens cause infections that are difficult to treat, which can have fatal outcomes and require lengthy hospital stays. As a result, we created butyl 2-bromoisonicotinate and tested its antibacterial effectiveness against the ESBL-E. coli ST 405 and MRSA pathogens. Natural product discovery is complemented by synthetic compound synthesis because of the latter’s potential for superior characteristics, target specificity, scalability, intellectual advantages, and chemical diversity. Because of this, the potential for discovering new medicinal compounds is increased, and the constraints placed on natural sources are overcome. Natural items are tough to obtain since they are hard to isolate and synthesize. Therefore, modern science is actively searching for small molecules as therapeutic agents by applying sustainable techniques that can be commercialized.Methods: Two patients’ blood samples were taken, and the BACTEC/Alert system was used to process them. On blood and MacConkey agar, the positive samples were subcultured and incubated aerobically at 37 °C. Using the VITEK 2 compact system, the isolates were subjected to isolate identification and MIC. MLST of the ESBL-E. coli was performed by PCR. Additionally, Fischer esterification was used to create butyl 2-bromoisonicotinate in excellent yields. A commercially available palladium catalyst was then used to arylate the compound, resulting in medium to good yields of arylated butyl 2-bromoisonicotinates. Using the agar well diffusion assay and the micro-broth dilution method, we assessed the in-vitro activities of the synthesized molecules (3, 5a-h) against clinically isolated ESBL-E. coli ST405, and MRSA. A molecular operating environment was used to carry out in silico validation of the synthesized compounds’ binding to the active site and to evaluate the stability of their molecular interactions with the target E. coli 2Y2T protein.Results: MRSA and ESBL-producing E. coli were identified as the two clinical isolates. While MRSA was also resistant to beta-lactam drugs and least resistant to vancomycin, ESBL-producing E. coli belonged to ST405 and was resistant to cephalosporins and sensitive to carbapenems. Good yields of the desired compounds were produced by our effective and economical synthesis. By using a micro-broth dilution assay, the Molecules (3, 5a, and 5d) were most effective against both resistant strains. The Molecules (3, 5a, 5b, and 5d) also displayed good binding energies.Conclusion: The butyl 2-bromoisonicotinate displayed antibacterial efficacy against ESBL-producing E. coli ST405 and MRSA strains. After the in-vivo trial, this substance might offer an alternative therapeutic option.Keywords: ESBL, MRSA, MLST, Fischer esterification, Suzuki-Miyaura, docking studies

A primary dataset capturing five distinct types of sheep activities in realistic settings was constructed at various resolutions and viewing angles, targeting the expansion of the domain knowledge for non-contact virtual fencing approaches. The present dataset can be used to develop non-invasive approaches for sheep activity detection, which can be proven useful for farming activities including, but not limited to, sheep counting, virtual fencing, behavior detection for health status, and effective sheep breeding. Sheep activity classes include grazing, running, sitting, standing, and walking. The activities of individuals, as well as herds of sheep, were recorded at different resolutions and angles to provide a dataset of diverse characteristics, as summarized in Table 1. Overall, a total of 149,327 frames from 417 videos (the equivalent of 59 minutes of footage) are presented with a balanced set for each activity class, which can be utilized for robust non-invasive detection models based on computer vision techniques. Despite a decent existence of noise within the original data (e.g., segments with no sheep present, multiple sheep in single frames, multiple activities by one or more sheep in single as well as multiple frames, segments with sheep alongside other non-sheep objects), we provide original videos and the original videos’ frames (with videos and frames containing humans omitted for privacy reasons). The present dataset includes diverse sheep activity characteristics and can be useful for robust detection and recognition models, as well as advanced activity detection models as a function of time for the applications.

Abstract Background Acute toxicity (LD50) test provides an indication on the range of doses of a substance that could be toxic to the body systems. This study was aimed at determining the LD50 of different extracts of Bombax costatum stem bark and its effects on the histology of liver and kidneys in rats. Results Phytochemical screening of n-hexane, ethanol and chloroform extracts of B. costatum stem bark has revealed the presence of alkaloids, flavonoid, saponins, steroid, terpenes, anthraquinones and cardiac glycosides. No mortality was recorded after testing all the different extracts on rats. However, mild body weaknesses and pilo erection were observed in the first hour of extracts’ administration. No statistically significant differences was observed in most of the serum levels of kidney function biomarkers, although a significant decrease (p 0.05) were observed in hematological parameters as well as in the liver functions biomarkers in all the 3 extract exposed groups when compared to the control, except significant decrease (p

Advancements in technology have enabled us to learn, adapt and exploit our skills and knowledge in new ways. Appreciating the potential of technology may yet give growth and enrich the process of language education, particularly through a student-centred mobile learning environment. Consequently, a constructivist approach to learning can create tremendous possibilities for both language learners and teachers. By exploiting the affordances of mobile technologies and the Internet, a new platform of learning or Mobile Assisted Language Learning (MALL) can be realised, through which learners truly learn to learn. Yet, while many of today's language institutions and places of learning have begun to understand the potential presented by mobile technology as a tool and resource to content and language development, apprehension may still exist among educational practitioners, learners and senior management. Such apprehension may stem from a lack of understanding in fully appreciating the opportunities and affordances posed by MALL in creating a support structure to learning and teaching excellence. This may be particularly true within developing countries such as those found in the Middle East. Consequently, set in a Saudi university context, the purpose of this study is to investigate learners' and teachers' perceptions towards the proposed implementation of a MALL programme, while exploring whether the promotion of mobile technologies could assist learning and become a viable support structure in teaching English as a second language. Interviews were conducted with three English instructors who teach on the first year Business English programme at the university. The study also analysed 191 student participants who completed a Likert scale questionnaire designed to explore their mobile learning experiences, attitudes and perceptions towards the proposed MALL programme in their educational context. The findings from the student questionnaire and teacher interviews generally highlight positive attitudes and a receptiveness towards mobile learning implementation. This is perhaps due to the possibility of new pedagogical scaffoldings being created through MALL in their learning/teaching environment. The results further highlighted opportunities MALL presented in facilitating communication and affording learners greater motivation to engage in independent learning, thus facilitating learner autonomy and allowing for greater collaboration within a richer learner environment. However, while analysis highlighted a proposed MALL implementation could be a useful tool to support language acquisition for ESP/ESL learners (English for Specific Purposes/English as a Second Language), technical issues, network sustainability and comprehensive training were still significant factors of concern.

Academic misconduct in many educational institutions in the Middle East is an inherent problem. This has been particularly true amongst the university student population. The proliferation of the Internet and the ownership of mobile and electronic devices, have, in part, witnessed rates of cheating, plagiarism and academic misconduct cases steadily increase across higher education contexts. Though the growth of the Internet as an information source and gateway to knowledge has increased substantially in recent years, it has, however, opened up a plethora of varying forms and rates of academic dishonesty. This study was conducted through an online Likert scale questionnaire. Its purpose was to investigate first year male undergraduate students' attitudes, experiences and perceptions towards plagiarism and cheating in a university located in Saudi Arabia. The study aimed at addressing themes in relation to the meaning, forms, source, frequency and reasons of cheating and plagiarism. The study indicates that cheating and plagiarism is common among students, while a need to address student awareness and clarify student expectations towards academic integrity was also identified. The study also proposes several recommendations to alleviate the levels of academic misconduct, be it cheating in exams or plagiarising content, in the Saudi university context.

The King Abdullah Public Education Development Project or the "Tatweer" education reforms were created to improve the quality of teaching and learning in Saudi Arabia. It was a response to develop generations of Saudis who would contribute to the economic well-being of the nation. The Saudi Tatweer education reforms have been important in highlighting questions about power, globalisation and divergence. Therefore, set against a background of neoliberal discourse, I will assess the influences and impact of the drivers within Tatweer--a seemingly market-driven set of education reforms. With reference to Foucauldian thought on power and governance, I will also raise some questions on whether the Tatweer reforms were rigorous and effective enough in improving the level of education within the Saudi context. Finally, I will consider the importance of Ball's (2003) perspective on performativity, and how market forces and international influence have impacted on Saudi education policy reform.

Engineering structures are often subjected to the influences of performance deterioration and multiple hazards during their service lives, and consequently may suffer from damage/failure as a result of external loads. Structural reliability and resilience assessment is a powerful tool for quantifying the structural ability to withstand these environmental or operational attacks. This paper proposes new formulas for structural time-dependent reliability and resilience analyses in the presence of multiple hazards, which are functions of the duration of the reference period of interest. The joint impacts of nonstationarities in multiple hazards due to a changing environment, as well as the deterioration of structural performance, are explicitly incorporated. The correlation between the structural resistances/capacities associated with different hazard types is modeled by employing a copula function. It is observed that, under the context of multiple hazards and aging effects, the time-dependent resilience takes a generalized form of time-dependent reliability. The proposed formulas can be used to guide the adaptive design of structures, where adaptive strategies are identified across a range of possible future service conditions. An example is presented to demonstrate the applicability of the proposed method for structural reliability and resilience analyses.