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Electrophysiological findings for peripheral nerves of lower limb in women with stress urinary incontinence and non-specific low back pain

Abstract Stress urinary incontinence (SUI) and nonspecific low back pain (NSLBP) are prevalent conditions that significantly affect women’s quality of life. Recent studies have identified a connection between these conditions and dysfunction in the peripheral nerves of the lower limb. This study aims to compile existing knowledge on the electrophysiological findings in the peripheral nerves of the lower limb in women experiencing SUI and NSLBP. This was a prospective observational study involving fifty healthy women and women suffering from SUI and NSLBP. The participants were aged between 25 and 35 and had a body mass index (BMI) ranging from 20 to 24. The primary outcome measures focused on evaluating lower-limb peripheral nerves in women with SUI and NSLBP. There was an increase in both distal and proximal latencies of the tibial nerve, along with a significant decrease in distal and proximal amplitudes and nerve conduction velocity (NCV) in group A when compared to group B (p < 0.001). For the peroneal nerve, group A showed a significantly higher distal latency and a significantly lower NCV (p < 0.001), while no significant differences were observed in proximal latency or the proximal and distal amplitudes between the two groups (p > 0.05). Regarding the sural nerve, group A had significantly higher onset latency and lower amplitude and NCV compared to group B (p < 0.01). These findings reveal statistically significant nerve conduction abnormalities in tibial, peroneal, and sural nerve conduction in women suffering from stress urinary incontinence (SUI) and non-specific low back pain. The tibial nerve displayed extended distal and proximal latencies, diminished amplitudes, and lower nerve conduction velocity (NCV). The peroneal nerve showed prolonged distal latency and a reduced NCV, while the sural nerve had an extended onset latency, decreased amplitude, and slower NCV. The tibial nerve showed the most changes, including extended latencies and lower conduction velocity, implying both axonal and demyelinating involvement. These changes, supported by large effect sizes, high thresholds commonly associated with clinically meaningful decline, and may have functional consequences.

Analysis and simulation of a novel stochastic RNA silencing model

Abstract The current work aims to investigate how random environmental fluctuations affect the dynamics of the RNA Silencing model. To capture this complexity, a novel stochastic RNA Silencing model is proposed by incorporating four distinct white noise terms into key system parameters. Unlike previous deterministic approaches, our model explicitly accounts for stochastic perturbations using Brownian motion processes. A comprehensive analysis of both the deterministic as well as the stochastic one are presented. Employing Lyapunov analysis, the stochastic system yields a unique global positive solution for any initial value, ensuring its biological relevance. Lastly, numerical simulations based on the Milstein’s higher-order-method are conducted for the two models. The findings highlight the significant influence of stochasticity on RNA silencing dynamics, offering new insights into the stability and behavior of gene regulatory processes under random fluctuations.

Correction: A rapid and high-yield method for nucleic acid extraction

Enhanced phenol removal from wastewater via sulfuric acid activated eggshell derived carbon

Abstract This research explores the development of an innovative activated carbon adsorbent (ACES) derived from waste eggshells through sulfuric acid activation to effectively remove phenol from simulated wastewater. Optimization of adsorption parameters was conducted using Design-Expert 13 software and response surface methodology (RSM). Under optimal conditions (initial phenol concentration of 25.015 mg/L, adsorbent dosage of 4.913 g/L, pH of 4.693, and temperature of 25.013 °C), ACES achieved an outstanding phenol removal efficiency of 99.87%. Characterization studies revealed a high BET surface area of 1034.775 m²/g and enhanced porosity, significantly contributing to adsorption performance. Mechanistic insights showed that electrostatic attraction, π–π interactions, and hydrogen bonding drove adsorption. The Langmuir model provided the best fit for phenol adsorption on ACES (R² = 0.9845), indicating monolayer adsorption on uniform sites. Kinetic analysis revealed that the adsorption followed pseudo-second-order kinetics, with a rate constant (k) of 0.0078 g·min⁻¹·mg⁻¹ and a high correlation coefficient (R² = 0.9886), pointing to chemisorption rather than physical adsorption. Thermodynamic analysis further confirmed that the process is spontaneous and exothermic, accompanied by increased randomness at the adsorbent-adsorbate interface. ACES exhibited good reusability, retaining 80% efficiency after four regeneration cycles. The findings of this research highlight a sustainable approach to utilizing waste eggshells for phenol removal, offering potential applications in wastewater treatment.

Disease entity impacts muscle wasting in the ICU with COVID-19 patients losing muscle nearly twice as fast

Abstract Muscle loss in critically ill patients, particularly during prolonged ICU stays, poses significant challenges to recovery and long-term outcomes. ICU-acquired weakness (ICUAW) manifests as severe muscle depletion, correlating with illness severity and hospitalization duration. This study aims to characterize long-term muscle loss trajectories in ICU patients with acute respiratory distress syndrome (ARDS) due to COVID-19 and severe acute pancreatitis (AP) and to explore contributing factors to elevated muscle decay. Retrospective cohort study including 154 ICU patients, 100 individuals suffering from AP and 54 from COVID-19 ARDS, who underwent a minimum of three CT scans during hospitalization, totaling 988 assessments. Sequential segmentation of psoas muscle area (PMA) was performed, and relative muscle loss per day for the entire monitoring period, as well as for the interval between each consecutive scan, was calculated. Bivariate and multivariate linear regression analyses were conducted to identify and evaluate the factors contributing to muscle loss. ICU patients experienced an average PMA decline of 46.0%, with a reduction of 41.8% observed in COVID-19 patients and 48.2% in AP patients. Notably, the long-term daily PMA loss was significantly greater in COVID-19 patients (1.88%) compared to AP patients (0.98%; p < 0.001). Linear regression analysis identified disease entity (p < 0.001), length of hospitalization (p < 0.001), and obesity as significant contributors to daily muscle deterioration. Patients admitted to the ICU for COVID-19 and severe AP can experience extreme muscle decay, reaching up to 48.2%. While decay rates vary considerably, COVID-19 patients experienced nearly twice the daily muscle loss compared to AP patients. Key factors contributing to muscle decay included disease entity, hospitalization duration, and obesity. These findings highlight the distinct impact of the underlying disease on muscle deterioration and emphasize the heightened risk for obese patients and those undergoing extended hospitalization.

Morphology and material composition of raptorial foreleg cuticles in praying mantises Gongylus gongylodes and Sphodromantis lineola

Abstract Praying mantises (Mantodea, Insecta) are capable of capturing larger invertebrates and also smaller vertebrates with their raptorial forelegs. Forelegs may exhibit morphological adaptations based on the type of prey they prefer (e.g., softer or harder invertebrates). Since the functionality of a structure is also influenced by its material composition, it could be tailored to match the prey as well. To pave the way for deeper studies on trophic adaptations in insects, this study investigates the morphology, material composition and mechanical properties of the raptorial forelegs of two species of praying mantises with different prey preferences. Sphodromantis lineola favours harder and larger invertebrates, such as cockroaches (Blattaria), whereas Gongylus gongylodes feeds on softer and smaller invertebrates like flies (Diptera). Both males and females were examined in both species to assess potential sexual dimorphism. The results suggest that the morphology of the raptorial forelegs, the arrangement and tiltability of the spines, and the material mechanical properties of the leg cuticle could potentially be related to the preferred prey type. Autofluorescence documentations reflected the presence of micro gradients in the spines, which are related to their mechanical properties. However, pigmentation in the structures of G. gongylodes seemed to corrupt the autofluorescence, complicating comparisons between the species. Elemental analysis confirmed the presence of traces of transition and alkaline earth metals in the raptorial forelegs, though no significant species-specific differences and no correlations to the mechanical properties were detected.

Study on demetallization of heavy crude oil using different zeolitic materials

Abstract Refiners downstream worldwide are devoting more attention to managing industrial impacts and environmental contamination as an approach of reducing the trace metal content of heavy crude oil and its refined products. The removal of Cr2+, Ni2+, V4+, and Zn2+ (the most significant metal ions) from Egyptian heavy crude oil at the Belayim Desert (BD) oil field has been studied in this work applying the adsorption demetallization technique. Two types of zeolitic materials have been used in the metal removal experiments. Different operating parameters such as contact time, adsorbent concentration and initial metal ions concentration (crude oil quantity) were investigated for their effects on metal removal efficiency. The experimental results of adsorption test showed that the optimal removal conditions using NZ occurred after 6 h contact time, when using 0.5 g of NZ stirred with 50 ml crude oil, these experimental conditions produced maximum V, Ni, and Cr removal efficiencies as 75.6, 73.9, and 81.8% respectively, that cleared at PXRD and EDX analysis to confirm the highly tendency of NZ to remove metal ions from BD crude oil. While the optimal removal conditions using SZ occurred after 12 h contact time, when using 2 g of SZ stirred with 50 ml crude oil, these experimental conditions produced maximum Zn and Cr removal efficiencies as 70.5, 69.17% respectively, but the maximum removal efficiency of V and Ni obtained after 26 h contact time, 2 g SZ with 50 ml crude oil reached to 68.9%. In addition to the results revealed that Natural Zeolite (NZ) is more efficient than Synthetic Zeolite (SZ) in extracting metal ions mainly Ni and V ions from BD crude oil. Also, the selectivity of NZ for heavy metals removal efficiencies are Cr > V > Ni > Zn. Whilst the selectivity of SZ for heavy metals removal efficiencies are Cr > Zn > V > Ni.

3D printed tooth for endodontic training in dental education

Abstract In dental education root canal treatment is primarily trained with natural teeth models or transparent acrylic blocks. A newly constructed, 3D printed practice tooth can be produced cost-efficiently and reproducibly while based on a natural root canal anatomy. Based on a micro-CT scan of an extracted tooth number 36, a dataset was reconstructed (Autodesk Inventor 2019, Autodesk Inc.) and subsequently 3D printed (Form 3B, Formlabs Inc.). In a hands-on course, dental students performed root canal treatments on 3D printed practice teeth. The practice teeth were prepared and filled. A reciprocating system (Reciproc, VDW GmbH) and a rotating preparation system (Pro Taper Next, Dentsply Sirona GmbH) were utilized. Students completed a questionnaire to evaluate practice materials, learning success, and learning process. The students rated suitability as a practice option ( p  < 0.001) and handling ( p  = 0.001) of the 3D printed practice tooth, as well as natural tooth models, significantly better than acrylic blocks.

Automated oil spill detection using deep learning and SAR satellite data for the northern entrance of the Suez Canal

Abstract Oil spills threaten marine ecosystems, demanding swift detection and response. The northern entrance of the Suez Canal, a critical maritime route, is increasingly at risk of frequent oil spill incidents. This study employs the DeepLabv3 + deep learning model to automatically detect oil spills in the study area based on Sentinel-1 Synthetic Aperture Radar imagery provided by the European Space Agency. The model was trained separately on two datasets: the European Maritime Safety Agency CleanSeaNet (EMSA-CSN) dataset, comprising 1100 oil spill incidents, and a localized dataset containing 1500 oil spill incidents that occurred at the Egyptian territorial waters. A comparative analysis between the two models was conducted using 30 oil spill test cases located within the study area. The model trained on Egyptian data outperformed the EMSA-CSN-data- trained model, achieving a loss of 0.0516, an accuracy of 98.14%, a mean Intersection over Union (MIoU) of 0.7872, and a significantly higher ROC area of 0.91, compared to a loss of 0.1152, an accuracy of 96.45%, a MIoU of 0.7161, and a ROC area of 0.76 for the EMSA-CSN model. In addition, the area prediction analysis confirmed the superior performance of the Egyptian-data-trained model, which estimated a total affected area of 421.20 km2, closely aligning with the ground truth of 425.20 km2, whereas the EMSA-CSN-data-trained model underestimated oil spills of around 323.98 km2. These results highlight the benefits of region-specific training in improving segmentation quality and reducing errors. This study emphasizes the potential of AI-driven models for real-time oil spill monitoring, with applications in environmental protection and emergency response.

No associations between environmental exposures and stroke severity in a low pollution area in Sweden

Abstract Mounting evidence supports associations between air pollution and noise exposure and cardiovascular events; however, the relationships at low exposure levels and for stroke outcomes remain uncertain. The aim was to investigate the associations between environmental exposures over 1-year and 10-year periods and both stroke severity and stroke type in a registry-based cohort including people with stroke residing in a low-pollution area of Sweden. Patients with stroke admitted to the Sahlgrenska University Hospital from 2014 to 2019 were included. Stroke severity was assessed with the National Institutes of Health Stroke Scale and stroke types were ischemic and hemorrhagic. Annual residential environmental exposures (road traffic noise (LAeq,24h), inhalable particulate matter (PM10), and nitrogen oxides (NOx)) were assigned from high-resolution dispersion models to participants one year and for ten years prior to stroke, respectively. Of 4066 patients, 1965 (48.3%) were women. The mean (± SD) age was 73.6 (14.0) years. A total of 1563 (28%) had moderate to severe stroke, and 3603 (88.6%) had ischemic stroke. We did not find significant associations between environmental exposures (LAeq,24h, NOx, PM10) and stroke severity nor stroke type. The generally low levels of exposure and low variance of these environmental factors might explain the lack of observed associations.

Extensive electronic investigation of BMBH structure and adsorption locator on graphene with molecular dynamics of human serum albumin interaction

Abstract A comprehensive electronic investigation of Bambuterol Hydrochloride (BMBH) was conducted to explore its structural properties, adsorption behavior on graphene, molecular docking interactions, and molecular dynamics perturbations. FT-IR and XRD characteristics were performed to support the structural identity. Geometry optimization and theoretical calculations were carried out to study the structural and electronic properties of BMBH. The nature of hydrogen and halogen bonding interactions was analyzed using natural bond orbital (NBO) analysis, atoms in molecules (AIM) theory, and Reduced Density Gradient (RDG) analysis. Additionally, electron localization function (ELF) analysis provided deeper insights into the chemical bonding characteristics of BMB. Adsorption locator modelling was involved to allow activated carbon-carriers for sustained and controlled drug release, which helps maintain therapeutic drug levels in the body over time, reducing the frequency of administration. Molecular docking analysis was performed to assess the interaction of BMBH with key biological targets, revealing its potential pharmacological relevance. The inhibitory interaction of BMB with the butyrylcholinesterase enzyme, which is a major cause of dementia and Alzheimer’s disease, has been investigated based on molecular modelling. In addition to that the interaction between BMB and Human Serum Albumin (HSA) was assessed using molecular Docking and Molecular dynamics studies to investigate its transportation and bioavailability. Additionally, molecular dynamics simulations were employed to evaluate the structural perturbations and dynamic behaviour of the BMBH/graphene and BMB/target complexes over time. The study offers a detailed understanding of the electronic and interactional properties of BMB, contributing to its potential applications in nanomaterial-based drug delivery and therapeutic interventions.

Production and biochemical characterization of Pleurotus ostreatus NRC 620 laccase and evaluation of its efficacy in apple juice clarification

Abstract The highest activity of the Pleurotus ostreatus NRC620 laccase enzyme occurred in the broth of mushroom growth after 25 days of incubation at 28 °C and static conditions. The optimum pH and temperature of the enzyme activity were revealed at pH 3.0, and 70 °C, respectively, and retained 68.33 and 59.61% of its activity after incubation at 40 and 50 °C for 2 h, respectively. The enzyme retained 100% activity after 2 h of incubation in citrate–phosphate buffer (pH 7.0). The addition of MgSO4 and CuSO4 with concentrations of 10 mM caused about 21% and 35% increase in enzyme activity, while NaCl, MnCl2, KCl, and CaCl2 inhibited the enzyme. The values of the kinetic parameters (K m and V max ) of the Pleurotus ostreatus NRC 620 laccase were 1.99 mM and 16,217 µmol Min−1 L−1 respectively, using ABTS as a substrate. The apple juice enzyme-treated sample showed a significant reduction in pH, as well as viscosity, after enzyme treatment along with storage time. Apple juice treatment with laccase exhibited slight degradation in total phenolic; however, this observation was not found in antioxidant activity.

VTGAN based proactive VM consolidation in cloud data centers using value and trend approaches

Abstract Reducing energy consumption and optimizing resource usage are essential goals for researchers and cloud providers managing large cloud data centers. Recent advancements have demonstrated the effectiveness of virtual machine consolidation and live migrations as viable solutions. However, many existing strategies are based on immediate workload fluctuations to detect host overload or underload and trigger migration processes. This approach can lead to frequent and unnecessary VM migrations, resulting in energy inefficiency, performance degradation, and service-level agreement (SLA) breaches. Moreover, traditional time series and machine learning models often struggle to accurately predict the dynamic nature of cloud workloads. This paper presents a consolidation strategy based on predicting resource utilization to identify overloaded hosts using novel hybrid value trend generative adversarial network (VTGAN) models. These models not only predict future workloads but also forecast workload trends (i.e., the upward or downward direction of the workload). Trend classification can simplify the decision-making process in resource management approaches. We perform simulations using real PlanetLab workloads on Cloudsim to assess the effectiveness of the proposed VTGAN approaches, based on value and trend, compared to the baseline algorithms. The experimental findings demonstrate that the VTGAN (Up current and predicted trends) approach significantly reduces SLA violations and the number of VM migrations by 79% and 56%, respectively, compared to THR-MMT-PBFD. Additionally, incorporating VTGAN into the VM placement algorithm to disregard hosts predicted to become overloaded further improves performance. After excluding these predicted overloaded servers from the placement process, SLA violations and the number of VM migrations are reduced by 84% and 76%, respectively, compared to THR-MMT-PBFD.

Outcomes of complex decongestive therapy in managing upper limb lymphedema in female breast cancer patients at a palliative care unit of a tertiary care hospital in Bangladesh

Background Lymphedema is a chronic condition that significantly affects both physical function and quality of life of breast cancer patients. Although there is no definitive cure, various treatment options exist to alleviate its symptoms. Among these, Complex Decongestive Therapy (CDT) is widely regarded as a primary approach. This study seeks to evaluate the effectiveness of CDT for breast cancer patients with upper limb lymphedema and aims to assess the benefits of this treatment despite the challenges and constraints in resource-limited settings. Methods This observational study was conducted among 42 female breast cancer patients with unilateral upper limb lymphedema attending the Lymphedema Clinic of the Department of Palliative Medicine at Bangladesh Medical University in Dhaka, Bangladesh. Limb volume, skin condition, and clinical signs and symptoms were assessed at baseline. All patients received the intensive phase of Complex Decongestive Therapy (CDT) for 6 weeks, with follow-up assessments conducted at the 3rd week and the 6th week. Result A significant reduction in the volume of the affected limbs was observed from baseline to the 6th week, as well as from the 3rd week to the 6th week. Although no statistically significant improvement in skin edema was recorded during this period, visible clinical improvement in skin texture was noted. After receiving CDT and proper skin care, 59.5% of patients regained normal skin on the affected limb. Additionally, there was a significant reduction in self-reported symptoms such as tightness, heaviness, and pain in the affected limb from baseline to the 6th week. Conclusion Lymphedema management using all components of Complex Decongestive Therapy (CDT) was found to be effective in reducing limb volume and alleviating the distressing symptoms of patients. Timely referral of lymphedema patients to specialized clinics and initiation of CDT can significantly reduce their ongoing suffering in Bangladesh.

A study on multi-objective optimization for the location selection of smart underground parking facilities in high-density urban areas of megacities: A case study of Jing’an district, Shanghai

The acceleration of global urbanization and the rapid growth of urban populations have intensified the complexity and urgency of parking demand. In megacities with limited land resources, efficiently addressing diverse parking needs has become a critical issue for sustainable urban development. Multi-objective optimization methods are widely applied to tackle such challenges, providing decision-makers with a set of optimal solutions that balance multiple objectives. However, existing studies often lack quantitative analyses of the relationships among these solutions, limiting their applicability in accommodating decision-makers with varying preferences. This study focuses on Jing’an District in Shanghai, a representative region of a Chinese megacity, to address this global issue. Based on real-world data, a multi-objective optimization model is constructed considering convenience, coverage, and cost-efficiency. The model is solved using an improved Non-dominated Sorting Genetic Algorithm II (NSGA-II), which dynamically adjusts crossover and mutation rates. Furthermore, the Pareto solution set is quantitatively analyzed from a cost-benefit perspective by integrating marginal benefit theory. This approach provides robust support for decision-makers seeking an optimal balance between cost and benefit, offering scenario-specific strategies. The findings of this study not only present an innovative, systematic, and flexible solution to the “parking dilemma” in high-density residential areas but also provide practical guidance and insights for other large cities in the planning and implementation of smart underground parking facilities.

PPARγ inhibitors enhance the efficacy of statin therapy for steroid-induced osteonecrosis of the femoral head by directly inhibiting apoptosis and indirectly modulating lipoprotein subfractions

Background Steroid-induced osteonecrosis of the femoral head (SONFH) is a serious bone disease commonly seen in patients on long-term glucocorticoid therapy. Although statins have shown some efficacy in improving lipid metabolism, their efficacy in the treatment of SONFH remains limited. PPARγ inhibitors may enhance the efficacy of statins through several mechanisms. This study aims to investigate how PPARγ inhibitors may enhance the effects of statins in the treatment of SONFH by directly inhibiting apoptosis and indirectly modulating lipoprotein subfractions. Methods We first treated osteoblasts in vitro with high concentrations of hormones to simulate the SONFH environment. We then treated the cells with either the PPARγ inhibitor GW9662, the statin lovastatin, or a combination of both. We assessed cell proliferation and apoptosis using CCK-8, flow cytometry and Western blotting. We then established a SONFH rabbit model using high doses of methylprednisolone and lipopolysaccharide. The rabbits were randomly divided into four groups: control group, lovastatin group, GW9662 group and combination therapy group. We observed hip joint MRI before treatment, after 4 weeks of treatment, and 4 weeks after stopping treatment. We performed hematoxylin-eosin staining of the femoral head and analysed serum lipoprotein subfractions using VAP technology. In addition, we used quantitative polymerase chain reaction (qPCR) to analyse the expression of genes related to lipid metabolism at week 3. Results In vitro experiments showed that both GW9662 and lovastatin effectively inhibited hormone-induced apoptosis. In the animal studies, imaging and pathological results showed that the progression of SONFH was slower in the combination therapy group than in the other groups. VAP analysis showed that the lovastatin group had disturbed lipoprotein subfractions at the fourth week after stopping treatment, while the combination therapy group had more stable lipoprotein subfractions. Conclusion PPARγ inhibitors significantly enhance the efficacy of statins in the treatment of SONFH by directly inhibiting apoptosis and indirectly modulating lipoprotein subfractions. These findings provide new insights into the clinical management of SONFH and suggest that combination therapy may be an effective strategy.

Enhancement of plant growth in lentil (Lens culinaris) under salinity stress by exogenous application or seed priming with salicylic acid and hydrogen peroxide

This study was conducted in order to test the effect of seed pretreatment or exogenous application through the rooting medium of 0.1 mM Salicylic Acid (SA) and 0.1 mM hydrogen peroxide (H2O2) on growth, nutritional behavior and some biochemical parameters (photosynthetic pigments, gas exchange parameters, oxidative stress indicators and antioxidant enzymes activities) of lentil plants (Lens culinaris) under 75 mM salt stress. Our results demonstrated that salt stress noticeably reduced shoot and root DWs by 39.01 and 42.81%, respectively, as compared to controls. This reduction was associated with a significant decrease in all photosynthetic parameters, including Chlorophyll (Chl) and carotenoid (Car), net assimilation of photosynthesis (A), stomatal conductance (gs), transpiration (E) and internal CO2 level (Ci), an accumulation of Na+ and Cl- and a decrease of K+ and Ca2+ concentrations in plant shoots and roots. In addition, relative to control plants, salt stress remarkably increased the malondialdehyde MDA and H2O2 contents especially in roots and increased GPOX and SOD activities, especially in plant shoots. Both methods of SA and H2O2 application recovered the plant growth, enhanced shoot and root DWs (increase of 67.65 and 82.36% in shoots and roots, respectively, as compared to salt-stressed plants) and increased all parameters that were reduced by NaCl treatment. Nevertheless, the most prominent effects of SA and H2O2 on plant growth were obtained with the seed priming method. Thus, SA and H2O2 applications, especially the H2O2 seed priming method, induced the antioxidant system, improved the membrane stability and ameliorated the gas exchange parameters. As compared to salt plant stressed, Na+ and Cl- contents were significantly decreased and K+ and Ca2+ were significantly increased in shoots and roots following SA and H2O2 applications, especially with the H2O2 seed priming method. Similarly, this method was more efficient in alleviating the adverse effects of salt stress on all photosynthetic pigment contents and measured gas exchange parameters. Compared to salt stressed plants, it significantly decreased the H2O2 and MDA contents and further stimulated GPOX and SOD activities. Our results indicated that the seed priming method, particularly with H2O2, could be recommended for obtaining better growth of lentil seedlings under salt-affected soil conditions.

Efficient endogenous protein labelling in Dictyostelium using CRISPR/Cas9 knock-in and split fluorescent proteins

Fluorescent protein tagging is a powerful technique for visualising protein dynamics; however, full-length fluorescent protein knock-in can be inefficient at certain genomic loci, making it challenging to achieve stable and uniform expression. To address this issue, we used CRISPR/Cas9-mediated knock-in strategies with split fluorescent proteins in Dictyostelium discoideum. This approach enabled efficient integration of the short mNeonGreen2 (mNG2) fragment, mNG211, particularly at functionally critical loci such as major histone h2bv3, where full-length tagging was unsuccessful. Our analysis revealed that inserting tandem repeats of mNG211 at the h2bv3 locus progressively impaired cell proliferation, indicating that functional disruption depends on insert size. These findings suggest that using short tags like mNG211 minimises functional interference and facilitates knock-in at sensitive loci. We further optimised the fluorescence intensity by fine-tuning the expression of the long fragment, mNG21–10, and introducing tandem repeats of mNG211. This approach provides a reliable method for precise and stable endogenous protein labelling, facilitating live-cell imaging and functional studies in D. discoideum.

Geography of transnational knowledge flows from China: Distance, Pipelines and Hierarchy?

With the rise of China’s innovation capacity and position in the global innovation system, an increasing number of scholars are paying attention to the knowledge diffusion from developed economies to China. However, there is less research looking into the destinations of transnational knowledge diffusion from China and their influencing factors from the dynamic perspective. This study uses USPTO data for the period 2003–2022 to illustrate the spatial pattern of Chinese transnational knowledge diffusion and estimates the impact of geographical distance, knowledge pipelines, and hierarchy in the global innovation system. We find that the global innovation system has been comparatively stable, but that China has successfully transitioned from the periphery to being a semi-peripheral and then a core country. The knowledge transfer from China occurred firstly to core and semi-peripheral countries, as the reversed knowledge flow, and then to developing countries along the “Belt and Road” initiative with an increasingly important role in “South-South Cooperation”. Regarding its influencing factors, geographical distance is significant across all periods, highlighting that distance remains an indispensable factor in innovation and knowledge flow. Knowledge pipelines and hierarchy in the global innovation system are conditionally influential. Knowledge pipelines were only significantly positive when China was a semi-peripheral country. Compared with the periphery, the knowledge flow from China increasingly tended towards semi-peripheral countries during its catching-up process, but the knowledge could be accepted by the core countries only during the time when China was a semi-peripheral country. Our research unpacks the complexity of pipelines and hierarchy as influencing factors from the dynamic perspective.

Nicotine alters cellular activity and mRNA expression of patterns of Astrocytes

Nicotine exposure during neural development presents a significant public health concern. Nicotine, the primary addictive component of tobacco, influences the central nervous system by interacting with various cell types, including the glial cell termed astrocytes. Astrocytes are cells that are critical for supporting neurons, regulating neurotransmitter balance, and managing neuroinflammation. This current study explored nicotine’s effects on astrocytes, examining cellular activity and gene expression within an acute exposure period. Murine C8D1A astrocytic (garnered as a cell line from postnatal day 8 tissue) cells were treated with nicotine (0–500 ng/mL) in vitro, with assays measuring cell viability and apoptosis at 12, 18, 24, and 48 hours to establish a critical concentration gradient for nicotine. Nicotine exposure increased astrocyte viability at later time points (24 and 48 hours), while apoptosis rose initially but declined over time allowing for the establishment of pharmacologically and clinically relevant nicotine concentrations of 25,50 and 100ng/ml for subsequent experiments. Real-time quantitative PCR revealed that nicotine influenced inflammatory signaling, with pro-inflammatory (A1) markers (IL-6, IFNγ, TNFα) increasing in a dose- and time-dependent manner, while anti-inflammatory (A2) markers (ARG1, IL-10, TGFβ) displayed a more complex pattern after nicotine exposures to astrocytes. These results suggest that nicotine disrupts astrocyte function and inflammatory balance, which may contribute to neurodevelopmental disruptions and heightened neuroinflammatory risks in adults. Further research is needed to investigate the prolonged impact of nicotine on brain health, addiction, and associated neurological conditions.