Effective oral care is important for maintaining a high quality of life. Therefore, plaque control can prevent the development and recurrence of periodontitis. Brushing with a toothbrush and toothpaste is a common way to remove plaque; however, excessive brushing or brushing with abrasive toothpaste can cause wear and tear on the dental crown. Hence, we aimed to quantitatively compare the plaque-removal efficiency and tooth wear of toothbrushes using the bioelectric effect (BE) with those of electric–mechanical toothbrushes. To generate the BE signal, an electronic circuit was developed and embedded in a toothbrush. Further, typodonts were coated with cultured artificial plaques and placed in a brushing simulator. A toothpaste slurry was applied, and the typodonts were eluted with tap water after brushing. The plaques of the typodonts were captured, and the images were quantified. For the tooth wear experiment, polymethyl methacrylate disk resin blocks were brushed twice a day, and the thickness of the samples was measured. Subsequently, statistical differences between the experimental toothbrushes and typical toothbrushes were analyzed. The BE toothbrush had a higher plaque-removal efficiency and could minimize tooth wear. This study suggests that the application of BE may be a new solution for oral care. Full article

Perfluroalkyl substances (PFASs) are persistent man-made chemicals considered to be emerging pollutants, with Perfluorooctanoic acid (PFOA), Perfluorooctanesulfonic acid (PFOS), and Perfluorohexanesulphonic acid (PFHxS) being linked to hepatotoxicity and steatosis. PFOA, PFOS, and PFHxS can undergo placental and lactational transfer, which results in PFOA, PFOS, and PFHxS distribution to the neonatal liver. Moreover, in pregnant dams, exposure to a PFAS mixture, in combination with a high fat diet, increased hepatic steatosis in offspring at postnatal day 21, but the mechanisms have not been elucidated. It was hypothesized that gestational/lactational PFAS exposure would alter the pup liver proteome and biochemical/signaling pathways. Timed-pregnant CD-1 dams were fed a standard chow or 60% kcal high-fat diet. From GD1 until PND20, dams were dosed via oral gavage with vehicle (0.5% Tween 20), individual doses of PFOA, PFOS, PFHxS at 1 mg/kg, or a mixture (1 mg/kg each, totaling 3 mg/kg). Livers were collected from PND21 offspring and SWATH-MS proteomics was performed. IPA analysis revealed PFAS exposure modified disease and biological function pathways involved in liver damage, xenobiotics, and lipid regulation in the PND21 liver. These pathways included lipid and fatty acid transport, storage, oxidation, and synthesis, as well as xenobiotic metabolism and transport, and liver damage and inflammation. This indicates the pup liver proteome is altered via maternal exposure and predisposes the pup to metabolic dysfunctions. Full article

Livelihood loss and lower income because of peatland mismanagement are crucial issues that must be resolved in peatland areas. Although many studies have assessed farmers’ livelihoods and income enhancement, progress in addressing these problems remains inadequate. To address this issue, this study aimed to analyze various existing alternative livelihoods in the peatland community in Ogan Komering Ilir District, South Sumatra Province, Indonesia, and analyze scenarios for creating livelihoods and increasing people’s incomes through changes in peat ecosystem management and peatland restoration programs. This study used a survey method conducted in South Sumatra Province’s OKI District, one of the four priority peat-restoration districts in the province. We used three sampling stages, while descriptive, tabulated, and mathematical methods were used for analysis. We analyzed the feasibility of livelihoods that used benefit-cost analysis. The results showed that Sonor cultivation of paddies and catching fish in Rawang (swamp) were the livelihoods of farmers in peatlands. The community has also been processing peatland commodities into other products, such as Purun woven, and Gula Puan (buffalo milk processing). Several alternative livelihood scenarios that are financially profitable and can be developed include salted and smoked fish, Purun woven handicrafts, paludiculture, and agrosilvofishery, which can provide farmers with short-, medium-, and long-term income opportunities. This study can contribute to policymaking by fully considering the role of peat resources in rural livelihoods. Full article

Industrial boilers cause significant energy wastage that could be mitigated with oxy-fuel combustion versus traditional air combustion. Despite several feasibility studies on oxy-fuel burners, they are widely avoided in industry due to major infrastructural challenges. This study measured the performance and heat transfer characteristics of each component in a 0.5 MW fire tube gas boiler after retrofitting it with an oxy-fuel burner. Comparisons were drawn across three combustion modes—air combustion, oxy-fuel combustion, and oxy-fuel flue gas recirculation (FGR). The Dittus–Boelter equation was employed to predict heat transfer in the fire tube for all combustion modes at full load (100%). Heat transfer in the latent heat section of the economizer was measured and compared with predictions using the Zukauskas equation. With this retrofit, oxy-fuel combustion improved the thermal efficiency by about 3–4%. In oxy-fuel combustion, the flow rate of exhaust gas decreased. When integrated into an existing fire tube boiler, the fire tube’s heat transfer contribution diminished greatly, suggesting the economic viability of a redesigned, reduced fire tube section. Additionally, a new design could address the notable increase in gas radiation from the fire tube in oxy-fuel and FGR, as well as aid in the efficient recovery of condensation heat from exhaust gases. Full article

This study explores an approach to design and prepare a multilayer scaffold mimicking interstratified natural tissue. This multilayer construct, composed of chitosan matrices with graded nanohydroxyapatite concentrations, was achieved through an in situ biomineralization process applied to individual layers. Three distinct precursor concentrations were considered, resulting in 10, 20, and 30 wt% nanohydroxyapatite content in each layer. The resulting chitosan/nanohydroxyapatite (Cs/n-HAp) scaffolds, created via freeze-drying, exhibited nanohydroxyapatite nucleation, homogeneous distribution, improved mechanical properties, and good cytocompatibility. The cytocompatibility analysis revealed that the Cs/n-HAp layers presented cell proliferation similar to the control in pure Cs for the samples with 10% n-HAp, indicating good cytocompatibility at this concentration, while no induction of apoptotic death pathways was demonstrated up to a 20 wt% n-Hap concentration. Successful multilayer assembly of Cs and Cs/n-HAp layers highlighted that the proposed approach represents a promising strategy for mimicking multifaceted tissues, such as osteochondral onse. Full article

Tumor cells in hypoxic conditions control cancer metabolism and angiogenesis by expressing HIF-1α. Tanshinone is a traditional Chinese medicine that has been shown to possess antitumor properties and exerts a therapeutic impact on angiogenesis. However, the precise molecular mechanism responsible for the antitumor activity of 3-Hydroxytanshinone (3-HT), a type of tanshinone, has not been fully understood. Therefore, our study aimed to investigate the mechanism by which 3-HT regulates the expression of HIF-1α. Our findings demonstrate that 3-HT inhibits HIF-1α activity and expression under hypoxic conditions. Additionally, 3-HT inhibits hypoxia-induced angiogenesis by suppressing the expression of VEGF. Moreover, 3-HT was found to directly bind to α-enolase, an enzyme associated with glycolysis, resulting in the suppression of its activity. This inhibition of α-enolase activity by 3-HT leads to the blockade of the glycolytic pathway and a decrease in glycolysis products, ultimately altering HIF1-α expression. Furthermore, 3-HT negatively regulates the expression of HIF-1α by altering the phosphorylation of AMP-activated protein kinase (AMPK). Our study’s findings elucidate the mechanism by which 3-HT regulates HIF-1α through the inhibition of the glycolytic enzyme α-enolase and the phosphorylation of AMPK. These results suggest that 3-HT holds promise as a potential therapeutic agent for hypoxia-related angiogenesis and tumorigenesis. Full article

Background and Objectives: Patella baja is a common complication after operative treatment for patellar fracture. This study aimed to investigate (1) the serial changes in patellar height and (2) the potential predictive factors for patellar height changes after tension band wiring (TBW) for patellar fractures. Materials and Methods: Forty-one patients who underwent TBW for patellar fracture between March 2019 and September 2022 were enrolled. To identify serial changes in patellar height, modified Blackburne–Peel index (mBPI) was assessed at just after surgery, at 3 months, at 6 months, at 1 year and at the final follow-up. Multiple regression analysis was conducted to identify factors correlated with mBPI difference between the contralateral side (considered as preoperative status) and injured side. Results: The postoperative mBPI exhibited a decline over time (mean mBPI immediately post operation/3 months/6 months/1 year/final follow-up: 0.69/0.63/0.63/0.62/0.61) Specifically, mBPI showed a significant reduction immediately post operation to 3 months (p < 0.001), although comparisons at other time points did not reveal significant differences. A lower position of the fracture was associated with a decrease in patellar height after surgery. Conclusions: Patellar height was mainly decreased from immediately post operation to 3 months. A fracture in a lower position of associated with decreased patellar height after the TBW of the transverse patellar fracture. Full article

Spin–orbit coupling in nanoscale optical fields leads to the emergence of a nontrivial spin angular momentum component, transverse to the orbital momentum. In this study, we initially investigate how this spin–orbit coupling effect influences the dynamics in gold monomers. We observe that localized surface plasmon resonance induces self-generated transverse spin, affecting the trajectory of the nanoparticles as a function of the incident polarization. Furthermore, we investigate the spin–orbit coupling in gold dimers. The resonant spin momentum distribution is characterized by the unique formation of vortex and anti-vortex spin angular momentum pairs on opposite surfaces of the nanoparticles, also affecting the particle motion. These findings hold promise for various fields, particularly for the precision control in the development of plasmonic thrusters and the development of metasurfaces and other helicity-controlled system aspects. They offer a method for the development of novel systems and applications in the realm of spin optics. Full article

This pilot study examined the long-term structural changes in the osteochondral unit of 20 patients with knee osteoarthritis (KOA) who underwent high tibial osteotomy (HTO) and received post-treatment with either platelet-rich plasma (PRP) or stromal vascular fraction (SVF). Ten patients were injected with autologous PRP (PRP subgroup), while another ten patients received autologous SVF (SVF subgroup) six weeks after surgery and were monitored for 18 months. Histological samples of bone and cartilage (2 mm in diameter and 2 cm long) were taken from tibial and femoral sites during surgery and 18-month post-HTO, and morphometric analyses were conducted using Mega-Morf12 software. Both post-treatment resulted in an increase in articular cartilage height at both sites (p < 0.001 in the tibia and femur), indicating positive outcomes. Significant improvements in subchondral and trabecular bone architecture were also observed, with SVF injection showing higher reparative capacity in terms of bone volume (p < 0.001 for the tibia and p = 0.004 for the femur), subchondral bone height (p < 0.001 for the tibia and p = 0.014 for the femur), trabecular bone volume (p < 0.001 for the femur), and intertrabecular space (p = 0.009 for the tibia and p = 0.007 for the femur). This pilot study, for the first time, demonstrates that HTO surgery combined with PRP and SVF post-treatments can lead to significant enhancements in knee articular cartilage and bone architecture in KOA patients, with SVF showing higher regenerative potential. These findings may contribute to improving treatment strategies for better clinical outcomes in HTO therapy for patients with KOA. Full article

Electric vehicle charging stations are essential to enable broad reception due to the rise in electric vehicles in the transportation industry because they will lessen range anxiety concerns about distance. The primary objective of this work is to design a microgrid that is effective and affordable for an electric vehicle charging station that combines a photovoltaic, wind, and utility grid energy system (optional) as a principal source of energy. The proposed study employs a four-phase inductor coupled interleaved boost converter which is compact and effective with high power output which results in charging a vehicle within 33 min. A perturb and observe MPPT approach based on DC converters is used along with the digital 2PI controller to increase the effectiveness and performance of distributed energy systems. To make the converter a hassle-free operation, an interleaving technique is applied to the developed converter which results in ripple reduction, which results in an increase in the output current and voltage gain, with high power density and efficiency. For better understanding, real-time data for 2W/3W/4W are acquired and tested for various conditions and the maximum state of charge for the battery is gained within one-third of the usual time. At present, the interleaved converter’s operation is theoretically examined, and the behavior of the converter and the charging conditions of several electric vehicle systems are compared and shown in the simulation analysis. Full article

The nonhydraulic minerals (Fe₃O₄, RO phase, Fe) in slag are important indicators for evaluating the pozzolanic activity and detecting the quality of the slag activation processing technology. Fe₃O₄ is an important characteristic mineral among the nonhydraulic minerals. In order to accurately assess the pozzolanic activity of steel slag powder and to monitor the quality of the activation process of steel slag powder for separate nonhydraulic minerals, it is imperative to precisely determine the nonhydraulic mineral content within the steel slag. Further refinement and enhancement are required for both the HNO₃ dissolution method used in determining Fe₃O₄ content in steel slag, as well as for the EDTA-DEA-TEA (ethylenediamine tetraacetate sodium-diethylamine-triethanolamine) dissolution method employed in determining total nonhydraulic minerals, due to potential deviations caused by challenging impurity separations. The results show that the content of Fe₃O₄ is determined by 10%HNO₃-20%NaOH-chemical analysis method, which solves the problem that the impurities of refractory materials (quartz, corundum, mullite) and amorphous phase affects the content determination in HNO₃ dissolution method. The total amount of nonhydraulic minerals (Fe₃O₄, RO phase, Fe) was determined by the EDTA-NaOH-TEA dissolution method, which solved the problem that the incomplete dissolution of C₂F in the EDTA-DEA-TEA dissolution method affected the content determination. The maximum error between the content determination value and the theoretical calculation value of the two methods is less than 0.50%. The improved Fe₃O₄ and total nonhydraulic mineral quantification methods are feasible and reliable. Full article

In this paper, the solid-state shear milling (S³M) strategy featuring a very strong three-dimensional shear stress field was adopted to prepare the high-performance polyoxymethylene (POM)/molybdenum disulfide (MoS₂) functional nanocomposite. The transmission electron microscope and Raman measurement results confirmed that the bulk MoS₂ particle was successfully exfoliated into few-layer MoS₂ nanoplatelets by the above simple S³M physical method. The polarized optical microscope (PLM) observation indicated the pan-milled nanoscale MoS₂ particles presented a better dispersion performance in the POM matrix. The results of the tribological test indicated that the incorporation of MoS₂ could substantially improve the wear resistance performance of POM. Moreover, the pan-milled exfoliated MoS₂ nanosheets could further substantially decrease the friction coefficient of POM. Scanning electron microscope observations on the worn scar revealed the tribological mechanism of the POM/MoS₂ nanocomposite prepared by solid-state shear milling. The tensile test results showed that the pan-milled POM/MoS₂ nanocomposite has much higher elongation at break than the conventionally melt-compounded material. The solid-state shear milling strategy shows a promising prospect in the preparation of functional nanocomposite with excellent comprehensive performance at a large scale. Full article

The residential sector of existing buildings has great potential in energy savings and the improvement of indoor conditions. The modernization of buildings is of particular concern to the policies of the European Union, local governments, and building users. The aim of this paper is to present an analysis of indoor parameters and energy consumption for heating for an apartment located in a pre-war tenement building before and after thermomodernization. The analysis was conducted for winter conditions and was based on measurements and simulations. Originally, the building had not undergone any thermomodernization actions since its reconstruction after WWII. Interior, exterior, and surface temperatures were recorded to describe the thermal conditions of the apartment, while gas meter readings were used to estimate energy consumption for heating purposes. WUFI Plus software (v.3.2.0.1) was used to estimate energy consumption and perform energy simulations for the apartment over an extended period of time. The best thermomodernization effect resulted from the replacement of windows and the inefficient heating system, avoiding surface condensation and reducing final energy consumption by more than 50%. The extended options resulted in energy savings higher than 70%. The presented analysis shows the importance of retrofit measures and proves that even a small improvement can bring significant benefits. Full article

Ancient glass artifacts were susceptible to weathering from the environment, causing changes in their chemical composition, which pose significant obstacles to the identification of glass products. Analyzing the chemical composition of ancient glass has been beneficial for evaluating their weathering status and proposing measures to reduce glass weathering. The objective of this study was to explore the optimal machine learning algorithm for glass type classification based on chemical composition. A set of glass artifact data including color, emblazonry, weathering, and chemical composition was employed and various methods including logistic regression and machine learning techniques were used. The results indicated that a significant correlation (p < 0.05) could only observed between surface weathering and the glass types (high-potassium and lead–barium). Based on the random forest and logistic regression models, the primary chemical components that signify glass types and weathering status were determined using PbO, K₂O, BaO, SiO₂, Al₂O₃, and P₂O₅. The random forest model presented a superior ability to identify glass types and weathering status, with a global accuracy of 96.3%. This study demonstrates the great potential of machine learning for glass chemical component estimation and glass type and weathering status identification, providing technical guidance for the appraisal of ancient glass artifacts. Full article

Malignant melanoma represents a form of skin cancer characterized by a bleak prognosis and heightened resistance to traditional therapies. Quercetin has demonstrated notable anti-carcinogenic, anti-inflammatory, anti-oxidant, and pharmacological effects across various cancer types. However, the intricate relationship between quercetin’s anti-cancer properties and ganglioside expression in melanoma remains incompletely understood. In this study, quercetin manifests specific anti-proliferative, anti-migratory, and cell-cycle arrest effects, inducing mitochondrial dysfunction and apoptosis in two melanoma cancer cell lines. This positions quercetin as a promising candidate for treating malignant melanoma. Moreover, our investigation indicates that quercetin significantly reduces the expression levels of ganglioside GD3 and its synthetic enzyme. Notably, this reduction is achieved through the inhibition of the FAK/paxillin/Akt signaling pathway, which plays a crucial role in cancer development. Taken together, our findings suggest that quercetin may be a potent anti-cancer drug candidate for the treatment of malignant melanoma. Full article

Integrating eye gaze data with chest X-ray images in deep learning (DL) has led to contradictory conclusions in the literature. Some authors assert that eye gaze data can enhance prediction accuracy, while others consider eye tracking irrelevant for predictive tasks. We argue that this disagreement lies in how researchers process eye-tracking data as most remain agnostic to the human component and apply the data directly to DL models without proper preprocessing. We present EyeXNet, a multimodal DL architecture that combines images and radiologists’ fixation masks to predict abnormality locations in chest X-rays. We focus on fixation maps during reporting moments as radiologists are more likely to focus on regions with abnormalities and provide more targeted regions to the predictive models. Our analysis compares radiologist fixations in both silent and reporting moments, revealing that more targeted and focused fixations occur during reporting. Our results show that integrating the fixation masks in a multimodal DL architecture outperformed the baseline model in five out of eight experiments regarding average Recall and six out of eight regarding average Precision. Incorporating fixation masks representing radiologists’ classification patterns in a multimodal DL architecture benefits lesion detection in chest X-ray (CXR) images, particularly when there is a strong correlation between fixation masks and generated proposal regions. This highlights the potential of leveraging fixation masks to enhance multimodal DL architectures for CXR image analysis. This work represents a first step towards human-centered DL, moving away from traditional data-driven and human-agnostic approaches. Full article

Background: Lockdowns and other health protective measures, such as social distancing, imposed during the COVID-19 pandemic nurtured unprecedented levels of stress and social isolation around the world. This scenario triggered an increase in suicide thoughts and self-harm behaviours among children and young people. However, the longer-term impact of the pandemic on children’s and adolescents’ mental health, especially with regard to self-harm, is still to be fully discovered. Methods: We carried out a retrospective study where we collected data related to suicide ideation and self-harm behaviours in all patients aged under 18 that required on-call psychiatric services at the General Hospital Accident and Emergency (A&E) department in Salamanca, Spain, during 2019 (pre-pandemic) and in both 2021 and 2022 to capture possible variation at different time points during the post-pandemic period. Results: A total of 316 patients aged under 18 were seen by on-call psychiatric services at the A&E department during the three time periods: 78 in 2019, 98 in 2021 and 140 in 2022. The mean age was 15.12 (SD 2.25) and females represented more than twice the number of males each year. More than half of all patients assessed during 2022 disclosed suicide thoughts, whilst in 2019, it was near 25%. This increase in suicide ideation rates was more marked among females (X² = 15.127; p = 0.001), those aged over 15 (X² = 16.437; p < 0.001) and/or those with a previous history of mental health problems (X² = 17.823; p < 0.001). We identified an increase in the proportion of males with suicide ideas, especially between 2021 and 2022 (X² = 8.396; p = 0.015). Conclusions: Our study suggests that children’s and adolescents’ demand for urgent mental healthcare and their clinical presentations in A&E departments with suicide thoughts and/or self-injuries do not seem to be declining after the pandemic but increasing over time. More research is warranted to understand possible factors involved in this sustained upward trend. Full article

Acetylsalicylic acid or aspirin is the most commonly used drug in the world and is taken daily by millions of people. There is increasing evidence that chronic administration of low-dose aspirin of about 75–100 mg/day can cause iron deficiency anaemia (IDA) in the absence of major gastric bleeding; this is found in a large number of about 20% otherwise healthy elderly (>65 years) individuals. The mechanisms of the cause of IDA in this category of individuals are still largely unknown. Evidence is presented suggesting that a likely cause of IDA in this category of aspirin users is the chelation activity and increased excretion of iron caused by aspirin chelating metabolites (ACMs). It is estimated that 90% of oral aspirin is metabolized into about 70% of the ACMs salicyluric acid, salicylic acid, 2,5-dihydroxybenzoic acid, and 2,3-dihydroxybenzoic acid. All ACMs have a high affinity for binding iron and ability to mobilize iron from different iron pools, causing an overall net increase in iron excretion and altering iron balance. Interestingly, 2,3-dihydroxybenzoic acid has been previously tested in iron-loaded thalassaemia patients, leading to substantial increases in iron excretion. The daily administration of low-dose aspirin for long-term periods is likely to enhance the overall iron excretion in small increments each time due to the combined iron mobilization effect of the ACM. In particular, IDA is likely to occur mainly in populations such as elderly vegetarian adults with meals low in iron content. Furthermore, IDA may be exacerbated by the combinations of ACM with other dietary components, which can prevent iron absorption and enhance iron excretion. Overall, aspirin is acting as a chelating pro-drug similar to dexrazoxane, and the ACM as combination chelation therapy. Iron balance, pharmacological, and other studies on the interaction of iron and aspirin, as well as ACM, are likely to shed more light on the mechanism of IDA. Similar mechanisms of iron chelation through ACM may also be implicated in patient improvements observed in cancer, neurodegenerative, and other disease categories when treated long-term with daily aspirin. In particular, the role of aspirin and ACM in iron metabolism and free radical pathology includes ferroptosis, and may identify other missing links in the therapeutic effects of aspirin in many more diseases. It is suggested that aspirin is the first non-chelating drug described to cause IDA through its ACM metabolites. The therapeutic, pharmacological, toxicological and other implications of aspirin are incomplete without taking into consideration the iron binding and other effects of the ACM. Full article

With climate change and urbanization expansion, wetlands, which are some of the largest carbon stocks in the world, are facing threats such as shrinking areas and declining carbon sequestration capacities. Wetland carbon stocks are at risk of being transformed into carbon sources, especially those of wetlands with strong land use–natural resource conservation conflict. Moreover, there is a lack of well-established indicators for evaluating the health of wetland carbon stocks. To address this issue, we proposed a novel framework for the safety assessment of wetland carbon stocks using the Super Slack-Based Measure (Super-SBM), and we then conducted an empirical study on the Quanzhou Bay Estuary Wetland (QBEW). This framework integrates the unexpected output indicator (i.e., carbon emissions), the expected output indicators, including the GDP per capita and carbon stock estimates calculated via machine learning (ML)-based remote sensing inversion, and the input indicators, such as environmental governance investigations, climate conditions, socio-economic activities, and resource utilization. The results show that the annual average safety assessment for carbon pools in the QBEW was a meager 0.29 in 2015, signaling a very poor state, likely due to inadequate inputs or excessive unexpected outputs. However, there has been a substantial improvement since then, as evidenced by the fact that all the safety assessments have exceeded the threshold of 1 from 2018 onwards, reflecting a transition to a “weakly effective” status within a safe and acceptable range. Moreover, our investigation employing the Super-SBM model to calculate the “slack variables” yielded valuable insights into optimization strategies. This research advances the field by establishing a safety measurement framework for wetland carbon pools that leverages efficiency assessment methods, thereby offering a quantitative safeguard mechanism that supports the achievement of the “3060” dual-carbon target. Full article

Recombinant adeno-associated virus (rAAV) has emerged as a prominent vector for in vivo gene therapy, owing to its distinct advantages. Accurate determination of the rAAV genome titer is crucial for ensuring the safe and effective administration of clinical doses. The evolution of the rAAV genome titer assay from quantitative PCR (qPCR) to digital PCR (dPCR) has enhanced accuracy and precision, yet practical challenges persist. This study systematically investigated the impact of various operational factors on genome titration in a single-factor manner, aiming to address potential sources of variability in the quantitative determination process. Our findings revealed that a pretreatment procedure without genome extraction exhibits superior precision compared with titration with genome extraction. Additionally, notable variations in titration results across different brands of dPCR instruments were documented, with relative standard deviation (RSD) reaching 23.47% for AAV5 and 11.57% for AAV8. Notably, optimal operations about DNase I digestion were identified; we thought treatment time exceeding 30 min was necessary, and there was no need for thermal inactivation after digestion. And we highlighted that thermal capsid disruption before serial dilution substantially affected AAV genome titers, causing a greater than ten-fold decrease. Conversely, this study found that additive components of dilution buffer are not significant contributors to titration variations. Furthermore, we found that repeated freeze–thaw cycles significantly compromised AAV genome titers. In conclusion, a comprehensive dPCR titration protocol, incorporating insights from these impact factors, was proposed and successfully tested across multiple serotypes of AAV. The results demonstrate acceptable variations, with the RSD consistently below 5.00% for all tested AAV samples. This study provides valuable insights to reduce variability and improve the reproducibility of AAV genome titration using dPCR. Full article

The escalation of mining activities in the karst regions of Guizhou Province has heightened the occurrence of water-inrush incidents in deep coal mines. This study focused on water-inrush phenomena within the Xinhua mining area of Jinsha County, Guizhou Province, aiming to investigate the sources of these incidents. The findings indicated that the overlying limestone of the Changxing Formation in the coal seam served as a vulnerable aquifer under certain conditions, leading to water inrushes. The analysis of the spatiotemporal distribution patterns of water-inrush incidents at the working face indicated that previous mining operations damaged the shallow Changxing Formation limestone, resulting in the accumulation of goaf water and the formation of numerous mining-induced fractures. These fractures served as rapid conduits for water inrushes from both atmospheric precipitation and underground sources at the deep working face. The examination of surface water and mine water quality demonstrated that both exhibited similar characteristics, predominantly featuring bicarbonate, sulfate, and sodium compositions. Investigation into the relationship between mine water inflow and atmospheric precipitation established that atmospheric precipitation influenced the mine water supply cycle, with a replenishment period of ~10 months during the operational phase of the Jinyuan Coal Mine and about one month post-closure. The fractures induced by mining activities within the Changxing Formation limestone facilitated water flow, with atmospheric precipitation serving as the primary water source for the mine. This study offered a valuable scientific foundation for addressing water-related damage resulting from atmospheric precipitation in mines susceptible to water inrushes under analogous hydrogeological conditions. Full article