To address the safety problems posed by the transportation of boar semen using LN, this study was conducted on the short-term storage of frozen boar semen in dry ice (−79 °C). Boar semen frozen in LN was transferred to dry ice, kept for 1 day, 3 days, 5 days, 7 days, or 8 days, and then moved back to LN. The quality of frozen semen stored in LN or dry ice was determined to evaluate the feasibility of short-distance transportation with dry ice. The results showed that 60 °C for 8 s was the best condition for thawing frozen semen stored in dry ice. No significant differences in spermatozoa motility, plasma membrane integrity, or acrosome integrity were observed in semen after short-term storage in dry ice compared to LN (p > 0.05). There were no significant changes in antioxidant properties between storage groups either (p > 0.05). In conclusion, dry ice could be used as a cold source for the short-term transportation of frozen boar semen for at least 7 days, without affecting sperm motility, morphological integrity, or antioxidant indices. Full article

The paper proposes a method for the calibration of spacing in dual-grating based on Multiple Improved Complete Ensemble Empirical Mode Decomposition with Adaptive Noise (ICEEMDAN) combined with Hilbert Transform (HT), referred to as Multiple ICEEMDAN-HT. This method addresses the potential impact of nonlinear factors on phase extraction accuracy, consequently on ranging precision in the homodyne interference of the dual-grating. Building upon the ICEEMDAN algorithm, the signal undergoes iterative decomposition and reconstruction using the sample entropy criterion. The intrinsic mode functions (IMFs) obtained from multiple iterations are then reconstructed to obtain the complete signal. Through a simulation and comparison with other signal decomposition methods, the repeatability and completeness of signal reconstruction by Multiple ICEEMDAN are verified. Finally, an actual dual-grating ranging system is utilized to calibrate the spacing of the planar grating. Experimental results demonstrate that the calibration relative error of the Multiple ICEEMDAN-HT phase unwrapping method can be reduced to as low as 0.07%, effectively enhancing the signal robustness and spacing calibration precision. Full article

Purpose: The purpose of this study was to analyze the prevalence of and complications resulting from temporal bone fractures in adult and pediatric patients evaluated for cranio-facial trauma in an emergency setting. Methods: A retrospective blinded analysis of CT scans of a series of 294 consecutive adult and pediatric patients with cranio-facial trauma investigated in the emergency setting was conducted. Findings were compared between the two populations. Preliminary reports made by on-call residents were compared with the retrospective analysis, which was performed in consensus by two experienced readers and served as reference standard. Results: CT revealed 126 fractures in 116/294 (39.5%) patients, although fractures were clinically suspected only in 70/294 (23.8%); p < 0.05. Fractures were longitudinal, transverse and mixed in 69.5%, 10.3% and 19.8% of cases, respectively. Most fractures were otic-sparing fractures (95.2%). Involvement of the external auditory canal, ossicular chain and the osseous structures surrounding the facial nerve was present in 72.2%, 8.7% and 6.3% of cases, respectively. Temporal bone fractures extended into the venous sinuses/jugular foramen and carotid canal in 18.3% and 17.5% of cases, respectively. Vascular injuries (carotid dissection and venous thrombosis) were more common in children than in adults (13.6% versus 5.3%); however, the observed difference did not reach statistical significance. 79.5% of patients with temporal bone fractures had both brain injuries and fractures of the facial bones and cranial vault. Brain injuries were more common in adults (90.4%) than in children (63.6%), p = 0.001. Although on-call residents reliably detected temporal bone fractures (sensitivity = 92.8%), they often missed trauma-associated ossicular dislocation (sensitivity = 27.3%). Conclusions: Temporal bone fractures and related complications are common in patients with cranio-facial trauma and need to be thoroughly looked for; the pattern of associated injuries is slightly different in children and in adults. Full article

Background: Technological advancements, particularly in Artificial Intelligence (AI), are revolutionizing operations management, especially in the domain of supply chain management. This paper delves into the application of AI in demand planning processes within the supply chain context. Drawing upon a comprehensive review of the existing literature, the main objective of this study is to analyze how AI is being applied and adopted in the demand planning process, identifying the resources needed to build the capacity of AI in the demand process, as well as the mechanisms and practices contributing to AI capability’s advancement and formation. Methodology: The approach was qualitative, and case studies of three different companies were conducted. Results: This study identified crucial resources necessary for fostering AI capabilities in demand planning. Our study extends the literature on AI capability in several ways. First, we identify the resources that are important in the formation of the capacity to implement AI in the context of demand planning. Conclusions: This study’s practical contributions underscore the multifaceted nature of AI implementation for demand planning, emphasizing the importance of resource allocation, human capital development, collaborative relationships, organizational alignment, and relational capital and AI. Full article

Houttuynia cordata Thunb is rich in active substances and has excellent antioxidant and anti-inflammatory activity. Scanning electron microscopy and gel permeation chromatography were used to analyze the molecular characteristics of the fermentation broth of Houttuynia cordata Thunb obtained through fermentation with Clavispora lusitaniae (HCT-f). The molecular weight of HCT-f was 2.64265 × 10⁵ Da, and the polydispersity coefficient was 183.10, which were higher than that of unfermented broth of Houttuynia cordata Thunb (HCT). By investigating the active substance content and in vitro antioxidant activity of HCT-f and HCT, the results indicated that HCT-f had a higher active substance content and exhibited a superior scavenging effect on 2,2-diphenyl-1-picrylhydrazyl radicals and hydroxyl radicals, with IC50 values of 11.85% and 9.01%, respectively. Our results showed that HCT-f could effectively alleviate the increase in the secretion of inflammatory factors and apoptotic factors caused by lipopolysaccharide (LPS) stimulation, and had a certain effect on repairing skin barrier damage. HCT-f could exert an anti-inflammatory effect by down-regulating signaling in the MAPK/NF-κB pathway. The results of erythrocyte hemolysis and chicken embryo experiments showed that HCT-f had a high safety profile. Therefore, this study provides a theoretical basis for the application of HCT-f as an effective ingredient in food and cosmetics. Full article

The impact of climate change has become increasingly severe in forests, where droughts and strong winds on the one hand and extreme rainfall events on the other hand can damage forest ecosystems. To mitigate the effects of drought and enhance soil water retention capacity, three types of soil conditioners (SCs), labeled SC_R, SC_CG, and SC_ZZC, were developed as part of the European project ONEforest. All the conditioners are based on Xanthan gum and have different types and amounts of fillers with diverse cellulose fiber lengths. These can offer the potential to optimize the SC characteristics, e.g., water absorption, water retention, and mechanical stability. This paper focuses on the influence of fillers in the SCs on the geotechnical properties of forest soils from Ljubelj in the Alpine part of Slovenia (S1), Catalonia, northeastern Spain (S2), and Heldburg, Germany (S3). The results show an increase of 53% to 100% in the water absorption of treated soil. A less favorable impact of the SCs was found on the drained shear strength and the compressibility. The drained shear strength of untreated forest soils in a saturated state was S1 c′ = 4.4 kPa, φ′ = 33.5°; S2 c′ = 1.4 kPa, φ′ = 30.0°; and S3 c′ = 12 kPa, φ′ = 28.0°. The addition of SCs results in a reduction in the drained shear strength of saturated mixtures. The reduction depends on the dosage of added SC—whether it is a low (L) or a high (H) dosage. For instance, when the soil S1 was treated with a low dosage of the soil conditioner SC_R, it demonstrated a cohesion (c′) of 11 kPa and a friction angle (φ′) of 27.0°. However, increasing the dosage of the SC_R led to a decrease in both the cohesion and the friction angle for the same soil (c′ = 7.7 kPa, φ′ = 25.0°). Additionally, the type of soil conditioner also impacts the drained shear strength. Among the mixtures with a high dosage of the SC_R, SC_CG, or SC_ZZC, those containing the SC_CG with the longest fibers stand out, demonstrating the highest friction angle. Therefore, longer fibers can be a promising component of the SCs to reduce the negative influence of XG on the mechanical properties of treated soils. Full article

Viruses are a real threat to every organism at any stage of life leading to extensive infections and casualties. N-heterocycles can affect the viral life cycle at many points, including viral entrance into host cells, viral genome replication, and the production of novel viral species. Certain N-heterocycles can also stimulate the host’s immune system, producing antiviral cytokines and chemokines that can stop the reproduction of viruses. This review focused on recent five- or six-membered synthetic N-heterocyclic molecules showing antiviral activity through SAR analyses. The review will assist in identifying robust scaffolds that might be utilized to create effective antiviral drugs with either no or few side effects. Full article

Previously, a class of regular and asymptotically flat gravitating scalar solitons (scalarons) has been constructed in the Einstein–Klein–Gordon (EKG) theory by adopting a phantom field with Higgs-like potential where the kinetic term has the wrong sign and the scalaron possesses the negative Arnowitt–Deser–Misner (ADM) mass as a consequence. In this paper, we demonstrate that the use of the phantom field can be avoided by inverting the Higgs-like potential in the EKG system when the kinetic term has a proper sign, such that the corresponding gravitating scalaron can possess the positive ADM mass. We systematically study the basic properties of the gravitating scalaron, such as the ADM mass, the energy conditions, the geodesics of test particles, etc. Moreover, we find that it can be smoothly connected to the counterpart hairy black hole solutions from our recent work in the small horizon limit. Full article

Sea buckthorn has garnered significant attention owing to its nutritional richness; however, it has a limited shelf life. In this study, the drying process of sea buckthorn was categorized into the first-, second-, and third-drying stages. Regression models were employed to examine the effects of the drying temperature, relative humidity of the medium, and prolonged high humidity retention on various parameters during the first- and second-drying stages. Comparative analysis revealed that the optimal drying conditions for the first-drying stage of sea buckthorn were a drying temperature of 80 °C, relative humidity of 28%, and high humidity retention time of 84 min. In the second-drying phase, the optimal conditions were a drying temperature of 78 °C, a relative humidity of 17%, and a high humidity retention time of 84 min. One-way optimization revealed that the optimal drying temperature for the third-drying stage was 70 °C. The implementation of temperature- and humidity-controlled infrared hot-air drying (TH-IRHAD) techniques considerably improved the outcomes. Specifically, the drying time, energy consumption, and degree of browning decreased by 34.43%, 36.29%, and 21.43%, respectively, whereas the brightness, rehydration ratio, total flavonoid content, and total phenol content increased by 8.94%, 16.99%, 20.57%, and 28.32%, respectively. Staged TH-IRHAD substantially reduced the drying duration, increased the efficiency, and enhanced the drying quality. Full article

Though often under-represented in the official and national narratives and in Canadian military historiography more broadly, the intimate and personal lived experiences of Canadian prisoners of war (POW) during the Second World War can be found in archives, photography collections, and collections of war art. In an attempt to see past the mythologised versions of POWs that appear in Hollywood films, best-selling monographs, and other forms of popular culture, it is through bits of ephemera—including wartime log books and the drawings carefully kept and sent home to loved ones along with handwritten letters—that the stories of non-combatant men and women who spent their war as POWs, can be told. Together, Canadian POWs created and curated community and fostered unconventional family ties, sometimes called “emotional communities”, through the collection and accumulation of drawings, illustrations, paintings, and other examples of war art on the pages of their wartime log books while living behind barbed wire. This article uncovers some of these stories, buried in the thousands of boxes in the George Metcalf Archival Collection—the textual archives—at the Canadian War Museum (CWM) in Ottawa, Ontario, Canada. Full article

During the Holocaust, poets went to extraordinary lengths to write their poems and transmit them. Poems that were written during those years were often buried in the ground, stitched into clothing, smuggled out of prisons, or graffitied onto walls. These object documents carried more than facts about these events; they carried the feeling of living through these events. This research explores the last poems of four Holocaust poets, Władysław Szlengel, Selma Meerbaum-Eisinger, Hannah Szenes, and Abramek Koplowicz, investigating not only the poems but their object-ness and their stories of transference. These poems, like urgent postcards, deliver messages to a family, to a community, to the world. They ask―what does it mean to write a poem as a last will and testament? Full article

Background: HIV case finding is an essential component for ending AIDS, but there is limited evidence on the effectiveness of such a strategy in the pediatric population. We sought to determine HIV positivity rates among children according to entry points in Cameroon. Methods: A facility-based survey was conducted from January 2015 to December 2019 among mother–child couples at various entry points of health facilities in six regions of Cameroon. A questionnaire was administered to parents/guardians. Children were tested by polymerase chain reaction (PCR). Positivity rates were compared between entry points. Associations were quantified using the unadjusted positivity ratio (PR) for univariate analyses and the adjusted positivity ratio (aPR) for multiple Poisson regression analyses with 95% confidence intervals (CIs). p-values < 0.05 were considered significant. Results: Overall, 24,097 children were enrolled. Among them, 75.91% were tested through the HIV prevention of mother-to-child transmission (PMTCT) program, followed by outpatient (13.27%) and immunization (6.27%) services. In total, PMTCT, immunization, and outpatient services accounted for 95.39% of children. The overall positivity was 5.71%, with significant differences (p < 0.001) between entry points. Univariate analysis showed that inpatient service (PR = 1.45; 95% CI: [1.08, 1.94]; p = 0.014), infant welfare (PR = 0.43; 95% CI: [0.28, 0.66]; p < 0.001), immunization (PR = 0.56; 95% CI: [0.45, 0.70]; p < 0.001), and PMTCT (PR = 0.41; 95% CI: [0.37, 0.46]; p < 0.001) were associated with HIV transmission. After adjusting for other covariates, only PMTCT was associated with transmission (aPR = 0.66; 95% CI: [0.51, 0.86]; p = 0.002). Conclusions: While PMTCT accounts for most tested children, high HIV positivity rates were found among children presenting at inpatient, nutrition, and outpatient services and HIV care units. Thus, systematic HIV testing should be proposed for all sick children presenting at the hospital who have escaped the PMTCT cascade. Full article

Attending university is a time of considerable change, and there are rising concerns about the mental health and well-being of university students, leading to calls for a whole-university approach to student support. Resilience interventions offer an opportunity to improve mental health and well-being, whilst also developing a much sought-after graduate attribute. We conducted a systematic review of interventions designed to increase university students’ levels of resilience and examined the impact of these on students’ mental health and well-being. Five databases identified 1377 unique records, 47 of which were eligible for inclusion. Eligible studies were drawn from a range of countries and adopted different designs, with the most common being the randomised controlled trial (RCT). Interventions were classified into mindfulness, skills-based, psychoeducation, and coaching, with delivery both online and face-to-face. The most common outcomes were depression, anxiety, stress, and well-being. The ‘Quality Assessment Tool for Before-After (Pre-/Post) Studies’ was used to assess the risk of bias with most studies rated as fair. Overall, the studies indicated that there is little evidence of a positive effect on depression, but stress and anxiety may be reduced following interventions. Well-being data were inconclusive. Interestingly, most interventions did not impact measures of resilience or mindfulness, despite the training targeting these constructs. The available research is currently limited and there is a need for more high-quality designs providing descriptions of interventions. Full article

In recent decades, there has been a growing interest in the study of emotions, demonstrating their importance in understanding diverse human and social dynamics. However, as this field of study develops and diversifies, a structural yet simple question remains virtually undiscussed: Is it really possible to say that societies possess emotions or a certain sensitivity akin to individual experiences? This exploratory study, based on documentary analysis, endeavours to identify and examine emotional patterns across six different periods, spanning from the Middle Ages to the present day. Among the eight secondary findings, three fundamental conclusions stand out: (1) throughout history, societies have experienced different emotional atmospheres, sometimes simultaneously; (2) although societies generally propose an emotional model to follow, the reality does not always conform to it; and (3) it is mainly through the culture that society creates a certain emotional harmony, allowing the social body to remain cohesive and develop, thus postponing or preventing its disintegration. This study aims to offer a modest contribution to the complex and under-explored discussion on the correlation between specific emotional climates and particular social contexts. Full article

Reducing the nitrogen (N) application rate and improving water-use efficiency (WUE) are extremely important for sustainable agricultural development in wheat–maize rotation systems in the North China Plain (NCP). We conducted a three-year experiment to investigate the effects of the N application rate on the water-use characteristics of wheat–maize rotation systems in the NCP. The experiment consisted of four N application rates: 250, 167, 84, and 0 kg hm⁻², denoted by N3, N2, N1, and N0, respectively. The results showed the following: For the 0–60 cm soil layer, N deficiency could lead to reduced soil water use (SWU) in wheat seasons, but in maize seasons, N deficiency showed no significant effects on SWU in the 0–60 cm layer. For the 60–140 cm soil layer, N deficiency could lead to reduced SWU in wheat seasons, but in maize seasons, the effects of N deficiency on SWU in the 60–140 cm layer varied with the SWC in the 0–60 cm layer. Throughout the three-year experiment, the evapotranspiration (ET), leaf area index (LAI), yield, and WUE of plants receiving low N treatments decreased with the growing season due to the negative effects of low N treatment (N1 and N0) on the soil. The LAI, total ET, grain yield, and WUE were all positively correlated with each other for both wheat and maize. Considering grain yield and WUE, a single-season N application rate of 167 kg hm⁻² (N2 treatment) in the NCP could meet the growth needs of the wheat–maize rotation system. 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

Neurodegenerative diseases are chronic conditions affecting the central nervous system (CNS). Alzheimer’s disease (AD) is a neurodegenerative disorder characterized by the accumulation of amyloid beta in the limbic and cortical brain regions. AD is presumed to result from genetic abnormalities or environmental factors, including viral infections, which may have deleterious, long-term effects. In this study, we demonstrate that the Venezuelan equine encephalitis virus (VEEV) commonly induces neurodegeneration and long-term neurological or cognitive sequelae. Notably, the effects of VEEV infection can persistently influence gene expression in the mouse brain, suggesting a potential link between the observed neurodegenerative outcomes and long-term alterations in gene expression. Additionally, we show that alphavirus encephalitis exacerbates the neuropathological profile of AD through crosstalk between inflammatory and kynurenine pathways, generating a range of metabolites with potent effects. Using a mouse model for β-amyloidosis, Tg2576 mice, we found that cognitive deficits and brain pathology were more severe in Tg2576 mice infected with VEEV TC-83 compared to mock-infected controls. Thus, during immune activation, the kynurenine pathway plays a more active role in the VEEV TC-83-infected cells, leading to increases in the abundance of transcripts related to the kynurenine pathway of tryptophan metabolism. This pathway generates several metabolites with potent effects on neurotransmitter systems as well as on inflammation, as observed in VEEV TC-83-infected animals. Full article

The search for DNA polymorphisms useful for the genetic improvement of dairy farm animals has spanned more than 40 years, yielding relevant findings in cattle for milk traits, where the best combination of alleles for dairy processing has been found in casein genes and in DGAT1. Nowadays, similar results have not yet been reached in river buffaloes, despite the availability of advanced genomic technologies and accurate phenotype records. The aim of the present study was to investigate and validate the effect of four single nucleotide polymorphisms (SNP) in the CSN1S1, CSN3, SCD and LPL genes on seven milk traits in a larger buffalo population. These SNPs have previously been reported to be associated with, or affect, dairy traits in smaller populations often belonging to one farm. A total of 800 buffaloes were genotyped. The following traits were individually recorded, monthly, throughout each whole lactation period from 2010 to 2021: daily milk yield (dMY, kg), protein yield (dPY, kg) and fat yield (dFY, kg), fat and protein contents (dFP, % and dPP, %), somatic cell count (SCC, 10³ cell/mL) and urea (mg/dL). A total of 15,742 individual milk test day records (2496 lactations) were available for 680 buffalo cows, with 3.6 ± 1.7 parities (from 1 to 13) and an average of 6.1 ± 1.2 test day records per lactation. Three out four SNPs in the CSN1S1, CSN3 and LPL genes were associated with at least one of analyzed traits. In particular, the CSN1S1 (AJ005430:c.578C>T) gave favorable associations with all yield traits (dMY, p = 0.022; dPY, p = 0.014; dFY, p = 0.029) and somatic cell score (SCS, p = 0.032). The CSN3 (HQ677596: c.536C>T) was positively associated with SCS (p = 0.005) and milk urea (p = 0.04). Favorable effects on daily milk yield (dMY, p = 0.028), fat (dFP, p = 0.027) and protein (dPP, p = 0.050) percentages were observed for the LPL. Conversely, the SCD did not show any association with milk traits. This is the first example of a confirmation study carried out in the Mediterranean river buffalo for genes of economic interest in the dairy field, and it represents a very important indication for the preselection of young bulls destined for breeding programs aimed at more sustainable dairy production. Full article

The application results of profile control and water plugging technology are highly related to the gelation time and strength of phenolic resin hydrogel. In this work, a hydrogel solution was prepared by fully mixing the prepared polymer solution with a crosslinker. The static gelation process of PFR hydrogel in ampoule bottles and porous media was analyzed by changes in the viscosity and residual resistance coefficient. Then, the dynamic gelation of the PFR hydrogel in porous media was tested using a circulating flow device, and the changes in viscosity and injection pressure were analyzed during the dynamic gelation process. Finally, the effects of the polymer concentration and crosslinker concentration on dynamic gelation were analyzed. The initial gelation time and final gelation time in porous media were 1–1.5 times and 1.5–2 times those in ampoule bottles under static conditions, respectively. The initial dynamic gelation time in porous media was 2–2.5 times and 1.5–2 times the initial static gelation times in ampoule bottles and porous media, respectively. The final dynamic gelation time was four times and two times the initial static gelation times in ampoule bottles and porous media, respectively. The production after dynamic gelation in porous media comprised hydrogel aggregates and water fluid, leading to a high injection pressure and low viscosity of the produced liquid. As the concentration of polymer and crosslinker increased, the dynamic gelation time was shortened and the gel strength was increased. In the dynamic gelation process in porous media, the phenol resin hydrogel could migrate deeply, but it was limited by the concentrations of the polymer and crosslinker. The results of subsequent water flooding showed that the polymer hydrogel had a good plugging ability after dynamic gelation. The deep reservoir could only be blocked off in the subsequent water flooding process when the migration of hydrogel happened in the dynamic gelation process. Full article

The capability of dielectric measurements was significantly increased with the development of capacitive one-side access physical sensors. Complete samples give no opportunity to study electric susceptibility at a partial coverage of the one-side access sensor’s active area; therefore, partial samples are proposed. The electric susceptibility at the partial coverage of a circular one-side access sensor with cylinders and shells is investigated for polyurethane materials. The implementation of the relative partial susceptibility permitted us to transform the calculated susceptibility data to a common scale of 0.0–1.0 and to outline the main trends for PU materials. The partial susceptibility, relative partial susceptibility, and change rate of relative partial susceptibility exhibited dependence on the coverage coefficient of the sensor’s active area. The overall character of the curves for the change rate of the relative partial susceptibility, characterised by slopes of lines and the ratio of the change rate in the centre and near the gap, corresponds with the character of the surface charge density distribution curves, calculated from mathematical models. The elaborated methods may be useful in the design and optimization of capacitive OSA sensors of other configurations of electrodes, independent of the particular technical solution. 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