Eye fatigue has a fatiguing effect on the eye muscles, and eye movement performance is a macroscopic response to the eye fatigue state. To detect and prevent the risk of eye fatigue in advance, this study designed an eye fatigue detection experiment, collected experimental data samples, and constructed experimental data sets. In this study, eye-tracking feature extraction was completed, and the significance difference of eye-tracking features under different fatigue states was discussed by two-way repeated-measures ANOVA (Analysis of Variance). The experimental results demonstrate the feasibility of eye fatigue detection from eye-tracking signals. In addition, this study considers the effects of different feature extraction methods on eye fatigue detection accuracy. This study examines the performance of machine learning algorithms based on manual feature calculation (SVM, DT, RM, ET) and deep learning algorithms based on automatic feature extraction (CNN, auto-encoder, transformer) in eye fatigue detection. Based on the combination of the methods, this study proposes the feature union auto-encoder algorithm, and the accuracy of the algorithm for eye fatigue detection on the experimental dataset is improved from 82.4% to 87.9%. Full article

Pregnancy induces significant changes in oral health because of hormonal fluctuations, making it a crucial period for preventive measures. Dental stem cells (DSCs), particularly those derived from the dental pulp and periodontal ligaments, offer promising avenues for regenerative therapies and, possibly, preventive interventions. While the use of DSCs already includes various applications in regenerative dentistry in the general population, their use during pregnancy requires careful consideration. This review explores recent advancements, challenges, and prospects in using DSCs to address oral health issues, possibly during pregnancy. Critical aspects of the responsible use of DSCs in pregnant women are discussed, including safety, ethical issues, regulatory frameworks, and the need for interdisciplinary collaborations. We aimed to provide a comprehensive understanding of leveraging DSCs to improve maternal oral health. Full article

The examination of connected speech may serve as a valuable tool for exploring speech output in both healthy speakers and individuals with language disorders. Numerous studies incorporate various fluency and silence measures into their analyses to investigate speech output patterns in different populations, along with the underlying cognitive processes that occur while speaking. However, methodological inconsistencies across existing studies pose challenges in comparing their results. In the current study, we introduce CSAP (Connected Speech Analysis Protocol), which is a specific methodological approach to investigate fluency metrics, such as articulation rate and speech rate, as well as silence measures, including silent pauses’ frequency and duration. We emphasize the importance of employing a comprehensive set of measures within a specific methodological framework to better understand speech output patterns. Additionally, we advocate for the use of distinct narrative tasks for a thorough investigation of speech output in different conditions. We provide an example of data on which we implement CSAP to showcase the proposed pipeline. In conclusion, CSAP offers a comprehensive framework for investigating speech output patterns, incorporating fluency metrics and silence measures in distinct narrative tasks, thus allowing a detailed quantification of connected speech in both healthy and clinical populations. We emphasize the significance of adopting a unified methodological approach in connected speech studies, enabling the integration of results for more robust and generalizable conclusions. Full article

Mycorrhizal symbiosis enhances host plant resistance to various unfavorable environmental stresses, but whether and how it also enhances waterlogging tolerance in cucumber plants is not known. The objective of this study was to analyze the effect of Paraglomus occultum inoculation on biomass production, osmolyte levels, and the expression of 12 heat shock protein 70 (Hsp70) genes and 14 plasma membrane intrinsic protein (PIP) genes in the roots of cucumber plants under a short-term waterlogging stress (WS) (5 days) condition. Although the short-term WS treatment significantly inhibited the arbuscular mycorrhizal fungal colonization of roots, the inoculation with arbuscular mycorrhizal fungi (AMFs) significantly increased leaf, stem, and root biomass under WS. AMF inoculation also significantly increased root glucose, sucrose, betaine, and proline contents, along with decreased fructose levels, compared with the uninoculated control. More CsHsp70 and CsPIP genes were up-regulated in AMF-inoculated plants than in AMF-uninoculated plants in response to WS. AMF inoculation showed no significant effect on the expression of any of the examined CsHsp70 genes under no-waterlogging stress, but it did raise the expression of 11 of 12 CsHsp70 genes under WS. AMF colonization also down-regulated or had no effect on CsPIP expression under no-waterlogging stress, whereas it up-regulated the expression of 12 of the 14 CsPIP genes under WS. It is concluded that AMF inoculation enhances waterlogging tolerance in cucumber plants by increasing osmolyte levels and stress-responsive gene (CsPIP and CsHsp70) expression. Full article

Horticultural plant material offers several advantages for isolating exosomes and other natural plant-derived exosome-like nanoparticles (PDENs) due to the accessibility and affordability of plant material for widespread applications. This study aims to explore the impacts of the tomato genotype (‘Admiro’, ‘Roma’, ‘Brooklyn’, ‘Marmande’ and ‘Betalux’) and the main cultivation parameters in controlled environment agriculture on the yield and properties of their PDENs for pharmaceutical and cosmeceutical applications. The PDEN yield, size distribution, and antioxidative properties of young tomato seedlings were evaluated. The ‘Betalux’ tomato was distinguished by a remarkably higher nanoparticle concentration and a uniform size distribution and was selected for further experiments. The impact of cultivation temperature (18, 22, and 26 °C), nitrogen nutrition (0, 250, and 500 mg L⁻¹), and the lighting photosynthetic photon flux density (PPFD; 150, 250, and 450 µmol m⁻² s⁻¹) on nanoparticle properties was investigated. Optimal conditions consisting of a temperature of 22 °C, 250 mg L⁻¹ nitrogen nutrition, and 250 µmol m⁻² s⁻¹ lighting PPFD were used as a reference. Optimal temperature, nitrogen nutrition, and lighting intensity resulted in the highest nanoparticle yield, the most uniform particle distribution, and the highest impact of PDEN preparations on keratinocyte metabolic activity. Deviation from optimal cultivation conditions reduced the tomato biomass and the PDEN protein and yield. Full article

Human Immunodeficiency Virus (HIV) is a stigmatizing disease that disproportionately affects African Americans and Latinos among people living with HIV (PLWH). Researchers are increasingly utilizing artificial intelligence (AI) to analyze large amounts of data such as social media data and electronic health records (EHR) for various HIV-related tasks, from prevention and surveillance to treatment and counseling. This paper explores the ethical considerations surrounding the use of AI for HIV with a focus on acceptability, trust, fairness, and transparency. To improve acceptability and trust towards AI systems for HIV, informed consent and a Federated Learning (FL) approach are suggested. In regard to unfairness, stakeholders should be wary of AI systems for HIV further stigmatizing or even being used as grounds to criminalize PLWH. To prevent criminalization, in particular, the application of differential privacy on HIV data generated by data linkage should be studied. Participatory design is crucial in designing the AI systems for HIV to be more transparent and inclusive. To this end, the formation of a data ethics committee and the construction of relevant frameworks and principles may need to be concurrently implemented. Lastly, the question of whether the amount of transparency beyond a certain threshold may overwhelm patients, thereby unexpectedly triggering negative consequences, is posed. Full article

Globe artichoke (Cynara cardunculus var. scolymus L.) is widely cultivated in the Mediterranean area and Italy is one of the largest producers. A great issue is represented by its high amount of by-product, mainly consisting of external bracts and stems, but also of residual leaves, stalks, roots, and seeds. Artichoke by-products are rich in nutrients (carbohydrates and proteins) and bioactive compounds (polyphenols and terpenes) and represent potential ingredients for foodstuffs, functional foods, and food supplements, due to their functional and biological properties. In fact, artichoke by-products’ components exhibit many beneficial effects, such as dyspeptic, prebiotic, antioxidant, anti-inflammatory, antiglycative, antimicrobial, anticarcinogenic, and hypolipidemic properties. Therefore, they can be considered potential food ingredients useful in reducing the risk of developing metabolic and age-related disorders. This work summarizes the economic and environmental impact of the recovery and valorization of artichoke by-products, focusing on rheological, physical, and biological properties of the different components present in each by-product and their different food applications. Full article

The condition assessment of water distribution pipes is of utmost importance for the prioritization of rehabilitation interventions. However, the application of available methodologies for condition assessment by water utilities with limited human, technological and financial resources is becoming increasingly complex. The current paper aims at the development and application of a methodology for the prediction of the physical condition of water distribution pipes without the need for visual inspection. The methodology includes the development and application of three different algorithms (heuristic, linear regression and support vector regression). The methodology is applied to a water distribution network located in the Algarve region, Portugal. The results obtained from each algorithm are compared with a well-known performance indicator, the ratio of useful life, and present significant differences in its overall pipe condition classification. Results have demonstrated the following: the ratio of useful life tends to distribute pipe classification more equally in the three classes (i.e., good, average and unsatisfactory); the heuristic algorithm classifies most pipes as average condition; and the linear regression algorithm classifies with unsatisfactory conditions. The support vector regression algorithm stands out as the main classifier for identifying pipes in good condition when compared to other algorithms. Full article

The purpose of this study was to investigate how students’ disabilities impacted their learning at the university and to measure their perceptions of universal design for learning (UDL). An online survey was administered at a large public research university in the south-central United States; 160 students with disabilities completed the survey. These students with disabilities described how their disabilities influenced their learning in the following aspects: attention issues, slow processing, absence, accessibility issues, reading and writing challenges, mental health challenges, and social interaction challenges. The results also revealed that some UDL teaching practices were not being fully utilized by instructors based on the perceptions of students with disabilities. We discussed those UDL practices which were not being used by instructors and highlighted the impact of using the practices on students’ learning. These practices include effective teaching methods such as helping students organize and summarize learning content, using technology to increase accessibility, providing flexibility in assessments and assignments, providing meaningful feedback, and recruiting students’ attention and engagement. Full article

Objective: This study evaluated the methodological quality of published systematic reviews on randomized and non-randomized clinical trials to synthesize evidence on the association between IL-6, immunosenescence, and aerobic and/or resistance exercise. Method: The Preferred Reporting Items for Overviews of Systematic Reviews (PRIO-harms) guideline was used, with registration number CRD42022346142-PROSPERO. Relevant databases such as Cochrane Library, PubMed, Web of Science, Scopus, and Google Scholar were searched using English Medical Subject Headings terms. Inclusion criteria were systematic reviews analyzing aerobic exercise, resistance exercise, or a combination of both and assessing IL-6 as a biomarker of cellular immunosenescence in humans. The Measurement Tool to Assess Systematic Reviews 2 (AMSTAR-2) was employed. Results: Out of 742 identified articles, 18 were eligible, and 13 were selected for analysis. Sample sizes ranged from 249 to 1421 participants, mostly female, with ages ranging from 17 to 95 years. Aerobic exercise was the most studied type (46.15%), followed by combined exercise (38.46%) and resistance exercise (15.38%). Aerobic exercise showed a statistically significant reduction in IL-6, C-reactive protein (CRP), and tumor necrosis factor-alpha (TNF-α) levels. Among the 13 reviews analyzed using AMSTAR-2, 8 were rated as critically low quality, and 5 were classified as low quality. Conclusion: Aerobic exercise has anti-inflammatory properties and the potential to modulate IL-6, CRP, and TNF-α levels in immunosenescence. However, the limited methodological quality of the analyzed systematic reviews highlights the urgent need for robust, high-quality studies to improve access to information and facilitate evidence-based decision-making in healthcare. Full article

Modeling of airborne virus transmission and protection against it requires knowledge of the amount of biofluid emitted into the atmosphere and its viral load. Whereas viral concentrations in biofluids are readily measured by quantitative PCR, the total volume of fluids aerosolized during speaking, as measured by different researchers using various technologies, differs by several orders of magnitude. We compared collection methods in which the aerosols first enter into a low-humidity chamber either by direct injection or via commonly used funnel and tubing arrangements, followed by standard optical particle sizer measurement. This “collect first, measure later” approach sacrifices the recording of the temporal correlation between aerosol generation and sound types such as plosives and vowels. However, the direct-injection mode prevents inertia deposition associated with the funnel arrangements and reveals far more intermediate-size (5–20 μm in diameter) particles that can dominate the total mass of ejected respiratory aerosol. The larger aerosol mass observed with our method partially reconciles the large discrepancy between the SARS-CoV-2 infectious dose estimated from superspreader event analyses and that from human challenge data. Our results also impact measures to combat airborne virus transmission because they indicate that aerosols that settle faster than good room ventilation rates can dominate this process. Full article

Ethanamizuril (EZL) is a new anticoccidial drug developed by our Shanghai Veterinary Research Institute. Since EZL is almost insoluble in water, we conducted a study to improve the solubility of EZL by forming inclusion complexes with β-cyclodextrin (β-CD) and hydroxypropyl-β-cyclodextrin (HP-β-CD). In this study, we performed molecular docking and then systematically compared the interactions of EZL with β-CD and HP-β-CD in both aqueous solution and the solid state, aiming to elucidate the solubilization effect and mechanism of cyclodextrins (CDs). The interactions were also examined in the solid state using DSC, PXRD, and FT-IR. The interactions of EZL with CDs in an aqueous solution were investigated using PSA, UV-vis spectroscopy, MS, ¹H NMR, and 2D ROESY. The results of phase solubility experiments revealed that both β-CD and HP-β-CD formed inclusion complexes with EZL in a 1:1 molar ratio. Among them, HP-β-CD exhibited higher K_f (stability constant) and CE (complexation efficiency) values as well as a stronger solubilization effect. Furthermore, the two cyclodextrins were found to interact with EZL in a similar manner. The results of our FT-IR and 2D ROESY experiments are in agreement with the theoretical results derived from molecular simulations. These results indicated that intermolecular hydrogen bonds existing between the C=O group on the triazine ring of EZL and the O-H group of CDs, as well as the hydrophobic interactions between the hydrogen on the benzene ring of EZL and the hydrogen of CDs, played crucial roles in the formation of EZL/CD inclusion complexes. The results of this study can lay the foundation for the future development of high-concentration drinking water delivery formulations for EZL. Full article

Forkhead box protein 3 (FoxP3) is a key transcription factor responsible for the development, maturation, and function of regulatory T cells (Tregs). The FoxP3 pre-mRNA is subject to alternative splicing, resulting in the translation of multiple splice variants. We have shown that Tregs from patients with amyotrophic lateral sclerosis (ALS) have reduced expression of full-length (FL) FoxP3, while other truncated splice variants are expressed predominantly. A correlation was observed between the reduced number of Tregs in the peripheral blood of ALS patients, reduced total FoxP3 mRNA, and reduced mRNA of its FL splice variant. Induction of FL FoxP3 was achieved using splice-switching oligonucleotides capable of base pairing with FoxP3 pre-mRNA and selectively modulating the inclusion of exons 2 and 7 in the mature mRNA. Selective expression of FL FoxP3 resulted in the induction of CD127^low, CD152, and Helios-positive cells, while the cell markers CD4 and CD25 were not altered. Such Tregs had an increased proliferative activity and a higher frequency of cell divisions per day. The increased suppressive activity of Tregs with the induced FL FoxP3 splice variant was associated with the increased synthesis of the pro-apoptotic granzymes A and B, and perforin, IL-10, and IL-35, which are responsible for contact-independent suppression, and with the increased ability to suppress telomerase in target cells. The upregulation of Treg suppressive and proliferative activity using splice-switching oligonucleotides to induce the predominant expression of the FoxP3 FL variant is a promising approach for regenerative cell therapy in Treg-associated diseases. Full article

Congenital Adrenal Hyperplasia (CAH) is an autosomal recessive disorder impairing cortisol synthesis due to reduced enzymatic activity. This leads to persistent adrenocortical overstimulation and the accumulation of precursors before the blocked enzymatic step. The predominant form of CAH arises from mutations in CYP21A2, causing 21-hydroxylase deficiency (21-OHD). Despite emerging treatment options for CAH, it is not always possible to physiologically replace cortisol levels and counteract hyperandrogenism. Moreover, there is a notable absence of an effective in vivo model for pre-clinical testing. In this work, we developed an animal model for CAH with the clinically relevant point mutation p.R484Q in the previously humanized CYP21A2 mouse strain. Mutant mice showed hyperplastic adrenals and exhibited reduced levels of corticosterone and 11-deoxycorticosterone and an increase in progesterone. Female mutants presented with higher aldosterone concentrations, but blood pressure remained similar between wildtype and mutant mice in both sexes. Male mutant mice have normal fertility with a typical testicular appearance, whereas female mutants are infertile, exhibit an abnormal ovarian structure, and remain in a consistent diestrus phase. Conclusively, we show that the animal model has the potential to contribute to testing new treatment options and to prevent comorbidities that result from hormone-related derangements and treatment-related side effects in CAH patients. Full article

This review comprehensively evaluates the effects of physical exercise on oxidative and nitrosative stress, mainly focusing on the role of antioxidants. Using a narrative synthesis approach, data from empirical studies, reviews, systematic reviews, and meta-analyses published between 2004 and 2024 were collated from databases like PubMed, EBSCO (EDS), and Google Scholar, culminating in the inclusion of 41 studies. The quality of these studies was rigorously assessed to ensure the clarity of objectives, coherence in arguments, comprehensive literature coverage, and depth of critical analysis. Findings revealed that moderate exercise enhances antioxidant defenses through hormesis, while excessive exercise may exacerbate oxidative stress. The review also highlights that while natural dietary antioxidants are beneficial, high-dose supplements could impede the positive adaptations to exercise. In conclusion, the review calls for more focused research on tailored exercise and nutrition plans to further understand these complex interactions and optimize the health outcomes for athletes and the general population. Full article

Many studies have shown that melatonin (an indoleamine) is an important molecule in plant physiology. It is known that this indoleamine is crucial during plant stress responses, especially by counteracting secondary oxidative stress (efficient direct and indirect antioxidant) and switching on different defense plant strategies. In this report, we present exogenous melatonin’s potential to protect lipid profile modification and membrane integrity in Nicotiana tabacum L. line Bright Yellow 2 (BY-2) cell culture exposed to lead. There are some reports of the positive effect of melatonin on animal cell membranes; ours is the first to report changes in the lipid profile in plant cells. The experiments were performed in the following variants: LS: cells cultured on unmodified LS medium—control; (ii) MEL: BY-2 cells cultured on LS medium with melatonin added from the beginning of culture; (iii) Pb: BY-2 cells cultured on LS medium with Pb²⁺ added on the 4th day of culture; (iv) MEL+Pb: BY-2 cells cultured on LS medium with melatonin added from the start of culture and stressed with Pb²⁺ added on the 4th day of culture. Lipidomic analysis of BY-2 cells revealed the presence of 40 different phospholipids. Exposing cells to lead led to the overproduction of ROS, altered fatty acid composition and increased PLD activity and subsequently elevated the level of phosphatidic acid at the cost of dropping the phosphatidylcholine. In the presence of lead, double-bond index elevation, mainly by higher quantities of linoleic (C18:2) and linolenic (C18:3) acids in the log phase of growth, was observed. In contrast, cells exposed to heavy metal but primed with melatonin showed more similarities with the control. Surprisingly, the overproduction of ROS caused of lipid peroxidation only in the stationary phase of growth, although considerable changes in lipid profiles were observed in the log phase of growth—just 4 h after lead administration. Our results indicate that the pretreatment of BY-2 with exogenous melatonin protected tobacco cells against membrane dysfunctions caused by oxidative stress (lipid oxidation), but also findings on a molecular level suggest the possible role of this indoleamine in the safeguarding of the membrane lipid composition that limited lead-provoked cell death. The presented research indicates a new mechanism of the defense strategy of plant cells generated by melatonin. Full article

Tillage practices govern crop quality and quantity through soil nutrient availability and crop root systems. A deeper knowledge of the impact of conservation tillage on soil chemical characteristics (such as pH, soil organic carbon, macro and micronutrient storage and distribution) is required for both the promotion of agricultural sustainability and environmental preservation. This study assesses the changes in soil features and properties in the context of a long-field experiment with different tillage systems and straw management practices. Research findings revealed that compared with conventional tillage (CT) conservative tillage with partial straw retention (MT) and no-tillage with straw mulching (NT) substantially boosted the organic carbon (OC) (by 6–19%), total nitrogen (TN) (by 2–12%), and available potassium content (AK) (by 2–5%), in 0–30 cm soil depth. However, the stratification trend was observed for available macro and micronutrient content (Zn, Fe, Mn) in both conservative management practices. The concentration of Cu indicates a constant pattern through a 0–30 cm soil profile with a higher concentration under MT (1.41 mg kg⁻¹) compared to NT (1.10 mg kg⁻¹). In particular, the results failed to establish if conservation tillage can increase the total phosphorus (TP) and potassium content (TK), where only in surface 0–10 cm an increase was observed. This research also suggested that the X-ray fluorescence analysis (XRF) of total micronutrient content (Zn, Cu, Fe, Mn) is minimal or unpredictable with no substantial differences between the tillage systems and straw return management practices. These findings suggest that conservation tillage in north-eastern Romania might be optimal to maintain soil quality status and sustain high yields. Full article

Growing butter markets, domestically and globally, provide opportunities for value-added variants of butter. Adding probiotics to butter could boosts its bioactivity; however, maintaining probiotic viability during storage is a major challenge. Mathematical analysis of probiotic population changes could help improve our understanding of how probiotics interact with butter and storage conditions. Two strains of probiotics in a 1:1 ratio as free cells or Whey Protein Hydrolysate–Maltodextrin (WPH-MD)-encapsulated cells, Lactobacillus acidophilus ATCC 4356 (LA5) and Bifidobacterium animalis ssp. lactis ATCC 27536 (BB12), were separately mixed into butter at 1% levels. Using analysis of covariance, a mathematical evaluation for probiotic population changes was performed by periodically determining viable counts, resulting in an adjusted R² value of 0.98 and demonstrating a strong relationship between the dependent variable (log₁₀ counts of probiotics) and independent variables (cell type, temperature of storage, and time of storage). After 21 days of storage, the number of free cells in butter dropped from 7.45 log₁₀ CFU/g to 0.56 log₁₀ CFU/g. On the other hand, it took 63 days for encapsulated cells to achieve 0.80 log₁₀ CFU/g at the same temperature. The same trend persisted at −18 °C, indicating that the WPH-MD encapsulant had a protective effect. Full article

Chronic obstructive pulmonary disease (COPD) is comprised of histopathological alterations such as pulmonary emphysema and peribronchial fibrosis. Matrix metalloproteinase 9 (MMP-9) is one of the key enzymes involved in both types of tissue remodeling during the development of lung damage. In recent studies, it was demonstrated that deflamin, a protein component extracted from Lupinus albus, markedly inhibits the catalytic activity of MMP-9 in experimental models of colon adenocarcinoma and ulcerative colitis. Therefore, in the present study, we investigated for the first time the biological effect of deflamin in a murine COPD model induced by chronic exposure to ozone. Ozone exposure was carried out in C57BL/6 mice twice a week for six weeks for 3 h each time, and the treated group was orally administered deflamin (20 mg/kg body weight) after each ozone exposure. The histological results showed that deflamin attenuated pulmonary emphysema and peribronchial fibrosis, as evidenced by H&E and Masson’s trichrome staining. Furthermore, deflamin administration significantly decreased MMP-9 activity, as assessed by fluorogenic substrate assay and gelatin zymography. Interestingly, bioinformatic analysis reveals a plausible interaction between deflamin and MMP-9. Collectively, our findings demonstrate the therapeutic potential of deflamin in a COPD murine model, and suggest that the attenuation of the development of lung tissue damage occurs by deflamin-regulated MMP-9 catalytic activity. Full article

While many countries across the world have had difficulty in providing continuous coronavirus disease (COVID-19) vaccine boosters, in Japan, the number of COVID-19 vaccinations has increased rapidly in the past two years to the point where the vaccination booster numbers are now at the highest level in the world (by March 2023). Against this backdrop, this study aimed to determine the characteristics of individuals who either refused to be vaccinated or who were vaccinated multiple (five) times during this period. We analyzed data from a sample of 3710 Japanese adults that were collected in a nationwide, cross-sectional Internet survey that was undertaken in March 2023. Several demographics (e.g., age, education level, marital status, job status), medical conditions, daily smoking, and mental health/psychological factors (i.e., perceived stress, anxiety symptoms, loneliness) were associated with not having been vaccinated. Although the most frequently vaccinated status was also associated with some demographics (e.g., age, employment status), medical conditions, and daily smoking, other predictors such as having a COVID-19 infection history were unique to this outcome. Moreover, age-stratified analyses showed that depression (OR: 2.05, 95%CI: 1.08–3.89) and anxiety (OR: 3.87, 95%CI: 1.80–8.31) symptoms were associated with higher odds of being unvaccinated while loneliness was associated with lower odds for the most frequently vaccinated status (OR: 0.72, 95%CI: 0.54–0.96) among older adults (aged ≥ 60 years). The findings of this study indicate that many of the same factors are associated with vaccine hesitancy and being vaccinated multiple times among adults in Japan but that among older individuals, worse mental/psychological health problems may be important for vaccine hesitancy/infrequent vaccine uptake in an age range where the majority of individuals (57.7%) had been vaccinated five times. Full article