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14 pages, 3646 KiB  
Article
Benzyldimethyldodecyl Ammonium Chloride-Doped Denture-Based Resin: Impact on Strength, Surface Properties, Antifungal Activities, and In Silico Molecular Docking Analysis
by Sarah Aldulaijan, Raghad Alruwili, Rawan Almulaify, Fatimah A. Alhassan, Yousif A. Al-Dulaijan, Faris A. Alshahrani, Lamia Mokeem, Mohammed M. Gad, Mary Anne S. Melo and Abdulrahman A. Balhaddad
J. Funct. Biomater. 2024, 15(10), 310; https://doi.org/10.3390/jfb15100310 (registering DOI) - 18 Oct 2024
Abstract
Candida albicans (C. albicans) adhering to denture-based resins (DBRs) is a known cause of denture stomatitis. A new approach to prevent denture stomatitis is to include antimicrobial substances within DBRs. Here, we examined the mechanical performance and antifungal properties of DBRs [...] Read more.
Candida albicans (C. albicans) adhering to denture-based resins (DBRs) is a known cause of denture stomatitis. A new approach to prevent denture stomatitis is to include antimicrobial substances within DBRs. Here, we examined the mechanical performance and antifungal properties of DBRs containing benzyldimethyldodecyl ammonium chloride (C12BDMA-Cl) as an antimicrobial compound. C12BDMA-Cl is a quaternary ammonium compound, and its antifungal properties have never been investigated when combined with dental acrylic resin. Therefore, we modified a commercially available heat-polymerized acrylic DBR to contain 3 and 5 wt.% of C12BDMA-Cl. Unmodified DBR was used as a control group. Specimens were prepared using the conventional heat processing method. The specimen’s flexural strength, elastic modulus, microhardness, and surface roughness were evaluated. C. albicans biofilm was grown on the specimens and assessed via colony-forming units (CFUs) and scanning electron microscopy (SEM). In silico molecular docking was applied to predict the potential C12BDMA-Cl inhibition activity as an antifungal drug. The 3% C12BDMA-Cl DBR demonstrated antifungal activities without a deterioration effect on the mechanical performance. SEM images indicated fewer colonies in DBR containing C12BDMA-Cl, which can be a potential approach to managing denture stomatitis. In conclusion, C12BDMA-Cl is a promising antifungal agent for preventing and treating denture stomatitis. Full article
(This article belongs to the Special Issue Advanced Dental Restorative Composite Materials)
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<p>Flowchart of the study design and specimen fabrication.</p>
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<p>The mechanical properties assessment of the C<sub>12</sub>BDMA-Cl DBR groups (n = 6, mean ± SD). (<b>A</b>) Flexural strength, (<b>B</b>) elastic modulus, and (<b>C</b>) microhardness. Dissimilar letters indicate a significant difference (<span class="html-italic">p</span> ≤ 0.05). (<b>D</b>) Representative optical images for the microhardness test. No significant differences were found among the groups concerning their microhardness.</p>
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<p>Surface roughness values of the C<sub>12</sub>BDMA-Cl DBR groups. (<b>A</b>) Average surface roughness, (<b>B</b>) maximum height of the profile, (<b>C</b>) maximum peak height, and (<b>D</b>) maximum valley depth (n = 10, mean ± SD). Dissimilar letters indicate a significant difference (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>The antifungal properties of the C<sub>12</sub>BDMA-Cl DBR groups (n = 9, mean ± SD). Dissimilar letters indicate a significant difference (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Scanning electron microscope (SEM) images of the C<sub>12</sub>BDMA-Cl -DBR groups (n = 1 per group). (<b>A</b>,<b>B</b>) The control DBR with no C<sub>12</sub>BDMA-Cl displays the growth of <span class="html-italic">Candida albicans</span> colonies (red arrows) over the specimens. (<b>C</b>,<b>D</b>) The 3 wt.% C<sub>12</sub>BDMA-Cl DBR and (<b>E</b>,<b>F</b>) the 5 wt.% C<sub>12</sub>BDMA-Cl DBR display reduced biofilm growth with fewer morphological yeast-to-hyphal transitions (yellow arrow) compared to the control, indicating that these colonies are less pathogenic.</p>
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<p>Three-dimensional (3D) structures of the binding positions between <span class="html-italic">Candida albicans</span> and C<sub>12</sub>BDMA-Cl. The C<sub>12</sub>BDMA-Cl compound (green) and the reference compounds (red) in the active site of (<b>A</b>) Dihydrofolate reductase (DHFR) enzyme and (<b>B</b>) Candidapepsin-1 (SAPs) enzyme. (<b>C</b>,<b>D</b>) Two-dimensional structures of the binding between C<sub>12</sub>BDMA-Cl compound and the two receptors; (<b>C</b>) Dihydrofolate reductase (DHFR) enzyme and (<b>D</b>) Candidapepsin-1 (SAPs) enzyme.</p>
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15 pages, 902 KiB  
Review
Instruments for Patient Safety Assessment: A Scoping Review
by Elisabete Nunes, Fernanda Sirtoli, Eliane Lima, Greyce Minarini, Filomena Gaspar, Pedro Lucas and Cândida Primo
Healthcare 2024, 12(20), 2075; https://doi.org/10.3390/healthcare12202075 (registering DOI) - 18 Oct 2024
Abstract
Background: Patient safety is an important component of healthcare service quality, and there are numerous instruments in the literature that measure patient safety. This scoping reviewaims to map the instruments/scales for assessing patient safety in healthcare services. Method: This scoping review follows the [...] Read more.
Background: Patient safety is an important component of healthcare service quality, and there are numerous instruments in the literature that measure patient safety. This scoping reviewaims to map the instruments/scales for assessing patient safety in healthcare services. Method: This scoping review follows the JBI methodology. The protocol was registered on the Open Science Framework. Eligibility criteria were defined based on studies that include instruments or scales for assessing patient safety in healthcare services, in any language, and without temporal restrictions. It adhered to all scoping review checklist items [PRISMA-ScR], with searches in the Embase, Lilacs, MedLine, and Scopus databases, as well as the repository of the Brazilian Digital Library of Theses and Dissertations. Two independent reviewers performed selection and data extraction in July 2023. Results: Of the 4019 potential titles, 63 studies reported on a total of 47 instruments/scales and 71 dimensions for patient safety assessment. The most-described dimensions were teamwork, professional satisfaction, safety climate, communication, and working conditions. Conclusion: The diversity of instruments and dimensions for patient safety assessment characterizes the multidimensionality and scope of patient safety. However, it hinders benchmarking between institutions and healthcare units. Full article
(This article belongs to the Special Issue Review Research on Healthcare Quality and Patient Safety)
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<p>Study selection flowchart—Prisma-SCR. Source: Adapted from Peters et al. [2020] [<a href="#B10-healthcare-12-02075" class="html-bibr">10</a>].</p>
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34 pages, 5078 KiB  
Systematic Review
Context-Aware Embedding Techniques for Addressing Meaning Conflation Deficiency in Morphologically Rich Languages Word Embedding: A Systematic Review and Meta Analysis
by Mosima Anna Masethe, Hlaudi Daniel Masethe and Sunday O. Ojo
Computers 2024, 13(10), 271; https://doi.org/10.3390/computers13100271 (registering DOI) - 17 Oct 2024
Viewed by 159
Abstract
This systematic literature review aims to evaluate and synthesize the effectiveness of various embedding techniques—word embeddings, contextual word embeddings, and context-aware embeddings—in addressing Meaning Conflation Deficiency (MCD). Using the PRISMA framework, this study assesses the current state of research and provides insights into [...] Read more.
This systematic literature review aims to evaluate and synthesize the effectiveness of various embedding techniques—word embeddings, contextual word embeddings, and context-aware embeddings—in addressing Meaning Conflation Deficiency (MCD). Using the PRISMA framework, this study assesses the current state of research and provides insights into the impact of these techniques on resolving meaning conflation issues. After a thorough literature search, 403 articles on the subject were found. A thorough screening and selection process resulted in the inclusion of 25 studies in the meta-analysis. The evaluation adhered to the PRISMA principles, guaranteeing a methodical and lucid process. To estimate effect sizes and evaluate heterogeneity and publication bias among the chosen papers, meta-analytic approaches were utilized such as the tau-squared (τ2) which represents a statistical parameter used in random-effects, H-squared (H2) is a statistic used to measure heterogeneity, and I-squared (I2) quantify the degree of heterogeneity. The meta-analysis demonstrated a high degree of variation in effect sizes among the studies, with a τ2 value of 8.8724. The significant degree of heterogeneity was further emphasized by the H2 score of 8.10 and the I2 value of 87.65%. A trim and fill analysis with a beta value of 5.95, a standard error of 4.767, a Z-value (or Z-score) of 1.25 which is a statistical term used to express the number of standard deviations a data point deviates from the established mean, and a p-value (probability value) of 0.2 was performed to account for publication bias which is one statistical tool that can be used to assess the importance of hypothesis test results. The results point to a sizable impact size, but the estimates are highly unclear, as evidenced by the huge standard error and non-significant p-value. The review concludes that although contextually aware embeddings have promise in treating Meaning Conflation Deficiency, there is a great deal of variability and uncertainty in the available data. The varied findings among studies are highlighted by the large τ2, I2, and H2 values, and the trim and fill analysis show that changes in publication bias do not alter the impact size’s non-significance. To generate more trustworthy insights, future research should concentrate on enhancing methodological consistency, investigating other embedding strategies, and extending analysis across various languages and contexts. Even though the results demonstrate a significant impact size in addressing MCD through sophisticated word embedding techniques, like context-aware embeddings, there is still a great deal of variability and uncertainty because of various factors, including the different languages studied, the sizes of the corpuses, and the embedding techniques used. These differences show how future research methods must be standardized to guarantee that study results can be compared to one another. The results emphasize how crucial it is to extend the linguistic scope to more morphologically rich and low-resource languages, where MCD is especially difficult. The creation of language-specific models for low-resource languages is one way to increase performance and consistency across Natural Language Processing (NLP) applications in a practical sense. By taking these actions, we can advance our understanding of MCD more thoroughly, which will ultimately improve the performance of NLP systems in a variety of language circumstances. Full article
(This article belongs to the Special Issue Natural Language Processing (NLP) and Large Language Modelling)
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<p>PRISMA flow diagram for the CAWE research [<a href="#B30-computers-13-00271" class="html-bibr">30</a>].</p>
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<p>Proposed Embeddings Taxonomy.</p>
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<p>Forest plot for distribution of effect size of meta-analysis summary of CAWE in <a href="#computers-13-00271-t003" class="html-table">Table 3</a> [<a href="#B29-computers-13-00271" class="html-bibr">29</a>,<a href="#B33-computers-13-00271" class="html-bibr">33</a>,<a href="#B34-computers-13-00271" class="html-bibr">34</a>,<a href="#B35-computers-13-00271" class="html-bibr">35</a>,<a href="#B36-computers-13-00271" class="html-bibr">36</a>,<a href="#B37-computers-13-00271" class="html-bibr">37</a>,<a href="#B38-computers-13-00271" class="html-bibr">38</a>,<a href="#B39-computers-13-00271" class="html-bibr">39</a>,<a href="#B40-computers-13-00271" class="html-bibr">40</a>,<a href="#B41-computers-13-00271" class="html-bibr">41</a>,<a href="#B42-computers-13-00271" class="html-bibr">42</a>,<a href="#B43-computers-13-00271" class="html-bibr">43</a>,<a href="#B44-computers-13-00271" class="html-bibr">44</a>,<a href="#B45-computers-13-00271" class="html-bibr">45</a>,<a href="#B46-computers-13-00271" class="html-bibr">46</a>,<a href="#B47-computers-13-00271" class="html-bibr">47</a>,<a href="#B48-computers-13-00271" class="html-bibr">48</a>,<a href="#B49-computers-13-00271" class="html-bibr">49</a>,<a href="#B50-computers-13-00271" class="html-bibr">50</a>,<a href="#B51-computers-13-00271" class="html-bibr">51</a>,<a href="#B52-computers-13-00271" class="html-bibr">52</a>,<a href="#B53-computers-13-00271" class="html-bibr">53</a>,<a href="#B54-computers-13-00271" class="html-bibr">54</a>,<a href="#B55-computers-13-00271" class="html-bibr">55</a>,<a href="#B56-computers-13-00271" class="html-bibr">56</a>].</p>
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<p>Forest plot with subgroups for distribution of effect size of meta-analysis summary of CAWE in <a href="#computers-13-00271-t003" class="html-table">Table 3</a> [<a href="#B29-computers-13-00271" class="html-bibr">29</a>,<a href="#B33-computers-13-00271" class="html-bibr">33</a>,<a href="#B34-computers-13-00271" class="html-bibr">34</a>,<a href="#B35-computers-13-00271" class="html-bibr">35</a>,<a href="#B36-computers-13-00271" class="html-bibr">36</a>,<a href="#B37-computers-13-00271" class="html-bibr">37</a>,<a href="#B38-computers-13-00271" class="html-bibr">38</a>,<a href="#B39-computers-13-00271" class="html-bibr">39</a>,<a href="#B40-computers-13-00271" class="html-bibr">40</a>,<a href="#B41-computers-13-00271" class="html-bibr">41</a>,<a href="#B42-computers-13-00271" class="html-bibr">42</a>,<a href="#B43-computers-13-00271" class="html-bibr">43</a>,<a href="#B44-computers-13-00271" class="html-bibr">44</a>,<a href="#B45-computers-13-00271" class="html-bibr">45</a>,<a href="#B46-computers-13-00271" class="html-bibr">46</a>,<a href="#B47-computers-13-00271" class="html-bibr">47</a>,<a href="#B48-computers-13-00271" class="html-bibr">48</a>,<a href="#B49-computers-13-00271" class="html-bibr">49</a>,<a href="#B50-computers-13-00271" class="html-bibr">50</a>,<a href="#B51-computers-13-00271" class="html-bibr">51</a>,<a href="#B52-computers-13-00271" class="html-bibr">52</a>,<a href="#B53-computers-13-00271" class="html-bibr">53</a>,<a href="#B54-computers-13-00271" class="html-bibr">54</a>,<a href="#B55-computers-13-00271" class="html-bibr">55</a>,<a href="#B56-computers-13-00271" class="html-bibr">56</a>].</p>
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<p>Galbraith Plot.</p>
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<p>Funnel Plot.</p>
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<p>Bubble Plot per Publication year.</p>
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<p>Bubble Plot for Dataset.</p>
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<p>Publications by Year.</p>
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<p>Publication per Year by Source.</p>
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<p>MCD by Country.</p>
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<p>Bibliometric Analysis on Network Visualization per cited Authors.</p>
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<p>Approaches per year.</p>
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<p>Models per year.</p>
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<p>Publisher per citations.</p>
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16 pages, 1516 KiB  
Article
Association of Ovocalyxin-32 Gene Variants with Egg Quality Traits in Indigenous Chicken Breeds
by Haitham A. Yacoub, Moataz M. Fathi, Ibrahim H. Al-Homidan, Moataz I. Badawy, Mohamed H. Abdelfattah, Mohamed F. Elzarei, Osama K. Abou-Emera and Gamal N. Rayan
Animals 2024, 14(20), 3010; https://doi.org/10.3390/ani14203010 (registering DOI) - 17 Oct 2024
Viewed by 204
Abstract
This study sought to evaluate the genetic variations of the ovocalyxin-32 gene and its association with egg quality traits in indigenous chicken populations, focusing on exons 1 and 6. Genotype frequencies of SNPs (G/T and A/G) within these exons were assessed for their [...] Read more.
This study sought to evaluate the genetic variations of the ovocalyxin-32 gene and its association with egg quality traits in indigenous chicken populations, focusing on exons 1 and 6. Genotype frequencies of SNPs (G/T and A/G) within these exons were assessed for their conformity to the Hardy–Weinberg equilibrium (HWE) across several strains. While most strains exhibited close adherence to HWE expectations, some like light-brown and gray strains indicated substantial discrepancies, particularly for the TT genotype, which points towards the possible effects of genetic drift as well as selection pressures. This study also analyzed the influence of such SNPs on egg quality parameters. A thinner eggshell, reduced shell weight, and decreased breaking strength were associated with the G/T SNP in exon 1, suggesting a likely negative effect on egg quality in T allele carriers. Conversely, the AG genotype displayed better performance in shell thickness, weight and egg weight in the A/G SNP in exon 1, whilst yolk height was best improved by the AA genotype compared to breaking strength. For instance, in exon 6, the A/G SNP enhanced the shell and yolk quality among AG genotypes, while the CC genotype resulted in better eggshell characteristics with enlarged yolks because the C/T SNP was linked. Nonetheless, there were no significant deviations from the HWE despite these associations, which suggested that most breeds had a stable genetic background. Further, considering SNPs’ additive and dominant effects in this research, it was indicated that additive effects account for phenotypic expressions given by the G/T SNP located at exon 1. In contrast, significant additive and dominant effects were observed under the A/G SNP situated at the exon. Generally, it therefore could be concluded from this study that specific SNPs within the ovocalyxin-32 gene may act as good markers for marker-assisted selection (MAS) that can improve desired characteristics—such as those of egg quality—in indigenous chicken breeds. This study demonstrated that both additive and dominance effects must be taken into account when performing genetic analyses, thereby emphasizing the complexity of phenotypic variation caused by genetic mechanisms in native chicken races. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>Genotyping of native chicken strains using DNA sequence of C/T SNP of exon 1 of <span class="html-italic">ovocalyxin-32</span> gene. Three different patterns were detected: (<b>a</b>) GG genotype, (<b>b</b>) TT genotype and (<b>c</b>) GT heterozygous. The arrows indicated the mutation position and type.</p>
Full article ">Figure 1 Cont.
<p>Genotyping of native chicken strains using DNA sequence of C/T SNP of exon 1 of <span class="html-italic">ovocalyxin-32</span> gene. Three different patterns were detected: (<b>a</b>) GG genotype, (<b>b</b>) TT genotype and (<b>c</b>) GT heterozygous. The arrows indicated the mutation position and type.</p>
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<p>Genotyping of native chicken strains using DNA sequence of A/G SNP of exon 1 of <span class="html-italic">ovocalyxin-32</span> gene. Three different genotypes were detected: (<b>a</b>) AA genotype, (<b>b</b>) GG genotype and (<b>c</b>) AG heterozygous. The arrows indicated the mutation position and type.</p>
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<p>Genotyping of native chicken strains using DNA sequence of A/G SNP of Exon 6 of <span class="html-italic">ovocalyxin-32</span> gene. Three different genotypes were detected: (<b>a</b>) GG genotype, (<b>b</b>) AA genotype and (<b>c</b>) AG heterozygous. The arrows indicated the mutation position and type.</p>
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<p>Genotyping of native chicken strains using DNA sequence of second SNP of Exon 6 of <span class="html-italic">ovocalyxin-32</span> gene. The three patterns were detected: (<b>a</b>) CC genotype, (<b>b</b>) TT genotype and (<b>c</b>) CT heterozygous. The arrows indicated the mutation position and type.</p>
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15 pages, 318 KiB  
Article
A Comparison of MLE for Some Index Distributions Based on Censored Samples
by Yunhan Liu, Changchun Gao, Xiaofeng Liu, Ping Luo and Jianguo Ren
Mathematics 2024, 12(20), 3264; https://doi.org/10.3390/math12203264 (registering DOI) - 17 Oct 2024
Viewed by 207
Abstract
This paper elucidates the prerequisites for maximum likelihood estimation (MLE) of parameters within the exponential and scale parameter families. Estimation of these parameters is predicated on data derived from censored samples and seeks to adhere to stochastic ordering principles. The study establishes that [...] Read more.
This paper elucidates the prerequisites for maximum likelihood estimation (MLE) of parameters within the exponential and scale parameter families. Estimation of these parameters is predicated on data derived from censored samples and seeks to adhere to stochastic ordering principles. The study establishes that for two independent normal distributions and a two-parameter exponential distribution discernible by the distinct parameter sets, the MLEs of the parameters evince a stochastically ordered relationship when evaluated using full datasets. Furthermore, this research is extended to corroborate the persistence of stochastic ordering in the MLEs of such parameters under conditions of fixed censoring of samples. Full article
(This article belongs to the Special Issue Decision Making under Uncertainty in Soft Computing)
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<p>Censoring with <math display="inline"><semantics> <mrow> <mi>r</mi> <mo>=</mo> <mn>35</mn> </mrow> </semantics></math>.</p>
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10 pages, 571 KiB  
Article
Total Neoadjuvant Therapy in Locally Advanced Rectal Cancer: Insights from the Western Australian Context
by Oliver Oey, Chak Pan Lin, Muhammad Adnan Khattak, Thomas Ferguson, Mary Theophilus, Siaw Sze Tiong, Sayed Ali and Yasir Khan
Diseases 2024, 12(10), 257; https://doi.org/10.3390/diseases12100257 (registering DOI) - 17 Oct 2024
Viewed by 222
Abstract
Background: Recent studies have associated total neoadjuvant therapy (TNT) with better treatment adherence, decreased toxicity, improved complete clinical response and anal sphincter preservation rates in patients with locally advanced rectal cancer (LARC). However, real-world experience with TNT in the management of LARC remains [...] Read more.
Background: Recent studies have associated total neoadjuvant therapy (TNT) with better treatment adherence, decreased toxicity, improved complete clinical response and anal sphincter preservation rates in patients with locally advanced rectal cancer (LARC). However, real-world experience with TNT in the management of LARC remains limited. Aim: This study aimed to evaluate the efficacy and safety outcomes of TNT for LARC in Western Australia. Methods: Patients with LARC (cT2-4 and/or cN1-2) who underwent induction chemotherapy followed by neoadjuvant chemoradiotherapy or neoadjuvant chemoradiotherapy followed by consolidation chemotherapy, followed by surgery were recruited from two hospitals in Western Australia. Efficacy outcomes assessed included clinical response (complete, partial, no response), and pathologic complete response (pCR) rate, R0 resection rate, and R1 resection rate were evaluated. Those patients who achieved clinical complete response following TNT were given the option of active surveillance. The safety and tolerability of TNT were assessed. Results: 32 patients with LARC were treated with TNT. In total, 17 patients (53%) received chemoradiotherapy followed by consolidation chemotherapy and 15 patients (47%) received induction chemotherapy followed by chemoradiotherapy. Nine (28%) of the patients with LARC treated with TNT had a complete clinical response, twenty-one (66%) patients had a partial clinical response, and two (6%) patients had no response to TNT. Of the 32 patients, 27 (84%) underwent surgery. There was a 100% R0 resection rate. The pCR rate was 15%. pCR, clinical response, and the R0 resection rate were similar between the two TNT regimens. TNT was well tolerated, with the majority of patients (88%) completing the chemotherapy course with grade 1 and 2 adverse effects. Conclusions: In conclusion, TNT emerges as a promising approach for the management of LARC. However, further research is warranted to refine the optimal TNT protocols, determine its long-term outcomes, and identify patient populations who would benefit the most from this innovative therapeutic strategy. Full article
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<p>CONSORT diagram demonstrating the efficacy of TNT.</p>
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12 pages, 846 KiB  
Article
Preimplantation Genetic Testing for Aneuploidy (PGT-A) in In-Vitro Fertilisation (IVF) Treatment: Study Protocol for Pilot Phase of a Randomised Controlled Trial
by Yusuf Beebeejaun, Kypros H. Nicolaides, Anastasia Mania, Ippokratis Sarris and Sesh K. Sunkara
J. Clin. Med. 2024, 13(20), 6192; https://doi.org/10.3390/jcm13206192 - 17 Oct 2024
Viewed by 202
Abstract
Introduction: Poor outcomes following IVF treatments are speculated to be due to the transfer of aneuploid embryos that cannot be identified based on morphological evaluation alone. This leads to patients requiring numerous embryo transfers and, consequently, a prolonged time interval before live [...] Read more.
Introduction: Poor outcomes following IVF treatments are speculated to be due to the transfer of aneuploid embryos that cannot be identified based on morphological evaluation alone. This leads to patients requiring numerous embryo transfers and, consequently, a prolonged time interval before live birth. Embryo selection following preimplantation genetic testing for aneuploidy (PGT-A) with next-generation sequencing (NGS) has been suggested as an intervention to shorten time to pregnancy in women undergoing in vitro fertilisation (IVF). Past studies assessing the clinical efficacy of PGT-A in improving clinical outcomes have been conflicting and the associated clinical pregnancy rates and live birth rates following the transfer of a mosaic embryos have yet to be determined. None of the existing studies solely included women of advanced reproductive age (ARA). The pilot study and proposed RCT will determine if, compared to morphological evaluation alone, the use of PGT-A through NGS is a more clinically effective, safer, and more cost-effective way to provide IVF treatment in women of advanced reproductive age. Method and Analysis: The proposed pilot study will aim to randomise 100 patients within a single-centre study to evaluate recruitment, randomisation, and adherence to study protocol and allocated trail arms by participating patients. The results of the pilot study will enable us to determine the sample size for a larger study to establish the effectiveness of PGT-A in ARA women. Ethics and Dissemination: The study (Integrated Research Application System Number 236067) received approval from the Health Research Authority and Health and Care Research Wales (HCRW) and the East Midlands—Leicester South Research Ethics Committee (20/EM/0290). The results will be made available to patients, the funders, the Reproductive Medicine societies, and other researchers. Trial registration: ClinicalTrials.gov Identifier: NCT05009745, n. Full article
(This article belongs to the Section Reproductive Medicine & Andrology)
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<p>Participant flow chart through PGT-A study.</p>
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12 pages, 2470 KiB  
Article
Triple Therapy in COPD in Real Life: Is It Better to Use Single or Multiple Inhalers?
by Bruno Sposato, Alberto Ricci, Leonardo Gianluca Lacerenza, Elisa Petrucci, Alberto Cresti, Pasquale Baratta, Antonio Perrella, Andrea Serafini and Marco Scalese
J. Clin. Med. 2024, 13(20), 6191; https://doi.org/10.3390/jcm13206191 - 17 Oct 2024
Viewed by 198
Abstract
Background: Today, single-inhaler ICS/LAMA/LABA (SITT) COPD therapies are available. It is unclear whether they are more effective than multiple-device triple therapies (MITT) in improving COPD outcomes. Methods: We retrospectively considered patients on SITT/MITT in 2019/2020 who were prescribed >7 packages of ICS/LABA/LAMA [...] Read more.
Background: Today, single-inhaler ICS/LAMA/LABA (SITT) COPD therapies are available. It is unclear whether they are more effective than multiple-device triple therapies (MITT) in improving COPD outcomes. Methods: We retrospectively considered patients on SITT/MITT in 2019/2020 who were prescribed >7 packages of ICS/LABA/LAMA or ICS/LAMA (+LAMA). The two treatments were compared concerning systemic corticosteroids, antibiotics, salbutamol, antifungal prescriptions, and number of emergency room visits/hospitalizations (ERV/Hs). We studied 292 MITT patients (Group A) during 2019, switching to SITT in 2020, and 366 subjects (Group B) who took SITT during 2019, and 206 MITT individuals (Group C) in 2020. Results: ICS/LABA + LAMA (MITT) package use was 8.2 ± 4.2 and 7.85 ± 4 in 2019, switching to 11.2 ± 3.2 when patients shifted to SITT in 2020 (p = 0.0001). Group A MITT package use was lower than in SITT patients in 2019 (9.31 ± 2.71, p = 0.0001; Group B). Throughout 2020, Group C (10.3 ± 6.1 packs) adherence to ICS/LABA was similar to SITT adherence in Group A (p = 0.270), whereas LAMA package use (9.1 ± 5) was lower (p = 0.0038). Patients using systemic corticosteroids/antibiotics were fewer in SITT in 2020, compared to the MITT results in 2019. Subjects with fewer ERV/Hs were observed with SITT rather than with MITT. Particularly, 13.8% of patients needed ≥2 ERV/Hs when treated with SITT, whereas 19.2% needed ≥2 ERV/Hs with MITT in 2019 (p = 0.08). Multivariate analysis, adjusted for all confounding factors including treatment adherence, showed that MITT (vs. SITT) can have an increased risk of at least one ERV/H (subjects receiving MITT during 2019 passing to SITT in 2020, OR: 1.718 [1.010–2.924], p = 0.046; Group A/MITT/2019 vs. Group B/SITT/2019, OR: 1.650 [0.973–3.153], p = 0.056; Group C/MITT/2020 vs. Group B/SITT/2019, OR: 1.908 [1.018–3.577], p = 0.044). Conclusions: SITT therapy may promote treatment adherence more effectively, therefore, reducing COPD exacerbations better than MITT. A possible synergistic effect of ICS/LABA/LAMA intake with a single device might be another cause of SITT’s greater efficacy. Full article
(This article belongs to the Section Pulmonology)
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<p>Study protocol. Patients treated with triple therapy with multiple devices (MITT) and with triple therapy with single devices (SITT). Inhaled corticosteroids (ICS), long-acting β2 agonist (LABA) and long-acting muscarinic antagonist (LAMA). SITT: ICS/LABA/LAMA (single inhaler); MITT: ICS/LABA + LAMA (triple therapy consisting of two devices, one for ICS/LABA and the other for LAMA).</p>
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<p>Adherence (number of packages/year) observed in the different groups treated with triple therapy with multiple devices (MITT) and with triple therapy with single devices (SITT) in 2019 or 2020.</p>
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<p>Percentages of subjects who had had at least one prescription of systemic corticosteroids, antibiotics, SABA (short-acting β2-agonist), and antifungal packages during 2019 and 2020 in the various groups considered.</p>
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<p>Percentages of subjects who had one or at least two emergency department visits/hospitalizations during one year of MITT or SITT therapy (2019 or 2020) in the various groups considered.</p>
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<p>Logistic regression models that compared the effects (on different outcomes) of the treatment with triple therapy with multiple inhalers performed in 2019 and triple therapy with single inhalers prescribed in 2020 in the same subjects (Group A); The figure shows the odds ratios with a 95% confidence interval (OR [95% CI]).</p>
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<p>Logistic regression models that compared the effects (on different outcomes) of the treatment with triple therapy with multiple inhalers prescribed in Group A in 2019 and triple therapy with single inhalers in Group B in 2019; The figure shows the odds ratios with a 95% confidence interval (OR [95% CI]).</p>
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<p>Logistic regression models that compared the effects (on different outcomes) of the treatment with triple therapy with single inhalers prescribed in Group B in 2019 and triple therapy with multiple inhalers in Group C in 2020; The figure shows the odds ratios with a 95% confidence interval (OR [95% CI]).</p>
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22 pages, 356 KiB  
Article
“The Battle for Men’s Minds”: Subliminal Message as Conspiracy Theory in Seventh-Day Adventist Discourse
by Allan Novaes
Religions 2024, 15(10), 1276; https://doi.org/10.3390/rel15101276 - 17 Oct 2024
Viewed by 243
Abstract
This article describes the presence of a subliminal thesis—with conspiratorial and apocalyptic content—in the discourse of the Seventh-day Adventist tradition based on a documentary analysis of Adventist publications from the 1900s to the 1990s. The history of the development of this thesis is [...] Read more.
This article describes the presence of a subliminal thesis—with conspiratorial and apocalyptic content—in the discourse of the Seventh-day Adventist tradition based on a documentary analysis of Adventist publications from the 1900s to the 1990s. The history of the development of this thesis is classified into three periods: (1) Proto-Adventist Subliminal Thesis, from 1900s to 1940s, with a discourse of anti-spiritualist emphasis; (2) Adventist Subliminal Thesis’ First Wave, from 1950s to 1960s, with a discourse of anti-media emphasis in the context of James Vicary’s experiments in the 1950s; and (3) Adventist Subliminal Thesis’ Second Wave, from 1970s to 1990s, with a discourse of conspiratorial emphasis in the context of the satanic panic of the 1980s and 1990s. The Adventist subliminal thesis is configured in a way of thinking that considers (1) the human being as a “mass-man” and culture as “mass culture”; (2) the media as having the power of manipulation and mental control; (3) adherence to moral panic phenomena as reactions to media threats to traditional values; and (4) the cosmic narrative of the Great Controversy as a worldview for understanding media messages and products as part of a satanic conspiracy. Full article
(This article belongs to the Special Issue Contemporary Religion, Media and Popular Culture)
9 pages, 1702 KiB  
Article
Sub-Optimal Compliance to Long-Term Inhalation Strategies and Poorer Health Care Outcomes Associated with Extended Tattoos in Adolescents with Mild-to-Moderate Bronchial Asthma
by Roberto W. Dal Negro, Paola Turco and Massimiliano Povero
Children 2024, 11(10), 1254; https://doi.org/10.3390/children11101254 - 17 Oct 2024
Viewed by 180
Abstract
Background: Patients’ compliance to inhalation therapy is usually sub-optimal in young asthmatics. Adolescents poorly compliant to regular anti-asthma treatments and those with tattoos (and the associated attitude) can share some personality traits and maladaptive behaviors. This relationship has not been investigated. Objective: To [...] Read more.
Background: Patients’ compliance to inhalation therapy is usually sub-optimal in young asthmatics. Adolescents poorly compliant to regular anti-asthma treatments and those with tattoos (and the associated attitude) can share some personality traits and maladaptive behaviors. This relationship has not been investigated. Objective: To assess if “extended” tattoos can predict long-term compliance to regular therapy of adolescents with mild-to-moderate asthma. Methods: A 12-month retrospective observational investigation was conducted on non-smoker asthmatic adolescents of both genders. Patients assuming <70% of prescribed vilanterol/fluticasone furoate o.d. were defined as “non-compliant”. Tattoo surfaces were defined as “mild” or “extended” if they were < or ≥400 cm2, respectively. The relationship between tattoos and compliance on the evolution of resources consumption at 6 and 12 months was assessed by generalized estimating equation (GEE) models at the first and second semester of the treatment period. Results: It was found that 13.2% of compliant adolescents had mild tattoos, while 47.2% of non-compliant adolescents had mild-to-extended tattoos (odds ratio (OR) 6.91, 95% CI 2.49 to 19.17, p < 0.001). The mean annual adherence to treatment was 57.8% ± 10.1 SD expected doses in non-compliant subjects with “mild tattoos” (54.8 cm2 ± 36.9 SD), but 38.6% ± 11.4 SD expected doses in those with “extended tattoos” (568.4 cm2 ± 111.6 SD, p < 0.001). Total cost proved to be a linear trend from the lowest values of compliant patients with no/mild tattoos (EUR 65.22 at 6 months and EUR 33.63 at 12 months) to the highest values of non-compliant adolescents with extended tattoos (EUR 330.75 at 6 months and EUR 297.34 at 12 months). Conclusions: Tattoo extension might be used as a reliable predictor of poor compliance and higher health care costs in adolescents with mild-to-moderate asthma. Patients characterized by poor compliance to a long-term therapeutic strategy and tattooing attitude likely share some aspects of their personality profile. Full article
(This article belongs to the Special Issue Asthma and Its Impact in Adolescent: 2nd Edition)
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<p>Distribution of tattoos among complaint and non-compliant adolescents.</p>
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<p>Time series for visits and hospitalizations cost (from baseline up to 12 months) stratified by compliance and tattoos prevalence.</p>
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<p>Time series for drugs cost (from baseline up to 12 months) stratified by compliance and tattoos prevalence.</p>
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<p>Time series for cost due to productivity loss (from baseline up to 12 months) stratified by compliance and tattoos prevalence.</p>
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<p>Time series for total cost (from baseline up to 12 months) stratified by compliance and tattoos prevalence.</p>
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16 pages, 989 KiB  
Article
A Prospective Study of the Medication Regimen Complexity Index and Hospitalization Due to Adverse Drug Reactions Among People Living with HIV
by Basavaraj Poojar, Ashwin Kamath, Sathish B. Rao, Sheetal Dinkar Ullal, John Ramapuram, Muralidhar B. Yadiyal and Ashok K. Shenoy
Medicina 2024, 60(10), 1705; https://doi.org/10.3390/medicina60101705 (registering DOI) - 17 Oct 2024
Viewed by 221
Abstract
Background and Objective: The complexity of antiretroviral therapy (ART) regimens in people living with HIV (PLHIV) poses significant challenges for medication management, impacting adherence and overall health outcomes. The Medication Regimen Complexity Index (MRCI) is a tool that quantifies regimen complexity, yet [...] Read more.
Background and Objective: The complexity of antiretroviral therapy (ART) regimens in people living with HIV (PLHIV) poses significant challenges for medication management, impacting adherence and overall health outcomes. The Medication Regimen Complexity Index (MRCI) is a tool that quantifies regimen complexity, yet its correlation with hospitalization rates and adverse drug reactions (ADRs) in PLHIV remains underexplored. Materials and Methods: This prospective study, which was conducted at a government-funded antiretroviral treatment center, investigated the relationships among MRCI scores, hospitalization due to ADRs, and the ADR rates in 285 PLHIV participants over 18 months. Results: The study revealed a significant association between higher baseline MRCI scores and hospitalization due to ADRs, with a threshold MRCI score of 8 indicating increased risk. There was no significant association between average MRCI scores and overall ADR rates or non-ADR-related hospitalizations. Conclusions: These findings emphasize the importance of monitoring medication regimen complexity in PLHIV, particularly in the context of preventing hospitalizations related to ADRs. Further research is needed to understand the multifactorial influences on ADR occurrence and to optimize ART regimens for better patient outcomes. Full article
(This article belongs to the Section Pharmacology)
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<p>Study flow chart.</p>
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<p>Scatter plot of the correlation between the average medication regimen complexity index and the total number of adverse drug reactions.</p>
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17 pages, 459 KiB  
Review
Iron Deficiency Anaemia in Pregnancy: A Narrative Review from a Clinical Perspective
by Chidi Obianeli, Khaled Afifi, Simon Stanworth and David Churchill
Diagnostics 2024, 14(20), 2306; https://doi.org/10.3390/diagnostics14202306 - 17 Oct 2024
Viewed by 284
Abstract
Anaemia in pregnancy is a global problem of significance in all settings. The most common cause is iron deficiency. Large numbers of women are affected, ranging up to 25–30% antenatally and 20–40% postnatally. It is associated with serious adverse outcomes for both the [...] Read more.
Anaemia in pregnancy is a global problem of significance in all settings. The most common cause is iron deficiency. Large numbers of women are affected, ranging up to 25–30% antenatally and 20–40% postnatally. It is associated with serious adverse outcomes for both the mother and her baby. The risk of low birth weight, preterm birth, postpartum haemorrhage, stillbirth, and neonatal death are all increased in the presence of anaemia. For the infants of affected pregnancies, complications may include neurocognitive impairment. Making an accurate diagnosis during pregnancy has its challenges, which include the choice of thresholds of haemoglobin below which a diagnosis of anaemia in each trimester of pregnancy can be made and, aligned with this question, which are the most appropriate biomarkers to use to define iron deficiency. Treatment with oral iron supplements increases the haemoglobin concentration and corrects iron deficiency. But high numbers of women fail to respond, probably due to poor adherence to medication, resulting from side effects. This has resulted in an increased use of more expensive intravenous iron. Doubts remain about the optimal regimen to of oral iron for use (daily, alternate days, or some other frequency) and the cost-effectiveness of intravenous iron. There is interest in strategies for prevention but these have yet to be proven clinically safe and effective. Full article
(This article belongs to the Special Issue Laboratory Medicine: Extended Roles in Healthcare Delivery)
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<p>With permission. The U-shaped curve of iron’s adequacy range compared with other nutrients. The blue line indicates the wide range for which most nutrients doses are considered to be adequate. The range for iron, shown by the red line, is narrower and can with much smaller variation in dose easily fall into deficient or toxic ranges [<a href="#B51-diagnostics-14-02306" class="html-bibr">51</a>].</p>
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19 pages, 486 KiB  
Article
Parents’ Knowledge and Attitudes Toward Pediatric Ophthalmic Disorders in Saudi Arabia: A Cross-Sectional Study
by Saja Radhi G. Alanazi, Haneen Wadi H. Alanazi, Wasan Ghathwan Alanazi, Nawal Surhuj Q. Alanazi, Dareen Owaid B. Alenezi, Maisa Al-Sweilem, Maram Hassan Alqattan, Iftikhar Lafi N. Alanazi, Jumanah Mohammed Tirksstani, Reem Saeed AlSarhan, Saleh Ghulaysi, Hanan A. Elgendy and Manal S. Fawzy
Pediatr. Rep. 2024, 16(4), 902-920; https://doi.org/10.3390/pediatric16040077 - 17 Oct 2024
Viewed by 218
Abstract
Recognizing pediatric eye health issues at an early stage, along with ensuring that parents are well informed, is crucial. This study measures parents’ knowledge and perspectives on pediatric ophthalmic disease. The study utilized a cross-sectional design, and participants included Saudi parents of children [...] Read more.
Recognizing pediatric eye health issues at an early stage, along with ensuring that parents are well informed, is crucial. This study measures parents’ knowledge and perspectives on pediatric ophthalmic disease. The study utilized a cross-sectional design, and participants included Saudi parents of children residing in Saudi Arabia. Data were collected using a pre-validated self-administered questionnaire with a scoring system. Sociodemographic characteristics and factors associated with knowledge/attitude were collected and analyzed. Initially, 425 respondents participated in this study. Excluding the participants whose work was related to healthcare and those with incomplete data yielded 370 responses on which the subsequent analyses were performed. The analysis revealed that only half of the parents recognized the need for annual eye examinations for children. Most of them showed gaps in knowledge regarding the frequency of routine eye exams and indicators of visual problems. Notably, parents with good/excellent knowledge were more proactive in their eye care practices, such as adherence to recommended eye examination schedules and accepting corrective measures like glasses for common conditions such as refractive errors and amblyopia. However, parental willingness to permit surgical interventions did not correlate significantly with their level of knowledge, signaling the influence of other factors. In conclusion, this study underscores the need for enhanced public health education to improve parental awareness of pediatric eye diseases in Saudi Arabia. Given the link between knowledge and proactive eye health practices, targeted interventions should distribute comprehensive, culturally sensitive information accessible to all demographics. Full article
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<p>The overall parents’ knowledge level of pediatric ophthalmic diseases. Data are presented as proportions (%).</p>
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Article
Interpretable Machine Learning: A Case Study on Predicting Fuel Consumption in VLGC Ship Propulsion
by Aleksandar Vorkapić, Sanda Martinčić-Ipšić and Rok Piltaver
J. Mar. Sci. Eng. 2024, 12(10), 1849; https://doi.org/10.3390/jmse12101849 (registering DOI) - 16 Oct 2024
Viewed by 437
Abstract
The integration of machine learning (ML) in marine engineering has been increasingly subjected to stringent regulatory scrutiny. While environmental regulations aim to reduce harmful emissions and energy consumption, there is also a growing demand for the interpretability of ML models to ensure their [...] Read more.
The integration of machine learning (ML) in marine engineering has been increasingly subjected to stringent regulatory scrutiny. While environmental regulations aim to reduce harmful emissions and energy consumption, there is also a growing demand for the interpretability of ML models to ensure their reliability and adherence to safety standards. This research highlights the need to develop models that are both transparent and comprehensible to domain experts and regulatory bodies. This paper underscores the importance of transparency in machine learning through a use case involving a VLGC ship two-stroke propulsion engine. By adhering to the CRISP-DM standard, we fostered close collaboration between marine engineers and machine learning experts to circumvent the common pitfalls of automated ML. The methodology included comprehensive data exploration, cleaning, and verification, followed by feature selection and training of linear regression and decision tree models that are not only transparent but also highly interpretable. The linear model achieved an RMSE of 23.16 and an MRAE of 14.7%, while the accuracy of decision trees ranged between 96.4% and 97.69%. This study demonstrates that machine learning models for predicting propulsion engine fuel consumption can be interpretable, adhering to regulatory requirements, while still achieving adequate predictive performance. Full article
(This article belongs to the Special Issue Smart Seaport and Maritime Transport Management)
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<p>Spearman correlations between reduced set of variables.</p>
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<p>Feature importance for reduced set of variables according to correlation, genetic, and ReliefF methods.</p>
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<p>Dots represent the average predicted fuel consumption over the 1000 bootstrapped models for each sample in the test set. Error bars represent the range that contains 95% predictions. Distance from the diagonal line indicates the prediction error.</p>
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<p>Classification tree that predicts fuel consumption classes a to i. Bold text represents the splitting criterion, letter represents the predicted class in each node and the numbers represent the number of training data samples belonging to the predicted class vs. all samples belonging to the node.</p>
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<p>Pruned J48/C4.5 tree for shaft revolutions and fuel consumption variables and classes a to i.</p>
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<p>Pruned J48/C4.5 tree for shaft power and fuel consumption variables (classes a to i).</p>
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<p>Pairwise parameter scatter plots and Pearson correlations, colors correspond to the dates when data was collected.</p>
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<p>Slip and shaft power vs. fuel consumption.</p>
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<p>Feature selection on all variables using correlation, genetic and RelieF methods. Figure shows that the simple correlation-based feature selection method fails in case of multiple correlated features (e.g., same parameter measured at each of the 6 engine cylinders) and that data understanding is the key for removing such redundant features (i.e., understanding which features should be removed and why). Furthermore, it demonstrates that advanced feature selection methods such as ReliefF can narrow down the set of useful features much better even when many redundant features are present.</p>
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Article
Micro RNA-175 Targets Claudin-1 to Inhibit Madin–Darby Canine Kidney Cell Adhesion
by Xiaoyun Li, Fangfang Ma, Siya Wang, Tian Tang, Liyuan Ma, Zilin Qiao, Zhongren Ma, Jiamin Wang and Zhenbin Liu
Genes 2024, 15(10), 1333; https://doi.org/10.3390/genes15101333 - 16 Oct 2024
Viewed by 272
Abstract
Background: The Madin–Darby canine kidney (MDCK) cell line constitutes a key component of influenza vaccine production, but its dependence on adherent growth limits cell culture density and hinders vaccine yield. There is evidence that the use of gene editing techniques to inhibit cell [...] Read more.
Background: The Madin–Darby canine kidney (MDCK) cell line constitutes a key component of influenza vaccine production, but its dependence on adherent growth limits cell culture density and hinders vaccine yield. There is evidence that the use of gene editing techniques to inhibit cell adhesion and establish an easily suspended cell line can improve vaccine yield; however, the mechanisms underlying MDCK cell adhesion are unclear. Methods: In this study, we used transcriptomics to analyse differentially expressed mRNAs and miRNAs in adherent and suspension cultures of MDCK cells. Results: We found that claudin-1 (CLDN1) expression was downregulated in the suspension MDCK cells and that CLDN1 promotes MDCK cell–extracellular matrix adhesion. Additionally, microRNA (miR)-175 expression was upregulated in the suspension MDCK cells. Importantly, we demonstrated that miR-175 inhibits MDCK cell adhesion by targeting the CLDN1 3′-untranslated region (UTR). These findings contribute to a more comprehensive understanding of the regulatory mechanisms modulating cell adhesion and provide a basis for establishing suspension-adapted, genetically engineered cell lines. Our work could also facilitate the identification of targets for tumour therapy. Full article
(This article belongs to the Special Issue RNAs in Biology)
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<p>Volcano plot and heatmap of differentially expressed mRNAs and miRNAs in XF04 compared with M60. (<b>A</b>) mRNA volcano plot: Light green spots indicate downregulated expression, grey spots indicate not differentially expressed, and red spots indicate upregulated expression. Total number of mRNAs: red column indicates upregulated expression and blue column indicates downregulated expression. (<b>B</b>) miRNA volcano plot: Light green spots indicate downregulated expression, dark spots indicate not differentially expressed, and red spots indicate upregulated expression. Total number of miRNAs: red column indicates upregulated expression and blue column indicates downregulated expression. (<b>C</b>) mRNA heatmap: Each column represents a sample, and each row represents an mRNA. mRNA expression levels in different samples are represented by different colours. Red indicates higher expression, whereas blue indicates lower expression. Light green denotes M60, and pink denotes XF04. (<b>D</b>) miRNA heatmap: Each column represents a sample, and each row represents an miRNA. miRNA expression levels in different samples are represented by different colours. Red indicates higher expression, whereas blue indicates lower expression. Light green denotes M60 (the adherent MDCK cell), and pink denotes XF04 (the suspension MDCK cell).</p>
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<p>GO and KEGG functional analyses of all differentially expressed genes. (<b>A</b>) GO functional annotation histogram of all differentially expressed genes. GO terms were ranked according to number of genes across biological processes (green), cellular components (red), and molecular functions (blue). (<b>B</b>) KEGG pathway assessment of all differentially expressed genes. Lower <span class="html-italic">p</span>-values indicate greater pathway enrichment. Black circles represent gene numbers; a larger circle indicates a higher number of genes. (<b>C</b>) GO functional annotation histogram of adhesion-related differentially expressed mRNAs. GO terms were ranked according to number of genes across biological processes (green), cellular components (red), and molecular functions (blue). (<b>D</b>) KEGG pathway assessment of adhesion-related differentially expressed mRNAs. Lower <span class="html-italic">p</span>-values indicate greater pathway enrichment. Black circles represent gene numbers; a larger circle indicates a higher number of genes.</p>
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<p>CLDN1 expression was downregulated in the suspension MDCK cells. A fluorescent reverse transcription–quantitative PCR was performed to detect the relative mRNA expression levels of adhesion-related genes identified via transcriptomics. (<b>A</b>,<b>B</b>) Genes with downregulated expression in XF04 (the suspension MDCK cell) compared with M60 (The adherent MDCK cell), and (<b>C</b>,<b>D</b>) genes with upregulated expression in XF04 compared with M60. (<b>E</b>) Western blotting detection of CLDN1 protein expression in XF04, and (<b>F</b>) relative expression of CLDN1 protein in XF04. Red, M60 control group; blue, XF04 experimental group. Data are expressed as ±standard deviation (SD). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.001, *** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>CLDN1 knockdown inhibits MDCK cell adhesion. (<b>A</b>) The RT-qPCR analysis to verify the expression of <span class="html-italic">CLDN1</span> mRNA in the sh-CLDN1 cells, and (<b>B</b>) the Western blotting analysis to verify the expression of CLDN1 protein in the sh-CLDN1 cells. (<b>C</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 knockdown on MDCK cell adhesion to the extracellular matrix protein fibronectin (FN); <b>middle</b>, fluorescence microscopy to detect the effect of CLDN1 knockdown on MDCK cell adhesion to FN; <b>right</b>, cell counting to detect the effect of CLDN1 knockdown on MDCK cell adhesion to FN. (<b>D</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 knockdown on MDCK cell adhesion to the extracellular matrix protein collagen I (CL); <b>middle</b>, fluorescence microscopy to detect the effect of CLDN1 knockdown on MDCK cell adhesion to CL; <b>right</b>, cell counting to detect the effect of CLDN1 knockdown on MDCK cell adhesion to CL. (<b>E</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 knockdown on MDCK cell adhesion to the extracellular matrix protein laminin (LN); <b>middle</b>, fluorescence microscopy to detect the effect of CLDN1 knockdown on MDCK cell adhesion to LN; <b>right</b>, cell counting to detect the effect of CLDN1 knockdown on MDCK cell adhesion to LN. Blue denotes CLDN1 knockdown cells (sh-CLDN1); red denotes empty vector control cells (sh-CON). Data are expressed as ±standard deviation. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>CLDN1 overexpression promotes MDCK cell adhesion. (<b>A</b>) The RT-qPCR analysis to verify the expression of <span class="html-italic">CLDN1</span> mRNA in the OE-CLDN1 cells, (<b>B</b>) lentiviral transfection for the fluorescence-based verification of expression, and (<b>C</b>) the Western blotting analysis to verify the expression of CLDN1 protein in the OE-CLDN1 cells. (<b>D</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 overexpression on MDCK cell adhesion to the extracellular matrix protein fibronectin (FN); <b>middle</b>, fluorescence microscopy to detect the effect of CLDN1 overexpression on MDCK cell adhesion to FN; <b>right</b>, cell counting to detect the effect of CLDN1 overexpression on MDCK cell adhesion to FN. (<b>E</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 overexpression on MDCK cell adhesion to the extracellular matrix protein collagen I (CL); <b>middle</b>, fluorescence microscopy assay to detect the effect of CLDN1 overexpression on MDCK cell adhesion to CL; <b>right</b>, cell counting to detect the effect of CLDN1 overexpression on MDCK cell adhesion to CL. (<b>F</b>) <b>Left</b>, CCK8 assay to detect the effect of CLDN1 overexpression on MDCK cell adhesion to the extracellular matrix protein laminin (LN); <b>middle</b>, fluorescence microscopy to detect the effect of CLDN1 overexpression on MDCK cell adhesion to LN; <b>right</b>, cell counting to detect the effect of CLDN1 overexpression on MDCK cell adhesion to LN. Blue denotes CLDN1 knockdown cells (OE-CLDN1); red denotes empty vector control cells (OE-CON). Data are expressed as ± standard deviation. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>CLDN1 promotes MDCK cell proliferation and migration. (<b>A</b>) Cell counting was performed to determine the effects of the <span class="html-italic">CLDN1</span> knockdown and overexpression on MDCK cell proliferation. (<b>B</b>) The effects of <span class="html-italic">CLDN1</span> knockdown and overexpression on MDCK cell colony formation were determined by colony formation assays. (<b>C</b>,<b>D</b>) The effects of <span class="html-italic">CLDN1</span> knockdown and overexpression on MDCK cell migration were detected by scratch assays. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>miR-175 directly targets the CLDN1 3′-UTR. (<b>A</b>) The RT-qPCR analysis to verify the expression of <span class="html-italic">CLDN1</span>-associated miRNAs. (<b>B</b>) The RT-qPCR analysis to verify the relative expression of miR-175. (<b>C</b>) The Western blotting analysis to detect miR-175 overexpression and relative protein expression of CLDN1 in functional rescue cells. (<b>D</b>) The RT-qPCR analysis to detect miR-175 overexpression and relative protein expression of CLDN1 in functionally rotating cells. (<b>E</b>) The schematic representation of the conserved miR-175 binding site within the <span class="html-italic">CLDN1</span> 3′-UTR and its potential interactions with the miR-175 sequence. (<b>F</b>) The dual luciferase reporter gene assay to detect the targeting relationship between miR-175 and <span class="html-italic">CLDN1</span>. miR-175 mimic or miR mimic control (mimic NC) were co-transfected with HEK293 cells for 48 h, and luciferase reporter gene activity was measured. All the measurements were performed in triplicate, and the experiment was repeated three times. Data are expressed as ±standard deviation. ns <span class="html-italic">p</span> &gt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>miR-175 inhibits MDCK cell adhesion by targeting <span class="html-italic">CLDN1</span>. (<b>A</b>) <b>Left</b>, 96-well plates were coated with fibronectin (FN) and the effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell adhesion were detected by CCK8 assays; centre, 24-well plates were coated with FN and adherent cells were detected by fluorescence microscopy; <b>right</b>, adherent cells were counted in 24-well plates coated with FN. (<b>B</b>) <b>Left</b>, 96-well plates were coated with collagen I (CL) and the effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell adhesion were detected by CCK8 assays; centre, 24-well plates were coated with CL and adherent cells were detected by fluorescence microscopy; <b>right</b>, 24-well plates were coated with CL and adherent cells were counted. (<b>C</b>) <b>Left</b>, 96-well plates were coated with laminin (LN) and the effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell adhesion were detected by CCK8 assays; centre, 24-well plates were coated with LN and adherent cells were detected by fluorescence microscopy; <b>right</b>, 24-well plates were coated with LN and adherent cells were counted. Orange, miR-175 overexpression control cells (miR-OECON); green, miR-175 overexpression cells (miR-175OE); blue, target gene CLDN1 rescue control cells (miR-175OE + CLDN1-OECON); purple, target gene CLDN1 rescue cells (miR-175OE + CLDN1-OE). Data are expressed as ± standard deviation. ns <span class="html-italic">p</span> &gt; 0.05, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>miR-175 inhibits MDCK cell proliferation and migration. (<b>A</b>) The effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell proliferation were determined by cell counting. (<b>B</b>) The effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell colony formation were determined by colony formation assays. (<b>C</b>) The effects of miR-175OE or miR-175OE + CLDN1-OE on MDCK cell migration were determined by cell scratch. Orange, miR-175 overexpression control cells (miR-OECON); green, miR-175 overexpression cells (miR-175OE); blue, target gene CLDN1 rescue control cells (miR-175OE + CLDN1-OECON); purple, target gene <span class="html-italic">CLDN1</span> rescue cells (miR-175OE + CLDN1-OE). Data are expressed as ± standard deviation. ns <span class="html-italic">p</span> &gt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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