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Search Results (1,509)

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7 pages, 203 KiB  
Editorial
Disclosing the Complexities of Childhood Neurodevelopmental Disorders
by Luigi Tarani and Marco Fiore
Children 2025, 12(1), 16; https://doi.org/10.3390/children12010016 - 25 Dec 2024
Viewed by 19
Abstract
Neurodevelopmental disorders represent an important and complex area of pediatric medicine, including a wide range of conditions affecting brain and nervous system functioning during development [...] Full article
(This article belongs to the Special Issue Neurodevelopmental Disorders in Pediatrics)
12 pages, 1673 KiB  
Article
Effects on Posture of a Two-Diopter Horizontal Prism Base Out on the Non-Dominant Eye
by Davide Marini, Giovanni Rubegni, Lorenzo Sarti, Alessandra Rufa, Marco Mandalà, Fabio Ferretti, Gian Marco Tosi and Mario Fruschelli
J. Clin. Med. 2024, 13(24), 7847; https://doi.org/10.3390/jcm13247847 - 23 Dec 2024
Viewed by 281
Abstract
Background/Objectives: Ocular proprioception is implicated in balance control and heterophoria is associated with abnormal posture, though previous research focused mainly on the role of vertical phoria and the use of vertical prisms. This study aims to evaluate whether ocular misalignment and prismatic [...] Read more.
Background/Objectives: Ocular proprioception is implicated in balance control and heterophoria is associated with abnormal posture, though previous research focused mainly on the role of vertical phoria and the use of vertical prisms. This study aims to evaluate whether ocular misalignment and prismatic correction of horizontal phoria affect posture. Methods: Sixty-nine (N = 69) young healthy subjects were included and equally divided by horizontal distance phoria: orthophoria (n = 23), esophoria (n = 23) and exophoria (n = 23). A prism of low power (two-diopter) was placed base out on the non-dominant eye, reducing misalignment in esophorics and increasing it in exophorics more than in orthophorics. Dynamic computerized posturography was performed with the sensory organization test protocol (SOT) of the EquiTest® NeuroCom® version 8 platform both without and with prism, always maintaining subjects unaware of prism use. A mixed model for repeated measures analysis of variance was run to evaluate the main effect of prism and the interaction effect of prism with baseline phoria. Results: Composite movement strategy score without prism was 88.1 ± 2.8% (ankle-dominant strategy) and slightly increased to 89.0 ± 3.1% with prism insertion (p = 0.004), further shifting toward ankle strategy. Composite equilibrium score without prism was 80.3 ± 6.5% and remained stable with prism insertion (81.3 ± 8.2%, p = 0.117), medio-lateral and antero-posterior projection of center of gravity did not displace significantly under prism insertion (p = 0.652 and p = 0.270, respectively). At baseline, posturographic parameters were statistically independent of individual phoria, and no significant interaction between prism insertion and individual phoria was documented for any parameters (p > 0.05 for all). Secondary analysis and pairwise comparisons confirmed that the effect of prism was strongly selective on condition SOT 5 (eyes-closed, platform sway-referenced) with improvement of equilibrium (70.4 ± 9.7% with prism vs. 65.7 ± 11.6% without) and more use of ankle strategy (81.6 ± 5.3% with prism vs. 78.2 ± 6.0% without), without any interaction of phoria and ocular dominance, while the other conditions were comparable with and without prism. Conclusions: A two-diopter prism base out on the non-dominant eye induces the body to use the ankle joint more independently of individual phoria, suggesting a small improvement in postural control, while maintaining oscillations of the center of gravity unaltered. Prism seems to enhance the function of vestibular system selectively. Phoria adjustments with prismatic correction enable intervention in postural behavior. Extraocular muscles could act as proprioceptors influencing postural stability. Full article
(This article belongs to the Section Ophthalmology)
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Figure 1
<p>Distributions of horizontal distance phoria (Δ, prismatic diopter) and stereoacuity (log arcsec) <span class="html-italic">without</span> and <span class="html-italic">with prism</span>. Column and bar respectively represent median and interquartile range of phoria, and mean and standard error of stereoacuity. Wilcoxon signed-rank test statistical significance (<span class="html-italic">P</span>) and effect size (<span class="html-italic">r</span>) showed a significant exophoric shift in all groups after prism insertion. Two-way ANOVA statistical significance (<span class="html-italic">P</span>) and partial effect size (η<sup>2</sup>) of main effect (<span class="html-italic">Prism</span>) and interaction (<span class="html-italic">Prism × Phoria</span>) showed a significant small increase in logarithm of stereoacuity seconds of arc for all groups.</p>
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<p>Composite equilibrium (CES) and composite movement strategy scores (CMS) <span class="html-italic">without</span> (at baseline) and <span class="html-italic">with prism</span>, sorted by <span class="html-italic">baseline phoria</span>. A higher CSS denotes a better performance (less sway); a higher CMS indicates a more predominant ankle strategy (versus hip). Dot and bar represent mean and standard error, respectively. Two-way ANOVA statistical significance (<span class="html-italic">P</span>) and partial effect size (η<sup>2</sup>) of main effect (<span class="html-italic">Prism</span>) and interaction (<span class="html-italic">Prism × Phoria</span>) showed only a significant shift of CMS toward ankle strategy for all groups.</p>
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<p>Medio-lateral (ML/Px) and antero-posterior (AP/Py) projection of center of gravity (COG) <span class="html-italic">without</span> (at baseline) and <span class="html-italic">with prism</span>, sorted by <span class="html-italic">baseline phoria</span>. Right and forward displacements are positive, left and backward negative. Dot and bar represent mean and standard error, respectively. Two-way ANOVA statistical significance (<span class="html-italic">P</span>) and partial effect size (η<sup>2</sup>) of main effect (<span class="html-italic">Prism</span>) and interaction (<span class="html-italic">Prism × Phoria</span>) showed no significant displacement of COG, though COG-Px displacement of exophoric subjects was divergent (rightward) from esophoric and orthophoric subjects (leftward).</p>
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8 pages, 730 KiB  
Article
Efficacy and Safety of Intra-Class Switching from Ixekizumab to Secukinumab in Patients with Plaque Psoriasis: A Multicenter Retrospective Study
by Nicoletta Bernardini, Annunziata Dattola, Raimondo Rossi, Gianluca Pagnanelli, Paolo Amerio, Laura Atzori, Cristina Mugheddu, Viviana Lora, Domenico Giordano, Lucia Finistauri Guacci, Severino Persechino, Antonio Giovanni Richetta, Nevena Skroza and Concetta Potenza
J. Pers. Med. 2024, 14(12), 1169; https://doi.org/10.3390/jpm14121169 - 22 Dec 2024
Viewed by 246
Abstract
Background: the present multicenter retrospective study aimed to evaluate the efficacy and safety of intra-class switching between interleukin-17A (IL-17A) inhibitors, specifically from ixekizumab to secukinumab, in patients with plaque psoriasis. Methods: this study included 11 patients (6 male, 5 female) who had previously [...] Read more.
Background: the present multicenter retrospective study aimed to evaluate the efficacy and safety of intra-class switching between interleukin-17A (IL-17A) inhibitors, specifically from ixekizumab to secukinumab, in patients with plaque psoriasis. Methods: this study included 11 patients (6 male, 5 female) who had previously received ixekizumab and then were switched to secukinumab. Patients’ PASI, DLQI, and pain VAS (in those with psoriatic arthritis) were evaluated at weeks 16, 24, 54, and 98. Results: PASI-90 was reached in 10 (91%) cases at week 24. One patient experienced secondary failure to secukinumab at week 98. No adverse events were reported. Conclusions: overall, the majority of patients experienced a favorable response to secukinumab, suggesting that it may be an effective treatment option for patients with an inadequate response or loss of efficacy to ixekizumab. Full article
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<p>Flowchart summarizing the allocation of patients throughout the various phases of the study. Anti-IL17: anti-interleukin 17; IXE: ixekizumab; SEC: secukinumab.</p>
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<p>Mean PASI, DLQI and pain VAS trend throughout the study period. μ: mean; PASI: psoriasis Area and Severity Index; DLQI: Dermatology Life Quality Index; VAS: Visual Analog Scale; SD: standard deviation.</p>
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10 pages, 281 KiB  
Article
Single-Leg Balance and Lower Limb Strength: Quantitative Analysis with the Balance Master System
by José Manuel Delfa-de-la-Morena, Pedro Pinheiro Paes, Débora Priscila Lima de Oliveira, Frederico Camarotti Júnior, Bruna Daniele Monteiro Lima, Miriam García-González, Juan-José Mijarra-Murillo and Víctor Riquelme-Aguado
J. Funct. Morphol. Kinesiol. 2024, 9(4), 282; https://doi.org/10.3390/jfmk9040282 - 21 Dec 2024
Viewed by 244
Abstract
Objective: This study investigates the relationship between lower limb strength and postural stability in single-leg stance using the Balance Master system. Methods: The research involved 64 participants divided into sedentary and physically active groups based on metabolic equivalents of task (METs) [...] Read more.
Objective: This study investigates the relationship between lower limb strength and postural stability in single-leg stance using the Balance Master system. Methods: The research involved 64 participants divided into sedentary and physically active groups based on metabolic equivalents of task (METs) values, normal weight, overweight, and obese according to body composition. Postural control was evaluated using the Sensory Organization Test. Results: The results showed that there were no significant differences in mean and maximum lower limb strength between the groups. Furthermore, postural stability in open and closed eyes conditions did not show significant differences between the groups. However, a significant positive correlation was observed between lower limb strength and stability in single-leg stance with eyes open. Conclusions: These findings suggest that lower limb muscle strength is essential for postural stability, especially when vision is available to aid balance. The study highlights the importance of interventions focused on strengthening muscles to improve physical functionality in adults. Full article
(This article belongs to the Section Kinesiology and Biomechanics)
16 pages, 906 KiB  
Review
Alcohol Consumption and Breast and Ovarian Cancer Development: Molecular Pathways and Mechanisms
by Francesca Fanfarillo, Brunella Caronti, Marco Lucarelli, Silvia Francati, Luigi Tarani, Mauro Ceccanti, Maria Grazia Piccioni, Loredana Verdone, Micaela Caserta, Sabrina Venditti, Giampiero Ferraguti and Marco Fiore
Curr. Issues Mol. Biol. 2024, 46(12), 14438-14452; https://doi.org/10.3390/cimb46120866 - 20 Dec 2024
Viewed by 265
Abstract
Alcohol consumption has been consistently linked to an increased risk of several cancers, including breast and ovarian cancer. Despite substantial evidence supporting this association, the precise mechanisms underlying alcohol’s contribution to cancer pathogenesis remain incompletely understood. This narrative review focuses on the key [...] Read more.
Alcohol consumption has been consistently linked to an increased risk of several cancers, including breast and ovarian cancer. Despite substantial evidence supporting this association, the precise mechanisms underlying alcohol’s contribution to cancer pathogenesis remain incompletely understood. This narrative review focuses on the key current literature on the biological pathways through which alcohol may influence the development of breast and ovarian cancer. Key mechanisms discussed include the modulation of estrogen levels, the generation of reactive oxygen species, the production of acetaldehyde, the promotion of chronic inflammation, and the induction of epigenetic changes. Alcohol’s impact on estrogenic signaling, particularly in the regulation of estrogen and progesterone, is explored in the context of hormone-dependent cancers. Additionally, the role of alcohol-induced DNA damage, mutagenesis, and immune system modulation in tumor initiation and progression is examined. Overall, this review emphasizes the importance of alcohol as a modifiable risk factor for breast and ovarian cancer and highlights the need for further research to clarify its role in cancer biology. Full article
(This article belongs to the Special Issue Latest Review Papers in Molecular Biology 2024)
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<p>Mechanisms of alcohol-mediated carcinogenesis. Alcohol metabolism produces acetaldehyde, which can bind to DNA and proteins, forming DNA adducts. Another mechanism involves the induction of oxidative stress and the generation of ROS, which can lead to lipid peroxidation and the production of aldehydes capable of binding to DNA and creating etheno-DNA-protein adducts. Furthermore, alcohol can disrupt retinoid metabolism by inhibiting the conversion of vitamin A to retinoic acid. Chronic alcohol consumption can also promote the recruitment of monocytes and macrophages into the tumor microenvironment, producing pro-inflammatory cytokines. Lastly, chronic alcohol consumption can affect the individual’s epigenetic profile by depleting intracellular methyl group reserves that support the epigenome. The short down pointing arrow indicates decrease.</p>
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<p>Alcohol consumption can increase the risk of breast cancer by affecting estrogen metabolism. Chronic alcohol intake increases circulating estrogen levels, promoting the growth of estrogen receptor (ER)-positive breast cancer cells. Alcohol also enhances the activity of aromatase, resulting in the conversion of androgens into estrogens, especially in peripheral tissues like adipose tissue. This results in higher local concentrations of estrogen in breast tissue, which further stimulates estrogen receptor signaling and cell proliferation. Additionally, alcohol can alter the metabolism of estrogen in the liver, leading to the production of 16α-hydroxyestrone, which can cause DNA damage and mutations, contributing to the development of breast cancer. The short pointing up/down arrows indicate increase/decrease.</p>
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12 pages, 292 KiB  
Article
Obsessive–Compulsive Symptoms in the General Population Under Stressful Conditions: Lessons Learned from the COVID-19 Pandemic
by Luca Pellegrini, Umberto Albert, Claudia Carmassi, Giuseppe Carrà, Francesca Cirulli, Bernardo Dell’Osso, Matteo Di Vincenzo, Mario Luciano, Maria Giulia Nanni, Maurizio Pompili, Gabriele Sani, Alfonso Tortorella, Umberto Volpe, Andrea Fiorillo and Gaia Sampogna
Brain Sci. 2024, 14(12), 1280; https://doi.org/10.3390/brainsci14121280 - 19 Dec 2024
Viewed by 514
Abstract
Introduction: The COVID-19 pandemic had a negative impact on mental health in the general population. The fear, stress, and uncertainty surrounding that traumatic period could have contributed to the aggravation or possible new onset of obsessive–compulsive symptoms. Methods: The COvid Mental hEalth [...] Read more.
Introduction: The COVID-19 pandemic had a negative impact on mental health in the general population. The fear, stress, and uncertainty surrounding that traumatic period could have contributed to the aggravation or possible new onset of obsessive–compulsive symptoms. Methods: The COvid Mental hEalth Trial (COMET) is a nationwide project organized by the University of Campania “Luigi Vanvitelli”, designed as an observational investigation that aimed to gather data from a representative sample of the Italian general population. The current study is a report from the main project and it focuses on obsessive–compulsive (OC) symptoms. Results: A total sample of N = 20,720 took part in the survey. N = 2332 individuals had a total Obsessive–Compulsive Inventory—Revised (OCI-R) score greater than or equal to 21 (11.3% of the entire sample), indicating the presence of clinically relevant obsessive–compulsive symptoms. By excluding patients with a history of previous mental illnesses, we still obtained a high number of individuals with an OCI-R score greater than or equal to 21 (N = 2024), representing 10.3% of the overall sample, possibly indicating a new incidence of OC symptoms during the pandemic. Discussion: Our study highlights a substantial new incidence of obsessive–compulsive symptoms in the general public. Risk factors or red flags such as being male, being of working age, living in a highly stressful environment such as one of the Italian regions most affected and severely hit by the pandemic, having higher levels of loneliness, and using substances to cope with stress, should be paid particular attention in order to prevent the development of OC symptoms during a critical and traumatic event such as the COVID-19 pandemic. Full article
(This article belongs to the Section Psychiatric Diseases)
13 pages, 3114 KiB  
Article
Assessment of the Peripheral and Central Auditory System in Infants Whose Mothers Tested Positive for COVID-19 During Pregnancy
by Jheniffer Queiroz Raimundo, Milaine Dominici Sanfins, Piotr Henryk Skarzynski, Magdalena Beata Skarżyńska and Maria Francisca Colella-Santos
Children 2024, 11(12), 1527; https://doi.org/10.3390/children11121527 - 16 Dec 2024
Viewed by 1597
Abstract
Introduction: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), a new member of the coronavirus family. While respiratory transmission is the main route, concerns have arisen regarding possible vertical transmission, which refers to [...] Read more.
Introduction: Coronavirus disease 2019 (COVID-19) is an infectious disease caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), a new member of the coronavirus family. While respiratory transmission is the main route, concerns have arisen regarding possible vertical transmission, which refers to the transmission of the virus from mother to fetus through the dissemination of viral particles in the amniotic fluid. Fetal viral infection via the placenta can affect the formation of the auditory system and lead to congenital hearing disorders. The aim of this research was to investigate the effects of vertical exposure to COVID-19 on the auditory system of newborns. Methodology: The study included a study group (SG) and a control group (CG). Selected during neonatal hearing screening, the SG consisted of 23 infants up to 1 year old whose mothers had been infected with SARS-CoV-2. The CG consisted of 15 infants whose mothers became pregnant after the end of the pandemic, had tested negative for COVID-19, and had no respiratory symptoms during pregnancy. The assessments for both groups were brainstem auditory evoked potentials (BAEPs), tympanometry, transient evoked otoacoustic emissions (TEOAEs), and distortion product otoacoustic emissions (DPOAEs). The research was divided into two studies, one cross-sectional and the other longitudinal. Results: All participants exhibited absolute latencies within the normal range for waves I, III, and V, although in the SG, there was a statistically significant increase in the latency of wave I in the left ear. In terms of OAEs, in the SG, there appeared to be a tendency for TEOAEs to be absent at high frequencies. Over several months, there was a general decrease in the amplitude of high-frequency responses in both TEOAEs and DPOAEs. Conclusion: No evidence was found that vertical exposure to COVID-19 causes hearing loss, although there were signs of possible deterioration in hair cell functioning. Full article
(This article belongs to the Section Pediatric Neonatology)
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Graphical abstract

Graphical abstract
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<p>Description of the call and stages of the study.</p>
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<p>Presence and absence of TEOAEs by frequency in the study group (SG). Key: RE, right ear; LE, left ear.</p>
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<p>Presence and absence of DPOAEs by frequency in the study group (SG). Key: RE, right ear; LE, left ear.</p>
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<p>Average difference between the response amplitudes from the initial evaluation and the second evaluation of TEOAEs.</p>
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<p>Average difference between the response amplitudes from the initial evaluation and the second evaluation of DPOAEs.</p>
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<p>Average difference between the measures (amplitude and absolute latency of waves I, III, and V, interpeak intervals I–III, III–V, and I–V, and amplitude ratio between wave V and wave I) from the initial evaluation and the second evaluation of the BAEPs. Key: amp = amplitude; Ip = interpeak interval; lat = latency.</p>
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19 pages, 667 KiB  
Article
Quality Assessment of Organic Kefirs Made with Kefir Grains and Freeze-Dried Starter Cultures
by Katarzyna Szkolnicka, Izabela Dmytrów, Anna Mituniewicz-Małek and Elżbieta Bogusławska-Wąs
Appl. Sci. 2024, 14(24), 11746; https://doi.org/10.3390/app142411746 - 16 Dec 2024
Viewed by 437
Abstract
Organic dairy products, including fermented milk, are gaining popularity among consumers. Traditionally, for kefir production, kefir grains are used; however, in the modern industry, freeze-dried cultures are commonly employed. This study aimed to analyze the quality of kefirs produced from organic cow milk [...] Read more.
Organic dairy products, including fermented milk, are gaining popularity among consumers. Traditionally, for kefir production, kefir grains are used; however, in the modern industry, freeze-dried cultures are commonly employed. This study aimed to analyze the quality of kefirs produced from organic cow milk with two fermentation times (12 and 24 h) with the use of kefir grains or freeze-dried culture. During a 3-week storage period, physicochemical properties, color, syneresis index, texture, sensory, and microbiological quality were evaluated. The results proved that organic cow milk was suitable for kefir production both with the use of kefir grains and freeze-dried culture. Using freeze-dried culture and a 24 h fermentation period resulted in kefir with the lowest syneresis tendency (3.65–9.62%), along with the best textural properties (the highest cohesiveness and viscosity index), and desired sensory characteristics. Kefir grains had a better acidification ability, and the resulting products had a higher count of yeasts, lactobacilli, and lactococci compared with kefirs obtained with freeze-dried culture. However, both the type of starter culture and the fermentation time influenced the product’s quality. The longer fermentation time of 24 h was more appropriate for the kefir production. Kefir grains, as a traditional form of kefir culture, may be preferred in organic kefir production. Full article
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<p>Scheme of experimental kefir production.</p>
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19 pages, 3835 KiB  
Article
Aged Garlic Extract (AGE) and Its Constituent S-Allyl-Cysteine (SAC) Inhibit the Expression of Pro-Inflammatory Genes Induced in Bronchial Epithelial IB3-1 Cells by Exposure to the SARS-CoV-2 Spike Protein and the BNT162b2 Vaccine
by Jessica Gasparello, Chiara Papi, Giovanni Marzaro, Alberto Macone, Matteo Zurlo, Alessia Finotti, Enzo Agostinelli and Roberto Gambari
Molecules 2024, 29(24), 5938; https://doi.org/10.3390/molecules29245938 - 16 Dec 2024
Viewed by 353
Abstract
Garlic (Allium sativum L.) is a species of the onion family (Alliaceae) widely used as a food and a folk medicine. The objective of this study was to determine the effects of AGE (aged garlic extract) on pro-inflammatory genes relevant [...] Read more.
Garlic (Allium sativum L.) is a species of the onion family (Alliaceae) widely used as a food and a folk medicine. The objective of this study was to determine the effects of AGE (aged garlic extract) on pro-inflammatory genes relevant to COVID-19. To this aim, we treated bronchial epithelial IB3-1 cells with SARS-CoV-2 spike protein (S-protein) or with the COVID-19 BNT162b2 vaccine in the absence or in the presence of AGE. The results obtained demonstrated that AGE is a potent inhibitor of the S-protein-induced expression of the IL-1β, IL-6 and IL-8 genes. Bio-Plex analysis demonstrated that AGE reduced release of IL-6 and IL-8, which were highly induced by S-protein. No inhibition of cells’ growth, toxicity and pro-apoptotic effects were found in AGE-treated cells. The effects of one of the major AGE constituents (S-allyl cysteine, SAC) were studied on the same experimental model systems. SAC was able to inhibit the S-protein-induced expression of IL-1β, IL-6 and IL-8 genes and extracellular release of IL-6 and IL-8, confirming that S-allyl-cysteine is one of the constituents of AGE that is responsible for inhibiting S-protein-induced pro-inflammatory genes. Docking experiments suggest that a possible mechanism of action of SAC is an interference with the activity of Toll-like receptors (TLRs), particularly TLR4, thereby inhibiting NF-κB- and NF-κB-regulated genes, such as IL-1β, IL-6 and IL-8 genes. These results suggest that both AGE and SAC deserve further experimental efforts to verify their effects on pro-inflammatory genes in SARS-CoV-2-infected cells. Full article
(This article belongs to the Section Bioorganic Chemistry)
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Figure 1
<p>Pictorial representation of the experimental model systems of the present study. A: human bronchial epithelial IB3-1 cells stimulated with the SARS-CoV-1 spike protein; B: IB3-1 cells stimulated with the BNT162b2 vaccine.</p>
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<p>GC-MS analysis of AGE and SAC. (<b>A</b>) Total ion current (TIC) chromatogram of the AGE sample derivatized with TBDMS; (<b>B</b>) total ion current (TIC) chromatogram of the SAC sample derivatized with TBDMS; (<b>C</b>) overlaid selected ion chromatograms (SIM) of AGE and SAC samples derivatized with TBDMS acquiring the following ions: <span class="html-italic">m</span>/<span class="html-italic">z</span> 239 (3,4-dimethoxybenzoic acid, internal standard) and <span class="html-italic">m</span>/<span class="html-italic">z</span> 332 (SAC); (<b>D</b>), electron impact mass spectrum of S-allyl-cysteine as di-TBDMS derivative (retention time 10.49 min). Both AGE (<b>A</b>–<b>C</b>) and the SAC (<b>D</b>–<b>F</b>) powders were freshly dissolved in complete RPMI-1640 medium prior to each experiment. Enlargements/details are shown in the sub-panels in (<b>B</b>,<b>E</b>).</p>
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<p>Representative examples of the effects of increasing amounts of AGE on S-protein-induced expression of IL-1β (<b>A</b>), IL-6 (<b>B</b>) and IL-8 (<b>C</b>) genes. IB3-1 cells were either untreated (white histograms) or exposed to SASRS-CoV-2 S-protein (5 nM) in the absence (black histograms) or in the presence of the indicated concentrations of AGE (grey histograms). After 72 h, total RNA was isolated and mRNA content analyzed by RT-qPCR. (<b>D</b>) Summary of experiments (<span class="html-italic">n</span> = 3) analyzing the relative content of IL-6 mRNA. Results represent the average ± S.D. Statistical significance was high: ** (<span class="html-italic">p</span> &lt; 0.01) and *** (<span class="html-italic">p</span> &lt; 0.001).</p>
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<p>Effects of AGE on cell vitality and apoptosis. IB3-1 cells were cultured in the absence or the presence of the indicated concentrations of AGE for 72 h. Vitality and apoptosis were determined using the Muse Annexin V &amp; Dead Cell Kit. (<b>A</b>–<b>D</b>). Representative annex V assays performed on control untreated IB3-1 cells (<b>A</b>) or IB3-1 cells treated for 72 h with AGE, used at 0.1 mg/mL (<b>B</b>), 0.5 mg/mL (<b>C</b>) and 1 mg/mL (<b>D</b>). A summary of the annexin V assay data is shown in panel (<b>E</b>).</p>
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<p>Representative examples of the effects of increasing amounts of SAC on S-protein-induced expression of IL-1β (<b>A</b>), IL-6 (<b>B</b>) and IL-8 (<b>C</b>) genes. IB3-1 cells were either untreated (white histograms) or exposed to SARS-CoV-2 S-protein (5 nM) in the absence (black histograms) or in the presence of the indicated concentrations of SAC (grey histograms). After 72 h, total RNA was isolated and mRNA content analyzed by RT-qPCR. (<b>D</b>) Summary of experiments (<span class="html-italic">n</span> = 3) analyzing the relative content of IL-6 mRNA. The results represent the average ± S.D. Statistical significance was as follows: * (<span class="html-italic">p</span> &lt; 0.05): significant; ** (<span class="html-italic">p</span> &lt; 0.01) and *** (<span class="html-italic">p</span> &lt; 0.001): highly significant.</p>
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<p>Effects of AGE and SAC on release of IL-6 (<b>A</b>) and IL-8 (<b>B</b>) by S-protein-stimulated IB3-1 cells. A panel of 27 cytokines/chemokines/growth factors was analyzed by Bio-Plex technology in IB3-1-infected cells 24 h after exposure to the S-protein. Results represent the average ± SD (<span class="html-italic">n</span> = 3). The results represent the average ± S.D. Statistical significance was as follows: * (<span class="html-italic">p</span> &lt; 0.05): significant; ** (<span class="html-italic">p</span> &lt; 0.01): highly significant.</p>
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<p>Effects of AGE and SAC on IL-8 gene expression induced in IB3-1 cells by treatment with BNT162b2 COVID-19 vaccine. The results represent the average ± S.D (<span class="html-italic">n</span> = 3). Statistical significance was as follows: * (<span class="html-italic">p</span> &lt; 0.05): significant; ** (<span class="html-italic">p</span> &lt; 0.01): highly significant.</p>
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<p>Simulation of SAC binding to TLR-4 through docking and molecular dynamics simulations. The docking studies were conducted with AutoDock Vina [<a href="#B59-molecules-29-05938" class="html-bibr">59</a>]. Molecular dynamics studies were conducted with Gromacs software (v. 2024,0). See the Materials and Methods section for further details. Binding mode predicted for SAC (<b>A</b>) with the TLR4-TIR dimer and details of interactions (<b>B</b>). TLR4 monomers are depicted in blue (chain A) and green (chain B), respectively; SAC is depicted as stick (magenta-colored carbon); hydrogen bonds are depicted as dashed black lines. Number of H bonds formed between chains A and B during 50 ns of MDs (<b>C</b>). Black line represents the simulations of apo-TLR-4; red line represents the simulation of SAC-bound TLR-4. Note a marked reduction in interchain interactions as a consequence of the interaction with SAC. Number of H bonds formed between SAC and TLR-4 chain A (black) and chain B (red) (<b>D</b>). Root-mean-squared deviation (RMSD, nm) of Cα atoms in SAC-bound TLR-4 (<b>E</b>). Black line represents the total RMSD; blue line represents the chain A RMSD; red line represents the chain B RMSD. Root-mean-squared deviation (RMSD, nm) of SAC heavy atoms with respect to Cα atoms (<b>F</b>). Black line represents the SAC RMSD relative to the dimer; blue line represents the SAC RMSD relative to chain A; red line represents the SAC RMSD relative to chain B. Note that the binding between SAC and chain A remains constant during the 50 ns of MDs.</p>
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<p>Proposed mechanism of action of SAC. Indirect inhibition of NF-κB might be caused by a direct inhibition of TLR4. The associated NF-κB inhibition is causative of the indirect inhibitory effects of SAC on NF-κB-regulated genes, such as IL-1κ, IL-6, IL-8 and others.</p>
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40 pages, 4412 KiB  
Review
Trends and Innovations in Wearable Technology for Motor Rehabilitation, Prediction, and Monitoring: A Comprehensive Review
by Pedro Lobo, Pedro Morais, Patrick Murray and João L. Vilaça
Sensors 2024, 24(24), 7973; https://doi.org/10.3390/s24247973 - 13 Dec 2024
Viewed by 606
Abstract
(1) Background: Continuous health promotion systems are increasingly important, enabling decentralized patient care, providing comfort, and reducing congestion in healthcare facilities. These systems allow for treatment beyond clinical settings and support preventive monitoring. Wearable systems have become essential tools for health monitoring, but [...] Read more.
(1) Background: Continuous health promotion systems are increasingly important, enabling decentralized patient care, providing comfort, and reducing congestion in healthcare facilities. These systems allow for treatment beyond clinical settings and support preventive monitoring. Wearable systems have become essential tools for health monitoring, but they focus mainly on physiological data, overlooking motor data evaluation. The World Health Organization reports that 1.71 billion people globally suffer from musculoskeletal conditions, marked by pain and limited mobility. (2) Methods: To gain a deeper understanding of wearables for the motor rehabilitation, monitoring, and prediction of the progression and/or degradation of symptoms directly associated with upper-limb pathologies, this study was conducted. Thus, all articles indexed in the Web of Science database containing the terms “wearable”, “upper limb”, and (“rehabilitation” or “monitor” or “predict”) between 2019 and 2023 were flagged for analysis. (3) Results: Out of 391 papers identified, 148 were included and analyzed, exploring pathologies, technologies, and their interrelationships. Technologies were categorized by typology and primary purpose. (4) Conclusions: The study identified essential sensory units and actuators in wearable systems for upper-limb physiotherapy and analyzed them based on treatment methods and targeted pathologies. Full article
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<p>The data flow of the systematic review.</p>
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<p>The figure overview of the technological groups identified throughout our review as well as the relationship between them.</p>
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<p>Study characteristics for camera studies [<a href="#B28-sensors-24-07973" class="html-bibr">28</a>,<a href="#B29-sensors-24-07973" class="html-bibr">29</a>,<a href="#B30-sensors-24-07973" class="html-bibr">30</a>,<a href="#B31-sensors-24-07973" class="html-bibr">31</a>,<a href="#B32-sensors-24-07973" class="html-bibr">32</a>,<a href="#B33-sensors-24-07973" class="html-bibr">33</a>,<a href="#B34-sensors-24-07973" class="html-bibr">34</a>,<a href="#B35-sensors-24-07973" class="html-bibr">35</a>,<a href="#B36-sensors-24-07973" class="html-bibr">36</a>,<a href="#B37-sensors-24-07973" class="html-bibr">37</a>,<a href="#B38-sensors-24-07973" class="html-bibr">38</a>,<a href="#B39-sensors-24-07973" class="html-bibr">39</a>,<a href="#B40-sensors-24-07973" class="html-bibr">40</a>,<a href="#B41-sensors-24-07973" class="html-bibr">41</a>,<a href="#B42-sensors-24-07973" class="html-bibr">42</a>].</p>
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<p>Study characteristics for complementary technologies studies [<a href="#B43-sensors-24-07973" class="html-bibr">43</a>,<a href="#B44-sensors-24-07973" class="html-bibr">44</a>,<a href="#B45-sensors-24-07973" class="html-bibr">45</a>].</p>
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<p>Study characteristics for other studies [<a href="#B46-sensors-24-07973" class="html-bibr">46</a>,<a href="#B47-sensors-24-07973" class="html-bibr">47</a>,<a href="#B48-sensors-24-07973" class="html-bibr">48</a>,<a href="#B49-sensors-24-07973" class="html-bibr">49</a>,<a href="#B50-sensors-24-07973" class="html-bibr">50</a>].</p>
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<p>Identifying hemiparesis using wrist-worn accelerometry, as presented in the work by S. Datta et al. [<a href="#B59-sensors-24-07973" class="html-bibr">59</a>].</p>
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<p>An illustration of the sensor network used for the reconstruction of upper-limb joints from the work of Meng et al. and the anatomical model of the entire upper limb with the definition of joint axes.</p>
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<p>A flowchart of the methodology for estimating task-specific ARAT scores using inertial sensors mounted on the wrist. Synthetic minority over-sampling technique (SMOTE).</p>
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<p>A wristband functions as a joystick for controlling a remote-controlled car within a maze; wrist flexion-extension controls the car’s forward and backward movement, while pronation–supination enables on-the-spot turning.</p>
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<p>Wearable multimodal rehabilitation utilizing serious games involves extracting kinematic data. Relevant features are identified and input into classification algorithms to predict movements that serve as inputs for the game.</p>
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<p>The stimulation system proposed in the work by Ferrari et al [<a href="#B169-sensors-24-07973" class="html-bibr">169</a>].</p>
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<p>Contributions from the conducted review, including the characterization of the pathologies explored in the studies as well as the main technologies addressed.</p>
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16 pages, 3131 KiB  
Article
Comparison of Untargeted and Markers Analysis of Volatile Organic Compounds with SIFT-MS and SPME-GC-MS to Assess Tea Traceability
by Marine Reyrolle, Valérie Desauziers, Thierry Pigot, Lydia Gautier and Mickael Le Bechec
Foods 2024, 13(24), 3996; https://doi.org/10.3390/foods13243996 - 11 Dec 2024
Viewed by 444
Abstract
Tea is one of the most consumed beverages in the world and presents a great aromatic diversity depending on the origin of the production and the transformation process. Volatile organic compounds (VOCs) greatly contribute to the sensory perception of tea and are excellent [...] Read more.
Tea is one of the most consumed beverages in the world and presents a great aromatic diversity depending on the origin of the production and the transformation process. Volatile organic compounds (VOCs) greatly contribute to the sensory perception of tea and are excellent markers for traceability and quality. In this work, we analyzed the volatile organic compounds (VOCs) emitted by twenty-six perfectly traced samples of tea with two analytical techniques and two data treatment strategies. First, we performed headspace solid-phase microextraction gas chromatography–mass spectrometry (HS-SPME-GC-MS) as the most widely used reference method for sanitary and quality controls of food. Next, we analyzed the samples with selected-ion flow-tube mass spectrometry (SIFT-MS), an emerging method for direct analysis of food products and aroma. We compared the performances of both techniques to trace the origin and the transformation processes. We selected the forty-eight most relevant markers with HS-SPME-GC-MS and evaluated their concentrations with a flame ionization detector (FID) on the same instrument. This set of markers permitted separation of the origins of samples but did not allow the samples to be differentiated based on the color. The same set of markers was measured with SIFT-MS instrument without success for either origin separation or color differentiation. Finally, a post-processing treatment of raw data signals with an untargeted approach was applied to the GC-MS and SIFT-MS dataset. This strategy allowed a good discrimination of origin and color with both instruments. Advantages and drawbacks of volatile profiles with both instruments were discussed for the traceability and quality assessment of food. Full article
(This article belongs to the Special Issue Tea: Processing Techniques, Flavor Chemistry and Health Benefits)
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<p>Individual plots of PLS-DA for country discrimination. (<b>A</b>) PC1 and PC2 plot for the GC-FID data. (<b>B</b>) PC2 and PC3 plot for the GC-FID data. (<b>C</b>) PC1 and PC2 plot for the SIFT-MS data. (<b>D</b>) PC2 and PC3 plot for the SIFT-MS data.</p>
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<p>Individual plots of sparse PLS-DA for color discrimination. (<b>A</b>) PC1 and PC2 plot for the GC-FID data. (<b>B</b>) PC1 and PC2 plot for the SIFT-MS data.</p>
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<p>Individual plots of sparse PLS-DA for country discrimination. (<b>A</b>) PC1 and PC2 plot for the volatiles profile obtained by GC-MS/FID. (<b>B</b>) PC2 and PC3 plot for the volatiles profile obtained by GC-MS/FID. (<b>C</b>) PC1 and PC2 plot for the volatile profile obtained by SIFT-MS. (<b>D</b>) PC2 and PC3 plot for the volatiles profile obtained by SIFT-MS.</p>
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<p>Individual plots of sparse PLS-DA for color discrimination. (<b>A</b>) PC1 and PC2 plot for the volatiles profile obtained by GC-MS/FID. (<b>B</b>) PC2 and PC3 plot for the volatiles profile obtained by GC-MS/FID. (<b>C</b>) PC1 and PC2 plot for the volatiles profile obtained by SIFT-MS. (<b>D</b>) PC1 and PC2 plot for the volatiles profile obtained by the SIFT-MS data.</p>
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10 pages, 1338 KiB  
Article
A Comparative Study of the Electroneurographic Findings in Amyloidotic Polyneuropathy in Patients with Light-Chain Amyloidosis and Glu54Gln Transthyretin Amyloidosis
by Mirela Drăghici, Sorina N. Bădeliță, Andreea Jercan, Oana Obrișcă, Crisanda Vîlciu, Monica Popescu, Adina Turcu-Stiolica and Daniel Coriu
Medicina 2024, 60(12), 2027; https://doi.org/10.3390/medicina60122027 - 9 Dec 2024
Viewed by 464
Abstract
Background and Objectives: Amyloidosis is a disorder characterized by the abnormal folding of proteins, forming insoluble fibrils that accumulate in tissues and organs. This accumulation disrupts normal tissue architecture and organ function, often with serious consequences, including death if left untreated. Light-chain [...] Read more.
Background and Objectives: Amyloidosis is a disorder characterized by the abnormal folding of proteins, forming insoluble fibrils that accumulate in tissues and organs. This accumulation disrupts normal tissue architecture and organ function, often with serious consequences, including death if left untreated. Light-chain amyloidosis (AL) and hereditary transthyretin-type amyloidosis (hATTR) are two of the most common types. In amyloidosis, peripheral nervous system involvement is a significant diagnostic feature, particularly when it manifests as polyneuropathy, carpal tunnel syndrome (CTS), and dysautonomia. These neurological symptoms often point to the involvement of amyloid deposits in the peripheral and autonomic nervous systems, which can help identify and differentiate between the various types of amyloidosis. Materials and Methods: This retrospective study focused on the evolution of electrophysiological parameters in two groups: AL (n = 22) and hATTR-Glu54Gln patients (n = 14), with mixed axonal polyneuropathy. Patients were followed for two consecutive years to assess disease progression. The PND scale (polyneuropathy disability) was also used to assess motor impairment for each patient. Results: In our study AL amyloidosis patients presented with mixed, axonal polyneuropathy associated with CTS in 63.6% of cases and cardiomyopathy (45.5%). Serial EMGs (electromyography) showed decreased motor amplitudes of the common peroneal and tibial nerves and sensory amplitude of the superficial peroneal nerve, with mostly preserved conduction velocities. The patients maintained stage I PND throughout the monitoring period. The entire hATTR group displayed mixed, axonal polyneuropathy and cardiomyopathy; 85.7% of them had CTS, and 42.9% had orthostatic hypotension. EMG data showed decreased motor amplitudes of the tibial and common peroneal nerves, decreased sensory amplitudes of the superficial peroneal nerve, and mildly reduced conduction velocities, with significant progression at 12 and 24 months. The patients displayed additional reduced muscle strength, some reaching stage 3A and 3B-PND at the end of the study. Conclusions: The amyloidotic polyneuropathy found in the groups was similar in its axonal, sensory-motor, and length-dependent characteristics, but the study showed significant differences in its progression, with more abrupt changes in the hATTR-Glu54Gln group. The amyloidosis AL patients remained in stage 1 PND, while the hATTR-Glu54Gln patients progressed to stage 3 PND at 24 months. Full article
(This article belongs to the Special Issue Neuromuscular Disorders: Diagnostical Approaches and Treatments)
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<p>Electrophysiological data of AL patients. (<b>A</b>), CMAP tibial nerve. (<b>B</b>), MCV tibial nerve. (<b>C</b>), CMAP common peroneal nerve. (<b>D</b>), MCV common peroneal nerve. (<b>E</b>), SNAP superficial peroneal nerve. (<b>F</b>), SCV superficial peroneal nerve.</p>
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<p>Electrophysiological measurements for hATTR-Glu54Gln patients. (<b>A</b>), CMAP of the tibial nerve. (<b>B</b>), MCV of the tibial nerve. (<b>C</b>), CMAP of the common peroneal nerve. (<b>D</b>), MCV of the common peroneal nerve. (<b>E</b>), SNAP of the superficial peroneal nerve. (<b>F</b>), SCV of the superficial peroneal nerve.</p>
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20 pages, 2939 KiB  
Article
From Waste to Taste: Coffee By-Products as Starter Cultures for Sustainable Fermentation and Improved Coffee Quality
by Anna María Polanía Rivera, Jhennifer López Silva, Laura Torres-Valenzuela and José Luis Plaza-Dorado
Sustainability 2024, 16(23), 10763; https://doi.org/10.3390/su162310763 - 8 Dec 2024
Viewed by 847
Abstract
Utilizing coffee by-products in the fermentation process of coffee offers a sustainable strategy by repurposing agricultural waste and enhancing product quality. This study evaluates the effect of applying a starter culture, derived from coffee residues, on the dynamics of reducing and total sugars [...] Read more.
Utilizing coffee by-products in the fermentation process of coffee offers a sustainable strategy by repurposing agricultural waste and enhancing product quality. This study evaluates the effect of applying a starter culture, derived from coffee residues, on the dynamics of reducing and total sugars during coffee fermentation, as well as the composition of aromatic compounds, organic acids, and the sensory profile of coffee inoculated with yeast (Saccharomyces cerevisiae) and lactic acid bacteria (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus), in comparison to a spontaneously fermented sample. Volatile compounds were identified and quantified using dynamic headspace gas chromatography-mass spectrometry (HS/GC-MS), with predominant detection of 2-furancarboxaldehyde, 5-methyl; 2-furanmethanol; and furfural—compounds associated with caramel, nut, and sweet aromas from the roasting process. A reduction in sugars (glucose, fructose, and sucrose) occurred over the 36 h fermentation period. Lactic acid (2.79 g/L) was the predominant organic acid, followed by acetic acid (0.69 g/L). The application of the inoculum improved the sensory quality of the coffee, achieving a score of 86.6 in evaluations by Q-graders, compared to 84 for the control sample. Additionally, descriptors such as red apple, honey, and citrus were prominent, contributing to a uniform and balanced flavor profile. These findings indicate that controlled fermentation with starter cultures derived from coffee by-products enhances sustainability in coffee production. It achieves this by supporting a circular economy, reducing reliance on chemical additives, and improving product quality. This approach aligns with sustainable development goals by promoting environmental stewardship, economic viability, and social well-being within the coffee industry. Full article
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<p>Behavior of sugars during the fermentation without inoculum (control).</p>
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<p>Behavior of sugars during the fermentation with inoculum.</p>
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<p>Behavior of organic acids during the fermentation without inoculum (control).</p>
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<p>Behavior of organic acids during the fermentation with inoculum.</p>
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<p>Roasting curve for fermented coffee with inoculum.</p>
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<p>Roasting curve for fermented coffee without inoculum.</p>
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<p>Chromatogram fermentation with inoculum.</p>
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<p>Chromatogram of coffee sample without inoculum.</p>
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17 pages, 4000 KiB  
Article
Development of an rpS6-Based Ex Vivo Assay for the Analysis of Neuronal Activity in Mouse and Human Olfactory Systems
by Emma Broillet-Olivier, Yaëlle Wenger, Noah Gilliand, Hugues Cadas, Sara Sabatasso, Marie-Christine Broillet and Julien Brechbühl
Int. J. Mol. Sci. 2024, 25(23), 13173; https://doi.org/10.3390/ijms252313173 - 7 Dec 2024
Viewed by 520
Abstract
Olfactory sensitivity to odorant molecules is a complex biological function influenced by both endogenous factors, such as genetic background and physiological state, and exogenous factors, such as environmental conditions. In animals, this vital ability is mediated by olfactory sensory neurons (OSNs), which are [...] Read more.
Olfactory sensitivity to odorant molecules is a complex biological function influenced by both endogenous factors, such as genetic background and physiological state, and exogenous factors, such as environmental conditions. In animals, this vital ability is mediated by olfactory sensory neurons (OSNs), which are distributed across several specialized olfactory subsystems depending on the species. Using the phosphorylation of the ribosomal protein S6 (rpS6) in OSNs following sensory stimulation, we developed an ex vivo assay allowing the simultaneous conditioning and odorant stimulation of different mouse olfactory subsystems, including the main olfactory epithelium, the vomeronasal organ, and the Grueneberg ganglion. This approach enabled us to observe odorant-induced neuronal activity within the different olfactory subsystems and to demonstrate the impact of environmental conditioning, such as temperature variations, on olfactory sensitivity, specifically in the Grueneberg ganglion. We further applied our rpS6-based assay to the human olfactory system and demonstrated its feasibility. Our findings show that analyzing rpS6 signal intensity is a robust and highly reproducible indicator of neuronal activity across various olfactory systems, while avoiding stress and some experimental limitations associated with in vivo exposure. The potential extension of this assay to other conditioning paradigms and olfactory systems, as well as its application to other animal species, including human olfactory diagnostics, is also discussed. Full article
(This article belongs to the Special Issue Molecular Mechanisms Subserving Taste and Olfaction Systems)
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<p>Development of an ex vivo assay based on the rpS6 signal for the assessment of neuronal activity in olfactory systems. (<b>a</b>) Schematic representation of the various steps involved in the ex vivo assay. Illustrations were created using BioRender.com (accessed first on 30 November 2022). (<b>b</b>) Left panel: representative immunostaining showing the rpS6 signal obtained with the anti-rpS6 antibody (α-rpS6, 1:5000, in red) from the Grueneberg ganglion (GG) of an OMP-GFP mouse, where GG neurons (in green) are visualized based on their endogenous GFP expression. Right panel: the merged view with a nuclear Dapi counterstain (in blue) is shown. (<b>c</b>) Negative control performed without the α-rpS6 (Cy3 signal in red). (<b>d</b>) Merged view of a double immunohistochemical analysis for rpS6 (α-rpS6, in red) and the OMP protein (α-OMP, in green) in a BL/6 mouse. (<b>e</b>,<b>f</b>) Assessment of the temperature effect on the rpS6 signal in the GG. Representative immunohistochemical investigations, performed on OMP-GFP mice, are illustrated in (<b>e</b>) at different temperatures (4 °C, 23 °C, 30 °C, and 37 °C), and the statistical analysis is shown in (<b>f</b>) for both OMP-GFP and BL/6 mice. Nasal cavities are indicated (nc; <b>b</b>–<b>e</b>). White arrowheads indicate the zoom-in regions (<b>b</b>) or the rpS6-related signal zoom-in regions (<b>c</b>,<b>d</b>). Scale bars: 20 µm (<b>b</b>–<b>e</b>). Data are expressed as a standardized percentage of the rpS6 signal intensity and represented as the mean ± SEM with aligned dot plots for a minimum of three GG sections per animal, from at least two mice per condition. Comparisons between conditions were performed using two-tailed Welch’s <span class="html-italic">t</span>-tests or Mann–Whitney <span class="html-italic">U</span>-tests, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Odorant-induced increases of the rpS6 signal in the GG at different conditioning temperatures. (<b>a</b>) Representative immunostaining for the rpS6 signal (in red) in the GG of OMP-GFP mice (OMP-GFP signal, in green) following conditioning at different temperatures (4 °C, 23 °C, 30 °C, and 37 °C) and odorant stimulations, with ACSF serving as the reference control (Ctrl), TMT, or SBT. The merged images include nuclear Dapi staining (in blue). (<b>b</b>) The statistical analysis comparing the effects of odorant stimulations calibrated at different conditioning temperatures is displayed. Stimulations with the Ctrl and odorants (TMT in left panels and SBT in right panels) are represented by white and gray bars, respectively. Nasal cavities (nc) are indicated in (<b>a</b>). White arrowheads highlight zoomed-in regions of the rpS6 signal (<b>a</b>). Scale bars: 20 μm (<b>a</b>). Data are presented as a standardized percentage of the rpS6 signal intensity, with the means ± SEM displayed using aligned dot plots. A minimum of three GG sections per animal from at least two mice per condition were analyzed. Comparisons between conditions were performed using two-tailed Mann–Whitney <span class="html-italic">U</span>-tests, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Investigation of rpS6-based odorant signals in the MOE and VNO of mice. (<b>a</b>) Here, representative immunostaining for the rpS6 signal (in red) in the MOE of OMP-GFP mice (OMP-GFP signal, in green) under non-stimulated control conditions (Ctrl, left panel) and after TMT stimulation (TMT, right panels) are shown here for a conditioning temperature of 4 °C. (<b>b</b>) Statistical analysis comparing the effects of TMT stimulations on GFP-positive OSNs in the MOE. Data were combined across the conditioning temperatures of 4 °C and 23 °C. (<b>c</b>) Here, representative immunostaining for the rpS6 signal (in red) in the VNO of OMP-GFP mice under non-stimulated conditions (Ctrl, left panel) and after SBT stimulation (SBT, right panels), are shown for the same conditioning temperature of 4 °C. The approximate boundaries between the vomeronasal type-1 receptor (V1R) and vomeronasal type-2 receptor (V2R) layers are marked with a white dashed line. Non-sensory GFP-negative cells expressing the rpS6 signal are indicated by white asterisks. (<b>d</b>) Statistical analysis comparing the effects of SBT stimulations on GFP-positive OSNs in the VNO. Data were combined across the conditioning temperatures of 4 °C and 23 °C. Nasal cavities (nc) in the MOE (<b>a</b>) and lumen (lu) in the VNO (<b>c</b>) are annotated. White arrowheads highlight regions where the rpS6 signal is zoomed in (<b>a</b>,<b>c</b>). Scale bars: 20 μm (<b>a</b>,<b>c</b>). Merged images include nuclear Dapi staining (in blue, (<b>a</b>,<b>c</b>)). Stimulations with the Ctrl and odorants are represented by white and gray bars, respectively (in (<b>b</b>,<b>d</b>)). A minimum of four tissue sections per animal from at least three mice per condition were analyzed. Comparisons between conditions were performed using two-tailed Mann–Whitney <span class="html-italic">U</span>-tests, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Investigation of rpS6-based signals in the posterosuperior region of the human olfactory system. (<b>a</b>) Negative control performed on the posterosuperior region of the human olfactory epithelium (OE) without primary antibodies, illustrating the endogenous signals related to Cy3 (in red) and FITC (in green). (<b>b</b>) Representative immunostaining showing the α-OMP antibody signal (in green) in the human OE and its apparent lack of neuronal specificity. (<b>c</b>) Representative immunostaining for the α-CK18 antibody signal (in red) performed in the same region, highlighting its specificity for the sustentacular supporting cells of the olfactory epithelium. (<b>d</b>) Labeling for the rpS6 signals (in red) in the posterosuperior region of the human OE. Nasal cavities (nc, (<b>a</b>–<b>d</b>)) are indicated. The white arrowhead highlights a zoomed-in region of the rpS6 signals, within which white and yellow asterisks indicate basal cells and sporadic cells of the sensory epithelium, respectively (<b>d</b>). Scale bars: 10 μm (<b>a</b>–<b>d</b>). Dapi staining is used as a nuclear marker (in blue, (<b>a</b>–<b>d</b>)).</p>
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15 pages, 1431 KiB  
Article
CD3+CD4-CD8- Double-Negative Lymphocytes Are Increased in the Aqueous Humor of Patients with Retinitis Pigmentosa: Their Possible Role in Mediating Inflammation
by Daniela Bacherini, Laura Maggi, Francesco Faraldi, Andrea Sodi, Lorenzo Vannozzi, Alessio Mazzoni, Manuela Capone, Gianni Virgili, Giulio Vicini, Benedetto Falsini, Lorenzo Cosmi, Pasquale Viggiano, Stanislao Rizzo, Francesco Annunziato, Fabrizio Giansanti and Francesco Liotta
Int. J. Mol. Sci. 2024, 25(23), 13163; https://doi.org/10.3390/ijms252313163 - 7 Dec 2024
Viewed by 482
Abstract
Recently, evidence has supported a significant role for immune and oxidative-mediated damage underlying the pathogenesis of different types of retinal diseases, including retinitis pigmentosa (RP). Our study aimed to evaluate the presence of immune cells and mediators in patients with RP using flow [...] Read more.
Recently, evidence has supported a significant role for immune and oxidative-mediated damage underlying the pathogenesis of different types of retinal diseases, including retinitis pigmentosa (RP). Our study aimed to evaluate the presence of immune cells and mediators in patients with RP using flow cytometric analysis of peripheral blood (PB) and aqueous humor (AH) samples. We recruited 12 patients with RP and nine controls undergoing cataract surgery. Flow cytometric analysis of PB and AH samples provided a membrane staining that targeted surface molecules (CD14, CD16, CD19, CD3, CD4, CD8, and CD161) identifying monocytes, natural killer (NK) cells, B cells, T cells, and T subpopulations, respectively. Moreover, lymphocytes were polyclonally stimulated to evaluate cytokine (CK) production at single-cell level. The circulating immune cell distribution was comparable between patients with RP and controls. Conversely, in the AH of controls we could detect no cells, while in the RP AH samples we found infiltrating leukocytes, consisting of T (CD3+), B (CD19+), NK (CD16+CD3-) cells, and monocytes (CD14+). In patients with RP, the frequency of most infiltrating immune cell populations was similar between the AH and PB. However, among T cell subpopulations, the frequency of CD3+CD4+ T cells was significantly lower in the RP AH compared to RP PB, whereas CD3+CD4-CD8- double-negative (DN) T cells were significantly higher in the RP AH compared to RP PB. Cytokine production analysis revealed a trend toward an increased frequency of CD3+CD8-CD161+IFN-ɣ-producing cells and a decreased frequency of CD3+CD8+IL-4-producing cells in the RP AH compared to RP PB. The detection of immune cells, particularly DN T cells, and a Th1-skewed phenotype in RP AH suggests immune-mediated and inflammatory mechanisms in the disease. Full article
(This article belongs to the Special Issue Emerging Roles of Inflammation in Retinal and Optic Nerve Pathologies)
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<p>Flow cytometry «counts» of cellular events in aqueous humor (AH) sample. (<b>A</b>) AH samples from patients with retinitis pigmentosa (RP) (n = 14) and from controls (n = 9) were analyzed using flow cytometry in order to identify the presence of cells in each sample. The columns represent the mean value and CI of the total number of events recorded in the P1 gate, defined in the plot of physical parameters scatter (FSC/SSC) as classic mononuclear cell gate. (<b>B</b>) Flow cytometric dot plots of physical parameters scatter (FSC/SSC) representative of one RP sample (left panel) and one control sample (right panel).</p>
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<p>Flow cytometry analysis of immune cell phenotypes at peripheral blood (PB) and aqueous umor (AH) levels in patients with retinitis pigmentosa (n = 14). PB (black columns) and AH (white columns) samples were analyzed using flow cytometry for surface marker expression to permit identification of T cells (CD3+), B cells (CD19+), natural killer cells (CD3–CD16), and monocytes (CD14) in the mononuclear cells (MNCs) (<b>A</b>), and of the different T cell subpopulations (CD3+CD4+, CD3+CD8+, and CD3+CD4-CD8-) in the T cells (<b>B</b>). Columns represent the mean values + CI of the frequency of the indicated cell population and lymphocyte sub-population.</p>
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<p>Flow cytometry analysis of intracellular cytokine production. Peripheral blood (PB, black columns) and aqueous humor (AH, white columns) samples from patients with RP (n = 5) were stimulated with phorbol 12-myristate 13-acetate and ionomycin for 6 h, the last four in the presence of Brefeldin A, then fixed and stained for intracellular cytokines associated with surface markers. The flow cytometry analysis was performed on CD3+CD8- (<b>A</b>) and CD3+CD8+ (<b>B</b>) gated cells. The columns represent the mean values + CI of the frequency of positive cells for the indicated cytokine.</p>
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