Biomedicines

16 pages, 1306 KiB

Open AccessArticle

Stress Hormone Corticosterone Controls Metabolic Mitochondrial Performance and Inflammatory Signaling of In Vitro Cultured Sertoli Cells

by Ana M. Silva, Carina T. Ribeiro, Raquel L. Bernardino, Ivana Jarak, Rui A. Carvalho, M. A. Pereira-Sampaio, Diogo B. de Souza, Marco G. Alves and Pedro F. Oliveira

Biomedicines 2022, 10(9), 2331; https://doi.org/10.3390/biomedicines10092331 - 19 Sep 2022

Cited by 3 | Viewed by 2896

Abstract

Stress, as a physiological response, is a major factor that affects several processes, including reproductive functions. The main hormonal players of stress are cortisol (humans) and corticosterone (rodents). Sertoli cells (SCs), as key contributors for the testicular homeostasis maintenance, are extensively challenged by [...] Read more.

Stress, as a physiological response, is a major factor that affects several processes, including reproductive functions. The main hormonal players of stress are cortisol (humans) and corticosterone (rodents). Sertoli cells (SCs), as key contributors for the testicular homeostasis maintenance, are extensively challenged by different hormones, with glucocorticoid corticosterone being the signaling modulator that may impact these cells at different levels. We aimed to characterize how corticosterone modulates SCs energy balance, putting the mitochondrial performance and signaling output in perspective as the cells can disperse to the surroundings. TM4 mouse SCs were cultured in the absence and presence of corticosterone (in nM: 20, 200, and 2000). Cells were assessed for extracellular metabolic fluxes, mitochondrial performance (cell respirometry, mitochondrial potential, and mitochondrial complex expressions and activities), and the expression of androgen and corticosteroid receptors, as well as interleukine-6 (IL-6) and glutathione content. Corticosterone presented a biphasic impact on the extracellular fluxes of metabolites. Low sub-physiological corticosterone stimulated the glycolytic activity of SCs. Still, no alterations were perceived for lactate and alanine production. However, the lactate/alanine ratio was decreased in a dose-dependent mode, opposite to the mitochondrial complex II activity rise and concurrent with the decrease of IL-6 expression levels. Our results suggest that corticosterone finely tuned the energetic profile of mouse SCs, with sub-physiological concentrations promoting glycolytic expenditure, without translating into cell redox power and mitochondrial respiratory chain performance. Corticosterone deeply impacted the expression of the pro-inflammatory IL-6, which may alter cell-to-cell communication in the testis, in the last instance and impact of the spermatogenic performance. Full article

(This article belongs to the Special Issue Hormonal Regulation of Male Reproductive System)

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Figure 1

Figure 1
Effect of corticosterone (20, 200, and 2000 nM) treatment in mouse SCs (TM4) Ar (panel (A)) and Nr3c1 (panel (B)) transcript levels. Corticosterone-free condition (CORT-free) was taken into account to normalize data. Ordinary one-way ANOVA, with a Tukey’s multiple comparison test, was used for statistical analysis. Results are expressed as box plots (with median, minimum, and maximal values, as well as total points and first and third quartiles) (n = 6 for each condition). Data with p < 0.05 were considered statistically different (* p < 0.05). Full article ">Figure 2
Effect of corticosterone (20, 200, and 2000 nM) treatment in mouse SCs (TM4) consumption and production of the 1H-NMR detectable metabolites glucose, glutamine, lactate, succinate, and alanine (Panels (A–E)), as well as lactate/alanine ratio (Panel (F)) and LDH protein expression (semi-quantitation–Panel (G); representative membrane–Panel (H)). Corticosterone-free condition (CORT-free) was considered to normalize data (Panel (G)). Ordinary one-way ANOVA, with a Tukey’s multiple comparison test, was used for statistical analysis. Results are expressed as box plots (with median, minimum, and maximal values, as well as total points and first and third quartiles)—panels (A–F) (n = 6 for each condition) and mean ± SEM-Panel (G) (n = 5 for each condition). Data with p < 0.05 were considered statistically different (* p < 0.05; ** p < 0.01; *** p < 0.001). Full article ">Figure 3
Effect of corticosterone (20, 200, and 2000 nM) treatment in mouse SCs (TM4) intact cell oxygen consumption rates (Panel (A)) and mitochondrial membrane potential (ΔΨ relative values by mean of JC-1 fluorescence aggregates/monomers ratio (Panel (B)). Mitochondrial complexes I and II enzymatic activity (Panels (C,D)) and total OXPHOS protein expression (semi-quantitation–Panel (E); representative membrane–Panel (F)). Corticosterone-free condition (CORT-free) was taken into account to normalize data in Panels (A,B,E). Ordinary one-way ANOVA, with a Tukey’s multiple comparison test, was used for statistical analysis. Results are expressed as mean ± SEM—Panels (A) (n = 4–9, different respiratory parameters) and (E) (n = 5 for each condition)—or box plots (with median, minimum, and maximal values, as well as total points and first and third quartiles)—Panels (B) (n = 3 for each condition, with 6 technical replicates), (C,D) (n = 4 for each condition). NDUFB8 (NADH dehydrogenase (ubiquinone) 1 beta subcomplex subunit 8), complex I; SDH8 (succinate dehydrogenase assembly factor 4), complex II; UQCRC2 (cytochrome b-c1complex subunit 2), complex III; MTCO-1 (mitochondria-encoded cytochrome c oxidase I), complex IV; and ATP5A (ATP synthase F1 subunit alpha), complex V. Data with * p < 0.05 were considered statistically different. Full article ">Figure 4
Effect of corticosterone (20, 200, and 2000 nM) treatment in mouse SCs (TM4) on reduced glutathione/oxidized glutathione ratio, as an indirect measurement of cell redox power (as outlined in the panel (A)). Corticosterone-free condition (CORT-free) was considered to normalize data (panel (B)). Ordinary one-way ANOVA, with a Tukey’s multiple comparison test, was used for statistical analysis. Results are expressed as box plots (with median, minimum, and maximal values, as well as total points and first and third quartiles)—(n = 3 for each condition, with 3–9 technical replicates). Data with p < 0.05 were considered statistically different. Full article ">Figure 5
Effect of corticosterone (20, 200, and 2000 nM) treatment in mouse SCs (TM4) Il6 transcript levels. Corticosterone-free condition (CORT-free) was taken into account to normalize data. Ordinary one-way ANOVA, with Tukey’s multiple box plots, was used for statistical analysis. Results are expressed as box plots (with median, minimum, and maximal values, as well as total points and first and third quartiles) (n = 6 for each condition). Data with * p < 0.05 were considered statistically different. Full article ">

24 pages, 1296 KiB

Open AccessSystematic Review

Investigational Treatments in Phase I and II Clinical Trials: A Systematic Review in Asthma

by Luigino Calzetta, Marina Aiello, Annalisa Frizzelli, Elena Pistocchini, Beatrice Ludovica Ritondo, Paola Rogliani and Alfredo Chetta

Biomedicines 2022, 10(9), 2330; https://doi.org/10.3390/biomedicines10092330 - 19 Sep 2022

Cited by 7 | Viewed by 3592

Abstract

Inhaled corticosteroids (ICS) remain the mainstay of asthma treatment, along with bronchodilators serving as control agents in combination with ICS or reliever therapy. Although current pharmacological treatments improve symptom control, health status, and the frequency and severity of exacerbations, they do not really [...] Read more.

Inhaled corticosteroids (ICS) remain the mainstay of asthma treatment, along with bronchodilators serving as control agents in combination with ICS or reliever therapy. Although current pharmacological treatments improve symptom control, health status, and the frequency and severity of exacerbations, they do not really change the natural course of asthma, including disease remission. Considering the highly heterogeneous nature of asthma, there is a strong need for innovative medications that selectively target components of the inflammatory cascade. The aim of this review was to systematically assess current investigational agents in Phase I and II randomised controlled trials (RCTs) over the last five years. Sixteen classes of novel therapeutic options were identified from 19 RCTs. Drugs belonging to different classes, such as the anti-interleukin (IL)-4Rα inhibitors, anti-IL-5 monoclonal antibodies (mAbs), anti-IL-17A mAbs, anti-thymic stromal lymphopoietin (TSLP) mAbs, epithelial sodium channel (ENaC) inhibitors, bifunctional M₃ receptor muscarinic antagonists/β₂-adrenoceptor agonists (MABAs), and anti-Fel d 1 mAbs, were found to be effective in the treatment of asthma, with lung function being the main assessed outcome across the RCTs. Several novel investigational molecules, particularly biologics, seem promising as future disease-modifying agents; nevertheless, further larger studies are required to confirm positive results from Phase I and II RCTs. Full article

(This article belongs to the Special Issue Pathogenesis and Novel Therapeutics in Asthma)

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14 pages, 3350 KiB

Open AccessArticle

Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay

by Aleksandra Anna Zasada, Ewa Mosiej, Marta Prygiel, Maciej Polak, Karol Wdowiak, Kamila Formińska, Robert Ziółkowski, Kamil Żukowski, Kasper Marchlewicz, Adam Nowiński, Julia Nowińska, Waldemar Rastawicki and Elżbieta Malinowska

Biomedicines 2022, 10(9), 2329; https://doi.org/10.3390/biomedicines10092329 - 19 Sep 2022

Cited by 18 | Viewed by 3071

Abstract

Rapid and accurate detection and identification of pathogens in clinical samples is essential for all infection diseases. However, in the case of epidemics, it plays a key role not only in the implementation of effective therapy but also in limiting the spread of [...] Read more.

Rapid and accurate detection and identification of pathogens in clinical samples is essential for all infection diseases. However, in the case of epidemics, it plays a key role not only in the implementation of effective therapy but also in limiting the spread of the epidemic. In this study, we present the application of two nucleic acid isothermal amplification methods—reverse transcription helicase dependent amplification (RT-HDA) and reverse transcription loop-mediated amplification (RT-LAMP)—combined with lateral flow assay as the tools for the rapid detection of SARS-CoV-2, the etiological agent of COVID-19, which caused the ongoing global pandemic. In order to optimize the RT-had, the LOD was 3 genome copies per reaction for amplification conducted for 10–20 min, whereas for RT-LAMP, the LOD was 30–300 genome copies per reaction for a reaction conducted for 40 min. No false-positive results were detected for RT-HDA conducted for 10 to 90 min, but false-positive results occurred when RT-LAMP was conducted for longer than 40 min. We concluded that RT-HDA combined with LFA is more sensitive than RT-LAMP, and it is a good alternative for the development of point-of-care tests for SARS-CoV-2 detection as this method is simple, inexpensive, practical, and does not require qualified personnel to perform the test and interpret its results. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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Agarose gel electrophoresis of PCR products: M—molecular DNA marker 100 bp; 1—PCR for gene RdRP (primers nCoV_IP4-14059Fw/Rv); 2—negative control for gene RdRP; 3—PCR for gene E (primers E_Sarbeco_F/R); 4—negative control for gene E; 5—PCR for gene N (primers 2019-nCoV_N2-F/R); and 6—negative control for gene N. Full article ">Figure 2
Detection of PCR products using LFA. 1—gene RdRP (primers nCoV_IP4-14059Fw/Rv); 2—negative control for gene RdRP; 3—gene E (primers E_Sarbeco_F/); 4—negative control for gene E; 5—gene N (primers 2019-nCoV_N2-F/R); and 6—negative control for gene E. Full article ">Figure 3
An example of RT-HDA combined with LFA performed using serial dilutions of the synthetic template and various incubation times. Full article ">Figure 4
An example of RT-HDA combined with LFA performed using serial dilutions of AMPLIRUN SARS-CoV-2 RNA control and various incubation times. cp/rx—copies/reactions; neg.—negative control. Full article ">Figure 5
An example of RT-LAMP combined with LFA performed using AMPLIRUN SARS-CoV-2 RNA control at concentration 300 copies/reactions and various incubation times. Full article ">Figure 6
Agarose gel electrophoresis of RT-LAMP for clinical samples conducted for 40 min. M—molecular DNA marker 100 bp; S—clinical sample; K (-)—negative control; and K (+)—AMPLIRUN SARS-CoV-2 RNA control at concentration 3000 copies/reactions. Full article ">

23 pages, 5297 KiB

Open AccessArticle

Dexamethasone-Induced Adipose Tissue Redistribution and Metabolic Changes: Is Gene Expression the Main Factor? An Animal Model of Chronic Hypercortisolism

by Flaviane de Fatima Silva, Ayumi Cristina Medeiros Komino, Sandra Andreotti, Gabriela Boltes Reis, Rennan Oliveira Caminhotto, Richardt Gama Landgraf, Gabriel Orefice de Souza, Rogerio Antonio Laurato Sertié, Sheila Collins, Jose Donato, Jr. and Fabio Bessa Lima

Biomedicines 2022, 10(9), 2328; https://doi.org/10.3390/biomedicines10092328 - 19 Sep 2022

Cited by 7 | Viewed by 2981

Abstract

Chronic hypercortisolism has been associated with the development of several metabolic alterations, mostly caused by the effects of chronic glucocorticoid (GC) exposure over gene expression. The metabolic changes can be partially explained by the GC actions on different adipose tissues (ATs), leading to [...] Read more.

Chronic hypercortisolism has been associated with the development of several metabolic alterations, mostly caused by the effects of chronic glucocorticoid (GC) exposure over gene expression. The metabolic changes can be partially explained by the GC actions on different adipose tissues (ATs), leading to central obesity. In this regard, we aimed to characterize an experimental model of iatrogenic hypercortisolism in rats with significant AT redistribution. Male Wistar rats were distributed into control (CT) and GC-treated, which received dexamethasone sodium phosphate (0.5 mg/kg/day) by an osmotic minipump, for 4 weeks. GC-treated rats reproduced several characteristics observed in human hypercortisolism/Cushing’s syndrome, such as HPA axis inhibition, glucose intolerance, insulin resistance, dyslipidemia, hepatic lipid accumulation, and AT redistribution. There was an increase in the mesenteric (meWAT), perirenal (prWAT), and interscapular brown (BAT) ATs mass, but a reduction of the retroperitoneal (rpWAT) mass compared to CT rats. Overexpressed lipolytic and lipogenic gene profiles were observed in white adipose tissue (WAT) of GC rats as BAT dysfunction and whitening. The AT remodeling in response to GC excess showed more importance than the increase of AT mass per se, and it cannot be explained just by GC regulation of gene transcription. Full article

(This article belongs to the Special Issue Animal Models of Human Pathology: Revision, Relevance and Refinements)

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Glucocorticoid continuous administration promoted inhibition of the HPA axis. (a) Serum corticosterone, (b) adrenal gland mass, and (c) histological sections; (d) spleen mass, and (e) Hsd11b1 gene expression in mesenteric—meWAT, perirenal—prWAT, retroperitoneal—rpWAT, epididymal—epWAT, and subcutaneous inguinal—scWAT of control (CT), and glucocorticoid-treated rats (GC). Data are mean ± SEM of 13 (CT) and 17 (GC) rats (a,b,d), and 7–12 (CT) 11-12 (GC) samples (e). * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 vs. CT (Unpaired Student t-test; Mann–Whitney test in a and d). (c) HE staining—100× and 400× magnification. G: Glomerulosa zone; F: Fasciculata zone; R: Reticularis zone, and M: Medulla. Full article ">Figure 2
Chronic iatrogenic hypercortisolism promotes glucose intolerance, insulin resistance, dyslipidemia, and increase of hepatic lipid content. (a) Blood glucose and (b) plasma insulin during oGTT test; (c) 12h fasted blood glucose and insulin; (d) HOMA-IR; serum (e) adiponectin, (f) leptin; (g) lipids-NEFA, triglycerides, total cholesterol, HDL, LDL, and VLDL-cholesterol fractions; (h) lactate, and the (i) transaminases ALT and AST; (j) liver mass, (k) triglycerides content, and histological sections of control (CT), and glucocorticoid-treated (GC) rats. HE staining; 100×, 200×, and 400× magnification; the black arrows indicates the central vein. Data are mean ± SEM of 13 (CT) and 17 (GC) rats/samples. * p < 0.05; ** p < 0.01; *** p < 0.001, and **** p < 0.0001 vs. CT (unpaired Student t-test, Mann–Whitney test in b-AUC, and c-fasting glucose). Full article ">Figure 3
Food intake and body changes in response to GC excess. (a) Daily food intake per week; (b) average 28 days daily food intake; (c) feed efficiency; (d) hypothalamic gene expression of Npy, Agrp, Pomc, Cartpt, and Lepr; (e) body weight change; (f) nasal–anal length; (g) gastrocnemius—G, soleus—S, and extensor digitorum longus—E muscle mass; and (h) tibia bone weight, length, and circumference of control (CT), and glucocorticoid-treated (GC) rats. Data are mean ± SEM of 13 (CT) and 17 (GC) rats/samples (a–c,e–h), and 6 (CT) and 6 (GC) samples (d). * p < 0.05; ** p < 0.01; *** p < 0.001 and **** p < 0.0001 vs. CT (unpaired Student t-test; Mann–Whitney test in h-length). (f) **** p < 0.0001 (Repeated Measures Two-Way ANOVA with Bonferroni’s post hoc test). Full article ">Figure 4
Adipose tissue redistribution after chronic glucocorticoid treatment. (a) adipose mass of mesenteric—meWAT, perirenal—prWAT, retroperitoneal—rpWAT, epididymal—epWAT, and subcutaneous inguinal—scWAT; (b) interscapular brown adipose tissue—BAT mass and histological sections; (c) WAT mass; (d) WAT + BAT mass; frequency distribution of adipocyte volume (pL) and histological sections of (e) meWAT, (f) prWAT, (g) rpWAT, (h) epWAT, and (i) scWAT of control (CT), and glucocorticoid-treated (GC) rats. HE staining; 100× magnification. Data are mean ± SEM of 13 (CT) and 17 (GC) rats/samples. * p < 0.05; ** p < 0.01, and **** p < 0.0001 vs. CT (unpaired Student t-test). Full article ">Figure 5
Gene expression of the lipolytic pathway in white adipose tissue. (a) mesenteric—meWAT; (b) perirenal—prWAT; (c) retroperitoneal—rpWAT; (d) epididymal—epWAT; and (e) subcutaneous inguinal—scWAT. Genes of beta-adrenergic receptors—Adrb1, Adrb2, and Adbr3; catalytic subunits of PKA—Prkaca and Prkacb; perilipin in white mature adipocytes—Plin1; lipase cofactors—Abdh5, G0s2, and Fabp4; lipases—Pnpla2, Lipe, and Mgll; lipolytic products channel and transporter—Aqp7 and Cd36—of control (CT; n = 7–12), and glucocorticoid-treated (GC; n = 11–12) rats. Data are mean ± SEM. * p < 0.05; ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. CT (unpaired Student t-test; Mann–Whitney test in (a) (Adrb2, Adrb3, Fabp4, Abdh5, Mgll, and Aqp7), (b) (Adrb3 and Fabp4), (c) (Adrb2 and Adrb3), (d) (Adrb2, Adrb3, and Prkacb), and e (all, except G0s2)). Full article ">Figure 6
Gene expression of the lipogenic pathway in white adipose tissue. (a) mesenteric—meWAT; (b) perirenal—prWAT; (c) retroperitoneal—rpWAT; (d) epididymal—epWAT; and (e) subcutaneous inguinal. Genes of lipoprotein lipase—Lpl; enzymes of fatty acids esterification—Gpam, Agpat1, Agpat2, Dgat1, and Dgat2; cytosolic NADPH recyclers—G6pd and Me1; enzymes of lipogenesis de novo—Acly, Acaca, and Fasn; fatty acid desaturases—Scd and Scd2; monocarboxylate transporters—Slc16a1 and Slc16a7; lactate dehydrogenase—Ldhb; the key enzyme of glyceroneogenesis—Pck1; glucose transporter 1—Slc2a1; the last enzyme of glycerol-3-phosphate generation glycolytic pathway and glyceroneogenesis—Gpd1; and the glycerol kinase—Gk—of control (CT; n = 7–12), and glucocorticoid-treated (GC; n = 11–12) rats. Data are mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001, and **** p < 0.0001 vs. CT (unpaired Student t-test; Mann–Whitney test in (a) (Gpam, G6pd, Me1, Acly, Acaca, Fasn, Scd, Scd2 and Gpd1), (b) (Agpat1, Dgat2, G6pd, Me1, Acly, Acaca, Scd, Slc16a7, and Gk), (c) (Me1, Fasn, Scd, and Slc2a1), (d) (Agpat2, G6pd, Acly, and Ldhb), and (e) (Lpl, Agpat2, Dgat1, Dgat2, Acaca, Scd2, Slc16a1, Slc16a7, Pck1, Slc2a1, and Gpd1)). Full article ">Figure 7
BAT gene and functional changes after chronic GC exposure. (a) Ucp1 gene expression; (b) UCP1 immunohistochemistry; (c) Ucp1 expression regulators—Adrb3, Prdm16, Ppargc1a, and Pparg; (d) brown adipocytes markers—Zic1, Tmem26, Pat2, P2rx5, Cidea, and Dio2; (e) citrate synthase maximal activity; oxidation of (f) [1-14C]-palmitic acid and (g) D-[U-14C]- glucose; (h) lipolysis pathway genes—Abdh5, G0s2, Fabp4, Pnpla2, Lipe, and Mgll; (i) incorporation of D-[U-C14]- glucose into lipids; (j) lipogenic pathway genes—Acly, Acaca, Fasn, and Dgat2; (k) Lpl expression; in vivo temperature analysis of (l) body and (m) BAT area on days 0 and 28th of control (CT), and glucocorticoid-treated (GC) rats. (b) UCP1 staining: 100× magnification. Data are mean ± SEM of 8 (CT) and 8 (GC) samples (a,c,d,h,j,k), 8 (CT) and 9 (GC) samples (e,f,g,i), and 10 (CT) and 11 (GC) rats (l,m). * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 vs. CT (unpaired Student t-test; Mann–Whitney test in (c)-Adrb3 and Ppargc1a). Full article ">

9 pages, 711 KiB

Open AccessArticle

Clinical Performance of Self-Collected Nasal Swabs and Antigen Rapid Tests for SARS-CoV-2 Detection in Resource-Poor Settings

by Nádia Sitoe, Júlia Sambo, Nédio Mabunda, Neuza Nguenha, Jorfélia Chilaúle, Júlio Rafael, Anésio Macicame, Imelda Chelene, Chishamiso Mudenyanga, Jillian Sacks, Sofia Viegas, Osvaldo Loquiha and Ilesh Jani

Biomedicines 2022, 10(9), 2327; https://doi.org/10.3390/biomedicines10092327 - 19 Sep 2022

Cited by 2 | Viewed by 2387

Abstract

Background: In resource-poor countries, antigen-based rapid tests (Ag-RDTs) performed at primary healthcare and community settings improved access to SARS-CoV-2 diagnostics. However, the technical skills and biosafety requirements inherent to nasopharyngeal and oropharyngeal (OP) specimens limit the scale-up of SARS-CoV-2 testing. The collection of [...] Read more.

Background: In resource-poor countries, antigen-based rapid tests (Ag-RDTs) performed at primary healthcare and community settings improved access to SARS-CoV-2 diagnostics. However, the technical skills and biosafety requirements inherent to nasopharyngeal and oropharyngeal (OP) specimens limit the scale-up of SARS-CoV-2 testing. The collection of nasal-swabs is programmatically viable, but its performance has not been evaluated in resource-poor settings. Methods: We first evaluated the performance of SteriPack self-collected nasal swabs for the detection of SARS-CoV-2 by real-time PCR in 1498 consecutively enrolled patients with suspected infection. Next, we evaluated the clinical performance of three nasal swab-based Ag-RDTs against real-time PCR on OP specimens. Results: The sensitivity of nasal swabs was 80.6% [95% CI: 75.3–85.2%] compared to OP specimens. There was a good correlation (r = 0.58; p < 0.0001) between Ct values of 213 positive cases obtained using nasal and OP swabs. Our findings show sensitivities of 79.7% [95% CI: 73.3–85.1%] for Panbio COVID-19 Ag-RDT, 59.6% [95% CI: 55.2–63.8%] for COVIOS Ag-RDT, and 78.0% [95% CI: 73.5–82.0%] for the LumiraDx SARS-CoV-2 Ag-RDT. Conclusions: In our setting, the COVIOS Ag-RDT did not meet WHO requirements. Nasal swab-based Ag-RDTs for SARS-CoV-2 detection constitute a viable and accurate diagnostic option in resource-poor settings. Full article

(This article belongs to the Special Issue High Sensitivity Lateral Flow Assays for SARS-CoV-2 and Other Infections)

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23 pages, 5211 KiB

Open AccessArticle

Small Molecules as Toll-like Receptor 4 Modulators Drug and In-House Computational Repurposing

by Lucía Pérez-Regidor, Joan Guzmán-Caldentey, Nils Oberhauser, Carmen Punzón, Balázs Balogh, José R. Pedro, Eva Falomir, Alessandra Nurisso, Péter Mátyus, J. Carlos Menéndez, Belén de Andrés, Manuel Fresno and Sonsoles Martín-Santamaría

Biomedicines 2022, 10(9), 2326; https://doi.org/10.3390/biomedicines10092326 - 19 Sep 2022

Cited by 9 | Viewed by 3510

Abstract

The innate immunity toll-like receptor 4 (TLR4) system is a receptor of paramount importance as a therapeutic target. Virtual screening following a “computer-aided drug repurposing” approach was applied to the discovery of novel TLR4 modulators with a non-lipopolysaccharide-like structure. We screened almost 29,000 [...] Read more.

The innate immunity toll-like receptor 4 (TLR4) system is a receptor of paramount importance as a therapeutic target. Virtual screening following a “computer-aided drug repurposing” approach was applied to the discovery of novel TLR4 modulators with a non-lipopolysaccharide-like structure. We screened almost 29,000 approved drugs and drug-like molecules from commercial, public, and in-house academia chemical libraries and, after biological assays, identified several compounds with TLR4 antagonist activity. Our computational protocol showed to be a robust approach for the identification of hits with drug-like scaffolds as possible inhibitors of the TLR4 innate immune pathways. Our collaborative work broadens the chemical diversity for inspiration of new classes of TLR4 modulators. Full article

(This article belongs to the Special Issue State-of-the-Art Drug Discovery and Development in Spain)

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Graphical abstract

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Full article ">Figure 1
(A) Full-atom 3D model of the agonist LPS-bound TLR4 dimer in a membrane environment [<a href="#B81-biomedicines-10-02326" class="html-bibr">81</a>]; ectodomain is framed in a red box. (B) X-ray structure of the TLR4/MD-2 dimer in complex with E. coli LPSs (from PDB ID 3FXI). (C) Three-dimensional model of TLR4/MD-2 in the antagonist conformation [<a href="#B66-biomedicines-10-02326" class="html-bibr">66</a>]. Full article ">Figure 2
Flowchart of the virtual screening, redocking, and biological assay protocol. Full article ">Figure 3
Redocked ID 5382 (orange sticks) in TLR4/MD-2. MD-2-interacting residues are highlighted with grey sticks. TLR4 is depicted in pale pink, MD-2—in yellow. Full article ">Figure 4
Redocked PM1090 (salmon sticks) compound in TLR4/MD-2. TLR4 is depicted in pale pink; MD-2—in yellow. MD-2 interacting residues are highlighted with grey sticks. Full article ">Figure 5
Redocked MS-21 ((A), green sticks) and MS-32 ((B), cyan sticks) in TLR4/MD2. TLR4 is depicted in pale pink; MD-2—in yellow. MD-2 interacting residues are highlighted with grey sticks. Full article ">Figure 6
HEK-BlueTM treatment with increasing concentrations of compounds, stimulated with LPSs from E. coli. The results are expressed in the percentage of TLR4 activation (positive control: E. coli LPSs, 20 ng/mL). Full article ">Figure 7
Expression of TNF-α secretion (black bars) and cell viability (white bars) in the J744 cell line upon treatment with the screened compounds B, F, I, J, X, Z, and M4. The results were normalized with the positive control (LPSs alone) and expressed as the mean percentage ± SD of at least three independent experiments. Full article ">

19 pages, 987 KiB

Open AccessReview

Trends and Perspectives of Biological Drug Approvals by the FDA: A Review from 2015 to 2021

by Alexander C. Martins, Mariana Y. Oshiro, Fernando Albericio, Beatriz G. de la Torre, Gustavo José V. Pereira and Rodrigo V. Gonzaga

Biomedicines 2022, 10(9), 2325; https://doi.org/10.3390/biomedicines10092325 - 19 Sep 2022

Cited by 21 | Viewed by 6366

Abstract

Despite belonging to a relatively new class of pharmaceuticals, biological drugs have been used since the 1980s, when they brought about a breakthrough in the treatment of chronic diseases, especially cancer. They conquered a large space in the pipeline of the pharmaceutical industry [...] Read more.

Despite belonging to a relatively new class of pharmaceuticals, biological drugs have been used since the 1980s, when they brought about a breakthrough in the treatment of chronic diseases, especially cancer. They conquered a large space in the pipeline of the pharmaceutical industry and boosted the innovation portfolio and arsenal of therapeutic compounds available. Here, we report on biological drug approvals by the US Food and Drug Administration (FDA) from 2015 to 2021. The number of drugs included in this class grew over this period, totaling 90 approvals, with an average of 13 authorizations per year. This figure contrasts with previous periods, which registered between 2 and 8 approvals per year. We highlight the great potential and advantages of biological drugs. In this context, these therapeutics show high efficacy and high selectivity, and they have brought about a significant increase in patient survival and a reduction of adverse reactions. The development and production of biopharmaceuticals pose a major challenge because these processes require cutting-edge technology, thereby making the drugs very expensive. However, we believe that, in the near future, biological medicines will be more accessible and new drugs belonging to this class will become available as new technologies emerge. Such advances will enhance the production of these biopharmaceuticals, thereby making the process increasingly profitable and less expensive, thereby bringing about greater availability of these drugs. Full article

(This article belongs to the Section Drug Discovery, Development and Delivery)

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13 pages, 1589 KiB

Open AccessArticle

The JAK1/2 Inhibitor Baricitinib Mitigates the Spike-Induced Inflammatory Response of Immune and Endothelial Cells In Vitro

by Amelia Barilli, Rossana Visigalli, Francesca Ferrari, Giulia Recchia Luciani, Maurizio Soli, Valeria Dall’Asta and Bianca Maria Rotoli

Biomedicines 2022, 10(9), 2324; https://doi.org/10.3390/biomedicines10092324 - 19 Sep 2022

Cited by 9 | Viewed by 2458

Abstract

The purpose of this study was to examine the effect of the JAK-STAT inhibitor baricitinib on the inflammatory response of human monocyte-derived macrophages (MDM) and endothelial cells upon exposure to the spike S1 protein from SARS-CoV-2. The effect of the drug has been [...] Read more.

The purpose of this study was to examine the effect of the JAK-STAT inhibitor baricitinib on the inflammatory response of human monocyte-derived macrophages (MDM) and endothelial cells upon exposure to the spike S1 protein from SARS-CoV-2. The effect of the drug has been evaluated on the release of cytokines and chemokines from spike-treated MDM, as well as on the activation of endothelial cells (HUVECs) after exposure to conditioned medium collected from spike-activated MDM. Results obtained indicate that, in MDM, baricitinib prevents the S1-dependent phosphorylation of STAT1 and STAT3, along with the induction of IP-10- and MCP-1 secretion; the release of IL-6 and TNFα is also reduced, while all other mediators tested (IL-1β, IL-8, RANTES, MIP-1α and MIP-1β) are not modified. Baricitinib is, instead, poorly effective on endothelial activation when HUVECs are exposed to supernatants from S1-activated macrophages; the induction of VCAM-1, indeed, is not affected by the drug, while that of ICAM-1 is only poorly inhibited. The drug, however, also exerts protective effects on the endothelium by limiting the expression of pro-inflammatory mediators, specifically IL-6, RANTES and IP-10. No effect of baricitinib has been observed on IL-8 synthesis and, consistently, on neutrophils chemiotaxis. Our in vitro findings reveal that the efficacy of baricitinib is limited, with effects mainly focused on the inhibition of the IL-6-mediated inflammatory loop. Full article

(This article belongs to the Special Issue Non-antiviral Agents for Treatment of COVID-19)

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Figure 1
Monocyte-derived macrophages (MDM) were left untreated (control) or incubated for 4 h with 5 nM spike S1 protein in the absence (S1) or in the presence of 1 µM baricitinib (S1+baricitinib). The expression of the indicated genes was measured by means of RT-qPCR and calculated relatively to that of the housekeeping gene RPL15, by employing the ∆∆Ct method. Bars are means ± SEM of eight independent determinations (single dots), each performed in duplicate. ** p < 0.01, *** p < 0.001 vs. untreated cells (control); $ p < 0.05, $$$ p < 0.001 vs. S1-treated cells with one-way ANOVA. Full article ">Figure 2
MDM were left untreated (control) or incubated for 24 h with 5 nM spike S1 protein in the absence (S1) or in the presence of 1 µM baricitinib (S1+baricitinib). Cytokines released in the medium were quantified with an ELISA assay, as described in Materials and Methods. Bars are means ± SEM of eight independent experiments (single dots). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. untreated cells (control); $ p < 0.05, $$ p < 0.01, $$$ p < 0.001 vs. S1-treated cells with one-way ANOVA. Full article ">Figure 3
Human polymorphonucleates (PMN, Panel (A)) and T lymphocytes (CD3+ cells, Panel (B)) were isolated and allowed to migrate through confluent HUVECs grown on transwell inserts towards CM_cont, CM_S1 or CM_S1_baricitinb. After 4 h (PMN) or 24 h (CD3+ cells), the amount of transmigrated cells in the lower compartment was determined as described in Materials and Methods. Bars are the mean ± SEM of three independent determinations (single dots). * p < 0.05 with one-way ANOVA. Full article ">Figure 4
Monocyte-derived macrophages (MDM) were left untreated (control) or incubated for the indicated times with 5 nM spike S1 in the absence (S1) or in the presence of 1 µM baricitinib (S1+baricitinib). The expression of the indicated proteins was assessed by means of western blot analysis, as detailed in Materials and Methods; representative blots are shown, along with the mean of the densitometric analyses (bars) of three different experiments (single dots). Full article ">Figure 5
HUVEC were incubated in MDM-conditioned medium obtained by incubating MDM for 24 h in the absence (CM_cont) or in the presence of 5 nM S1 (CM_S1); baricitinib was added to CM_S1 1 h before the experiment and maintained during the whole experiment. After 4 h, the expression of the indicated genes (left panels) was measured by means of RT-qPCR and calculated relatively to CM_cont (=1; dotted line) upon normalization for the housekeeping gene RPL15, by employing the 2∆Ct method. Bars are means ± SEM of four experiments (single dots), each performed in duplicate. After 6 h, the amount of the corresponding proteins (right panels) was assessed by means of Western Blot analysis (see Materials and Methods); representative blots are shown, along with the mean of the densitometric analyses (bars) of three different experiments (single dots). Full article ">Figure 6
HUVECs were incubated for 4 h as described in <a href="#biomedicines-10-02324-f005" class="html-fig">Figure 5</a>. The expression of the indicated genes was measured by means of RT-qPCR and calculated relatively to CM_cont (=1; dotted line) upon normalization for the housekeeping gene RPL15, by employing the 2∆Ct method. Bars are means ± SEM of four experiments (single dots), each performed in duplicate. $ p < 0.05, $$ p < 0.01 vs. S1-treated cells with with Student’s t test. Full article ">

11 pages, 1332 KiB

Open AccessArticle

Deep Learning for Bone Mineral Density and T-Score Prediction from Chest X-rays: A Multicenter Study

by Yoichi Sato, Norio Yamamoto, Naoya Inagaki, Yusuke Iesaki, Takamune Asamoto, Tomohiro Suzuki and Shunsuke Takahara

Biomedicines 2022, 10(9), 2323; https://doi.org/10.3390/biomedicines10092323 - 19 Sep 2022

Cited by 28 | Viewed by 5642

Abstract

Although the number of patients with osteoporosis is increasing worldwide, diagnosis and treatment are presently inadequate. In this study, we developed a deep learning model to predict bone mineral density (BMD) and T-score from chest X-rays, which are one of the most common, [...] Read more.

Although the number of patients with osteoporosis is increasing worldwide, diagnosis and treatment are presently inadequate. In this study, we developed a deep learning model to predict bone mineral density (BMD) and T-score from chest X-rays, which are one of the most common, easily accessible, and low-cost medical imaging examination methods. The dataset used in this study contained patients who underwent dual-energy X-ray absorptiometry (DXA) and chest radiography at six hospitals between 2010 and 2021. We trained the deep learning model through ensemble learning of chest X-rays, age, and sex to predict BMD using regression and T-score for multiclass classification. We assessed the following two metrics to evaluate the performance of the deep learning model: (1) correlation between the predicted and true BMDs and (2) consistency in the T-score between the predicted class and true class. The correlation coefficients for BMD prediction were hip = 0.75 and lumbar spine = 0.63. The areas under the curves for the T-score predictions of normal, osteopenia, and osteoporosis diagnoses were 0.89, 0.70, and 0.84, respectively. These results suggest that the proposed deep learning model may be suitable for screening patients with osteoporosis by predicting BMD and T-score from chest X-rays. Full article

(This article belongs to the Special Issue Artificial Intelligence in Biological and Biomedical Imaging 2.0)

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15 pages, 1755 KiB

Open AccessBrief Report

Retinol Binding Protein, Sunlight Hours, and the Influenza Virus-Specific Immune Response

by Nehali Patel, Rhiannon R. Penkert, Robert E. Sealy, Sherri L. Surman, Bart G. Jones, Karen Ringwald-Smith, A. Catharine Ross and Julia L. Hurwitz

Biomedicines 2022, 10(9), 2322; https://doi.org/10.3390/biomedicines10092322 - 19 Sep 2022

Cited by 4 | Viewed by 2743

Abstract

Healthy pediatric immune responses depend on adequate vitamin A and D levels. Relationships between solar ultraviolet B (UVB) radiation and vitamin D are well understood, while relationships between sunlight, vitamin A, and its serum escort, retinol binding protein (RBP), are not. A pediatric [...] Read more.

Healthy pediatric immune responses depend on adequate vitamin A and D levels. Relationships between solar ultraviolet B (UVB) radiation and vitamin D are well understood, while relationships between sunlight, vitamin A, and its serum escort, retinol binding protein (RBP), are not. A pediatric clinical study enrolled 2–8-year-old children at various times between September 2016 and March 2017, inclusive, in Memphis, Tennessee. A serum sample from each child was then assayed to examine the influence of season on vitamin levels. We found that RBP and RBP/retinol molar ratios decreased in winter months and RBP/retinol ratios correlated positively with the average daily sunlight hours per month. A food frequency questionnaire given to parents/guardians indicated a shift in dietary intake from plant-based foods to animal-based foods by children between winter and spring months. This translated to higher retinol and zinc (integral to RBP–transthyretin–retinol complexes) in the spring, perhaps explaining the seasonal influence on RBP/retinol. RBP and retinol were associated positively with IgG/IgM and IgA/IgM ratios. RBP and retinol, but not 25(OH)D, also correlated positively with influenza virus-specific antibodies. Retinol correlated negatively, while 25(OH)D correlated positively, with certain serum cytokine/chemokine levels. Significant differences in 25(OH)D, immunoglobulin ratios, and cytokines/chemokines were observed between black and white children. In sum, seasonal changes in dietary foods rich in retinol and zinc may have influenced RBP levels, which in turn influenced innate and adaptive immune responses. Results encourage routine monitoring and reporting of season, RBP, and vitamin levels in future clinical studies, as seasons may affect sunlight exposures, diet, vitamin levels, and immune protection against infectious disease. Full article

(This article belongs to the Special Issue Molecular Research in Infectious Diseases)

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The 25(OH)D levels are shown as categorized by month for (A) all study participants (solid and open symbols represent black and white participants, respectively), (B) black participants alone, and (C) white participants alone. The month with the lowest mean value is highlighted in yellow. Means (also recorded by number above symbols) with standard deviations are shown. (D) 25(OH)D levels were plotted against Julian day of the year with solid and open symbols representing black and white participants, respectively. A linear regression line and a quadratic regression curve are shown (R2 = 0.01, p = 0.44 for the linear regression; R2 = 0.07 for the quadratic regression). Full article ">Figure 2
Seasonal changes in retinol/RBP ratios. (A) Individual RBP values (μmol/L) and means are shown per month of study enrollment. The month with the lowest mean value is highlighted in yellow. (B) Individual retinol values (μmol/L) and means are shown per month of study enrollment. (C) RBP/retinol molar ratios and means are shown per month. The month with the lowest mean ratio is highlighted in yellow. (D) RBP:retinol molar ratios were correlated with Julian days of the year. A linear regression line and a quadratic regression curve are shown (R2 = 0.001, p = 0.81 for the linear regression; R2 = 0.20 for the quadratic regression). Solid and open symbols represent black and white participants, respectively. Each symbol represents a different study participant for (A–D). (E) Mean RBP:retinol molar ratios were correlated with mean sunlight hours per day as measured in Nashville, Tennessee, each month (Pearson correlation). Full article ">Figure 3
Dietary intake changes with season. Parents/guardians of 30 children completed an FFQ. FFQ results reported in the winter (December (2016), January (2017), and February (2017)) were compared to results reported in the spring (March to May 2017). A portion of variables are shown, including (A–D) food items, and (E–H) calculated intakes of vitamins and zinc. Each symbol represents a different child with medians shown. There were seven foods (from approximately 90 tested food groups) that were significantly different between winter and spring, as revealed both by t-tests and Mann–Whitney tests. These were peas, vegetable stews, peanut butter, butter, and pretzels (more prevalent in the winter), and fried fish and burgers (more prevalent in the spring). Results of Mann–Whitney tests are shown. Significant differences between black and white participants were not observed for these variables. Full article ">Figure 4
Vitamin levels correlate with serum immunoglobulin isotype patterns. (A–I) RBP and retinol, but not 25(OH)D, correlate positively with isotype ratios, indicative of immunoglobulin class switch from IgM to non-IgM isotypes. Each symbol represents a different participant. A simple linear regression line is plotted. Spearman correlation test results are shown. (J–L) Black and white children differed significantly in isotype ratios. Medians and results from Mann–Whitney tests are shown. Full article ">Figure 5
RBP and retinol, but not 25(OH)D, correlate with influenza virus B-specific binding antibodies in pediatric human sera. (A) RBP, (B) retinol, and (C) 25(OH)D are each plotted as a function of baseline scores for influenza virus B-specific binding antibodies. Each symbol represents a different participant. A simple linear regression line is plotted. Spearman correlation test results are shown. Full article ">Figure 6
Vitamin and cytokine/chemokine levels. Significant correlations are shown between cytokine/chemokine (A,B) and retinol (C–J) or 25(OH)D levels. Each symbol represents a different individual. Spearman correlation test results are shown. Full article ">Figure 7
Cytokine/chemokine levels in black and white children. Black and white children were compared for the nine cytokines/chemokines shown in <a href="#biomedicines-10-02322-f006" class="html-fig">Figure 6</a>. In five cases, white children exhibited higher values than black children, consistent with their higher serum 25(OH)D levels. Each symbol represents a different individual. Median and Mann–Whitney results are shown. Levels of IL-1β, IL-2, MCP-3, and TGFα did not differ significantly between the black and white populations. Full article ">

19 pages, 8013 KiB

Open AccessArticle

Level of Amyloid-β (Aβ) Binding Leading to Differential Effects on Resting State Functional Connectivity in Major Brain Networks

by Eva Y. W. Cheung, Anson C. M. Chau, Yat-Fung Shea, Patrick K. C. Chiu, Joseph S. K. Kwan and Henry K. F. Mak

Biomedicines 2022, 10(9), 2321; https://doi.org/10.3390/biomedicines10092321 - 19 Sep 2022

Cited by 5 | Viewed by 2353

Abstract

Introduction: Amyloid-β protein (Aβ) is one of the biomarkers for Alzheimer’s disease (AD). The recent application of interhemispheric functional connectivity (IFC) in resting-state fMRI has been used as a non-invasive diagnostic tool for early dementia. In this study, we focused on the level [...] Read more.

Introduction: Amyloid-β protein (Aβ) is one of the biomarkers for Alzheimer’s disease (AD). The recent application of interhemispheric functional connectivity (IFC) in resting-state fMRI has been used as a non-invasive diagnostic tool for early dementia. In this study, we focused on the level of Aβ accumulated and its effects on the major functional networks, including default mode network (DMN), central executive network (CEN), salience network (SN), self-referential network (SRN) and sensory motor network (SMN). Methods: 58 participants (27 Hi Aβ (HiAmy) and 31 low Aβ (LowAmy)) and 25 healthy controls (HC) were recruited. [¹⁸F]flutemetamol PET/CT was performed for diseased groups, and MRI scanning was done for all participants. Voxel-by-voxel correlation analysis was done for both groups in all networks. Results: In HiAmy, IFC was reduced in all networks except SN. A negative correlation in DMN, CEN, SRN and SMN suggests high Aβ related to IFC reduction; However, a positive correlation in SN suggests high Aβ related to an increase in IFC. In LowAmy, IFC increased in CEN, SMN, SN and SRN. Positive correlation in all major brain networks. Conclusion: The level of Aβ accumulated demonstrated differential effects on IFC in various brain networks. As the treatment to reduce Aβ plaque deposition is available in the market, it may be an option for the HiAmy group to improve their IFC in major brain networks. Full article

(This article belongs to the Special Issue State of the Art: Prerequisites, Prevention and Treatment of Neurodegenerative Diseases)

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13 pages, 325 KiB

Open AccessReview

The Influence of SARS-CoV-2 Infection on Lipid Metabolism—The Potential Use of Lipid-Lowering Agents in COVID-19 Management

by Klaudia Kowalska, Zofia Sabatowska, Joanna Forycka, Ewelina Młynarska, Beata Franczyk and Jacek Rysz

Biomedicines 2022, 10(9), 2320; https://doi.org/10.3390/biomedicines10092320 - 18 Sep 2022

Cited by 18 | Viewed by 3972

Abstract

Several studies have indicated lipid metabolism alterations during COVID-19 infection, specifically a decrease in high-density lipoprotein (HDL) and low-density lipoprotein (LDL) concentrations and an increase in triglyceride (TG) levels during the infection. However, a decline in triglycerides can also be observed in critical [...] Read more.

Several studies have indicated lipid metabolism alterations during COVID-19 infection, specifically a decrease in high-density lipoprotein (HDL) and low-density lipoprotein (LDL) concentrations and an increase in triglyceride (TG) levels during the infection. However, a decline in triglycerides can also be observed in critical cases. A direct correlation can be observed between a decrease in serum cholesterol, HDL-C, LDL-C and TGs, and the severity of the disease; these laboratory findings can serve as potential markers for patient outcomes. The transmission of coronavirus increases proportionally with rising levels of cholesterol in the cell membrane. This is due to the fact that cholesterol increases the number of viral entry spots and the concentration of angiotensin-converting enzyme 2 (ACE2) receptor, crucial for viral penetration. Studies have found that lower HDL-C levels correspond with a higher susceptibility to SARS-CoV-2 infection and infections in general, while higher HDL-C levels were related to a lower risk of developing them. However, extremely high HDL-C levels in serum increase the risk of infectious diseases and is associated with a higher risk of cardiovascular events. Low HDL-C levels are already accepted as a marker for risk stratification in critical illnesses, and higher HDL-C levels prior to the infection is associated with a lower risk of death in older patients. The correlation between LDL-C levels and disease severity is still unclear. However, TG levels were significantly higher in non-surviving severe patients compared to those that survived; therefore, elevated TG-C levels in COVID-19 patients may be considered an indicator of uncontrolled inflammation and an increased risk of death. Full article

(This article belongs to the Special Issue Cardiovascular Diseases and COVID-19)

22 pages, 2925 KiB

Open AccessSystematic Review

Quantitative Measurement of Swallowing Performance Using Iowa Oral Performance Instrument: A Systematic Review and Meta-Analysis

by Raffaella Franciotti, Erica Di Maria, Michele D’Attilio, Giuseppe Aprile, Federica Giulia Cosentino and Vittoria Perrotti

Biomedicines 2022, 10(9), 2319; https://doi.org/10.3390/biomedicines10092319 - 18 Sep 2022

Cited by 14 | Viewed by 6904

Abstract

Swallowing is a complex but stereotyped motor activity aimed at serving two vital purposes: alimentary function and the protection of upper airways. Therefore, any impairment of the swallowing act can represent a significant clinical and personal problem that needs an accurate diagnosis by [...] Read more.

Swallowing is a complex but stereotyped motor activity aimed at serving two vital purposes: alimentary function and the protection of upper airways. Therefore, any impairment of the swallowing act can represent a significant clinical and personal problem that needs an accurate diagnosis by means of reliable and non-invasive techniques. Thus, a systematic review and meta-analysis was performed to investigate the reliability of the Iowa Oral Pressure Instrument (IOPI) in distinguishing healthy controls (HC) from patients affected by swallowing disorders or pathologies and conditions that imply dysphagia. A comprehensive search was conducted following the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines and using PubMed, Scopus, Web of Science, Cochrane, and Lilacs databases. Overall, 271 articles were identified and, after a three-step screening, 33 case-control and interventional studies reporting IOPI measurements were included. The methodological quality of the retrieved studies resulted in being at a low risk of bias. The meta-analysis on case-control studies showed that maximum tongue pressure (MIP) values were always higher in HC than in patients, with an overall effect of the MIP difference of 18.2 KPa (17.7–18.7 KPa CI). This result was also confirmed when the sample was split into adults and children, although the MIP difference between HC and patients was greater in children than in adults (21.0 vs. 15.4 KPa in the MIP mean difference overall effect, respectively). Tongue endurance (TE) showed conflicting results among studies, with an overall effect among studies near zero (0.7 s, 0.2–1.1 s CI) and a slight tendency toward higher TE values in HC than in patients. Among the intervention studies, MIP values were higher after treatment than before, with a better outcome after the experimental tongue training exercise than traditional treatments (the MIP mean difference overall effect was 10.8 and 2.3 KPa, respectively). In conclusion, MIP values can be considered as a reliable measure of swallowing function in adults and in children, with a more marked MIP difference between HC and patients for the children population. MIP measures in patients are also able to detect the best outcome on the tongue function after the training exercise compared to traditional training. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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14 pages, 322 KiB

Open AccessReview

Transcriptomic Harmonization as the Way for Suppressing Cross-Platform Bias and Batch Effect

by Nicolas Borisov and Anton Buzdin

Biomedicines 2022, 10(9), 2318; https://doi.org/10.3390/biomedicines10092318 - 18 Sep 2022

Cited by 11 | Viewed by 2969

Abstract

(1) Background: Emergence of methods interrogating gene expression at high throughput gave birth to quantitative transcriptomics, but also posed a question of inter-comparison of expression profiles obtained using different equipment and protocols and/or in different series of experiments. Addressing this issue is challenging, [...] Read more.

(1) Background: Emergence of methods interrogating gene expression at high throughput gave birth to quantitative transcriptomics, but also posed a question of inter-comparison of expression profiles obtained using different equipment and protocols and/or in different series of experiments. Addressing this issue is challenging, because all of the above variables can dramatically influence gene expression signals and, therefore, cause a plethora of peculiar features in the transcriptomic profiles. Millions of transcriptomic profiles were obtained and deposited in public databases of which the usefulness is however strongly limited due to the inter-comparison issues; (2) Methods: Dozens of methods and software packages that can be generally classified as either flexible or predefined format harmonizers have been proposed, but none has become to the date the gold standard for unification of this type of Big Data; (3) Results: However, recent developments evidence that platform/protocol/batch bias can be efficiently reduced not only for the comparisons of limited transcriptomic datasets. Instead, instruments were proposed for transforming gene expression profiles into the universal, uniformly shaped format that can support multiple inter-comparisons for reasonable calculation costs. This forms a basement for universal indexing of all or most of all types of RNA sequencing and microarray hybridization profiles; (4) Conclusions: In this paper, we attempted to overview the landscape of modern approaches and methods in transcriptomic harmonization and focused on the practical aspects of their application. Full article

(This article belongs to the Special Issue Multiomics Approaches for Translational Medicine)

16 pages, 2144 KiB

Open AccessHypothesis

Therapeutic Neuromodulation toward a Critical State May Serve as a General Treatment Strategy

by Simon Arvin, Keisuke Yonehara and Andreas Nørgaard Glud

Biomedicines 2022, 10(9), 2317; https://doi.org/10.3390/biomedicines10092317 - 18 Sep 2022

Viewed by 3631

Abstract

Brain disease has become one of this century’s biggest health challenges, urging the development of novel, more effective treatments. To this end, neuromodulation represents an excellent method to modulate the activity of distinct neuronal regions to alleviate disease. Recently, the medical indications for [...] Read more.

Brain disease has become one of this century’s biggest health challenges, urging the development of novel, more effective treatments. To this end, neuromodulation represents an excellent method to modulate the activity of distinct neuronal regions to alleviate disease. Recently, the medical indications for neuromodulation therapy have expanded through the adoption of the idea that neurological disorders emerge from deficits in systems-level structures, such as brain waves and neural topology. Connections between neuronal regions are thought to fluidly form and dissolve again based on the patterns by which neuronal populations synchronize. Akin to a fire that may spread or die out, the brain’s activity may similarly hyper-synchronize and ignite, such as seizures, or dwindle out and go stale, as in a state of coma. Remarkably, however, the healthy brain remains hedged in between these extremes in a critical state around which neuronal activity maneuvers local and global operational modes. While it has been suggested that perturbations of this criticality could underlie neuropathologies, such as vegetative states, epilepsy, and schizophrenia, a major translational impact is yet to be made. In this hypothesis article, we dissect recent computational findings demonstrating that a neural network’s short- and long-range connections have distinct and tractable roles in sustaining the critical regime. While short-range connections shape the dynamics of neuronal activity, long-range connections determine the scope of the neuronal processes. Thus, to facilitate translational progress, we introduce topological and dynamical system concepts within the framework of criticality and discuss the implications and possibilities for therapeutic neuromodulation guided by topological decompositions. Full article

(This article belongs to the Special Issue Neuromodulation from Theory to Therapy)

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The small-world topology. (A) By randomly rewiring an ordered lattice, it transitions into a disordered graph. Through this transition, the small-world arrangement defines a critical state. x and y demarcate two arbitrary nodes on the graph, connected by a red line through the shortest path length between the nodes. (B) With increasing disorder, or randomness, the separation between the network’s nodes rapidly decreases (red), while clustering remains practically unchanged (green). The small-world topology corresponds to the mid area marked by high clustering and low separation properties. The x-axis is logarithmic. (C) Criticality defines the state of a system undergoing a phase transition, a classic example being a sandpile on the verge of collapse [<a href="#B35-biomedicines-10-02317" class="html-bibr">35</a>]. As sand is poured onto the pile, it tends to the critical state hedged between stability and ruin before finally collapsing. Despite the quantity poured onto the pile, it keeps orbiting the critical point, cycling through the same three phases: stability, criticality, and collapse. This is a special type of system behavior, which the brain is thought to possess, called self-organized criticality (SOC). Here, the system, by intrinsic means, tends to the critical state. Figure adapted from the authors’ previous work [<a href="#B31-biomedicines-10-02317" class="html-bibr">31</a>], and the original depiction of small-world graphs by Watts and Strogatz [<a href="#B34-biomedicines-10-02317" class="html-bibr">34</a>]. Full article ">Figure 2
Neuronal dynamics. (A) Resonance of the neuronal membrane potential facilitates the selective transfer of information. The traces show a Hodgkin–Huxley neuron receiving step current pulses at two different frequencies (green: low frequency—red: high frequency). When the pulses do not resonate with the neuron’s underlying resonance tendency, the signals are poorly integrated (green). When pulses do resonate, the neuron integrates and transmits the signals onward (red). The inset shows spontaneous subthreshold oscillations. (B) In the “communication through coherence”-framework, neuronal inputs systematically arrive at moments of high receptiveness, or input gain. (C) Oscillation-based neuronal communication differentiated by frequency enables the multiplexing of information. The figure is based on a model originally described by Akam and Kullmann [<a href="#B23-biomedicines-10-02317" class="html-bibr">23</a>]. Two neuronal populations with differing stimulus sensitivities (“tuning curves”) connect to the same post-synaptic neuronal receiver. The mixed neuronal code is decoded via the receiver’s input gain that matches the target stimulus input train. Red: target. Green: distraction. Full article ">Figure 3
Roles of neuronal connectivity. (A) Brain system analogy. The system’s state corresponds to the vertical position of the ball, and the system’s “control parameter” corresponds to the ball’s horizontal position. When the slope steepens, changes to the control parameter cause a greater change in the system’s state (dy/dx increases). The system has destabilized. (B) Overview of the topological model. The long-range connectivity is the control parameter that defines the state of the system, represented by a point along the curve. Short-range connections modulate the system dynamic, i.e., the shape of the curve. The curve’s steepness reflects the system’s stability. Sub-critical states are marked by fragmented activity, such as disorders of consciousness (DoC), while super-critical states favor global activity, such as seizures. Near the critical state, the system fluctuates between the local and global modes of operation via self-organized criticality (SOC), counter-balancing the intrinsic topological propensities that pull the system to its extremes (red arrowheads). The brain’s computational power peaks near criticality, mirrored by the peak in the long-range temporal correlation. The long-range temporal correlation curve (green dashes) and disease mappings (black, in-figure text) were adapted with permission from Zimmern (2020) [<a href="#B30-biomedicines-10-02317" class="html-bibr">30</a>]. (C) Stability heat-maps based on artificial neural network simulations. Notice the change in the stability map as short-range connectivity decreases. Importantly, as short-range connectivity weakens, the critical regime is destabilized in favor of especially sub-critical system states. Figure adapted from the authors’ previous work [<a href="#B31-biomedicines-10-02317" class="html-bibr">31</a>]. Full article ">Figure 4
Summary of the hypothesis. (A) Under normal conditions, the SOC mechanisms perfectly balance out the system’s topological propensities (see inset). Thus, the system manages to maneuver the critical state to facilitate local (sub-) and global (super-) neurocomputations. (B) When long-range connections are impaired, such as by brainstem shear stress, the system’s SOC mechanisms are acutely voided (see inset). This shifts the system toward sub-criticality to an extent that corresponds to the severity of the long-range neuronal injury. Transient loss of consciousness occurs in complete, but reversible, disruption of the long-range neuronal connectivity. In irreversible cases, the extent of long-range injury corresponds to the depth of unconsciousness, ranging from minimally conscious to fully comatose states. Rehabilitation strategies include thalamic neuromodulation, e.g., through DBS or neurostimulant pharmacotherapy, to augment the thalamocortical loop interactions. (C) When short-range connections are impaired, such as by a neuro-traumatic cortico-dysfunction, the system’s topological propensities are intensified (see insets). In mild cases, this makes the system fluctuate between extreme dynamical regimes because the SOC mechanisms fail to stabilize near the critical state. In severe cases, the system is effectively trapped in a sub-critical trough in which SOC is completely blocked by the system’s now intense topological propensities. Potential therapeutic strategies include repeated TMS to potentiate cortical short-range connectivity or expedited acute rehabilitation to reduce neuroplastic synaptic depression in neuro-traumatic patients. S, SOC mechanisms; D, the system’s topological propensities; rTMS, repeated transcranial magnetic stimulation; VNS, vagus nerve stimulation; DBS, deep brain stimulation; ADHD, attention deficit hyperactivity disorder. Insets show the balances between the system’s SOC mechanisms and its topological propensities. Full article ">

15 pages, 1869 KiB

Open AccessReview

Chloride Ions, Vascular Function and Hypertension

by Kenichi Goto and Takanari Kitazono

Biomedicines 2022, 10(9), 2316; https://doi.org/10.3390/biomedicines10092316 - 18 Sep 2022

Cited by 11 | Viewed by 5595

Abstract

Blood pressure is determined by cardiac output and systemic vascular resistance, and mediators that induce vasoconstriction will increase systemic vascular resistance and thus elevate blood pressure. While peripheral vascular resistance reflects a complex interaction of multiple factors, vascular ion channels and transporters play [...] Read more.

Blood pressure is determined by cardiac output and systemic vascular resistance, and mediators that induce vasoconstriction will increase systemic vascular resistance and thus elevate blood pressure. While peripheral vascular resistance reflects a complex interaction of multiple factors, vascular ion channels and transporters play important roles in the regulation of vascular tone by modulating the membrane potential of vascular cells. In vascular smooth muscle cells, chloride ions (Cl⁻) are a type of anions accumulated by anion exchangers and the anion–proton cotransporter system, and efflux of Cl⁻ through Cl⁻ channels depolarizes the membrane and thereby triggers vasoconstriction. Among these Cl⁻ regulatory pathways, emerging evidence suggests that upregulation of the Ca²⁺-activated Cl⁻ channel TMEM16A in the vasculature contributes to the increased vascular contractility and elevated blood pressure in hypertension. A robust accumulation of intracellular Cl⁻ in vascular smooth muscle cells through the increased activity of Na⁺–K⁺–2Cl⁻ cotransporter 1 (NKCC1) during hypertension has also been reported. Thus, the enhanced activity of both TMEM16A and NKCC1 could act additively and sequentially to increase vascular contractility and hence blood pressure in hypertension. In this review, we discuss recent findings regarding the role of Cl⁻ in the regulation of vascular tone and arterial blood pressure and its association with hypertension, with a particular focus on TMEM16A and NKCC1. Full article

(This article belongs to the Special Issue Animal Models for Study of Pathophysiological Mechanisms of Hypertension and Its Complications)

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Figure 1
Acetylcholine (ACh)-evoked depolarization in mesenteric arteries of spontaneously hypertensive rats (SHRs). (a) A hidden depolarization emerged after blockade of endothelium-dependent hyperpolarization (EDH) with apamin (0.5 μmol/L, a small-conductance Ca2+-sensitive K+ channel (KCa) inhibitor) plus TRAM34 (100 nmol/L, an intermediate-conductance KCa inhibitor) in mesenteric arteries of SHRs. All recordings were from the same cell. (b) ACh-evoked depolarization in the presence of apamin (0.5 μmol/L) plus charybdotoxin (60 nmol/L, a large and intermediate-conductance KCa inhibitor) was larger in amplitude and faster in time course in SHRs than in Wistar Kyoto (WKY) rats. Each paired recording was from the same preparation. Indomethacin (10 μmol/L) and Nω-nitro-L-arginine methyl ester (100 μmol/L) were present throughout. Arrows, application of ACh. Modified from Goto et al. [<a href="#B87-biomedicines-10-02316" class="html-bibr">87</a>]. Full article ">Figure 2
Effects of niflumic acid on acetylcholine-induced, endothelium-dependent hyperpolarization (EDH) in mesenteric arteries of spontaneously hypertensive rats (SHRs) and Wistar Kyoto (WKY) rats. Niflumic acid (50 μmol/L) improved EDH in SHRs but not in WKY rats. Each paired recording was from the same preparation. Indomethacin (10 μmol/L) and Nω-nitro-L-arginine methyl ester (100 μmol/L) were present throughout. Full article ">Figure 3
Upregulation of endothelial TMEM16A impairs endothelial function in hypertension. In hypertension, the expression and function of vascular endothelial Ca2+-activated Cl— channel TMEM16A are increased. Endothelial stimulation with agonists and shear stress increases the intracellular Ca2+concentration, which subsequently activates endothelial small-conductance Ca2+-activated K+ channels (SKCas), intermediate-conductance KCa (IKCa) and TMEM16A simultaneously. The endothelium-dependent hyperpolarization (EDH) through the activation of both SKCa and IKCa is reduced by the opposing membrane depolarization evoked by the activation of TMEM16A. In addition, activation of TMEM16A may facilitate the generation of reactive oxygen species (ROS) through Nox2-containing NADPH oxidase, leading to reduced bioavailability of nitic oxide (NO). Impaired EDH and/or NO could be at least partly responsible for the blood pressure rise in hypertension. Full article ">Figure 4
Upregulation of smooth muscle Na+–K+–2Cl− cotransporter 1 (NKCC1) and TMEM16A additively and sequentially increases vascular contractility in hypertension. In hypertension, the intracellular concentration of Cl− is increased in vascular smooth muscle cells because of the increased activity of NKCC1. The increase in the intracellular Cl− concentration then increases the driving force for Cl− efflux via the Ca2+-activated Cl− channel TMEM16A when TMEM16A is activated by intracellular Ca2+ rise upon stimulation with vasoconstricting agonists, which in turn induces membrane depolarization. TMEM16A might be regulated by a local Ca2+ increase that could be generated by IP3R channels on the sarcoplasmic reticulum (SR) and/or transient receptor potential (TRP) channels. The membrane depolarization would then enhance the open probability of voltage-gated L-type Ca2+ channels, leading to an increase in vascular contractility and blood pressure. Full article ">

13 pages, 2672 KiB

Open AccessArticle

Benzalkonium Chloride, Even at Low Concentrations, Deteriorates Intracellular Metabolic Capacity in Human Conjunctival Fibroblasts

by Yuri Tsugeno, Tatsuya Sato, Megumi Watanabe, Masato Furuhashi, Araya Umetsu, Yosuke Ida, Fumihito Hikage and Hiroshi Ohguro

Biomedicines 2022, 10(9), 2315; https://doi.org/10.3390/biomedicines10092315 - 18 Sep 2022

Cited by 6 | Viewed by 2895

Abstract

The objective of this study was to clarify the effects of benzalkonium chloride (BAC) on two-dimensional (2D) and three-dimensional (3D) cultures of human conjunctival fibroblast (HconF) cells, which are in vitro models replicating the epithelial barrier and the stromal supportive functions of the [...] Read more.

The objective of this study was to clarify the effects of benzalkonium chloride (BAC) on two-dimensional (2D) and three-dimensional (3D) cultures of human conjunctival fibroblast (HconF) cells, which are in vitro models replicating the epithelial barrier and the stromal supportive functions of the human conjunctiva. The cultured HconF cells were subjected to the following analyses in the absence and presence of 10⁻⁵% or 10⁻⁴% concentrations of BAC; (1) the barrier function of the 2D HconF monolayers, as determined by trans-endothelial electrical resistance (TEER) and FITC dextran permeability, (2) real-time metabolic analysis using an extracellular Seahorse flux analyzer, (3) the size and stiffness of 3D HconF spheroids, and (4) the mRNA expression of genes that encode for extracellular matrix (ECM) molecules including collagen (COL)1, 4 and 6, and fibronectin (FN), α-smooth muscle actin (α-SMA), ER stress related genes including the X-box binding protein-1 (XBP1), the spliced XBP1 (sXBP1) glucose regulator protein (GRP)78, GRP94, and the CCAAT/enhancer-binding protein homologous protein (CHOP), hypoxia inducible factor 1α (HIF1α), and Peroxisome proliferator-activated receptor gamma coactivator 1α (PGC1α). In the presence of BAC, even at low concentrations at 10⁻⁵% or 10⁻⁴%, the maximal respiratory capacity, mitochondrial respiratory reserve, and glycolytic reserve of HconF cells were significantly decreased, although the barrier functions of 2D HconF monolayers, the physical properties of the 3D HconF spheroids, and the mRNA expression of the corresponding genes were not affected. The findings reported herein highlight the fact that BAC, even such low concentrations, may induce unfavorable adverse effects on the cellular metabolic capacity of the human conjunctiva. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Responses to Low-Intensity Exposures)

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Effects of benzalkonium chloride (BAC) on barrier functions of HconF 2D monolayers. Barrier function based on TEER and FITC dextran permeability measurements were made on HconF cell 2D monolayers in the absence or presence of 10−5% or 10−4% BAC. Plots of the electric resistance (Ωcm2) by TEER and the absorbance of the amounts of permeated fluorescein are shown in (A) and (B), respectively. Experiments were repeated in triplicate (n = 5 each). All data are expressed; the mean ± the standard error of the mean (SEM). Statistical significance was evaluated by ANOVA followed by a Tukey’s multiple comparison test. Full article ">Figure 2
Effects of BAC on the sizes and hardness of the HconF 3D spheroids. Analysis of the size and hardness of 3D HconF spheroids in the absence or presence of 10−5% or 10−4% BAC. The mean sizes (μm) and the force required to compress a single spheroid to the semidiameter (μN/μm) within 20 seconds are plotted in (A) and (B), respectively. Experiments were repeated in triplicate using fresh preparations (n = 16 spheroids each). All data are expressed; the mean ± the standard error of the mean (SEM). Statistical significance was evaluated by ANOVA followed by a Tukey’s multiple comparison test. Full article ">Figure 3
Effects of BAC on the mitochondrial and glycolytic functions in HconF 2D cells. A Seahorse real-time metabolic function analysis of HconF cells in the absence (blue dots, NT) or presence of BAC at concentrations of 10−5% (green dots) or 10−4% (brown dots) BAC. OCR (A) and ECAR (B) were measured at baseline and those with injections of sequential supplementation with a complex V inhibitor, oligomycin (Oligo), a protonphore, Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP), complex I/III inhibitors, rotenone/antimycin A (R/A), and a hexokinase inhibitor, 2-deoxyglucose (2DG). (C) Indicates the parameters of mitochondrial function. Basal OCR was calculated as the difference in OCR at the baseline and after the addition of R/A. ATP-linked respiration was calculated as the difference in OCR at the baseline and after the addition of Oligo. Proton leak was calculated as the difference between OCR after the addition of Oligo and OCR after the addition of R/A. Maximal respiration was calculated as the difference between OCR after the addition of FCCP and after the addition of R/A. Mitochondrial reserve capacity was calculated as the difference in OCR at baseline and after the addition of FCCP. (D) Indicates parameters of glycolytic function. Basal ECAR was calculated as the difference in ECAR at baseline and after the addition of 2DG. Glycolytic capacity was calculated as the difference between ECAR after the addition of Oligo and ECAR after the addition of 2DG. Glycolytic reserve was calculated as the difference in ECAR at baseline and after the addition of Oligo. (E) Energy map for cells in the absence or presence of 10−5% and 10−4% BAC. Experiments were performed using fresh preparations (n = 3). Data are expressed; the mean ± the standard error of the mean (SEM). * p < 0.05; ANOVA followed by a Tukey’s multiple comparison test. Full article ">Figure 4
Effects of BAC on the gene expression of ECMs of HconF cells. 2D and 3D HconF cells were subjected to qPCR analysis for ECMs including COL1, COL4, COL6, FN, and aSMA in the absence or presence of 10−5% or 10−4% BAC. Experiments were repeated in triplicate using 3 different confluent 6-well dishes (2D) or 15 freshly prepared 3D HconF spheroids (3D) in each experimental condition. All data are expressed; the mean ± the standard error of the mean (SEM). Statistical significance was evaluated by ANOVA followed by a Tukey’s multiple comparison test. Full article ">Figure 5
Effects of BAC on the gene expression of ER stress related factors of HconF cells. 2D and 3D HconF cells were subjected to qPCR analysis of ER stress-related genes in the absence or presence of 10−5% or 10−4% BAC; the glucose regulator protein (GRP)78, GRP94, the X-box binding protein-1 (XBP1), spliced XBP1 (sXBP1), and CCAAT/enhancer-binding protein homologous protein (CHOP). Experiments were repeated in triplicate using 3 different confluent 6-well dishes (2D) or 15 freshly prepared 3D HconF spheroids (3D) in each experimental condition. All data are expressed; the mean ± the standard error of the mean (SEM). Statistical significance was evaluated by ANOVA followed by a Tukey’s multiple comparison test. Full article ">Figure 6
Effects of BAC on the gene expression of HIF1 and PGC1α of HconF cells. 2D and 3D HconF cells were subjected to qPCR analysis in HIF1α and PGC1α and 14 in the absence or presence of 10−5% or 10−4% BAC. Experiments were repeated in triplicate using 3 different confluent 6-well dishes (2D) or 15 freshly prepared 3D HconF spheroids (3D) in each experimental condition. All data are expressed; the mean ± the standard error of the mean (SEM). Statistical significance was evaluated by ANOVA followed by a Tukey’s multiple comparison test. Full article ">

10 pages, 4519 KiB

Open AccessArticle

Choroidal and Retinal Thicknesses in Type 2 Diabetes Mellitus with Moderate Diabetic Retinopathy Measured by Swept Source OCT

by Guisela Fernández-Espinosa, Elvira Orduna-Hospital, Ana Boned-Murillo, Maria Dolores Diaz-Barreda, Ana Sanchez-Cano, María Sopeña-Pinilla and Isabel Pinilla

Biomedicines 2022, 10(9), 2314; https://doi.org/10.3390/biomedicines10092314 - 18 Sep 2022

Cited by 6 | Viewed by 2473

Abstract

Background: To study choroidal thickness (CT) in type 2 diabetes mellitus (DM2) patients with moderate diabetic retinopathy (DR) and to correlate with changes in retinal thickness (RT) with swept-source OCT (SS-OCT) compared to healthy subjects. Methods: Fifty-four DM2 patients with moderate DR without [...] Read more.

Background: To study choroidal thickness (CT) in type 2 diabetes mellitus (DM2) patients with moderate diabetic retinopathy (DR) and to correlate with changes in retinal thickness (RT) with swept-source OCT (SS-OCT) compared to healthy subjects. Methods: Fifty-four DM2 patients with moderate DR without diabetic macular edema (DME) and 73 age-matched healthy subjects were evaluated using SS-OCT to measure changes in total RT and CT in the nine areas of the Early Treatment Diabetic Retinopathy Study (ETDRS) macular grid. Results: The mean age was 64.06 ± 11.98 years and 60.79 ± 8.62 years in the diabetic and control groups, respectively. Total RT showed statistically significant differences in the temporal inner area, with higher values in the DM2 group (p = 0.010). CT did not show differences between the groups. There was a significant negative correlation between RT and age in all of the outer ETDRS areas and a positive significant correlation in the central area for the DM2 group. There was also a negative significant correlation between CT and age in all of the ETDRS areas except for the inferior inner area. In the DM2 group, a negative correlation was observed between RT and CT in the central area (p = 0.039) and in both horizontal parafoveal areas (temporal inner, p = 0.028; nasal inner, p= 0.003). Conclusion: DM2 patients with moderate DR have no changes with regard to CT. Both CT and RT decreased with age in DM2, showing a negative correlation between these factors in the central and horizontal parafoveal areas of the ETDRS grid. Full article

(This article belongs to the Special Issue Pathological Mechanisms in Diabetes 2.0)

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Total retinal thickness is between the red lines (A), choroidal thickness is between the green lines (B), and the nine areas of the ETDRS grid (C) measured by deep range imaging (DRI) Triton swept source (SS)-OCT for a right eye. (Abbreviations C, Central; SE, Superior External; TE, Temporal External; NE, Nasal External; IE, Inferior External; SI, Superior Internal; TI, Temporal Internal; NI, Nasal Internal; II, Inferior Internal). Full article ">Figure 2
Mean and standard deviation (SD) of retinal and choroidal thicknesses measured using DRI-Triton SS-OCT in patients with type 2 diabetes mellitus (DM2) and in healthy controls and their comparison (p-value): (A) The measurements are represented in the nine areas of the early treatment diabetic retinopathy study (ETDRS) grid. The left quadrants represent the temporal quadrants, and the right quadrants represent the nasal quadrants. Differences that reached statistical significance (p <0.05) are shown in red with a grey background. (B) Retinal and choroidal thicknesses are represented in a bar chart. Statistically significant differences (p < 0.05) are marked with (*). Full article ">Figure 3
Correlation coefficients and statistical significance (p-value) of retinal and choroidal thicknesses with age, time of DM evolution, and glycosylated hemoglobin (HbA1c) levels (%) in DM2 patients. They are represented in the nine areas of the early treatment diabetic retinopathy study (ETDRS) grid. The left areas represent the temporal quadrants, and the right areas represent the nasal quadrants, as if a right eye was represented. The values that reached statistical significance (p < 0.05) are shown in red with a grey background and marked with (*) when p < 0.05 and with (**) when p < 0.01. Full article ">Figure 4
(A) Correlation coefficients and statistical significance (p-value) between choroidal thickness and retinal thickness in DM2 patients in the nine areas of the early treatment diabetic retinopathy study (ETDRS) grid. Left areas represent the temporal quadrants, and right areas represent the nasal quadrants, as if the figure represents a right eye. The values that reached statistical significance (p <0.05) are shown in red with a purple background and marked with (*). (B) Scatter plot with regression lines correlating retinal and choroidal thicknesses in temporal internal (TI), central (C), and nasal internal (NI) areas. Full article ">

22 pages, 1110 KiB

Open AccessReview

Biodegradation of Dental Resin-Based Composite—A Potential Factor Affecting the Bonding Effect: A Narrative Review

by Xinwei Guo, Yiyan Yu, Shang Gao, Zhimin Zhang and Hongyan Zhao

Biomedicines 2022, 10(9), 2313; https://doi.org/10.3390/biomedicines10092313 - 18 Sep 2022

Cited by 24 | Viewed by 4697

Abstract

In recent years, although resin composite has played an important role in the restoration of tooth defects, it still has several disadvantages, including being biodegraded by saliva, bacteria and other enzymes in the oral cavity, which may result in repair failure. This factor [...] Read more.

In recent years, although resin composite has played an important role in the restoration of tooth defects, it still has several disadvantages, including being biodegraded by saliva, bacteria and other enzymes in the oral cavity, which may result in repair failure. This factor is not conducive to the long-term survival of the prosthesis in the mouth. In this article, we review the causes, influencing factors and prevention methods of resin biodegradation. Biodegradation is mainly caused by esterase in saliva and bacteria, which breaks the ester bond in resin and causes the release of monomers. The mechanical properties of the prosthesis can then be affected. Meanwhile, cathepsin and MMPs are activated on the bonding surface, which may decompose the dentin collagen. In addition, neutrophils and residual water on the bonding surface can also aggravate biodegradation. Currently, the primary methods to prevent biodegradation involve adding antibacterial agents to resin, inhibiting the activity of MMPs and enhancing the crosslinking of collagen fibers. All of the above indicates that in the preparation and adhesion of resin materials, attention should be paid to the influence of biodegradation to improve the prosthesis’s service life in the complex environment of the oral cavity. Full article

(This article belongs to the Section Biomedical Engineering and Materials)

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15 pages, 1504 KiB

Open AccessArticle

Photoreactive Coating Material as an Effective and Durable Antimicrobial Composite in Reducing Bacterial Load on Surfaces in Livestock

by Ádám Kerek, Mátyás Sasvári, Ákos Jerzsele, Zoltán Somogyi, László Janovák, Zsolt Abonyi-Tóth and Imre Dékány

Biomedicines 2022, 10(9), 2312; https://doi.org/10.3390/biomedicines10092312 - 17 Sep 2022

Cited by 1 | Viewed by 1951

Abstract

Titanium dioxide (TiO₂) is a well-known photocatalytic compound that can be used to effectively reduce the presence of pathogens in human and animal hospitals via ROS release. The aim of this study was to investigate the efficacy of a polymer-based composite [...] Read more.

Titanium dioxide (TiO₂) is a well-known photocatalytic compound that can be used to effectively reduce the presence of pathogens in human and animal hospitals via ROS release. The aim of this study was to investigate the efficacy of a polymer-based composite layer containing TiO₂ and zinc oxide (ZnO) against Escherichia coli (E. coli) of animal origin. We showed that the photocatalyst coating caused a significant (p < 0.001) reduction in pathogen numbers compared to the control with an average reduction of 94% over 30 min. We used six light sources of different wattages (4 W, 7 W, 9 W, 12 W, 18 W, 36 W) at six distances (35 cm, 100 cm, 150 cm, 200 cm, 250 cm, 300 cm). Samples (n = 2160) were taken in the 36 settings and showed no significant difference in efficacy between light intensity and distance. We also investigated the influence of organic contaminant that resulted in lower activity as well as the effect of a water jet and a high-pressure device on the antibacterial activity. We found that the latter completely removed the coating from the surface, which significantly (p < 0.0001) reduced its antibacterial potential. As a conclusion, light intensity and distance does not reduce the efficacy of the polymer, but the presence of organic contaminants does. Full article

(This article belongs to the Special Issue Drug Discovery for Infectious Diseases)

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The efficiency of a 4 W light source at each treatment distance compared to the untreated surface. The rate of decrease in the CFU count of a photocatalyst coating-treated surface as a function of time for a 4 W LED lamp at different distances compared to the average CFU count of untreated (Control) surfaces. Starting from 90 min, the bacterial count reduction is 99.5%. Full article ">Figure 2
Effect of organic contaminant on photocatalyst layer treated and untreated surface at a distance of 300 cm under illumination with 4 W and 7 W lamps. For samples taken after 15 min, the CFU reduction was not yet significant (78.8% after 30 min for 4 W and 79.5% for 7 W). Full article ">Figure 3
Washing test on the treated and untreated surfaces from a distance of 300 cm. In the case of normal water jet washing, the paint-treated area had only 300 CFU at minute 15, which further decreased to 1 CFU by minute 30, and by minute 120, the bacterial count reduction was 100%. After washing with high pressure equipment, there was no CFU reduction in samples taken from the treated surface even after 480 min. Full article ">Figure 4
Comparison of control and treated surfaces by random forest method as a function of light intensity and illumination distance. When examining luminance and distance, the p-value of all variables was less than 0.001 at the first branching, but overall, the effects of wattage, distance, and time were more moderate on the model hit rate compared to the treatment. Full article ">

10 pages, 1690 KiB

Open AccessArticle

Identification of Hub Genes and Potential Biomarkers for Childhood Asthma by Utilizing an Established Bioinformatic Analysis Approach

by Ichtiarini Nurullita Santri, Lalu Muhammad Irham, Gina Noor Djalilah, Dyah Aryani Perwitasari, Yuniar Wardani, Yohane Vincent Abero Phiri and Wirawan Adikusuma

Biomedicines 2022, 10(9), 2311; https://doi.org/10.3390/biomedicines10092311 - 16 Sep 2022

Cited by 11 | Viewed by 3214

Abstract

Childhood asthma represents a heterogeneous disease resulting from the interaction between genetic factors and environmental exposures. Currently, finding reliable biomarkers is necessary for the clinical management of childhood asthma. However, only a few biomarkers are being used in clinical practice in the pediatric [...] Read more.

Childhood asthma represents a heterogeneous disease resulting from the interaction between genetic factors and environmental exposures. Currently, finding reliable biomarkers is necessary for the clinical management of childhood asthma. However, only a few biomarkers are being used in clinical practice in the pediatric population. In the long run, new biomarkers for asthma in children are required and would help direct therapy approaches. This study aims to identify potential childhood asthma biomarkers using a genetic-driven biomarkers approach. Herein, childhood asthma-associated Single Nucleotide Polymorphisms (SNPs) were utilized from the GWAS database to drive and facilitate the biomarker of childhood asthma. We uncovered 466 childhood asthma-associated loci by extending to proximal SNPs based on r² > 0.8 in Asian populations and utilizing HaploReg version 4.1 to determine 393 childhood asthma risk genes. Next, the functional roles of these genes were subsequently investigated using Gene Ontology (GO) term enrichment analysis, a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and a protein–protein interaction (PPI) network. MCODE and CytoHubba are two Cytoscape plugins utilized to find biomarker genes from functional networks created using childhood asthma risk genes. Intriguingly, 10 hub genes (IL6, IL4, IL2, IL13, PTPRC, IL5, IL33, TBX21, IL2RA, and STAT6) were successfully identified and may have been identified to play a potential role in the pathogenesis of childhood asthma. Among 10 hub genes, we strongly suggest IL6 and IL4 as prospective childhood asthma biomarkers since both of these biomarkers achieved a high systemic score in Cytohubba’s MCC algorithm. In summary, this study offers a valuable genetic-driven biomarker approach to facilitate the potential biomarkers for asthma in children. Full article

(This article belongs to the Special Issue Allergy and Asthma: From Pathogenesis to Molecular Understanding of Therapies)

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11 pages, 938 KiB

Open AccessArticle

Beneficial Effects of Table Grape Use on Serum Levels of Omega-3 Index and Liver Function: A Randomized Controlled Clinical Trial

by Maria Notarnicola, Valentina De Nunzio, Tamara Lippolis, Valeria Tutino, Anna Maria Cisternino, Palma Aurelia Iacovazzi, Rosa Anna Milella, Marica Gasparro, Roberto Negro, Maurizio Polignano and Maria Gabriella Caruso

Biomedicines 2022, 10(9), 2310; https://doi.org/10.3390/biomedicines10092310 - 16 Sep 2022

Cited by 3 | Viewed by 2584

Abstract

This clinical trial was aimed to investigate the effects of fresh table grape intake on the serum levels of the Omega-3 index, defined as the sum of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) levels. Forty consecutive healthy subjects were randomly assigned to [...] Read more.

This clinical trial was aimed to investigate the effects of fresh table grape intake on the serum levels of the Omega-3 index, defined as the sum of eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA) levels. Forty consecutive healthy subjects were randomly assigned to the control group, receiving only dietary recommendations, and the grape group receiving a daily dose of 5 g of fresh table grape per kg of body weight, for 21 days. Compared with baseline, the grape treatment produced no significant difference in the serum levels of glucose, liver transaminase, and triglycerides, with the exception of cholesterol value, which was significantly reduced in both control and grape group (180.5 ± 20.32 vs. 196.1 ± 30.0 and 181.4 ± 21.9 vs. 194.3 ± 37.5, respectively). After 4 weeks from the end of grape treatment, the analysis of single fatty acids showed a significant increase in oleic acid content (14.15 ± 1.8 vs. 12.85 ± 1.6, p < 0.05) and a significant induction of the Omega-3 index (8.23 ± 1.9 vs. 6.09 ± 1.2, p < 0.05), associated with increased serum levels of adiponectin (24.09 ± 1.08 vs. 8.8 ± 0.7, p < 0.001). In contrast, the expression of fibroblast growth factor 21 (FGF21), a molecule associated with metabolic syndrome and liver disease, was significantly reduced (37.9 ± 6.8 vs. 107.8 ± 10.1, p < 0.001). The data suggest that the intake of fresh grape improves the Omega-3 index in the serum and exerts beneficial effects on liver function through the overexpression of adiponectin and the reduction in FGF21 levels. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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13 pages, 907 KiB

Open AccessArticle

The Role of Intraamygdaloid Oxytocin and D2 Dopamine Receptors in Reinforcement in the Valproate-Induced Autism Rat Model

by Kristóf László, Dávid Vörös, Orsolya Kiss, Bettina Réka László, Tamás Ollmann, László Péczely, Kitti Mintál, Attila Tóth, Anita Kovács, Olga Zagoracz, Erika Kertes, Veronika Kállai, Beáta Berta, Zoltán Karádi and László Lénárd

Biomedicines 2022, 10(9), 2309; https://doi.org/10.3390/biomedicines10092309 - 16 Sep 2022

Cited by 3 | Viewed by 2510

Abstract

Background: autism spectrum disorder (ASD) is a neurodevelopmental disorder affecting around 1 out of 68 children and its incidence shows an increasing tendency. There is currently no effective treatment for ASD. In autism research, the valproate (VPA)-induced autism rodent model is widely accepted. [...] Read more.

Background: autism spectrum disorder (ASD) is a neurodevelopmental disorder affecting around 1 out of 68 children and its incidence shows an increasing tendency. There is currently no effective treatment for ASD. In autism research, the valproate (VPA)-induced autism rodent model is widely accepted. Our previous results showed that intraamygdaloid oxytocin (OT) has anxiolytic effects on rats showing autistic signs under the VPA-induced autism model. Methods: rats were stereotaxically implanted with guide cannulae bilaterally and received intraamygdaloid microinjections. In the present study, we investigated the possible role of intraamygdaloid OT and D2 dopamine (DA) receptors on reinforcement using VPA-treated rats in a conditioned place preference test. OT and/or an OT receptor antagonist or a D2 DA antagonist were microinjected into the central nucleus of the amygdala (CeA). Results: valproate-treated rats receiving 10 ng OT spent significantly longer time in the treatment quadrant during the test session of the conditioned place preference test. Prior treatment with an OT receptor antagonist or with a D2 DA receptor antagonist blocked the positive reinforcing effects of OT. The OT receptor antagonist or D2 DA antagonist in themselves did not influence the time rats spent in the treatment quadrant. Conclusions: Our results show that OT has positive reinforcing effects under the VPA-induced autism rodent model and these effects are OT receptor-specific. Our data also suggest that the DAergic system plays a role in the positive reinforcing effects of OT because the D2 DA receptor antagonist can block these actions. Full article

(This article belongs to the Special Issue Neuropeptides, Dopamine and Their Interactions in Neuroscience)

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Figure 1
Illustration of reconstructed injection sites. Correct bilateral injection placements are indicated by closed gray and black circles in the CeA in panel (A) (n = 70). Incorrect injection placements are indicated in panel (B) (n = 9). Brain structure diagrams of coronal sections are adapted from the stereotaxic atlas of Paxinos and Watson. The numbers refer to anterior–posterior distances from the bregma in mm. Identical symbols in panel (B) indicate coherent injection sites of bilateral injections. Numbers above marked sites in panels (A,B) indicate numbers of animals. Full article ">Figure 2
Effects of OT and the OT receptor antagonist injected into the CeA of autistic rats on the conditioned place preference (CPP) test. Columns represent mean time spent in the treatment quadrant (±S.E.M.) during Habituation and Test sessions. Control: neurotypical/sham male Wistar rats receiving intraamygdaloid PBS (n = 6); VPA: intrauterine VPA-treated rats showing autistic-like behavior (n = 7); VPA + 10 ng OT: animals showing autistic-like behavior, microinjected with 10 ng OT (n = 7); VPA + ANT + OT: animals with autistic-like behavior, microinjected with 20 ng OT receptor antagonist and 10 ng OT (n = 7); VPA + ANT: animals with autistic-like behavior, microinjected with 20 ng OT receptor antagonist (n = 7); * p < 0.05; for more explanation, see the text. Full article ">Figure 3
Effects of D2 DA antagonist pretreatment in the CeA of autistic rats on the conditioned place preference (CPP) test. Columns represent mean time spent in the treatment quadrant (±S.E.M.) during Habituation and Test sessions. Control: neurotypical/sham male Wistar rats receiving intraamygdaloid PBS (n = 8); VPA: intrauterine VPA-treated rats showing autistic-like behavior (n = 7); VPA + 10 ng OT: animals showing autistic-like behavior, microinjected with 10 ng OT (n = 7); VPA + D2 ANT + OT: animals with autistic-like behavior, microinjected with 10 ng OT and pretreated with 4 µg Sulpiride (n = 7); VPA + D2 ANT: animals with autistic-like behavior, treated with 4 µg Sulpiride (n = 7); * p < 0.05, for more explanations, see the text. Full article ">

13 pages, 1286 KiB

Open AccessArticle

High Prevalence of Primary Aldosteronism in Patients with Type 2 Diabetes Mellitus and Hypertension

by Ernestini Tyfoxylou, Nick Voulgaris, Chris Gravvanis, Sophia Vlachou, Athina Markou, Labrini Papanastasiou, Nikolaos Tentolouris, Eva Kassi, Gregory Kaltsas, George P. Chrousos and George P. Piaditis

Biomedicines 2022, 10(9), 2308; https://doi.org/10.3390/biomedicines10092308 - 16 Sep 2022

Cited by 4 | Viewed by 2419

Abstract

Primary aldosteronism (PA) is the most common cause of endocrine hypertension. The prevalence of hypertension is higher in patients with diabetes mellitus-2 (DM-2). Following the limited existing data, we prospectively investigated the prevalence of aldosterone excess either as autonomous secretion (PA) or as [...] Read more.

Primary aldosteronism (PA) is the most common cause of endocrine hypertension. The prevalence of hypertension is higher in patients with diabetes mellitus-2 (DM-2). Following the limited existing data, we prospectively investigated the prevalence of aldosterone excess either as autonomous secretion (PA) or as a hyper-response to stress in hypertensive patients with DM-2 (HDM-2). A total of 137 HDM-2 patients and 61 non-diabetics with essential hypertension who served as controls (EH-C) underwent a combined, overnight diagnostic test, the Dexamethasone–captopril–valsartan test (DCVT) used for the diagnosis of PA and an ultralow dose (0.3 μg) ACTH stimulation test to identify an exaggerated aldosterone response to ACTH stimulation. Twenty-three normotensive individuals served as controls (NC) to define the normal response of aldosterone (ALD) and aldosterone-to-renin ratio (ARR) to the ultralow dose ACTH test. Using post-DCVTALD and ARR from the EH-C, and post-ACTH peak ALD and ARR from the NC, 47 (34.3%) HDM-2 patients were found to have PA, whereas 6 (10.4%) HDM-2 patients without PA (DCVT-negative) exhibited an exaggerated aldosterone response to stress—a prevalence much higher than ever reported. Treatment with mineralocorticoid receptor antagonists (MRAs) induced a significant and permanent reduction of BP in all HDM-2 patients. Early diagnosis and targeted treatment of PA is crucial to prevent any aggravating effect on chronic diabetic complications. Full article

(This article belongs to the Special Issue Recent Advances in Primary Aldosteronism: From Bench to Clinic)

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44 pages, 3300 KiB

Open AccessReview

A Comprehensive Review on Collagen Type I Development of Biomaterials for Tissue Engineering: From Biosynthesis to Bioscaffold

by Ibrahim N. Amirrah, Yogeswaran Lokanathan, Izzat Zulkiflee, M. F. Mohd Razip Wee, Antonella Motta and Mh Busra Fauzi

Biomedicines 2022, 10(9), 2307; https://doi.org/10.3390/biomedicines10092307 - 16 Sep 2022

Cited by 145 | Viewed by 22631

Abstract

Collagen is the most abundant structural protein found in humans and mammals, particularly in the extracellular matrix (ECM). Its primary function is to hold the body together. The collagen superfamily of proteins includes over 20 types that have been identified. Yet, collagen type [...] Read more.

Collagen is the most abundant structural protein found in humans and mammals, particularly in the extracellular matrix (ECM). Its primary function is to hold the body together. The collagen superfamily of proteins includes over 20 types that have been identified. Yet, collagen type I is the major component in many tissues and can be extracted as a natural biomaterial for various medical and biological purposes. Collagen has multiple advantageous characteristics, including varied sources, biocompatibility, sustainability, low immunogenicity, porosity, and biodegradability. As such, collagen-type-I-based bioscaffolds have been widely used in tissue engineering. Biomaterials based on collagen type I can also be modified to improve their functions, such as by crosslinking to strengthen the mechanical property or adding biochemical factors to enhance their biological activity. This review discusses the complexities of collagen type I structure, biosynthesis, sources for collagen derivatives, methods of isolation and purification, physicochemical characteristics, and the current development of collagen-type-I-based scaffolds in tissue engineering applications. The advancement of additional novel tissue engineered bioproducts with refined techniques and continuous biomaterial augmentation is facilitated by understanding the conventional design and application of biomaterials based on collagen type I. Full article

(This article belongs to the Special Issue Biomaterial Modifications and Improvement of Their Biocompatibility)

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(a) Collagen type I sources and approximate content in each tissue. * Cartilage percentage is based on all collagens in their dry weight, as it is predominantly collagen type II. (b) Collagen fibre structure, fibrils, triple helices of alpha chains as tropocollagen, and amino acid residues. (c) Triple-helix and single-chain collagen type I molecular structure (PDB 1CAG) [<a href="#B24-biomedicines-10-02307" class="html-bibr">24</a>] with gly-x-y repeats; x-y typically represents proline and hydroxyproline, respectively. Full article ">Figure 2
Overview of collagen type I biosynthesis from (a) intracellular transcription to (b) formation of extracellular complex macromolecular structures and what happens in between during (c) post-translational modifications. Full article ">Figure 3
Steps of the collagen isolation process: an overview. Examples of methods from Mogan et al. [<a href="#B73-biomedicines-10-02307" class="html-bibr">73</a>]. Different tissues require different optimisation in the reagent’s concentration, pH, processing time, and temperature. Full article ">Figure 4
Parametric selection for the most suitable biomaterial. Full article ">Figure 5
Illustration of different techniques for crosslinking collagen type I scaffolds. Physical crosslinking includes physical alterations, usually from DHT heat treatment or UV irradiation. Chemical crosslinking refers to chemical alterations using aldehydes such as glutaraldehyde, epoxy compounds, commonly used EDC/NHS crosslinking, and DPPA. Biological crosslinking uses materials derived from biological sources such as genipin, a plant-based crosslinker, plant polyphenols, and enzymes. Full article ">Scheme 1
Schematic illustration of crosslinking mechanisms of type I collagen with a physical crosslinker, (a) dehydrothermal treatment (DHT), and chemical crosslinkers, (b) aldehydes, (c) epoxy compounds, (d) NHS-activated carboxylic esters, (e), EDC and EDC/NHS, and (f) DPPA. These figures were adapted from Zhang et al. 2020 [<a href="#B112-biomedicines-10-02307" class="html-bibr">112</a>]. Full article ">Scheme 2
Biological crosslinking mechanisms of type I collagen for (a) genipin, (b) transglutaminase, and (c) plant polyphenols [<a href="#B112-biomedicines-10-02307" class="html-bibr">112</a>]. Full article ">

21 pages, 1106 KiB

Open AccessReview

Dipeptidyl Peptidase 4 (DPP4) as A Novel Adipokine: Role in Metabolism and Fat Homeostasis

by Ilaria Barchetta, Flavia Agata Cimini, Sara Dule and Maria Gisella Cavallo

Biomedicines 2022, 10(9), 2306; https://doi.org/10.3390/biomedicines10092306 - 16 Sep 2022

Cited by 25 | Viewed by 5361

Abstract

Dipeptidyl peptidase 4 (DPP4) is a molecule implicated in the regulation of metabolic homeostasis and inflammatory processes, and it exerts its main action through its enzymatic activity. DPP4 represents the enzyme most involved in the catabolism of incretin hormones; thus, its activity impacts [...] Read more.

Dipeptidyl peptidase 4 (DPP4) is a molecule implicated in the regulation of metabolic homeostasis and inflammatory processes, and it exerts its main action through its enzymatic activity. DPP4 represents the enzyme most involved in the catabolism of incretin hormones; thus, its activity impacts appetite, energy balance, and the fine regulation of glucose homeostasis. Indeed, DPP4 inhibitors represent a class of antidiabetic agents widely used for the treatment of Type 2 diabetes mellitus (T2DM). DPP4 also acts as an adipokine and is mainly secreted by the adipose tissue, mostly from mature adipocytes of the visceral compartment, where it exerts autocrine and paracrine activities. DPP4 can disrupt insulin signaling within the adipocyte and in other target cells and tissues, where it also favors the development of a proinflammatory environment. This is likely at the basis of the presence of elevated circulating DPP4 levels in several metabolic diseases. In this review, we summarize the most recent evidence of the role of the DPP4 as an adipokine-regulating glucose/insulin metabolism and fat homeostasis, with a particular focus on clinical outcomes associated with its increased secretion in the presence of adipose tissue accumulation and dysfunction. Full article

(This article belongs to the Special Issue 10th Anniversary of Biomedicines—Recent Advances on Adipokines)

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10 pages, 1015 KiB

Open AccessArticle

Prognostic Role of Post-Induction Fecal Calprotectin Levels in Patients with Inflammatory Bowel Disease Treated with Biological Therapies

by Antonio Facciorusso, Daryl Ramai, Cristina Ricciardelli, Rosa Paolillo, Marcello Maida, Saurabh Chandan, Babu P. Mohan, Viktor Domislovic and Rodolfo Sacco

Biomedicines 2022, 10(9), 2305; https://doi.org/10.3390/biomedicines10092305 - 16 Sep 2022

Cited by 7 | Viewed by 2416

Abstract

Background: There is currently scarce knowledge about markers of early therapeutic response in patients with inflammatory bowel disease (IBD) treated with biologics. The aim of this study was to evaluate the role of fecal calprotectin (FC) as an early predictor of mucosal healing [...] Read more.

Background: There is currently scarce knowledge about markers of early therapeutic response in patients with inflammatory bowel disease (IBD) treated with biologics. The aim of this study was to evaluate the role of fecal calprotectin (FC) as an early predictor of mucosal healing and clinical remission. Methods: Data from a multicenter series of 172 IBD patients treated with biologics between 2017 and 2020 were analyzed. Treatment outcomes were mucosal healing and clinical remission assessed at 2 years. FC levels were assessed at 14 weeks (post-induction), at 6 months, and yearly. The receiver operating characteristic (ROC) curve analysis was performed to calculate the best cut-off in % change of FC levels between post-induction and baseline predicting treatment outcomes. Sensitivity, specificity, and accuracy for several post-induction FC cut-off points were also calculated. Results: At 2 years, mucosal healing was noted in 77 patients (44.7%), of whom were 41 Crohn’s disease (CD) and 36 ulcerative colitis (UC) patients, whereas 106 patients experienced clinical remission (61.6%), of whom were 59 CD and 47 UC patients. Both baseline and post-induction FC levels were significantly higher in non-responders as compared to responders. On the other hand, FC decrease was less pronounced in non-responders. Similar results were observed in all subgroups, namely according to disease (CD vs. UC), or treatment used (TNF-inhibitors vs. vedolizumab). The best cut-off points were −86% in % change in FC levels to predict mucosal healing and −83% for clinical remission. Conclusions: The current study suggests a predictive role of post-induction FC assessment to predict treatment response in IBD patients treated with biologics. Full article

(This article belongs to the Section Cell Biology and Pathology)

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18 pages, 1247 KiB

Open AccessReview

Dynamic Regulation of NF-κB Response in Innate Immunity: The Case of the IMD Pathway in Drosophila

by Alexandre Cammarata-Mouchtouris, Adrian Acker, Akira Goto, Di Chen, Nicolas Matt and Vincent Leclerc

Biomedicines 2022, 10(9), 2304; https://doi.org/10.3390/biomedicines10092304 - 16 Sep 2022

Cited by 21 | Viewed by 5401

Abstract

Metazoans have developed strategies to protect themselves from pathogenic attack. These preserved mechanisms constitute the immune system, composed of innate and adaptive responses. Among the two kinds, the innate immune system involves the activation of a fast response. NF-κB signaling pathways are activated [...] Read more.

Metazoans have developed strategies to protect themselves from pathogenic attack. These preserved mechanisms constitute the immune system, composed of innate and adaptive responses. Among the two kinds, the innate immune system involves the activation of a fast response. NF-κB signaling pathways are activated during infections and lead to the expression of timely-controlled immune response genes. However, activation of NF-κB pathways can be deleterious when uncontrolled. Their regulation is necessary to prevent the development of inflammatory diseases or cancers. The similarity of the NF-κB pathways mediating immune mechanisms in insects and mammals makes Drosophila melanogaster a suitable model for studying the innate immune response and learning general mechanisms that are also relevant for humans. In this review, we summarize what is known about the dynamic regulation of the central NF-κB-pathways and go into detail on the molecular level of the IMD pathway. We report on the role of the nuclear protein Akirin in the regulation of the NF-κB Relish immune response. The use of the Drosophila model allows the understanding of the fine-tuned regulation of this central NF-κB pathway. Full article

(This article belongs to the Special Issue Roles of NF-κB in Cancer and Their Therapeutic Approaches 2.0)

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The NF-κB IMD pathway in Drosophila. IMD is activated through the recognition of Gram-negative bacteria-derived meso diaminopymelic-type (DAP-type) peptidoglycan (PGN) and tracheal cytotoxin (TCT) by the Peptidoglycan recognition (PGRP) domain of Peptidoglycan recognition protein -LC and -LE (PGRP LC, -LE). PGRP-LC and -LE death-domains recruit immune deficiency (IMD), FAS-associated death domain (FADD) and death-related ced-3/Nedd2-like protein (Dredd). A ubiquitin-ligase complex formed by the E3 ubiquitin ligase Drosophila inhibitor of apoptosis 2 (DIAP2) and the E2 ubiquitin conjugating Ubiquitin conjugating enzyme variant 1A (Uev1a), Bendless and Effete activates Dredd by K63-linked poly ubiquitinylation. Activated Dredd cleaves IMD N-terminal domain, leading to the recruitment of transforming growth factor beta (TGF-β)-activated kinase 1 (TAK1) and TAK1-associated binding protein 2 (TAB2). TAK1 is able to activate the inhibitor of NF-κB (IκB) kinase (IKK) complex formed of IKKβ and IKKγ subunits. Phosphorylated IKKβ is sumoylated by Leswright and phosphorylates the N-terminal portion of the NF-κB factor Relish to enable its transcriptional activity. Relish is separated from its IκB-like C-terminal ankyrin repeat region by Dredd through proteolytic cleavage. The NLS-containing N-terminal portion of Relish (Rel-68) is then imported to the nucleus while the IκB-like C-terminal portion (Rel-49) remains in the cytoplasm. Rel68 homodimers bind their cognate κB Response element, the consensus sequence 5′-GGGGATTYYY-3′ (Y: C or T) and activate IMD-pathway target genes with the help of the nuclear protein Akirin, which needs to be ubiquitinated by the E3-ligase Hyd beforehand. Dotted arrows indicate the activity of key cleaved proteins of the pathway (IMD, Relish), while continuous arrows are used for the other proteins. Negative regulators are highlighted in red. Full article ">Figure 2
Fine-tuning characterizes NF-κB IMD pathway expression. Following the stimulation of the IMD pathway, a non-identified epigenetic-related protein will deposit an acetyl group on the lysine 4 of histone 3 (H3K4Ac), nearby genes mostly coding for effectors (anti-microbial peptides). After being K63 ubiquitinilated by the E3-ligase Hyd, the conserved nuclear protein Akirin orchestrates a NF-κB transcriptional selectivity through the recruitment of the Osa-containing-SWI/SNF-like Brahma complex (BAP). The N-terminal portion of Relish (Rel-68) will then be recruited to the Akirin complex formed, which will lead to the expression of mostly effector genes of the pathway (anti-microbial peptides). In the case that the H3K4Ac mark is not deposited, Akirin will not be recruited. Rel68 will still bind to the consensus sequence and activate the expression of a second subset of genes, comprised of mostly negative regulators and some anti-microbial peptides. In brief, Akirin is a NF-κB co-factor acting as a molecular selector, required for the activation of a specific subset of Relish-dependent genes that correlates with the presence of H3K4Ac epigenetic marks. Akirin specifies the choice between subsets of NF-κB target genes, allowing Drosophila to modulate its innate immune response. Full article ">

9 pages, 744 KiB

Open AccessCommunication

Granulocyte Colony-Stimulating Factor Ameliorates Endothelial Activation and Thrombotic Diathesis Biomarkers in a Murine Model of Hind Limb Ischemia

by Angeliki Valatsou, Panagiotis Theofilis, Spyridon Simantiris, Georgia Vogiatzi, Alexandros Briasoulis, Marios Sagris, Evangelos Oikonomou, Alexios S. Antonopoulos, Alkistis Pantopoulou, Narjes Nasiri-Ansari, Elizabeth Fragopoulou, Despoina Perrea, Konstantinos Tsioufis and Dimitris Tousoulis

Biomedicines 2022, 10(9), 2303; https://doi.org/10.3390/biomedicines10092303 - 16 Sep 2022

Viewed by 2047

Abstract

Novel therapies in peripheral arterial disease, such as granulocyte colony-stimulating factor (GCSF) administration, might result in anti-atherosclerotic effects. In this study, we used 10-week-old male ApoE^−/− mice, which were fed an atherosclerosis-inducing diet for four weeks. At the end of the four [...] Read more.

Novel therapies in peripheral arterial disease, such as granulocyte colony-stimulating factor (GCSF) administration, might result in anti-atherosclerotic effects. In this study, we used 10-week-old male ApoE^−/− mice, which were fed an atherosclerosis-inducing diet for four weeks. At the end of the four weeks, hind limb ischemia was induced through left femoral artery ligation, the atherosclerosis-inducing diet was discontinued, and a normal diet was initiated. Mice were then randomized into a control group (intramuscular 0.4 mL normal saline 0.9% for 7 days) and a group in which GCSF was administrated intramuscularly in the left hind limb for 7 days (100 mg/kg). In the GCSF group, but not in the control group, we observed significant reductions in the soluble adhesion molecules (vascular cell adhesion molecule-1 (sVCAM-1) and intercellular adhesion molecule-1 (sICAM-1)), sE-Selectin, and plasminogen activator inhibitor (PAI)-1 when they were measured through ELISA on the 1st and the 28th days after hind limb ischemia induction. Therefore, GCSF administration in an atherosclerotic mouse model of hind limb ischemia led to decreases in the biomarkers associated with endothelial activation and thrombosis. These findings warrant further validation in future preclinical studies. Full article

(This article belongs to the Special Issue Advances in Cardiovascular Pharmacology: The Era of Targeted Therapies in Cardiology)

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11 pages, 859 KiB

Open AccessArticle

Sex-Differences in Pain and Opioid Use Disorder Management: A Cross-Sectional Real-World Study

by Mónica Escorial, Javier Muriel, César Margarit, Laura Agulló, Domingo Morales and Ana M. Peiró Peiró

Biomedicines 2022, 10(9), 2302; https://doi.org/10.3390/biomedicines10092302 - 16 Sep 2022

Cited by 5 | Viewed by 3103

Abstract

(1) Background: It is essential to focus attention on sex-specific factors which are clinically relevant in pain management, especially with regards to opioid use disorder (OUD) risk. The aim of this study was to explore potential sex-differences in chronic non-cancer pain (CNCP) outpatients. [...] Read more.

(1) Background: It is essential to focus attention on sex-specific factors which are clinically relevant in pain management, especially with regards to opioid use disorder (OUD) risk. The aim of this study was to explore potential sex-differences in chronic non-cancer pain (CNCP) outpatients. (2) Methods: An observational cross-sectional study was conducted under CNCP outpatients with long-term prescribed opioids (n = 806), wherein 137 patients had an OUD diagnosis (cases, 64% females) and 669 did not (controls, 66% females). Socio-demographic, clinical, and pharmacological outcomes were analyzed. (3) Results: Female controls presented an older age and less intensive pain therapy but higher psychotropic prescriptions and emergency department visits compared to male controls. Meanwhile, cases demonstrated a younger age, higher work disability, double morphine equivalent daily dose, and benzodiazepine use compared with controls. Here, female cases showed an 8% greater substance use disorder (OR 2.04 [1.11–3.76]) and 24% lower tramadol use, while male cases presented a 22% higher fentanyl use (OR 2.97 [1.52–5.81]) and reported the highest number of adverse drug reactions (24%, OR 2.40 [1.12–5.16]) compared with controls. (4) Conclusions: An OUD individual risk profile was evidenced with sex-differences to take into consideration to design equal prevention programs. Full article

(This article belongs to the Special Issue Gender Medicine and Pharmacology)

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