[go: up one dir, main page]

 
 
Sign in to use this feature.

Years

Between: -

Subjects

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Journals

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Article Types

Countries / Regions

remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline
remove_circle_outline

Search Results (6,837)

Search Parameters:
Keywords = lactation

Order results
Result details
Results per page
Select all
Export citation of selected articles as:
12 pages, 3455 KiB  
Article
Impact of Calcium Lactate Pretreatment on Enamel Fluoride Uptake: A Comparative In Vitro Study of Different Fluoride Types and Concentrations
by Fjolla Kullashi Spahija, Ivana Šutej, Kresimir Basic, Kreshnik Spahija and Kristina Peros
J. Funct. Biomater. 2024, 15(9), 269; https://doi.org/10.3390/jfb15090269 - 16 Sep 2024
Viewed by 72
Abstract
(1) Background: This study aimed to establish the effect of calcium lactate enamel pretreatment related to different fluoride types and concentrations on the enamel uptake of alkali-soluble fluorides. (2) Materials: In a blind and randomized in vitro study, a total of 60 teeth [...] Read more.
(1) Background: This study aimed to establish the effect of calcium lactate enamel pretreatment related to different fluoride types and concentrations on the enamel uptake of alkali-soluble fluorides. (2) Materials: In a blind and randomized in vitro study, a total of 60 teeth are used. The first 30 teeth were cut and randomly allocated into one of the following treatments: (A) calcium lactate pretreatment followed by three different fluoride solutions; (B) the “Fluoride only” group, with slabs treated with three different fluoride solutions; (C) the “Calcium only” group, with slabs treated with calcium lactate solution; (D) slabs treated with deionized water (negative control group). The next 30 teeth underwent all the above described group procedures but were treated with lower fluoride concentrations. Fluoride was extracted from enamel using 1 M KOH solution and analyzed using a fluoride ion-specific electrode. (3) Results: The findings revealed that slabs treated with NaF following calcium lactate pretreatment exhibited significantly greater enamel uptake of alkali-soluble fluoride compared to other substrates. This significant effect was not observed at lower fluoride concentrations. (4) Conclusion: The study demonstrates that pretreatment with calcium lactate followed by treatment with NaF at 226 ppm F significantly enhances the uptake of alkali-soluble fluoride in enamel compared to other fluoride types. Full article
(This article belongs to the Special Issue Property, Evaluation and Development of Dentin Materials)
Show Figures

Figure 1

Figure 1
<p>Uptake of alkali soluble fluoride during treatment with low concentration of F free ions.</p>
Full article ">Figure 2
<p>Uptake of alkali soluble fluoride during treatment with high concentration of F free ions.</p>
Full article ">
17 pages, 13879 KiB  
Article
Sirt2 Regulates Liver Metabolism in a Sex-Specific Manner
by Alexandra V. Schmidt, Sivakama S. Bharathi, Keaton J. Solo, Joanna Bons, Jacob P. Rose, Birgit Schilling and Eric S. Goetzman
Biomolecules 2024, 14(9), 1160; https://doi.org/10.3390/biom14091160 - 15 Sep 2024
Viewed by 236
Abstract
Sirtuin-2 (Sirt2), an NAD+-dependent lysine deacylase enzyme, has previously been implicated as a regulator of glucose metabolism, but the specific mechanisms remain poorly defined. Here, we observed that Sirt2−/− males, but not females, have decreased body fat, moderate hypoglycemia upon fasting, and perturbed [...] Read more.
Sirtuin-2 (Sirt2), an NAD+-dependent lysine deacylase enzyme, has previously been implicated as a regulator of glucose metabolism, but the specific mechanisms remain poorly defined. Here, we observed that Sirt2−/− males, but not females, have decreased body fat, moderate hypoglycemia upon fasting, and perturbed glucose handling during exercise compared to wild type controls. Conversion of injected lactate, pyruvate, and glycerol boluses into glucose via gluconeogenesis was impaired, but only in males. Primary Sirt2−/− male hepatocytes exhibited reduced glycolysis and reduced mitochondrial respiration. RNAseq and proteomics were used to interrogate the mechanisms behind this liver phenotype. Loss of Sirt2 did not lead to transcriptional dysregulation, as very few genes were altered in the transcriptome. In keeping with this, there were also negligible changes to protein abundance. Site-specific quantification of the hepatic acetylome, however, showed that 13% of all detected acetylated peptides were significantly increased in Sirt2−/− male liver versus wild type, representing putative Sirt2 target sites. Strikingly, none of these putative target sites were hyperacetylated in Sirt2−/− female liver. The target sites in the male liver were distributed across mitochondria (44%), cytoplasm (32%), nucleus (8%), and other compartments (16%). Despite the high number of putative mitochondrial Sirt2 targets, Sirt2 antigen was not detected in purified wild type liver mitochondria, suggesting that Sirt2’s regulation of mitochondrial function occurs from outside the organelle. We conclude that Sirt2 regulates hepatic protein acetylation and metabolism in a sex-specific manner. Full article
(This article belongs to the Special Issue Molecular Mechanisms Underlying Liver Diseases)
Show Figures

Figure 1

Figure 1
<p>(<b>a</b>) Mass spectrometry was used to quantify acetylated peptides from male <span class="html-italic">Sirt2+/+</span> (WT) and <span class="html-italic">Sirt2−/−</span> (KO) livers. Peptides with significant changes in acetylation are rendered in red. Blue lines indicate the mean log2 fold-change. A total of 2452 acetylated peptides were quantified. The mean acetylation level significantly increased in 317 peptides (<span class="html-italic">p</span> &lt; 0.01, FC &gt; 1.5) due to Sirt2 ablation. (<b>b</b>) Mass spectrometry was used to quantify acetylated peptides from female <span class="html-italic">Sirt2+/+</span> and <span class="html-italic">Sirt2−/−</span> livers. There was no appreciable change in acetylation levels across putative Sirt2-targets. (<b>c</b>) Volcano plot of <span class="html-italic">Sirt2+/+</span> female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (<b>d</b>) Volcano plot of <span class="html-italic">Sirt2−/−</span> female to male ratios at each of the 2452 putative Sirt2 targets, expressed as Log2 FC. (<b>e</b>) The mean Log2 FC in <span class="html-italic">Sirt2+/+</span> female/male acetylation was 4.2; the mean Log2 FC in <span class="html-italic">Sirt2−/−</span> female/male acetylation was 1.3. Quantification and comparison of the Log2 FC ratios using a Student’s two-sided <span class="html-italic">t</span>-test showed that <span class="html-italic">Sirt2−/−</span> males had significant decrease in hyperacetylation in 2452 putative sites compared to wild type counterparts (<span class="html-italic">p</span> &lt; 0.0001). (<b>f</b>) Immunoblotting for Sirt2 in livers from wild type mice indicated no sex difference in the expression of the Sirt2 protein. Note that there are multiple bands due to multiple isoforms of Sirt2. **** <span class="html-italic">p</span> &lt; 0.0001 by Student’s two-sided <span class="html-italic">t</span>-test.</p>
Full article ">Figure 2
<p>(<b>a</b>) The 306 putative Sirt2 target sites mapped to proteins spread across multiple cellular compartments but were most abundant in the mitochondria and cytosol. (<b>b</b>) A plot of the Log2 ratio (FC &gt; 1.5) of <span class="html-italic">Sirt2−/−</span> vs. <span class="html-italic">Sirt2+/+</span> by cellular compartment revealed that the mitochondria and cytosol had the largest FCs in lysine acetylation. (<b>c</b>) The Log2 Ratio of putative mitochondrial Sirt2 targets from this study was compared to known Sirt3 targets within the mitochondria. Of the 140 Sirt2 targets represented in this plot, 19% are also Sirt3 targets. (<b>d</b>) Preparations of <span class="html-italic">Sirt2+/+</span> nuclear and cytosolic fractions show that Sirt2 is present in both compartments, confirmed with the cytosolic marker α-tubulin and the nuclear marker, Lamin A/C (1 and 2 denote matched, replicate samples). Sirt2 is present in both compartments, confirming acetylomics data. (<b>e</b>) Sirt2 was not detected in mitochondria prepared from <span class="html-italic">Sirt2+/+</span> hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (<b>f</b>) Sirt2 was not detected in peroxisomes prepared from <span class="html-italic">Sirt2+/+</span> hepatic tissue lysates (N = 3). 1–3 denotes replicate samples. (<b>g</b>) The ultrapure subcellular fractionation technique used completely separated mitochondria and peroxisomes, confirmed with the peroxisomal marker, ACOX1, and the mitochondrial marker, TIMM23 (N = 3). 1–3 denotes matched, replicate samples. Membranes exposed at the same time, cropped for clarity.</p>
Full article ">Figure 3
<p>(<b>a</b>,<b>b</b>) Total body weight and lean body mass were the same between <span class="html-italic">Sirt2−/−</span> (N = 11) and <span class="html-italic">Sirt2+/+</span> (N = 8) males, but <span class="html-italic">Sirt2−/−</span> males had significantly lower fat mass compared to <span class="html-italic">Sirt2+/+</span> males (<span class="html-italic">p</span> &lt; 0.05). (<b>b</b>) Total body weight, fat mass, and lean mass were not significantly different between <span class="html-italic">Sirt2−/−</span> (N = 6) and <span class="html-italic">Sirt2+/+</span> females (N = 8). Body composition was calculated for each mouse by dividing fat mass by lean mass. Only <span class="html-italic">Sirt2−/−</span> males had a significantly reduced fat/lean mass body composition (<span class="html-italic">p</span> &lt; 0.001) compared to controls; female <span class="html-italic">Sirt2−/−</span> did not. (<b>c</b>) Blood glucose levels in <span class="html-italic">Sirt2−/−</span> vs. <span class="html-italic">Sirt2+/+</span> males or females in both a fed and fasted state. In a fed state, there was no difference in blood glucose levels between <span class="html-italic">Sirt2−/−</span> and <span class="html-italic">Sirt2+/+</span> in either males (N = 8) or females (N = 8). Upon fasting, only <span class="html-italic">Sirt2−/−</span> males had significantly reduced blood glucose levels (N = 8; <span class="html-italic">p</span> &lt; 0.05) while <span class="html-italic">Sirt2−/−</span> females showed no significant reduction compared to controls (N = 8). (<b>d</b>) H&amp;E staining of <span class="html-italic">Sirt2−/−</span> vs. <span class="html-italic">Sirt2+/+</span> livers indicated no obvious morphological changes or tissue damage (N = 4). (<b>e</b>) Male <span class="html-italic">Sirt2−/−</span> mice (N = 3) had significantly less triglycerides in hepatic tissue compared to <span class="html-italic">Sirt2+/+</span> males (N = 3; <span class="html-italic">p</span> &lt; 0.05). Comparisons made with a Student’s nonparametric <span class="html-italic">t</span>-test; * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001.</p>
Full article ">Figure 4
<p>(<b>a</b>) <span class="html-italic">Sirt2−/−</span> and <span class="html-italic">Sirt2+/+</span> males ran the same distance (N = 5) before reaching exhaustion. (<b>b</b>) Blood glucose levels were normal prior to the treadmill assay in male <span class="html-italic">Sirt2−/−</span> and <span class="html-italic">Sirt2+/+</span> mice. Upon exhaustion, male <span class="html-italic">Sirt2−/−</span> mice (N = 5) had significantly lower blood glucose levels than <span class="html-italic">Sirt2+/+</span> counterparts (N = 5; <span class="html-italic">p</span> &lt; 0.01). (<b>c</b>) <span class="html-italic">Sirt2−/−</span> females (N = 6) ran significantly further than <span class="html-italic">Sirt2+/+</span> females (N = 7; <span class="html-italic">p</span> &lt; 0.005) before reaching exhaustion. (<b>d</b>) There was no significant difference in blood glucose levels between <span class="html-italic">Sirt2−/−</span> (N = 6) and <span class="html-italic">Sirt2+/+</span> females (N = 7) post-exhaustion. Comparisons were made using a Student’s nonparametric <span class="html-italic">t</span>-test; ns = no difference; ** <span class="html-italic">p</span> &lt; 0.01.</p>
Full article ">Figure 5
<p>(<b>a</b>) Male <span class="html-italic">Sirt2−/−</span> mice had a blunted response to a bolus lactate injection, with significantly decreased glucose production 15-min post-injection (N = 5; <span class="html-italic">p</span> &lt; 0.05). (<b>b</b>) <span class="html-italic">Sirt2−/−</span> females had a non-significantly different response to bolus lactate injection compared to <span class="html-italic">Sirt2+/+</span> females (N = 5). (<b>c</b>) Ldha activity in <span class="html-italic">Sirt2−/−</span> fasted males (N = 3) was significantly reduced compared to <span class="html-italic">Sirt2+/+</span> male mice (N = 3; <span class="html-italic">p</span> &lt; 0.05). Ldha activity in fed <span class="html-italic">Sirt2−/−</span> mice was significantly higher than in <span class="html-italic">Sirt2+/+</span> male mice (N = 3; <span class="html-italic">p</span> &lt; 0.05). (<b>d</b>) Male <span class="html-italic">Sirt2−/−</span> mice (N = 5) had a significantly reduced response to a pyruvate tolerance test than male <span class="html-italic">Sirt2+/+</span> mice (N=) 75 (<span class="html-italic">p</span> &lt; 0.0005), 85 (<span class="html-italic">p</span> &lt; 0.005), and 100 min (<span class="html-italic">p</span> &lt; 0.05) post-injection. (<b>e</b>) <span class="html-italic">Sirt2−/−</span> males (N = 5) had significantly less glucose production at 30-, 60-, and 120-min post-glycerol injection (<span class="html-italic">p</span> &lt; 0.05). All comparisons were made at each time point using a Student’s parametric <span class="html-italic">t</span>-test; * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.001, *** <span class="html-italic">p</span> &lt; 0.0001.</p>
Full article ">Figure 6
<p>Schematic of glycolytic enzymes (red) and gluconeogenic enzymes (purple) that contain putative Sirt2 targets. Each significantly altered residue (<span class="html-italic">p</span> &lt; 0.01; FC &gt; 1.5) is identified with FC reported. Non-targets are in blue.</p>
Full article ">Figure 7
<p>(<b>a</b>) Hepatocytes isolated from a <span class="html-italic">Sirt2−/−</span> liver showed reduced OCR compared to <span class="html-italic">Sirt2+/+</span> hepatocytes during a mitochondrial stress test using a Seahorse Extracellular Flux Analyzer (<span class="html-italic">p</span> &lt; 0.005). (<b>b</b>) Hepatocytes isolated from <span class="html-italic">Sirt2−/−</span> liver showed significantly reduced OCR compared to <span class="html-italic">Sirt2+/+</span> isolated hepatocytes when given media containing only pyruvate as an energy source (<span class="html-italic">p</span> &lt; 0.00001). (<b>c</b>) <span class="html-italic">Sirt2−/−</span> liver tissue lysates (N = 4) had a significantly reduced FAO response to palmitate (16C chain fatty acid) compared to <span class="html-italic">Sirt2+/+</span> liver lysate (N = 4) during a radio-labeled redox assay (<span class="html-italic">p</span> &lt; 0.05). (<b>d</b>) <span class="html-italic">Sirt2-/-</span> liver lysates (N = 6) had a significantly reduced response to octanoate (8C chain fatty acid) compared to liver lysate prepared from <span class="html-italic">Sirt2+/+</span> mice (N = 6; <span class="html-italic">p</span> &lt; 0.05). (<b>e</b>) The maximal mitochondrial capacity was significantly lower in mitochondria isolated from male <span class="html-italic">Sirt2−/−</span> mouse hepatocytes (N = 3) compared to <span class="html-italic">Sirt2+/+</span> isolated mitochondria (N = 3; <span class="html-italic">p</span> &lt; 0.05). (<b>f</b>) The maximal mitochondrial capacity was relatively the same in mitochondria isolated from female <span class="html-italic">Sirt2−/−</span> mouse hepatocytes (N = 3) compared to <span class="html-italic">Sirt2+/+</span> isolated mitochondria (N = 3; <span class="html-italic">p</span> &gt; 0.05). All comparisons were performed with a Student’s nonparametric <span class="html-italic">t</span>-test; * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.0001, **** <span class="html-italic">p</span> &lt; 0.00001.</p>
Full article ">
10 pages, 1212 KiB  
Article
Prognostic Significance of Lactate Clearance in Cardiogenic Pulmonary Edema in the Emergency Department
by Mehmet Göktuğ Efgan, Ejder Saylav Bora, Ahmet Kayalı, Umut Payza, Tutku Duman Şahan and Zeynep Karakaya
Medicina 2024, 60(9), 1502; https://doi.org/10.3390/medicina60091502 - 14 Sep 2024
Viewed by 152
Abstract
Background and Objectives: Acute cardiorespiratory failure disrupts the delicate balance of energy supply, demand, and consumption, with elevated lactate levels and decreased blood pH serving as crucial indicators. Acute cardiogenic pulmonary edema (ACPO), a common cause of acute respiratory failure, poses a [...] Read more.
Background and Objectives: Acute cardiorespiratory failure disrupts the delicate balance of energy supply, demand, and consumption, with elevated lactate levels and decreased blood pH serving as crucial indicators. Acute cardiogenic pulmonary edema (ACPO), a common cause of acute respiratory failure, poses a substantial mortality risk. Lactate, a byproduct of pyruvate reduction, is a pertinent marker in perfusion assessment. Lactate clearance (LC) has proven prognostic efficacy in various conditions but lacks consensus on its predictive power in acute cardiogenic pulmonary edema. Materials and Methods: This prospective observational study, conducted in a metropolitan area’s third-level emergency department, involved patients with cardiogenic pulmonary edema from May 2021 to August 2023. The inclusion criteria specified acute cardiogenic pulmonary edema, excluding patients with incomplete data or other respiratory conditions. Lactate clearance, calculated at presentation and after 6 h, served as the primary outcome predictor. Our data analysis employed logistic regression, the ROC curve, and statistical tests. Results: The cohort of 106 patients revealed that a lactate clearance below 14.29% was significantly associated with mortality. While 51.6% of survivors were discharged, LC’s predictive success for discharge was inconclusive. Logistic regression underscored the significance of lactate clearance, with a one-unit increase yielding a 5.55-fold probability of survival. The AUC for LC was 0.759. Conclusions: This study pioneers the exploration of lactate clearance in patients with acute cardiogenic pulmonary edema. LC below 14.29% signifies a poor prognosis, emphasizing its potential as an early treatment initiation marker. While acknowledging this study’s limitations, we advocate for further multicenter research to refine the understanding of lactate clearance in this context. Full article
(This article belongs to the Section Emergency Medicine)
Show Figures

Figure 1

Figure 1
<p>Consultation diagram.</p>
Full article ">Figure 2
<p>ROC curve for LC measurement performance in predicting outcome.</p>
Full article ">Figure 3
<p>ROC analysis for LC success in predicting discharge.</p>
Full article ">
13 pages, 3650 KiB  
Article
Sirtuin 1 Inhibits Fatty Acid Synthesis through Forkhead Box Protein O1-Mediated Adipose Triglyceride Lipase Expression in Goat Mammary Epithelial Cells
by Qiuya He, Weiwei Yao, Li Lv, Xuelin Zhang, Jiao Wu and Jun Luo
Int. J. Mol. Sci. 2024, 25(18), 9923; https://doi.org/10.3390/ijms25189923 (registering DOI) - 14 Sep 2024
Viewed by 160
Abstract
Sirtuin 1 (SIRT1) is a key upstream regulator of lipid metabolism; however, the molecular mechanisms by which SIRT1 regulates milk fat synthesis in dairy goats remain unclear. This study aimed to investigate the regulatory roles of SIRT1 in modulating lipid metabolism in goat [...] Read more.
Sirtuin 1 (SIRT1) is a key upstream regulator of lipid metabolism; however, the molecular mechanisms by which SIRT1 regulates milk fat synthesis in dairy goats remain unclear. This study aimed to investigate the regulatory roles of SIRT1 in modulating lipid metabolism in goat mammary epithelial cells (GMECs) and its impact on the adipose triglyceride lipase (ATGL) promoter activity using RNA interference (RNAi) and gene overexpression techniques. The results showed that SIRT1 is significantly upregulated during lactation compared to the dry period. Additionally, SIRT1 knockdown notably increased the expressions of genes related to fatty acid synthesis (SREBP1, SCD1, FASN, ELOVL6), triacylglycerol (TAG) production (DGAT2, AGPAT6), and lipid droplet formation (PLIN2). Consistent with the transcriptional changes, SIRT1 knockdown significantly increased the intracellular contents of TAG and cholesterol and the lipid droplet abundance in the GMECs, while SIRT1 overexpression had the opposite effects. Furthermore, the co-overexpression of SIRT1 and Forkhead box protein O1 (FOXO1) led to a more pronounced increase in ATGL promoter activity, and the ability of SIRT1 to enhance ATGL promoter activity was nearly abolished when the FOXO1 binding sites (FKH1 and FKH2) were mutated, indicating that SIRT1 enhances the transcriptional activity of ATGL via the FKH element in the ATGL promoter. Collectively, our data reveal that SIRT1 enhances the transcriptional activity of ATGL through the FOXO1 binding sites located in the ATGL promoter, thereby regulating lipid metabolism. These findings provide novel insights into the role of SIRT1 in fatty acid metabolism in dairy goats. Full article
(This article belongs to the Special Issue Sirtuins as Players in Cell Metabolism and Functions)
Show Figures

Figure 1

Figure 1
<p>SIRT1 mRNA expression in goat tissues. (<b>A</b>) SIRT1 was expressed across multiple goat tissues. Lowercase letters indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05), while the same letters indicate no significant differences. (<b>B</b>) SIRT1 expression was significantly higher in the mammary gland during peak lactation compared to the dry period. The data were normalized to the dry period. The values are shown as the mean ± standard error of the mean (SEM) for three biological replicates. *, <span class="html-italic">p</span> &lt; 0.05; **, <span class="html-italic">p</span> &lt; 0.01.</p>
Full article ">Figure 2
<p>Effect of SIRT1 knockdown on the expressions of genes related to fatty acid metabolism in GMECs. (<b>A</b>) The mRNA levels of SIRT1 after infection with siRNA-SIRT1. (<b>B</b>,<b>C</b>) Protein level and quantification of SIRT1 in cells transfected with siRNA-SIRT1. The effect of SIRT1 knockdown on the expressions of genes involved in (<b>D</b>) fatty acid synthesis (SREBP1, SCD1, FASN, and ELOVL6), (<b>E</b>) TAG synthesis (DGAT1, DGAT2, GPAM, and AGPAT6), and (<b>F</b>) lipid droplet formation (PLIN2 and XDH). The RT-qPCR data were calculated using the 2<sup>−ΔΔCt</sup> method. Data shown are mean ± SEM. Statistically significant differences are indicated: ** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05. Note: SIRT1, Sirtuin 1; GMECs, goat mammary epithelial cells; SREBP1, Sterol Regulatory Element-Binding Proteins 1; SCD1, stearoyl-coenzyme A desaturase; FASN, fatty acid synthase; ELOVL6, elongase of very long chain fatty acid 6; DGAT1/2, Diacylglycerol-O-Acyltransferase; GPAM, glycerol-3-phosphate acyltransferase; AGPAT6, 1-Acylglycerol-3-Phosphate O-Acyltransferase 6; PLIN2, Perilipin 2; XDH, xanthine dehydrogenase.</p>
Full article ">Figure 3
<p>Effect of SIRT1 overexpression on the expressions of genes related to lipid metabolism in GMECs. (<b>A</b>) The SIRT1 mRNA expression in GMECs that were transfected with SIRT1 overexpression vector. (<b>B</b>,<b>C</b>) Protein abundance of SIRT1 after transfection of the pcDNA3.1-SIRT1 in GMECs (N = 3). The expressions of genes involved in lipid metabolism after SIRT1 overexpression including fatty acid synthesis (SREBP1, PPARG, FASN, and ELOVL6; panel (<b>D</b>)), TAG synthesis (DGAT2, GPAM, AGPAT6, and LIPIN1; panel (<b>E</b>)), and lipid droplet formation (PLIN2 and XDH; panel (<b>F</b>)). Data shown are mean ± SEM. Statistically significant differences are indicated: ** <span class="html-italic">p</span> &lt; 0.01; * <span class="html-italic">p</span> &lt; 0.05. Note: SIRT1, Sirtuin 1; SREBP1, Sterol Regulatory Element-Binding Proteins 1; PPARG, peroxisome proliferator-activated receptor gamma; FASN, fatty acid synthase; ELOVL6, elongase of very long chain fatty acid 6; DGAT2, Diacylglycerol-O-Acyltransferase 2; GPAM, glycerol-3-phosphate acyltransferase; AGPAT6, 1-Acylglycerol-3-Phosphate O-Acyltransferase 6; PLIN2, Perilipin 2; XDH, xanthine dehydrogenase.</p>
Full article ">Figure 4
<p>The effect of SIRT1 knockdown on the contents of triglycerides, cholesterol, and lipid droplets. (<b>A</b>) The content of triglycerides in cells transfected with siRNA-SIRT1. (<b>B</b>) Cellular cholesterol levels after knockdown of SIRT1 in GMECs. (<b>C</b>,<b>D</b>) Lipid droplets in SIRT1 knockdown cells were detected by BODIPY 493/503 staining. Scale bar = 200 μm.Values are shown as mean ± SEM. *, <span class="html-italic">p</span> &lt; 0.05. Note: SIRT1, Sirtuin 1.</p>
Full article ">Figure 5
<p>SIRT1 overexpression in GMECs regulates lipid metabolism. (<b>A</b>) SIRT1 overexpression significantly decreased the content of triacylglycerol (TAG) in GMECs. (<b>B</b>) SIRT1 overexpression reduced the cellular cholesterol levels. (<b>C</b>) Overexpression of SIRT1 inhibited the accumulation of lipid droplets. Cellular nuclei were stained with a blue signal, while the lipid droplets were labeled with a green signal. Scale bar = 200 μm. (<b>D</b>) The relative fluorescence intensity of the lipid droplets. Data shown are mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05.</p>
Full article ">Figure 6
<p>SIRT1 promotes ATGL transcription activity through FOXO1. (<b>A</b>,<b>B</b>) The expression of ATGL after SIRT1 overexpression or inhibition. (<b>C</b>) SIRT1 knockdown significantly decreased the ATGL promoter activity. (<b>D</b>) Effects of SIRT1 and FOXO1 on ATGL promoter activity. GMECs were co-transfected with different lengths of the ATGL promoter and plasmid expressing SIRT1 or empty vector for 48 h. Cells were harvested and luciferase activity assay was performed to detect the effect of SIRT1 on the ATGL promoter activity. In addition, to detect the effect of FOXO1 on the ATGL promoter activity, cells were co-transfected with different lengths of the ATGL promoter and plasmid expressing SIRT1 and FOXO1 for 48 h. (<b>E</b>) SIRT1 increases ATGL promoter activity in a FOXO1-dependent manner. GMECs were co-transfected with wild-type ATGL promoter (−882 + 216 bp) or constructs containing individually or simultaneously mutated FOXO1 binding sites and pcDNA3.1-SIRT1 for 48 h. Data are presented as mean ± SEM; *, <span class="html-italic">p</span> &lt; 0.05. Note: SIRT1, Sirtuin 1; FOXO1, Forkhead box protein O1; ATGL, Adipose Triglyceride Lipase; FKH, FOXO1 binding sites; Red cross, Mutation site.</p>
Full article ">
11 pages, 1982 KiB  
Article
Paper-Based Microfluidic Device for Extracellular Lactate Detection
by Yan Nan, Peng Zuo and Bangce Ye
Biosensors 2024, 14(9), 442; https://doi.org/10.3390/bios14090442 - 14 Sep 2024
Viewed by 266
Abstract
Lactate is a critical regulatory factor secreted by tumors, influencing tumor development, metastasis, and clinical prognosis. Precise analysis of tumor-cell-secreted lactate is pivotal for early cancer diagnosis. This study describes a paper-based microfluidic chip to enable the detection of lactate levels secreted externally [...] Read more.
Lactate is a critical regulatory factor secreted by tumors, influencing tumor development, metastasis, and clinical prognosis. Precise analysis of tumor-cell-secreted lactate is pivotal for early cancer diagnosis. This study describes a paper-based microfluidic chip to enable the detection of lactate levels secreted externally by living cells. Under optimized conditions, the lactate biosensor can complete the assay in less than 30 min. In addition, the platform can be used to distinguish lactate secretion levels in different cell lines and can be applied to the screening of antitumor drugs. Through enzymatic chemical conversion, this platform generates fluorescent signals, enabling qualitative assessment under a handheld UV lamp and quantitative analysis via grayscale intensity measurements using ImageJ (Ver. 1.50i) software. The paper-based platform presented in this study is rapid and highly sensitive and does not necessitate other costly and intricate instruments, thus making it applicable in resource-constrained areas and serving as a valuable tool for investigating cell lactate secretion and screening various anti-cancer drugs. Full article
(This article belongs to the Special Issue Application of Microfluidics in Cell Manipulation and Biosensing)
Show Figures

Figure 1

Figure 1
<p>Principle of the paper-based microfluidic platform. This paper-based microfluidic device combines the function of both analysis and detection.</p>
Full article ">Figure 2
<p>Validation of the feasibility of lactate detection using the paper-based platform. (<b>A</b>) The fluorescence images of the products from the reactions of different reagent combinations after 30 min (shot under a handheld UV lamp). (<b>B</b>) Fluorescence measurements of different combinations of the reagents with a microplate reader. Combination 1: the blank control; 2: LDH; 3: NAD<sup>+</sup>; 4: LA+LDH; 5: LA+NAD<sup>+</sup>; 6: LDH+NAD<sup>+</sup>; 7: LA+LDH+NAD<sup>+</sup>. All solutions were in Tris-HCl buffer.</p>
Full article ">Figure 3
<p>Optimization of the fluorescence assay for the detection of lactate. (<b>A</b>) Optimization of LDH concentration. (<b>B</b>) Optimization of NAD<sup>+</sup> concentration. (<b>C</b>) Optimization of buffer pH value. (<b>D</b>) Optimization of reaction time for the detection system. Each group includes fluorescence images taken under various conditions, alongside corresponding histograms showing grayscale signals. All the evaluations were carried out in triplicate (n = 3).</p>
Full article ">Figure 4
<p>Sensitivity of lactate detection based on the paper-based platform. Fluorescence photographs of lactate detection at different concentrations (i.e., 100 mM, 200 mM, 300 mM, 400 mM, 500 mM, and 600 mM) on the paper-based platform are shown. The calibration curve by plotting the average grayscale values against the different lactate concentration is presented. Error bars: standard deviation from different experiments (n = 3).</p>
Full article ">Figure 5
<p>Applications for the paper-based lactate detection platform. (<b>A</b>) Cultivation of different types of cells on the paper-based platform and lactate detection: fluorescence images were taken after the reaction of the lactate detection system 24 h after cell culture; (<b>B</b>) Lactate secretion of MCF-7 cells under different drug treatments (doxorubicin and paclitaxel at a concentration of 10 µg mL<sup>−1</sup>, as well as without treatment) were examined on the paper-based platform. Fluorescence images (arranged left to right: doxorubicin, paclitaxel, and no treatment) were used to derive grayscale values for the histograms. Error bar: standard deviation from different experiments (n = 3).</p>
Full article ">
12 pages, 891 KiB  
Article
Effect of Exogenous Melatonin on Performance and Mastitis in Dairy Cows
by Yunmeng Li, Zhiqiang Cheng, Wenting Ma, Yaqi Qiu, Tuo Liu, Bingyu Nan, Mengfei Li, Long Sun, Wentao Liu, Haina Yin, Caidie Wang, Xiaobin Li and Changjiang Zang
Vet. Sci. 2024, 11(9), 431; https://doi.org/10.3390/vetsci11090431 - 13 Sep 2024
Viewed by 302
Abstract
Mastitis is an important factor affecting the health of cows that leads to elevated somatic cell counts in milk, which can seriously affect milk quality and result in huge economic losses for the livestock industry. Therefore, the aim of this trial was to [...] Read more.
Mastitis is an important factor affecting the health of cows that leads to elevated somatic cell counts in milk, which can seriously affect milk quality and result in huge economic losses for the livestock industry. Therefore, the aim of this trial was to investigate the effect of melatonin on performance and mastitis in dairy cows. Forty-eight Holstein cows with a similar body weight (470 ± 10 kg), parity (2.75 ± 1.23), number of lactation days (143 ± 43 days), BCS (3.0–3.5), milk yield (36.80 ± 4.18 kg), and somatic cell count (300,000–500,000 cells/mL) were selected and randomly divided into four groups: control (CON group), trial Ⅰ (T80 group), trial Ⅱ (T120 group), and trial Ⅲ (T160 group). Twelve cows in trial groups I, II, and III were pre-dispensed 80, 120, and 160 mg of melatonin in edible glutinous rice capsules along with the basal ration, respectively, while the control group was fed an empty glutinous rice capsule along with the ration. The trial period was 37 days, which included a 7-day adaptive phase followed by a 30-day experimental period. At the end of the trial period, feeding was ended and the cows were observed for 7 days. Milk samples were collected on days 0, 7, 14, 21, 28, and 37 to determine the somatic cell number and milk composition. Blood samples were collected on days 0, 15, 30, and 37 of the trial to determine the serum biochemical indicators, antioxidant and immune indicators, and the amount of melatonin in the blood. The results showed that the somatic cell counts of lactating cows in the CON group were lower than those in the T120 group on days 14 (p < 0.05) and 28 (p < 0.01) at 1 week after melatonin cessation. The milk protein percentage and milk fat percentage of cows in the T120 group were higher than those in the CON group (p < 0.01). The total protein and globulin content in the T120 group were higher than those in the CON group (p < 0.01). In terms of antioxidant capacity and immunity, the cows 1 week after melatonin cessation showed higher superoxide dismutase activity and interleukin-10 contents (p < 0.01) compared with the CON group and lower malondialdehyde and tumor necrosis factor-alpha contents (p < 0.01) compared with the T120 group. The melatonin content in the T120 group was increased relative to that in the other groups. In conclusion, exogenous melatonin can increase the content of milk components, reduce the somatic cell count, and improve the antioxidant capacity and immune responses to a certain extent. Under the experimental conditions, 120 mg/day melatonin is recommended for mid- to late-lactation cows. Full article
(This article belongs to the Special Issue Effects of Nutrition on Ruminants Production Performance and Health)
Show Figures

Figure 1

Figure 1
<p>Trial design and grouping diagram.</p>
Full article ">Figure 2
<p>Effect of melatonin on somatic cell count in dairy cows.T80, trial 80 mg/day; T120, trial 120 mg/day; T160, trial 160 mg/day; SEM, standard error of mean; SCC, somatic cell count; CON, control with no MT; T80, T120, and T160, fed basal ration and 80 mg/day, 120 mg/day, or 160 mg/day of MT (Senrise Technology Co., Ltd., Anqing, China) per cow, respectively. In the figure, the same letter indicates no significant difference (<span class="html-italic">p</span> &gt; 0.05) and different lowercase or uppercase letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05; <span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">
17 pages, 2734 KiB  
Article
Effects of a Combined Chinese Herbal Medicine on Growth Performance, Intestinal Barrier Function, Immune Response, and Cecal Microflora in Broilers Infected with Salmonella enteritidis
by Changzhi Zou, Xin Xing, Shunxi Li, Xuelong Zheng, Jinshan Zhao and Huawei Liu
Animals 2024, 14(18), 2670; https://doi.org/10.3390/ani14182670 - 13 Sep 2024
Viewed by 206
Abstract
This study investigated the effects of CCHM in drinking water on broilers infected with Salmonella enteritidis. One-day-old male Cobb 500 broilers (n = 300) were randomly assigned to five groups: a control (NC) group, a Salmonella enteritidis challenge (SE) group, an antibiotic [...] Read more.
This study investigated the effects of CCHM in drinking water on broilers infected with Salmonella enteritidis. One-day-old male Cobb 500 broilers (n = 300) were randomly assigned to five groups: a control (NC) group, a Salmonella enteritidis challenge (SE) group, an antibiotic (AB) group, a low dose of CCHM (CL) group, and a high dose of CCHM (CH) group. Each group had six replicate cages with ten broilers per cage. The broilers in the NC and SE groups were given normal drinking water. From days 12 to 18, the AB group received water treated with ciprofloxacin lactate injection (1 mL/L), while the CL and CH groups received water containing CCHM at doses of 5 mL/L and 10 mL/L, respectively. Broilers in all groups except the NC group were orally given Salmonella enteritidis daily from days 9 to 11. The experimental period was 28 days. The results showed that, compared with the SE group, the CL and CH groups showed improved growth performance; increased immune organ indices, expressions of ileal occludin and ZO-1 proteins, jejunal and ileal villus heights (except at day 19), and cecal Lactobacillus counts on days 19 and 28 (p < 0.05); and decreased jejunal and ileal lesion scores, ileal interleukin 1β (IL-1β) (except at day 19), interferon-γ (IFN-γ), interleukin 6 (IL-6) (except at day 19), secretory immunoglobulin A (slgA) and tumor necrosis factor α (TNF-α) (except at day 19) levels, serum D-lactic acid and diamine oxidase (DAO) (except at day 19) contents, jejunal and ileal crypt depths (except at day 19), and cecal Salmonella and Escherichia coli counts on days 19 and 28 (p < 0.05). On day 28, except for the levels of ileal interleukin 10 (IL-10), TNF-α, slgA, and serum D-lactic acid content, there were no differences among the NC, AB, and CL groups (p > 0.05). In conclusion, drinking water supplemented with CCHM alleviated the intestinal damage caused by Salmonella enteritidis infection and improved growth performance and cecal microbiota in broilers. The optimal addition rate of CCHM was 5 mL/L. Full article
(This article belongs to the Section Poultry)
Show Figures

Figure 1

Figure 1
<p>Effect of CCHM on the immune-related indices of the ileal mucosa of broilers infected with <span class="html-italic">Salmonella enteritidis</span> on day 19 (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge.</p>
Full article ">Figure 1 Cont.
<p>Effect of CCHM on the immune-related indices of the ileal mucosa of broilers infected with <span class="html-italic">Salmonella enteritidis</span> on day 19 (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge.</p>
Full article ">Figure 2
<p>Effect of CCHM on the immune-related indices of the ileal mucosa of broilers infected with <span class="html-italic">Salmonella enteritidis</span> on day 28. (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge.</p>
Full article ">Figure 2 Cont.
<p>Effect of CCHM on the immune-related indices of the ileal mucosa of broilers infected with <span class="html-italic">Salmonella enteritidis</span> on day 28. (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge.</p>
Full article ">Figure 3
<p>Effect of CCHM on <span class="html-italic">D</span>-lactic acid and diamine oxidase in broilers infected with <span class="html-italic">Salmonella enteritidis</span> (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge.</p>
Full article ">Figure 4
<p>Effect of CCHM on the tight junction protein expressions of the ileal mucosa of broilers infected with <span class="html-italic">Salmonella enteritidis</span> (<span class="html-italic">n</span> = 6 per experimental group). <sup>a,b,c,d</sup> Different letters indicate differences (<span class="html-italic">p</span> &lt; 0.05). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge (<a href="#app1-animals-14-02670" class="html-app">Figure S2</a>).</p>
Full article ">Figure 5
<p>Effect of CCHM on intestinal morphology in broilers infected with <span class="html-italic">Salmonella enteritidis</span> (<span class="html-italic">n</span> = 6 per experimental group). (<b>A</b>,<b>B</b>) Jejunum and ileum tissues stained with hematoxylin and eosin on days 19 and 28 (scale bar: 200 μm). NC, normal drinking water; SE, normal drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; AB, ciprofloxacin lactate injection (1 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CL, CCHM (5 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge; CH, CCHM (10 mL/L)-treated drinking water + <span class="html-italic">Salmonella enteritidis</span> challenge. Villus height: black line; Crypt depth: blue line. Images of the transverse section were photographed under 50× magnification.</p>
Full article ">
21 pages, 12979 KiB  
Article
Lactate-Induced HBEGF Shedding and EGFR Activation: Paving the Way to a New Anticancer Therapeutic Opportunity
by Valentina Rossi, Alejandro Hochkoeppler, Marzia Govoni and Giuseppina Di Stefano
Cells 2024, 13(18), 1533; https://doi.org/10.3390/cells13181533 - 13 Sep 2024
Viewed by 331
Abstract
Cancer cells can release EGF-like peptides, acquiring the capacity of autocrine stimulation via EGFR-mediated signaling. One of these peptides (HBEGF) was found to be released from a membrane-bound precursor protein and is critically implicated in the proliferative potential of cancer cells. We observed [...] Read more.
Cancer cells can release EGF-like peptides, acquiring the capacity of autocrine stimulation via EGFR-mediated signaling. One of these peptides (HBEGF) was found to be released from a membrane-bound precursor protein and is critically implicated in the proliferative potential of cancer cells. We observed that the increased lactate levels characterizing neoplastic tissues can induce the release of uPA, a protease promoting HBEGF shedding. This effect led to EGFR activation and increased ERK1/2 phosphorylation. Since EGFR-mediated signaling potentiates glycolytic metabolism, this phenomenon can induce a self-sustaining deleterious loop, favoring tumor growth. A well characterized HBEGF inhibitor is CRM197, a single-site variant of diphtheria toxin. We observed that, when administered individually, CRM197 did not trigger evident antineoplastic effects. However, its association with a uPA inhibitor caused dampening of EGFR-mediated signaling and apoptosis induction. Overall, our study highlights that the increased glycolytic metabolism and lactate production can foster the activated state of EGFR receptor and suggests that the inhibition of EGFR-mediated signaling can be attempted by means of CRM197 administered with an appropriate protease inhibitor. This attempt could help in overcoming the problem of the acquired resistance to the conventionally used EGFR inhibitors. Full article
(This article belongs to the Special Issue Cell Biology: State-of-the-Art and Perspectives in Italy 2024)
Show Figures

Figure 1

Figure 1
<p>mRNA levels of EGFR and of proteins involved in its activation, assessed by RT–PCR. (<b>A</b>) A preliminary evaluation was carried out to verify whether the different DMEM formulations could affect the proliferation dynamics of cell cultures. *, a statistically significant difference with <span class="html-italic">p</span> &lt; 0.05 was found in cultures maintained in High-Glc DMEM vs those maintained in the Low-Glc medium (ANOVA followed by Tukey’s post-test). (<b>B</b>) mRNA levels in MDA-MB-231 cells grown in Low-Glc DMEM were compared to those detected in High-Glc DMEM maintained cells and in cells exposed to Low-Glc DMEM + lactate. For genes’ selection, a threshold at ≥50% increase was set (dotted line). Furthermore, only genes showing no statistically significant difference between High-Glc DMEM grown cells and those exposed to Low-Glc DMEM + lactate were considered. These criteria were met by uPA, ERR-alpha, and GPER1 (@). The selected genes (@) were also studied in HT-29 cells, cultured in Low-Glc DMEM and exposed to lactate (<b>C</b>), together with MMP9 and MMP2 (two proteases involved in HBEGF shedding). In these cells, RT–PCR analysis substantially confirmed the data obtained in MDA-MB-231 cultures. The statistical evaluations applied to the data shown in (<b>B</b>,<b>C</b>) are detailed in the text. (<b>D</b>) Comparison of HBEGF mRNA levels between the two cell cultures.</p>
Full article ">Figure 2
<p>The results of RT–PCR experiments (<a href="#cells-13-01533-f001" class="html-fig">Figure 1</a>) were validated by the immunoblotting detection of proteins. (<b>A</b>) Images of the protein bands and of the used internal standard (Actin). (<b>B</b>) The densitometric reading of bands, normalized on Actin levels, was used to calculate the % increase of protein levels in lactate-exposed cells vs. control cultures, maintained in Low-Glc DMEM. Because of their limited extent, the increases of ERR-alpha (in both cell lines) and GPER1 (in HT-29 cultures) were not further considered. The results concerning uPA (in both cell cultures) and GPER1 (only in MDA-MB-231 cells) were analyzed by one-sample <span class="html-italic">t</span>-tests; * and ** indicate a statistically significant increase compared to the control cultures, with <span class="html-italic">p</span> &lt; 0.05 and 0.01, respectively.</p>
Full article ">Figure 3
<p>(<b>A</b>) Effects caused by BC11 (a uPA inhibitor) on the proliferation of control (grown in Low-Glc DMEM) and lactate-exposed MDA-MB-231 and HT-29 cells, at 24 h. Lactate was found to drastically reduce the toxic effects of BC11 in MDA-MB-231 culture. No significant difference was observed between the two lactate-exposed cultures at all the tested doses of BC11. (<b>B</b>) Detection of released HBEGF (24 h) in control (Low-Glc DMEM) cultures and in cells exposed to lactate or lactate + BC11 (100 µM). In MDA-MB-231 cultures, lactate significantly increased HBEGF shedding (*, <span class="html-italic">p</span> &lt; 0.05, assessed by ANOVA followed by Dunnett’s post-test). In control HT-29 cells, released HBEGF was undetectable (ND), but reached the limit of detectability in lactate-exposed cells. In both lactate-exposed cultures, BC11 supplementation reduced the level of released HBEGF and no statistically significant difference was observed between control cells and cells exposed to lactate/BC11.</p>
Full article ">Figure 4
<p>(<b>A</b>) Immunoblotting detection of EGFR and ERK1/2 phosphorylation. The signal intensity ratios (phospho-protein/protein) were calculated and the obtained values were used to assess the % increase in phospho-EGFR and phospho-ERK1/2 observed in lactate-exposed cells (<b>B</b>). For both phospho-EGFR/EGFR and phospho-ERK1/2/ERK1/2 immunoblotting analyses, the same sample was used and was run in parallel experiments; gels and blots were processed in parallel. The data shown in (<b>B</b>) were statistically evaluated by one-sample <span class="html-italic">t</span>-tests. In both cell cultures the increased phosphorylation of EGFR reached the level of statistical significance; on the contrary, phospho-ERK1/2 was significantly increased only in MDA-MB-231 cells. **, <span class="html-italic">p</span> &lt; 0.01.</p>
Full article ">Figure 5
<p>Effect of cisplatin (CPL) in control and lactate-exposed cultures. (<b>A</b>) In MDA-MB-231 cells grown in Low-Glc DMEM, the antiproliferative effect of 50 µM CPL was increased by CRM197, given at 32 nM (* <span class="html-italic">p</span> &lt; 0.05, as assessed by <span class="html-italic">t</span>-test). This effect was not observed in lactate-exposed cells. (<b>B</b>) Lactate-exposed cells were exposed to CPL to evaluate the effect of CRM197 and BC11 on the drug response. Data were analyzed by ANOVA, followed by Tukey’s post-test. @: a statistically significant difference was observed between cell samples treated with BC11 and those exposed to BC11+CRM197 (<span class="html-italic">p</span> &lt; 0.05). *: BC11 significantly increased the effect of CPL (<span class="html-italic">p</span> &lt; 0.001). (<b>C</b>) In HT-29 cells grown in Low-Glc DMEM, the antiproliferative effect of 50 µM CPL was not modified by CRM197. Lactate-exposed cells showed a reduced response to CPL and, again, this effect was not modified by CRM197. (<b>D</b>) The experiments shown in (<b>B</b>) were replicated in HT-29 cultures. #: no statistically significant difference was observed between cell samples treated with BC11 and those exposed to BC11 + CRM197. §: the increased antiproliferative effect observed in cell samples exposed to CPL/CRM197/BC11 did not reach the level of statistical significance, when compared to the single CPL treatment. In these experiments, no difference in the proliferation rate was observed between cells maintained in Low Glc DMEM and those cultured in Low Glc DMEM + lactate.</p>
Full article ">Figure 6
<p>(<b>A</b>) Glycolysis inhibition, assessed by quantifying the released lactate. Data were analyzed by ANOVA followed by Dunnett’s post-test; a statistically significant reduction of lactate release was observed in cell samples exposed to the combined CRM197/BC11 treatment, with <span class="html-italic">p</span> &lt; 0.01. (<b>B</b>) According to the method described in [<a href="#B51-cells-13-01533" class="html-bibr">51</a>], the antiproliferative effect caused by CRM197/BC11 suggests synergism by the two compounds. (<b>C</b>) Immunoblotting evaluation of EGFR-mediated signaling shutdown and of apoptosis induction (PUMA). Phospho-EGFR band intensities were normalized on the corresponding EGFR signal; for this immunoblotting analysis the same sample was used and was run in parallel experiments; gels and blots were processed in parallel. PUMA band intensities were normalized on the corresponding Actin level. The bar graph shows the effects caused by the two compounds, given individually or in association. Data were analyzed by one-sample t-tests. The combination CRM197/BC11 significantly reduced phospho-EGFR, which became barely detectable, and markedly increased the level of PUMA. * and ** indicate a statistically significant difference compared to the control cultures, with <span class="html-italic">p</span> &lt; 0.05 and 0.01, respectively.</p>
Full article ">Figure 7
<p>(<b>A</b>) Representative pictures of MDA-MB-231 cultures. The limits of the wound area have been outlined in red; repopulation was evaluated with the aid of the ImageJ software, as described in <a href="#sec2dot7-cells-13-01533" class="html-sec">Section 2.7</a>, and the percentage of healed wound over time is reported in (<b>B</b>). Data were analyzed by ANOVA followed by Tukey’s post-test. *: a statistically significant difference was observed between control and lactate-exposed cultures at all the considered time intervals (<span class="html-italic">p</span> &lt; 0.01). #: CRM197/BC11 significantly reduced the effects of lactate at 20 and 30 h (<span class="html-italic">p</span> &lt; 0.01).</p>
Full article ">Figure 8
<p>(<b>A</b>) Representative pictures of colonies formed by MDA-MB-231 cells, stained with CV. (<b>B</b>) Colorimetric evaluation of colonies. Data were evaluated by ANOVA, followed by Tukey’s post-test. *: a statistically significant difference was observed between lactate-exposed cells compared to controls and to all the applied treatments, with <span class="html-italic">p</span> values &lt; 0.01–0.001. (<b>C</b>) High magnification pictures of colonies, showing the morphology changes induced by CRM197 (60×) and the immunostaining of E-cadherin (E-CAD) (600×). The green fluorescence indicative of E-CAD positive cells was clearly evident only in cells exposed to CRM197.</p>
Full article ">
15 pages, 1673 KiB  
Article
Effects of Dietary Supplementation with Cocrystals of Thymol and Carvacrol on Quality, Nutrient Composition, and Oxidative Stability of Broiler Meat
by Yang Li, Changjin Li, Yunlong Zhang, Nadia Everaert, Luke Comer, Libo Huang, Ning Jiao, Xuejun Yuan, Weiren Yang and Shuzhen Jiang
Foods 2024, 13(18), 2899; https://doi.org/10.3390/foods13182899 - 13 Sep 2024
Viewed by 351
Abstract
Consumer demand for high-quality meat has increased. This study aimed to investigate the potential application of cocrystals of thymol and carvacrol in broilers for high-quality meat production. Eight hundred 1-day-old chicks were assigned to four groups fed diets supplemented with 0, 40, 60, [...] Read more.
Consumer demand for high-quality meat has increased. This study aimed to investigate the potential application of cocrystals of thymol and carvacrol in broilers for high-quality meat production. Eight hundred 1-day-old chicks were assigned to four groups fed diets supplemented with 0, 40, 60, and 80 mg/kg of Crystal EO® (CEO), containing 25% cocrystals of thymol and carvacrol in a 42-d feeding trial. The results showed that dietary CEO supplementation decreased the muscle fiber diameter and increased the muscle fiber density, glycogen content, L*45 min value, and proportion of α-linolenic acid in the breast muscle; dietary 40 and 60 mg/kg of CEO decreased the lactate content, MDA concentration, cooking loss, shear force, and thrombogenicity index and increased the proportion of lauric acid in the breast muscle; dietary 60 and 80 mg/kg of CEO increased the glucose content, total superoxide dismutase, and total antioxidant capacity levels of breast muscle. Citrate synthase activity, free radical scavenging capacity, pH24 h and a*45 min values, and the cystine content in the breast muscle were especially higher in the 60 mg/kg CEO group compared to the control group. Collectively, dietary CEO supplementation improved meat quality and nutritional values and enhanced the antioxidant capacity of broiler meat, with 60 mg/kg of CEO having the greatest effect. Full article
Show Figures

Figure 1

Figure 1
<p>Effects of dietary supplementation with cocrystals of essential oils (CEO) on antioxidant capacity of the breast muscle in broilers. (<b>A</b>) T-SOD; (<b>B</b>) GSH-Px; (<b>C</b>) T-AOC; (<b>D</b>) MDA; (<b>E</b>) ABTS; (<b>F</b>) DPPH. CEO0, CEO40, CEO60, and CEO80 were the basal diet supplemented with 0, 40, 60, and 80 mg/kg cocrystals of thymol and carvacrol, respectively. Values are expressed as the mean and standard error of the mean (SEM). <span class="html-italic">n</span> = 8. <sup>a,b,c</sup> Means with different lowercase letters differ significantly among the four treatments (<span class="html-italic">p</span> &lt; 0.05).</p>
Full article ">Figure 2
<p>Effects of dietary supplementation with cocrystals of essential oils (CEO) on breast muscle fiber characteristics in broilers. (<b>A</b>) Hematoxylin and eosin photomicrographs of breast muscle (40× magnification); (<b>B</b>) Muscle fiber diameter; (<b>C</b>) Muscle fiber density. CEO0, CEO40, CEO60, and CEO80 indicate the basal diet supplemented with 0, 40, 60, and 80 mg/kg cocrystals of thymol and carvacrol, respectively. Values are expressed as the mean and standard error of the mean (SEM). <span class="html-italic">n</span> = 8. <sup>a,b,c</sup> Means with different lowercase letters differ significantly among the four treatments (<span class="html-italic">p</span> &lt; 0.05).</p>
Full article ">Figure 3
<p>Effects of dietary supplementation with cocrystals of essential oils (CEO) on glucose metabolism in the breast muscle in broilers. (<b>A</b>) Glycogen; (<b>B</b>) Citrate synthase; (<b>C</b>) Glucose-6-P; (<b>D</b>) Lactate; (<b>E</b>) Citrate synthase. CEO0, CEO40, CEO60, and CEO80 indicate the basal diet supplemented with 0, 40, 60, and 80 mg/kg cocrystals of thymol and carvacrol, respectively. Values are expressed as the mean and standard error of the mean (SEM). <span class="html-italic">n</span> = 8. <sup>a,b,c</sup> Means with different lowercase letters differ significantly among the four treatments (<span class="html-italic">p</span> &lt; 0.05).</p>
Full article ">
23 pages, 10968 KiB  
Article
Ginkgo Biloba Bioactive Phytochemicals against Age-Related Diseases: Evidence from a Stepwise, High-Throughput Research Platform
by Yuming Yuan, Xiaoyan Xiang, Xuejun Jiang, Yingju Liu, Ming Zhang, Luyang Lu, Xinping Zhang, Xinyi Liu, Qunyou Tan and Jingqing Zhang
Antioxidants 2024, 13(9), 1104; https://doi.org/10.3390/antiox13091104 - 12 Sep 2024
Viewed by 299
Abstract
The seeds of ginkgo biloba L (GB) have been widely used worldwide. This study investigated the bioefficacies of whole GB seed powder (WGP) retaining the full nutrients of ginkgo against aging, atherosclerosis, and fatigue. The experimental results indicated that WGP lowered brain monoamine [...] Read more.
The seeds of ginkgo biloba L (GB) have been widely used worldwide. This study investigated the bioefficacies of whole GB seed powder (WGP) retaining the full nutrients of ginkgo against aging, atherosclerosis, and fatigue. The experimental results indicated that WGP lowered brain monoamine oxidase and serum malondialdehyde levels, enhanced thymus/spleen indexes, and improved learning ability, and delayed aging in senescent mice. WGP regulated lipid levels and prevented atherosclerosis by reducing triglycerides, lowering low-density lipoprotein cholesterol, increasing high-density lipoprotein cholesterol, and decreasing the atherosclerosis index. WGP improved exercise performance by reducing blood lactate accumulation and extending exhaustive swimming and climbing times, improved energy storage by increasing muscle/liver glycogen levels, and relieved physical fatigue. Network pharmacology analysis revealed 270 potential targets of WGP that play roles in cellular pathways related to inflammation inhibition, metabolism regulation, and anti-cellular senescence, etc. Protein-protein interaction analysis identified 10 hub genes, including FOS, ESR1, MAPK8, and SP1 targets. Molecular docking and molecular dynamics simulations showed that the bioactive compounds of WGP bound well to the targets. This study suggests that WGP exerts prominent health-promoting effects through multiple components, targets, and pathways. Full article
Show Figures

Figure 1

Figure 1
<p>Schematic diagram depicting the experimental studies and network pharmacological analysis to assess the multifunction efficacy of whole ginkgo biloba L powder.</p>
Full article ">Figure 2
<p>Effects of whole ginkgo powder (WGP) on (<b>A</b>–<b>G</b>) motion map and (<b>H</b>) numbers to cross the virtual platform, (<b>I</b>) time in the first quadrant, (<b>J</b>) distance in the first quadrant in the water maze test for D-galactose (D-gal)-induced aging mice. Data are expressed means ± standard deviations (SDs) (<span class="html-italic">n</span> = 10). * and <sup>#</sup>, <span class="html-italic">p</span> &lt; 0.05 compared with the adult control (Group B) and old control (Group C), respectively; <sup>##</sup>, <span class="html-italic">p</span> &lt; 0.01 compared with e old control (Group C). Motion map of different groups A–G: Group A, young control; Group B, adult control, non-aging; Group C, old control, non-treated; Group D, vitamin E (VE) (0.1 g/kg)-treated, aging; Group E, WGP (2 g/kg)-treated, aging; Group F, WGP (4 g/kg)-treated, aging; Group G, WGP (6 g/kg)-treated, aging.</p>
Full article ">Figure 3
<p>Effects of WGP on (<b>A</b>) spleen index and (<b>B</b>) thymus index in different treatment groups. Data are expressed means ± SDs (<span class="html-italic">n</span> = 10).</p>
Full article ">Figure 4
<p>Effects of WGP of (<b>A</b>) superoxide dismutase (SOD) levels, (<b>B</b>) serum malondialdehyde (MDA), and (<b>C</b>) brain monoamine oxidase (MAO). Data are expressed means ± SDs (<span class="html-italic">n</span> = 10). * and <sup>#</sup>, <span class="html-italic">p</span> &lt; 0.05 compared with the adult control (Group B) and the old control (Group C), respectively; **, <span class="html-italic">p</span> &lt; 0.01 compared with the adult control (Group B) and the old control (Group C).</p>
Full article ">Figure 5
<p>Effects of WGP on the liver, lung, and kidney of D-gal-induced aging mice. The histological changes are revealed via hematoxylin and eosin (H&amp;E) staining (400×). Scale bar = 50 µm. Control 1, 60-d mice; Control 2, 300-d mice.</p>
Full article ">Figure 6
<p>Effects of WGP on (<b>A</b>–<b>D</b>) serum biochemical lipid parameters and (<b>E</b>) atherosclerosis index (AI) in rats. Data are expressed means ± standard deviations (SDs) (<span class="html-italic">n</span> = 10). * and <sup>#</sup>, <span class="html-italic">p</span> &lt; 0.05 compared with the negative control (Group A) and high-fat diet (HFD)-fed mice (Group B), respectively; ** and <sup>##</sup>, <span class="html-italic">p</span> &lt; 0.01 compared with the negative control (Group A) and high-fat diet (HFD)-fed mice (Group B). Group A: negative control, old; Group B: model control, HFD-fed; Group C: treatment, treated with 1 g of simvastatin (Sim); Group D: treatment, treated with 0.5 g of WGP; Group E: treatment, treated with 1 g of WGP; Group F: treatment, treated with 1.5 g of WGP.</p>
Full article ">Figure 7
<p>Effects of WGP on coronary artery and abdominal aorta. The histological examinations were performed via H&amp;E staining. The lipid plaque (LP) and endothelial injury (EI) were marked in different groups (400×). Scale bar = 50 μm.</p>
Full article ">Figure 8
<p>Effects of WGP on the (<b>A</b>) climbing time and (<b>B</b>) exhaustive swimming time of mice. Data are expressed as means ± standard deviations (SDs) (<span class="html-italic">n</span> = 10). * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 compared with the negative control (Group A). Group B: treatment, treated with 2 g of WGP; Group C: treatment, treated with 4 g of WGP; Group D: treatment, treated with 6 g of WGP.</p>
Full article ">Figure 9
<p>Effects of WGP on blood and tissue parameters. (<b>A</b>) Blood lactic acid (BLA); (<b>B</b>) blood urea nitrogen (BUN); (<b>C</b>) muscle glycogen (MG); (<b>D</b>) liver glycogen (LG) in mice. Data are expressed as means ± standard deviations (SD) (<span class="html-italic">n</span> = 10). * <span class="html-italic">p</span> &lt; 0.05 and ** <span class="html-italic">p</span> &lt; 0.01 compared with the negative control (Group A). Group B: treatment, treated with 2 g of WGP; Group C: treatment, treated with 4 g of WGP; Group D: treatment, treated with 6 g of WGP.</p>
Full article ">Figure 10
<p>Relationships of active components and disease targets. The WGP compounds-targets-aging networks include the WGP compounds-targets-aging network (<b>A</b>), WGP compounds-targets-atherosclerosis network (<b>B</b>), and WGP compounds-targets-fatigue network (<b>C</b>). The green hexagon represents the disease, the round blue rectangle represents the target of the WGP compound, the red circle represents the WGP active ingredient compound, the edges represent the interactions between the nodes, and the dark pink diamond represents the WGP formula.</p>
Full article ">Figure 11
<p>Network pharmacological analysis of WGP. (<b>A</b>–<b>C</b>) Venn diagram of the action targets of WGP and the targets of aging (<b>A</b>), atherosclerosis (<b>B</b>), and fatigue (<b>C</b>). (<b>D</b>–<b>F</b>) Protein-protein interaction network reflected in cytohubba-MCC identified hub genes in the gene lists for WGP-aging (<b>D</b>), WGP-atherosclerosis (<b>E</b>), and WGP-fatigue (<b>F</b>). Node size and color in the network graph are positively correlated with the degree value. Redder colors and larger nodes represent larger degree value.</p>
Full article ">Figure 12
<p>GO enrichment analysis and KEGG pathway analysis. (<b>A</b>–<b>D</b>) GO enrichment analysis and KEGG pathway of intersection target of WGP and atherosclerosis (FDR ≤ 0.05). The biological processes (top 10, (<b>A</b>)), the cellular components (top 10, (<b>B</b>)), the molecular functions (top 10, (<b>C</b>)), and the KEGG pathways (top 10, (<b>D</b>)). (<b>E</b>–<b>H</b>) GO enrichment analysis and KEGG pathway analysis for WGP-Aging (FDR ≤ 0.05). The biological processes (top 10, (<b>E</b>)), the cellular components (top 10, (<b>F</b>)), the molecular functions (top 10, (<b>G</b>)), and the KEGG pathways (top 10, (<b>H</b>)) analysis for WGP-aging. The bubble size represents the number of targets in the pathway. The bubble color indicates the magnitude of the −log<sub>10</sub>(<span class="html-italic">p</span>) values.</p>
Full article ">Figure 13
<p>Molecular docking to model the interaction between FOS and three active ingredients of WGP (quercetin, EGCG, or kaempferol). (<b>A</b>–<b>C</b>) Interaction of FOS and quercetin. (<b>A</b>) Binding conformation of FOS-quercetin complex. (<b>B</b>,<b>C</b>) Electrical interactions of residue of FOS with quercetin. (<b>D</b>–<b>F</b>) Interaction of FOS and EGCG. (<b>D</b>) Binding conformation of FOS-EGCG complex. (<b>E</b>,<b>F</b>) Electrical interactions of residue on FOS with EGCG. (<b>G</b>–<b>I</b>) Interaction of FOS and kaempferol. (<b>G</b>) Binding conformation of FOS-kaempferol complex. (<b>H</b>,<b>I</b>) Electrical interactions of residue on FOS with kaempferol.</p>
Full article ">
15 pages, 4584 KiB  
Article
Interactions of Saccharomyces cerevisiae and Lactiplantibacillus plantarum Isolated from Light-Flavor Jiupei at Various Fermentation Temperatures
by Pu Yang, Bo Xi, Ying Han, Jiayang Li, Lujun Luo, Chaofan Qu, Junfang Li, Shuai Liu, Le Kang, Baoqing Bai, Ben Zhang, Shaojie Zhao, Pan Zhen and Lizhen Zhang
Foods 2024, 13(18), 2884; https://doi.org/10.3390/foods13182884 - 12 Sep 2024
Viewed by 287
Abstract
Chinese Baijiu is a famous fermented alcoholic beverage in China. Interactions between key microorganisms, i.e., Saccharomyces cerevisiae and Lactiplantibacillus plantarum, have recently been reported at specific temperatures. However, empirical evidence of their interactions at various temperatures during fermentation is lacking. The results [...] Read more.
Chinese Baijiu is a famous fermented alcoholic beverage in China. Interactions between key microorganisms, i.e., Saccharomyces cerevisiae and Lactiplantibacillus plantarum, have recently been reported at specific temperatures. However, empirical evidence of their interactions at various temperatures during fermentation is lacking. The results of this study demonstrated that S. cerevisiae significantly suppressed the viability and lactic acid yield of L. plantarum when they were cocultured above 15 °C. On the other hand, L. plantarum had no pronounced effect on the growth and ethanol yield of S. cerevisiae in coculture systems. S. cerevisiae was the main reducing sugar consumer. Inhibition of lactic acid production was also observed when elevated cell density of L. plantarum was introduced into the coculture system. A proteomic analysis indicated that the enzymes involved in glycolysis, lactate dehydrogenase, and proteins related to phosphoribosyl diphosphate, ribosome, and aminoacyl-tRNA biosynthesis in L. plantarum were less abundant in the coculture system. Collectively, our data demonstrated the antagonistic effect of S. cerevisiae on L. plantarum and provided insights for effective process management in light-flavor Baijiu fermentation. Full article
(This article belongs to the Section Food Microbiology)
Show Figures

Figure 1

Figure 1
<p>Microbial biomass in monoculture and coculture systems at various temperatures: (<b>a</b>) 30 °C; (<b>b</b>) 27 °C; (<b>c</b>) 24 °C; (<b>d</b>) 21 °C; (<b>e</b>) 18 °C; (<b>f</b>) 15 °C; (<b>g</b>) 12 °C; and (<b>h</b>) 9 °C.</p>
Full article ">Figure 2
<p>pH changes in monoculture and coculture systems at various temperatures: (<b>a</b>) 30 °C; (<b>b</b>) 27 °C; (<b>c</b>) 24 °C; (<b>d</b>) 21 °C; (<b>e</b>) 18 °C; (<b>f</b>) 15 °C; (<b>g</b>) 12 °C; and (<b>h</b>) 9 °C.</p>
Full article ">Figure 3
<p>Lactic acid yield of <span class="html-italic">Lactiplantibacillus plantarum</span> R2 in monoculture and coculture systems at various temperatures: (<b>a</b>) 30 °C; (<b>b</b>) 27 °C; (<b>c</b>) 24 °C; (<b>d</b>) 21 °C; (<b>e</b>) 18 °C; (<b>f</b>) 15 °C; (<b>g</b>) 12 °C; and (<b>h</b>) 9 °C.</p>
Full article ">Figure 4
<p>Ethanol production by <span class="html-italic">Saccharomyces cerevisiae</span> Y28 in monoculture and coculture systems at various temperatures: (<b>a</b>) 30 °C; (<b>b</b>) 27 °C; (<b>c</b>) 24 °C; (d) 21 °C; (<b>e</b>) 18 °C; (<b>f</b>) 15 °C; (<b>g</b>) 12 °C; and (<b>h</b>) 9 °C.</p>
Full article ">Figure 5
<p>Utilization of reducing sugars in monoculture and coculture systems at various temperatures: (<b>a</b>) 30 °C; (<b>b</b>) 27 °C; (<b>c</b>) 24 °C; (<b>d</b>) 21 °C; (<b>e</b>) 18 °C; (<b>f</b>) 15 °C; (<b>g</b>) 12 °C; and (<b>h</b>) 9 °C.</p>
Full article ">Figure 6
<p>Microbial biomass (<b>a</b>–<b>c</b>), pH changes (<b>d</b>–<b>f</b>), lactic acid yield (<b>g</b>–<b>i</b>), ethanol yield (<b>j</b>–<b>l</b>), and utilization of reducing sugars (<b>m</b>–<b>o</b>) in monoculture and coculture systems under 30 °C (<b>a</b>,<b>d</b>,<b>g</b>,<b>j</b>,<b>m</b>), 21 °C (<b>b</b>,<b>e</b>,<b>h</b>,<b>k</b>,<b>n</b>), and 15 °C (<b>c</b>,<b>f</b>,<b>i</b>,<b>l</b>,<b>o</b>).</p>
Full article ">Figure 7
<p>General features of the proteomic pattern of <span class="html-italic">L. plantarum</span>. (<b>a</b>) PCA analysis of the protein expression patterns of <span class="html-italic">L. plantarum</span> in the presence or absence of <span class="html-italic">S. cerevisiae</span>. (<b>b</b>) Number of differentially expressed proteins. (<b>c</b>) Venn map of the downregulated proteins in the coculture system between 14 h and 16 h. (<b>d</b>) KEGG pathways of the downregulated proteins in the coculture system shared at 14 h and 16 h. Co14, sampled at 14 h in the coculture system; Co16, sampled at 16 h in the coculture system; Mo14, sampled at 16 h in the monoculture system; and Mo16, sampled at 16 h in the monoculture system.</p>
Full article ">Figure 8
<p>Changes in expression levels of proteins associated with lactic acid fermentation and phosphoribosyl diphosphate biosynthesis pathways.</p>
Full article ">
12 pages, 1145 KiB  
Article
Impact of Point-of-Care Lactate Testing for Sepsis on Bundle Adherence and Clinical Outcomes in the Emergency Department: A Pre–Post Observational Study
by Sukyo Lee, Juhyun Song, Sungwoo Lee, Su Jin Kim, Kap Su Han and Sijin Lee
J. Clin. Med. 2024, 13(18), 5389; https://doi.org/10.3390/jcm13185389 - 12 Sep 2024
Viewed by 318
Abstract
Background: The early diagnosis and prompt treatment of sepsis can enhance clinical outcomes. This study aimed to assess the relationship between point-of-care testing (POCT) for lactate levels and both adherence to the Surviving Sepsis Campaign (SSC) guidelines and mortality rates among sepsis [...] Read more.
Background: The early diagnosis and prompt treatment of sepsis can enhance clinical outcomes. This study aimed to assess the relationship between point-of-care testing (POCT) for lactate levels and both adherence to the Surviving Sepsis Campaign (SSC) guidelines and mortality rates among sepsis patients in the emergency department (ED). We hypothesized that bedside lactate POCT would lead to better clinical outcomes. Methods: We conducted a pre–post observational study utilizing data from a prospectively collected sepsis registry. Following the introduction of lactate POCT, lactate levels were determined using both the central laboratory pathway and a POCT device. We then compared the characteristics and clinical outcomes between the periods before and after the introduction of POCT lactate measurement. Results: The analysis included a total of 1191 patients. The introduction of bedside lactate POCT led to a significant reduction in the time taken to obtain lactate results (from 53 to 33 min) and an increase in the rate of subsequent lactate measurements (from 82.1% to 88.2%). Lactate POCT did not significantly affect adherence to the overall SSC guidelines bundle (47.5% vs. 45.0%) or reduce 30-day mortality rates (31.1% vs. 31.4%). However, bedside lactate POCT could decrease extremely delayed lactate measurements. Conclusions: Bedside lactate POCT successfully reduced the time to obtain lactate results. Although lactate POCT did not lead to improved adherence to the overall SSC guidelines bundle or affect short-term mortality rates in sepsis patients, it may have an advantage in a specific situation such as overcrowded ED where there are subsequent or multiple measurements required. Full article
(This article belongs to the Section Emergency Medicine)
Show Figures

Figure 1

Figure 1
<p>Flowchart of the study population.</p>
Full article ">Figure 2
<p>(<b>A</b>) Boxplots of time to lactate results administration in overall patients. (<b>B</b>) Boxplots of time to lactate results administration in patients with septic shock. The box contains the inter-quartile range and the black line in the middle of the box represents the median. The asterisks mean outliers.</p>
Full article ">Figure 3
<p>(<b>A</b>) Boxplots of time to intravenous (IV) fluid administration within first 3 h in overall patients with sepsis. (<b>B</b>) Boxplots of time to IV fluid administration within first 3 h in patients with septic shock. The box contains the inter-quartile range and the black line in the middle of the box represents the median. The tiny circles mean outliers.</p>
Full article ">Figure 4
<p>(<b>A</b>) Boxplots of time to antibiotics administration in overall patients with sepsis. (<b>B</b>) Boxplots of time to antibiotics administration in patients with septic shock. The box contains the inter-quartile range and the black line in the middle of the box represents the median. The tiny circles and asterisks mean outliers.</p>
Full article ">Figure 5
<p>(<b>A</b>) Scatterplot of point-of-care test and central laboratory lactates (r = 0.99); (<b>B</b>) Bland–Altman plot of central laboratory and point-of-care test lactates. Mean difference between lactates is 0.5.</p>
Full article ">
14 pages, 2002 KiB  
Article
Evolutive Study of Dietary Aspects and Intestinal Microbiota of Pediatric Cohort with Cow’s Milk Protein Allergy
by Ana M. Castro, Sandra Navarro, Ignacio Carvajal, Agueda García, Marta Suárez, Paula Toyos, Silvia Rodríguez, Santiago Jimenez, David González, Cristina Molinos, David Pérez-Solís, Porifirio Fernández, Abelardo Margolles, Juan J. Díaz-Martín, Isabel Gutiérrez-Díaz and Susana Delgado
Children 2024, 11(9), 1113; https://doi.org/10.3390/children11091113 - 12 Sep 2024
Viewed by 280
Abstract
Background: One of the most common food allergies in the pediatric population is allergy to cow’s milk protein (CMPA). Treatment consists of avoiding cow’s milk proteins in lactating mothers and/or using therapeutic formulas based on hydrolysates or vegetable formulas. In infants with CMPA [...] Read more.
Background: One of the most common food allergies in the pediatric population is allergy to cow’s milk protein (CMPA). Treatment consists of avoiding cow’s milk proteins in lactating mothers and/or using therapeutic formulas based on hydrolysates or vegetable formulas. In infants with CMPA at diagnosis, a different gut microbial profile has been found compared to healthy children, with a reduction in beneficial bacteria. The aim of this study was to evaluate changes in the gut microbiota profile and its metabolites, dietary patterns and anthropometric variables in a pediatric cohort with CMPA after six months on a restrictive diet compared to healthy controls. Methods: In total, 21 patients diagnosed with CMPA and a control group of 24 healthy infants participated in this study. The fecal microbiota of all participants were investigated by metataxonomic analysis of 16S rDNA amplicons, and fecal short-chain fatty acids were measured by gas chromatography. Epidemiological assessment and dietary questionnaires were carried out for both groups. Results: Regarding growth, no significant differences were found, but differences in dietary intake of some macro- and micronutrients were observed. Patients who were breastfed at six months had higher bifidobacteria and lipid intakes than patients fed with hydrolyzed formulas. Conclusions: Although the growth of CMPA infants fed with therapeutic formula is similar to breastfed CMPA infants, there are differences in microbiota composition and macronutrient intake that underline the importance of continued breastfeeding in CMPA cases. Full article
(This article belongs to the Special Issue Insights in Children with Allergy)
Show Figures

Figure 1

Figure 1
<p>Growth performance in terms of weight and height at birth, diagnosis and 6 months’ follow-up for controls and patients.</p>
Full article ">Figure 2
<p>Profile of the lipid intake according to the type of feeding of the patients. SFAs, saturated fatty acids. MUFAs, monounsaturated fatty acids. PUFAs, polyunsaturated fatty acids. Asterisks ** indicate significant statistical differences (<span class="html-italic">p</span> value ≤ 0.01).</p>
Full article ">Figure 3
<p>Relative abundance of the <span class="html-italic">Actinobacteria</span> phylum as a function of patient diet.</p>
Full article ">Figure 4
<p>Evolutionary fecal microbiota profile between patients and controls at phylum level. Asterisks indicate significant statistical differences; *: <span class="html-italic">p</span> value ≤ 0.05, **: <span class="html-italic">p</span> value ≤ 0.01.</p>
Full article ">Figure 5
<p>Changes in fecal short-chain fatty acids concentrations over time in controls and patients. Asterisks indicate significant statistical differences; *: <span class="html-italic">p</span> value ≤ 0.05, **: <span class="html-italic">p</span> value ≤ 0.01.</p>
Full article ">
10 pages, 249 KiB  
Article
Effects of High-Intensity Training on Complete Blood Count, Iron Metabolism, Lipid Profile, Liver, and Kidney Function Tests of Professional Water Polo Players
by Frane Bukvić, Domagoj Marijančević, Helena Čičak, Ana-Maria Šimundić, Daria Pašalić and Lora Dukić
Diagnostics 2024, 14(18), 2014; https://doi.org/10.3390/diagnostics14182014 - 11 Sep 2024
Viewed by 203
Abstract
AIM: Our goal was to examine the effect of high-intensity physical activity on changes in the lipid profile, complete blood count (CBC), iron metabolism, and kidney and liver function tests of professional water polo players. METHODS: This study included twenty professional male water [...] Read more.
AIM: Our goal was to examine the effect of high-intensity physical activity on changes in the lipid profile, complete blood count (CBC), iron metabolism, and kidney and liver function tests of professional water polo players. METHODS: This study included twenty professional male water polo players. Blood sampling was carried out at the beginning of the season and during periods of high-intensity training. CBCs were determined with a Siemens Advia 2120i hematology analyzer. A Beckman CoulterAU680 chemistry analyzer was used to determine the serum concentrations/activities of lipid profiles and liver and kidney function test analytes. The lipid athlete scores were also determined. RESULTS: The mean corpuscular volume (p = 0.006), platelet count (p = 0.008), and mean platelet volume (p < 0.001) significantly decreased during the high-intensity period, compared with the beginning of the season. The total iron-binding capacity increased (p = 0.001), and ferritin concentrations significantly declined (p = 0.017). The lipid profiles revealed a significant difference between phases, with slight increases in serum total (p = 0.025) and LDL cholesterol (p = 0.002) levels and a decrease in triglyceride concentrations (p = 0.040) in the high-intensity period. During the high-intensity period, the liver and kidney function tests showed a substantial positive effect on lactate dehydrogenase levels (p < 0.001), aspartate aminotransferase (p = 0.028) serum activity, and total protein concentrations (p = 0.033), compared with the beginning of the season. CONCLUSIONS: Water polo players might exhibit a decrease in some CBC parameters, an increase in LDL cholesterol, and a decrease in liver function biomarkers due to intense training at the peak of the competitive season. Kidney function biomarkers remain unchanged. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
9 pages, 3873 KiB  
Article
Osteochondral Tissue-On-a-Chip: A Novel Model for Osteoarthritis Research
by Irene González-Guede, Daniel Garriguez-Perez and Benjamin Fernandez-Gutierrez
Int. J. Mol. Sci. 2024, 25(18), 9834; https://doi.org/10.3390/ijms25189834 - 11 Sep 2024
Viewed by 246
Abstract
The existing in vitro and in vivo models for studying osteoarthritis have significant limitations in replicating the complexity of joint tissues. This research aims to validate a Tissue-On-a-Chip system for osteoarthritis research. Osteochondral tissues obtained from knee replacement surgeries of patients with osteoarthritis [...] Read more.
The existing in vitro and in vivo models for studying osteoarthritis have significant limitations in replicating the complexity of joint tissues. This research aims to validate a Tissue-On-a-Chip system for osteoarthritis research. Osteochondral tissues obtained from knee replacement surgeries of patients with osteoarthritis were cultured in an Organ-On-a-Chip system. This system was designed to supply oxygen and glucose to the cartilage from the bone. The distribution of oxygen and glucose was evaluated by fluorescence using Image-iT Green Hypoxia and 2-NBDG, respectively. Cytotoxicity was measured using lactate dehydrogenase (LDH) levels in chip cultures compared to plate cultures (12 tissues per method). Glycosaminoglycans (GAGs), alkaline phosphatase (ALP), Coll2-1, and procollagen type II N-terminal propeptide (PIINP) were measured in the perfused medium of the Tissue-On-a-Chip over a period of 70 days. Fluorescence of Image-iT Green Hypoxia was observed only in the cartilage area, while 2-NBDG was distributed throughout the tissue. An increase in LDH levels was noted in the plate cultures on day 24 and in the Tissue-On-a-Chip cultures on day 63. Compared to the start of the culture, GAG content increased on day 52, while ALP showed variations. A notable increase in GAG, ALP, and Coll2-1 levels was observed on day 59. PIINP levels remained stable throughout the experiment. The validated osteochondral Tissue-On-a-Chip system can replicate the joint microenvironment, with hypoxic conditions in cartilage and normoxic conditions in bone. Tissue survival and component stability were maintained for approximately two months. This platform is a useful tool for evaluating new drugs and represents a viable alternative to animal models. Full article
(This article belongs to the Special Issue Recent Development in Scaffolds for Tissue Engineering)
Show Figures

Figure 1

Figure 1
<p>Osteochondral tissues cultured in a Tissue-On-a-Chip device. Hypoxic regions were labeled with Image-iT Green Hypoxia, as shown in (<b>A</b>,<b>B</b>), where fluorescence was observed in the cartilage layer. Bone cells were stained only with DAPI (images captured with 4× and 10× objectives, respectively). Glucose supply was assessed using 2-NBDG; fluorescence was observed in both cartilage and bone, as shown in (<b>C</b>). In (<b>D</b>), a detailed view of the cartilage is presented (images captured with 4× and 10× objectives, respectively).</p>
Full article ">Figure 2
<p>LDH levels in osteochondral tissue cultured on plates and in Tissue-On-a-Chip. Femur and tibia explants, both lesioned and non-lesioned, from the knees of three patients were used (n = 12 per culture method). A one-way ANOVA test was used to compare each time point with the baseline at day 3: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
Full article ">Figure 3
<p>GAG, ALP, Coll2-1, and PIINP content in the perfused medium of the osteochondral Tissue-On-a-Chip determined by ELISA over 70 days. Femur and tibia explants, both lesioned and non-lesioned, from the knees of three patients were used (n = 12). A one-way ANOVA test was used to compare each time point with the baseline at day 3: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001. All bar graphs are presented as mean ± SEM.</p>
Full article ">Figure 4
<p>Osteochondral Tissue-On-a-Chip Setup. (<b>A</b>) Diagram of the osteochondral Tissue-On-a-Chip. The Organ-On-a-Chip materials used are gas tight, so oxygen is supplied solely from the culture medium. A syringe perfuses the culture medium at a flow rate of 2 µL/min. The subchondral bone of the osteochondral tissue is placed on the membrane, distributing glucose and oxygen to the cartilage, mimicking physiological conditions. Created with BioRender.com. (<b>B</b>) Organ-On-a-Chip model. The chip is composed of cyclic olefin polymer (COP) and cyclic olefin copolymer (COC). PET inlet and outlet tubes, connected with connectors, facilitate perfusion. (<b>C</b>) Assembly of the osteochondral Tissue-On-a-Chip. The photograph shows multiple chips connected to syringes inside an incubator, with tubes collecting the culture medium. (<b>D</b>) Cutting of osteochondral explants. The explants are sectioned into 3 × 3 × 4 mm pieces using an acrylic cutting block and carbon steel blades.</p>
Full article ">
Back to TopTop