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11 pages, 893 KiB  
Article
Persistent Elevation in Urinary Neutrophil Gelatinase-Associated Lipocalin Levels Can Be a Predictor of Urinary Tract Infection Recurrence or Persistence in Women
by Min-Ching Liu, Yuan-Hong Jiang, Jia-Fong Jhang, Tien-Lin Chang, Chia-Cheng Yang and Hann-Chorng Kuo
Int. J. Mol. Sci. 2024, 25(23), 12670; https://doi.org/10.3390/ijms252312670 (registering DOI) - 26 Nov 2024
Abstract
Women commonly experience urinary tract infection (UTI) recurrence. However, there is no effective tool for predicting recurrent UTI after the first UTI episode. Hence, this study aimed to investigate potential urinary inflammatory biomarkers and specific biomarkers for predicting UTI recurrence or persistence after [...] Read more.
Women commonly experience urinary tract infection (UTI) recurrence. However, there is no effective tool for predicting recurrent UTI after the first UTI episode. Hence, this study aimed to investigate potential urinary inflammatory biomarkers and specific biomarkers for predicting UTI recurrence or persistence after antibiotic treatment in women. Forty women who had a history of recurrent UTI within 1 year after the initial episode and acute bacterial cystitis were treated with broad-spectrum antibiotics for 1 week. To measure inflammatory biomarker levels, urine samples were collected at the baseline and after 1 week, 1 month, and 3 months. The levels of urinary pro-inflammatory proteins such as neutrophil gelatinase-associated lipocalin (NGAL), nerve growth factor, CXC-motif chemokine ligand (CXCL)-1, interleukin-8, CXCL-10, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha were measured using commercial kits. Seven healthy age-matched women were included as controls. The changes in urinary biomarker levels at the baseline and various time points were compared between women with and without UTI recurrence within 1 month or within 3 months after the initial antibiotic therapy. At the baseline, patients with a higher urinary white blood cell count had a significantly higher NGAL level than the controls and those with a low white blood cell count. Of the 40 patients with a history of recurrent UTI, 12 presented with UTI persistence or recurrence within 1 month and 19 within 3 months after the initial antibiotic treatment. Among the 28 patients without UTI recurrence at 1 month after treatment, 7 had UTI recurrence within 3 months. Compared with patients without UTI recurrence, those with UTI recurrence had significantly higher urinary NGAL levels at 1 week, 1 month, and 3 months after the initial treatment. This study concludes that persistent elevation in urinary NGAL levels after the initial antibiotic treatment indicated persistent bladder inflammation. Further, it could be a predictor of UTI persistence or recurrence within 1 or 3 months after the initial antibiotic treatment. Patients with a history of recurrent UTI and high urinary NGAL levels after antibiotic treatment might require a longer treatment duration to completely eradicate or prevent UTI recurrence. Full article
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<p>The changes in urine levels of NGAL, NGF, IL-8, and TNF-α from the baseline to different time points (1 week, 1 month, and 3 months) in the 40 patients with a history of recurrent urinary tract infection and comparison with the baseline values in controls. * indicates a significant difference with the baseline data. Data are expressed as mean ± standard error.</p>
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<p>The urinary biomarker levels at the baseline and different time points of 12 patients with and 28 without UTI persistence or recurrence within 1 month after the initial treatment. * indicates a significant difference with the baseline data. # indicates a significant difference between groups. Data are expressed as mean ± standard error.</p>
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<p>The changes in urinary cytokines NGAL, CXCL-1, IL-8, and CXCL-10 among patients with a history of recurrent UTI, who had persistent UTI (n = 9), recurrent UTI (n = 10), and no UTI recurrence (n = 21) within 3 months after the first UTI episode and antibiotics treatment. * significant change in urinary biomarker at a time point compared with the baseline in the same subgroup. # significant difference among the three subgroups at each time point.</p>
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17 pages, 2925 KiB  
Article
Identification of the P24 Gene Family Related to Vesicular Transport in Cyclocarya paliurus and Their Expression Analysis Under Salt Stress
by Yuxin Pan, Chunpeng Fei, Xin Liu, Fayin He, Jiana Zhu, Hui Yu, Fusheng Zhao and Zhengting Yang
Forests 2024, 15(12), 2081; https://doi.org/10.3390/f15122081 - 25 Nov 2024
Abstract
Vesicle transport is a fundamental mechanism for intracellular substance transfer and signal transduction, involving the formation, transport, fusion with target membranes, and release of vesicle contents within cells. Issues such as disruption of water balance, difficulty in regulating osmotic pressure, and oxidative stress [...] Read more.
Vesicle transport is a fundamental mechanism for intracellular substance transfer and signal transduction, involving the formation, transport, fusion with target membranes, and release of vesicle contents within cells. Issues such as disruption of water balance, difficulty in regulating osmotic pressure, and oxidative stress caused by salt stress can lead to disturbances in the secretion system of plants, thus affecting plant growth and development. Cyclocarya paliurus (C. paliurus), widely used in traditional Chinese medicine, has not been previously reported in terms of how the vesicle transport P24 gene regulates its adaptation to salt stress. In this research project, a total of eight CpP24 genes were successfully identified. Upon examination of gene architecture and conserved sequence elements, the CpP24 genes exhibited a variation in exon count, ranging from 4 to 6. Moreover, the CpP24 gene family’s reaction to salt stress and specific stressors including methyl jasmonate (MeJA), sodium hydrosulfide (NaHS), and sodium nitroprusside (SNP) was further explored in our study. Comprehensive analysis of the expression patterns of CpP24 genes under various conditions showed that salt stress induced the expression of these genes, and the combined treatment of salt stress with specific stresses caused changes in their expression. This study lays a theoretical foundation for further probing into the physiological functions of C. paliurus and the underlying mechanisms regarding its response to environmental stress. Full article
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<p>Phylogenetic tree of <span class="html-italic">CpP24</span> gene family members. The tree was constructed in MEGA 11 using a 1000-guided neighborhood joining method, and different branches were labeled with lines of different colors: δ subfamily I (red), δ subfamily II (purple), β subfamily (orange).</p>
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<p>Analysis of the structure and conserved motifs of the P24 gene family in <span class="html-italic">C. paliurus</span>.</p>
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<p>Genomic analysis and syntenic relationships of <span class="html-italic">P24</span> genes in <span class="html-italic">C.paliurus</span> with <span class="html-italic">A. thaliana</span> and <span class="html-italic">O. sativa</span>. (<b>A</b>) Chromosomal mapping of <span class="html-italic">P24</span> genes in <span class="html-italic">C. paliurus</span> illustrates locations, synteny blocks, and duplicated gene pairs within the genome. (<b>B</b>) Syntenic relationships of <span class="html-italic">P24</span> genes between <span class="html-italic">C. paliurus</span> and <span class="html-italic">A. thaliana</span>. (<b>C</b>) Syntenic relationships of <span class="html-italic">P24</span> genes between <span class="html-italic">C. paliurus</span> and <span class="html-italic">O. sativa</span>. Background gray lines depict collinear blocks across the genomes, with syntenic connections highlighted in red.</p>
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<p>Predicted structural and interaction analysis of <span class="html-italic">CpP24</span> proteins in <span class="html-italic">C. paliurus</span>. (<b>A</b>) Predicted secondary structure of <span class="html-italic">CpP24</span> proteins. (<b>B</b>) Tertiary structure prediction plot of <span class="html-italic">CpP24</span> proteins. (<b>C</b>) Protein interaction network diagram of the eight P24 proteins identified in <span class="html-italic">C. paliurus</span> and the related homologous genes in <span class="html-italic">A. thaliana</span>.</p>
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<p>Analysis of <span class="html-italic">cis</span>-acting elements of <span class="html-italic">CpP24</span> genes. The number in the heat map box indicates the number of <span class="html-italic">cis</span>-acting elements.</p>
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<p>Differential expression of <span class="html-italic">CpP24</span> genes under various stress treatments. (<b>A</b>) Expression heatmap of <span class="html-italic">CpP24</span> genes under control conditions (CK) and 0.4% NaCl treatment. The color scale indicates log2-transformed FPKM+1 values, with red denoting high expression and blue denoting low expression. (<b>B</b>) Heatmap showing expression patterns of <span class="html-italic">CpP24</span> genes under 0.2 mM methyl jasmonate (MeJA) alone and combined with 0.4% NaCl. This comparison highlights the modulatory effects of NaCl on MeJA-induced gene expression changes. (<b>C</b>) Heatmap of <span class="html-italic">CpP24</span> gene expression under 0.5 mM sodium hydrosulfide (NaHS) treatment, alone and in combination with 0.4% NaCl. The heatmap illustrates the interaction between NaHS and salt stress in influencing gene expression. (<b>D</b>) Heatmap depicting changes in <span class="html-italic">CpP24</span> gene expression under 0.25 mM sodium nitroprusside (SNP) treatment, alone and in combination with 0.4% NaCl. This panel shows how SNP responses are affected by concurrent salt stress. (<b>E</b>) Bar graphs validating expression profiles for selected <span class="html-italic">CpP24</span> genes (<span class="html-italic">CpP24-1</span>, <span class="html-italic">CpP24-2</span>, <span class="html-italic">CpP24-7</span>) under the different stress treatments. The bars represent mean expression values, and error bars indicate standard errors from three biological and technical replicates. Letters above bars denote statistically significant differences at <span class="html-italic">p</span> &lt; 0.05.</p>
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23 pages, 5062 KiB  
Article
Intermediate and Transitory Inflammation Mediate Proper Alveolar Bone Healing Outcome in Contrast to Extreme Low/High Responses: Evidence from Mice Strains Selected for Distinct Inflammatory Phenotypes
by Priscila Maria Colavite, Michelle de Campos Soriani Azevedo, Carolina Fávaro Francisconi, Angélica Cristina Fonseca, André Petenucci Tabanez, Jéssica Lima Melchiades, Daniela Carignatto Passadori, Andrea Borrego, Marcelo De Franco, Ana Paula Favaro Trombone and Gustavo Pompermaier Garlet
Biology 2024, 13(12), 972; https://doi.org/10.3390/biology13120972 - 25 Nov 2024
Abstract
Alveolar bone healing is influenced by various local and systemic factors, including the local inflammatory response. This study aimed to evaluate the role of inflammatory responsiveness in alveolar bone healing using 8-week-old male and female mice (N = 5/time/group) strains selected for maximum [...] Read more.
Alveolar bone healing is influenced by various local and systemic factors, including the local inflammatory response. This study aimed to evaluate the role of inflammatory responsiveness in alveolar bone healing using 8-week-old male and female mice (N = 5/time/group) strains selected for maximum (AIRmax) or minimum (AIRmin) acute inflammatory response carrying distinct homozygous RR/SS Slc11a1 genotypes, namely AIRminRR, AIRminSS, AIRmaxRR, and AIRmaxSS mice. After upper right incisor extraction, bone healing was analyzed at 0, 3, 7, and 14 days using micro-computed tomography, histomorphometry, birefringence, immunohistochemistry, and PCRArray analysis. AIRmaxSS and AIRminRR presented the highest and lowest inflammatory readouts, respectively, associated with lowest repair levels in both strains, while intermediate inflammatory phenotypes observed in AIRminSS and AIRmaxRR were associated with higher repair levels in such strains. The better healing outcomes are associated with intermediate inflammatory cell counts, a balanced expression of pro- and anti-inflammatory cytokines and chemokines, increased expression of growth and osteogenic factors and MSCs markers. Our results demonstrate that extreme high and low inflammatory responses are not ideal for a proper bone repair outcome, while an intermediate and transitory inflammation is associated with a proper alveolar bone healing outcome. Full article
17 pages, 5349 KiB  
Article
DNA Methylation Changes and Phenotypic Adaptations Induced Repeated Extreme Altitude Exposure at 8848 Meters
by Shixuan Zhang, La Yang, Zhuoma Duoji, Danzeng Qiangba, Xiaoxi Hu, Zeyu Jiang, Dandan Hou, Zixin Hu and Zhuoma Basang
Int. J. Mol. Sci. 2024, 25(23), 12652; https://doi.org/10.3390/ijms252312652 - 25 Nov 2024
Abstract
Repeated extreme environmental training (RET) enhances adaptability and induces lasting methylation modifications. We recruited 64 participants from a high-altitude region (4700 m), including 32 volunteers with repeated high-altitude exposure, reaching up to 8848 m and as many as 11 exposures. By analyzing 741,489 [...] Read more.
Repeated extreme environmental training (RET) enhances adaptability and induces lasting methylation modifications. We recruited 64 participants from a high-altitude region (4700 m), including 32 volunteers with repeated high-altitude exposure, reaching up to 8848 m and as many as 11 exposures. By analyzing 741,489 CpG loci and 39 phenotypes, we identified significant changes in 13 CpG loci (R2 > 0.8, ACC > 0.75) and 15 phenotypes correlated with increasing RET exposures. The phenotypic Bayesian causal network and phenotypic-CpG interaction networks showed greater robustness (node correlation) with more RET exposures, particularly in systolic blood pressure (SP), platelet count (PLT), and neutrophil count (NEUT). Six CpG sites were validated as significantly associated with hypoxia exposure using the GEO public da-taset (AltitudeOmics). Furthermore, dividing the participants into two groups based on the number of RET exposures (n = 9 and 4) revealed six CpG sites significantly corre-lated with PLT and red cell distribution width-standard deviation (RDW.SD). Our findings suggest that increased RET exposures strengthen the interactions between phenotypes and CpG sites, indicating that critical extreme adaptive states may alter methylation patterns, co-evolving with phenotypes such as PLT, RDW.SD, and NEUT. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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<p>(<b>A</b>) The overall research strategy is divided into two parts: First, we explore the relationship between climbing age (CA) and the “memory” of DNA methylation modifications. Second, we categorize all samples into high and low adaptation groups based on SpO<sub>2</sub> levels and investigate the phenotypic associations of key CpGs within these different adaptive groups. (<b>B</b>) The 13 CpGs of 11 genes were significantly and positively correlated with RET perturbations (R<sup>2</sup><sub>OLS</sub> ≥ 0.8). (<b>C</b>) The 15 phenotypes were significantly correlated with RET perturbations (P Spearman &lt; 0.05). (<b>D</b>) Correlation between CA and Phenotypes.</p>
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<p>Bayesian causal networks to explore the network properties of organ systems and MP-SSN in different times of RET environments. (<b>A</b>) The study constructed Bayesian causal networks for seven organ systems in which 15 physiological phenotypes were located. (<b>a</b>–<b>c</b>) The networks of RET are constructed at 0, 5, and 8, while the networks of other RETs are not inherently connected. (<b>B</b>) Small-world networks were used to evaluate 15 phenotypes of networks with different RET counts, which suggested enhanced network connectivity. (<b>C</b>) The network between Phenotype and CpG in different RET frequencies. The green line represents “CpG -&gt; Phenotype”; the blue line represents “Phenotype -&gt; CpG”. (<b>a</b>) MP-SSN Bayesian causal network for 0th RET perturbation population. (<b>b</b>) MP-SSN Bayesian causal network for 5th RET perturbation population. (<b>c</b>) MP-SSN Bayesian causal network for 8th RET perturbation population. (<b>d</b>) Small-world networks were used to evaluate MP-SSN networks with different RET counts, which suggested enhanced network connectivity. (<b>e</b>) The number of Bayesian causal network connections between CpG and phenotype in the MP-SSN network. “All” in the figure: the total number of connections between the two; “Phenotype -&gt; CpG”: the direction of the Bayesian causal network is that phenotype affects CpG.</p>
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<p>(<b>A</b>–<b>F</b>) Status of LIPN, PLCH1, EMR1, STX5 genes in the AltitudeOmics. Linear correlation between CpG and gene expression at different stages of hypoxia exposure. (<b>G</b>) The values in the graph represent the linear correlation coefficients between each CpG site and gene expression in the A-plot (Coef.). In the figure, * indicates <span class="html-italic">p</span> &lt; 0.05, ** indicates <span class="html-italic">p</span> &lt; 0.01, *** indicates <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>(<b>A</b>) A linear relationship between blood oxygen saturation and CA; Number of high and low adaptive populations distributed in different CAs. (<b>B</b>) The 13 CpGs are related to 32 Phenotypes in High and Low. All phenotypically unrelated CpGs were excluded from the figure. In addition, the part of the graph without color means no significant correlation (<span class="html-italic">p</span> &lt; 0.05, |Coef.| &gt; 0.3).</p>
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<p>Possible speculation on the effect of <span class="html-italic">LIPN</span> and <span class="html-italic">PLCH1</span> on blood oxygen saturation.</p>
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11 pages, 1801 KiB  
Article
Correlation of Red Blood Cell Morphology with Serum Cobalamin and Folate Concentration in Dogs with Chronic Diarrhea: A Retrospective Study
by Argyrios Ginoudis, Anna Maria Ioannidou, Dimitra Pardali, Asteria Tsikna and Zoe Polizopoulou
Metabolites 2024, 14(12), 657; https://doi.org/10.3390/metabo14120657 - 25 Nov 2024
Abstract
Background/Objectives: Chronic diarrhea in dogs is a prevalent condition that significantly impacts canine health, often leading to weight loss, dehydration, and malnutrition. Diagnosing and treating chronic diarrhea is challenging due to its multifactorial nature, necessitating collaboration among veterinarians across various specialties. Measuring [...] Read more.
Background/Objectives: Chronic diarrhea in dogs is a prevalent condition that significantly impacts canine health, often leading to weight loss, dehydration, and malnutrition. Diagnosing and treating chronic diarrhea is challenging due to its multifactorial nature, necessitating collaboration among veterinarians across various specialties. Measuring cobalamin and folate levels is a crucial diagnostic step for all dogs with chronic diarrhea. The role of these vitamins in erythropoiesis is well-documented in human medicine, where deficiencies are linked to erythropoietic disorders and megaloblastic anemia. This study explores the relationship between cobalamin and folate concentrations with hematologic parameters in dogs with chronic diarrhea to develop novel diagnostic methods that facilitate timely decision making. Methods: Forty-seven adult dogs with a history of chronic diarrhea (2019–2023) were included in the study. Upon presentation, complete blood count and measurement of cobalamin and folate concentrations were performed. The correlation of cobalamin and folate levels with erythrocytic parameters, including hematocrit (HCT), hemoglobin concentration (HGB), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red cell distribution width (RDW), and reticulocyte count, as well as morphological changes in the blood smear were examined. Results: Serum cobalamin was significantly correlated with RBC (p = 0.032), HGB (p = 0.006), HCT (p = 0.005), and MCV (p = 0.022). Anisocytosis was significantly correlated with hypocobalaminemia (p = 0.002), while acanthocytosis correlated with normal cobalamin levels (p = 0.046). No correlation was found between serum folate and erythrocytic parameters or morphological changes. Conclusions: These findings emphasize cobalamin’s potential role in canine erythropoiesis, highlighting the need for routine evaluation and supplementation when necessary. Conversely, the lack of association with folate suggests it plays a less significant role in this species. These results underscore the importance of complete blood count in the diagnostic investigation of dogs with chronic diarrhea. Full article
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and red blood cell count (RBC, ×10<sup>3</sup>/µL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the RBC concentration. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and RBC.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and hemoglobin concentration (HGB, g/dL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the HGB concentration. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and HGB.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and hematocrit (HCT, %). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the HCT. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and HCT.</p>
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<p>Scattergram illustrating the relationship between serum cobalamin concentration (B12, pmol/L) and mean corpuscular volume (MCV, fL). Each point represents an individual measurement, where the x-axis corresponds to the B12 concentration and the y-axis corresponds to the MCV. The dotted line represents the trend line (linear regression) indicating the correlation between B12 and MCV.</p>
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18 pages, 579 KiB  
Article
Effect of a Slow-Release Urea Nanofertilizer on Soil Microflora and Yield of Direct Seeded Rice (Oryza sativa L.)
by Yashika Sehgal, Anu Kalia, Buta Singh Dhillon and Gurmeet Singh Dheri
Nitrogen 2024, 5(4), 1074-1091; https://doi.org/10.3390/nitrogen5040069 (registering DOI) - 25 Nov 2024
Abstract
Nitrogen fertilizers have a significant impact on the growth of rice. The overuse and inappropriate application of nitrogen fertilizers have resulted in environmental pollution, in addition to subjecting both humans and livestock to negative health hazards. Finding a viable substitute for traditional nitrogen [...] Read more.
Nitrogen fertilizers have a significant impact on the growth of rice. The overuse and inappropriate application of nitrogen fertilizers have resulted in environmental pollution, in addition to subjecting both humans and livestock to negative health hazards. Finding a viable substitute for traditional nitrogen fertilizers is crucial and essential to help improve crop yield and minimize environmental damage. Nano-nitrogen fertilizers offer a possible alternative to traditional fertilizers due to a slow/controlled release of nitrogen. The present work aimed to study the effect of a slow-release urea nanofertilizer on soil ammonical (NH4-N) and nitrate-N (NO3-N) content, culturable soil microflora, and soil enzyme activities in three different soil samples procured from Ludhiana and Patiala districts through a soil column study. Seven treatments, including 0, 50 (75 kg/ha N), 75 (112.5 kg/ha N), and 100% (150 kg/ha N) of the recommended dose (RD) of conventional urea and nano-urea fertilizer were applied. The leachate samples collected from nano-urea treatment exhibited NH4-N for the first two weeks, followed by NO3-N appearance. The higher NH4-N and NO3-N contents in the leachate were recorded for light-textured soil as compared to medium- and heavy-textured soil samples. The soil microbial counts and enzyme activities were recorded to be maximum in light-textured soils. Therefore, this slow-release formulation could be more useful for light-textured soils to decrease applied N-fertilizer losses, as well as for improving the soil microbial viable cell counts and soil enzyme activities. The effect of urea nanofertilizer on the growth and yield of direct-seeded rice (Oryza sativa L.) was also evaluated under field conditions. Both studies were performed independently. Numerically, the highest shoot height, fresh and dry shoot weight, and significantly maximum total chlorophyll, carotenoid, and anthocyanins were recorded in the T2 (100% RDF through nano-urea) treatment. The yield-attributing traits, including the number of filled grains and thousand-grain weight, were also recorded to have increased in T2 treatment. A numerical increase in NPK for plant and grain of rice at 100% RDN through nano-urea was recorded. The soil application of the product exhibited no negative effect on the soil microbial viable cell count on different doses of nano-urea fertilizer. The soil nitrogen fixer viable counts were rather improved in nano-urea treatments. The results reflect that nano-urea fertilizer could be considered as a possible alternative to conventional fertilizer. Full article
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<p>Effect of different N-fertilizer levels of conventional and nano-urea formulation on the grain nitrogen, phosphorus, and potassium content. Mean values with different alphabetic scripting on bars depict statistically significant differences.</p>
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17 pages, 7822 KiB  
Article
Bovine NMRAL2 Protein Blunts Nitric Oxide Production and Inflammatory Response in Mycobacterium bovis Infected Bovine Lung Epithelial Cells
by Yongchong Peng, Shiying Zhou, Qin Sun, Xinjun Zhou, Chao Wang, Zijian Wang, Tahira Iftakhar, Yifan Zhu, Shengsong Xie, Xi Chen, Lei Zhang, Changmin Hu, Yingyu Chen and Aizhen Guo
Cells 2024, 13(23), 1953; https://doi.org/10.3390/cells13231953 - 24 Nov 2024
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Abstract
Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis (M. tb) and Mycobacterium bovis (M. bovis), remains the leading cause of death from a single infectious agent globally. Intracellular survival is crucial for their virulence; yet, the underlying mechanisms are not fully [...] Read more.
Tuberculosis (TB), primarily caused by Mycobacterium tuberculosis (M. tb) and Mycobacterium bovis (M. bovis), remains the leading cause of death from a single infectious agent globally. Intracellular survival is crucial for their virulence; yet, the underlying mechanisms are not fully understood. This study aimed to demonstrate the significance of a previously unannotated bovine gene ENSBTAG00000011305 in M. bovis intracellular survival. This gene was termed NMRAL2_Bovine due to its inclusion of the NmrA domain which has a relation to nitric oxide (NO) production. We used CRISPR/Cas9 to knock out NMRAL2_Bovine in bovine lung epithelial cells and observed a significant decrease in M. bovis-induced cell death and the intracellular bacterial count, alongside increased NO levels. A transcriptome analysis revealed the upregulation of pathways linked to NO, IL-6, and TNF-α production, which was confirmed by the increased expression of iNOS, IL-6, and TNF-α. Correspondingly, Western blotting indicated that key signaling pathways, including NF-κB and MAPK, were activated. In conclusion, our findings determined that NMRAL2_Bovine functions as a negative regulator of the inflammatory response induced by M. bovis infection at the cellular level and, thereby, provide a novel insight into TB pathogenesis and a potential target for developing novel host-directed therapies against TB. Full article
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<p>Bioinformatic analysis on domain and amino acid similarity of NMRAL2 with other NmrA_like proteins. (<b>a</b>) The schematic illustration depicts the NmrA-like domain and its conserved motifs within the NMRAL2 protein and its homologs. The analysis of conserved domains in NMRAL2 and its homologs was performed using the NCBI Conserved Domain Database (CDD) search tool. Additionally, the identification of conserved motifs within these protein sequences was carried out using the MEME Suite motif search tool. (<b>b</b>) A multiple sequence alignment of protein homologs related to NMRAL2 was performed using Clustal Omega, and the results were visualized with ESPript 3.0. The red boxes represent identity, and the blue boxes indicate partial similarity.</p>
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<p>The effect of NMRAL2 on cell survival during <span class="html-italic">M. bovis</span> infection determined with CCK8 assay. (<b>a</b>) DNA sequence analysis verified the mutation in clonal <span class="html-italic">NMRAL2</span> KO cells. In the KO cells, deleted bases are highlighted in red, while PAM sites are enclosed in a red box. (<b>b</b>) RT-qPCR analysis confirmed deficiency of <span class="html-italic">NMRAL2</span> expression in knockout cell lines. (<b>c</b>,<b>d</b>) The impact of <span class="html-italic">NMRAL2</span> on EBL cell proliferation was assessed through the CCK-8 assay and the EdU assay. The blue fluorescence represents the nucleus stained by DAPI and the green fluorescence represents EdU-positive cells. (<b>e</b>,<b>f</b>) The impact of <span class="html-italic">NMRAL2</span> knockout and knock-down on cell viability following infection with <span class="html-italic">M. bovis</span>. Cell survival of WT, NMRAL2-KO, and si-NMRAL2 EBL cells post <span class="html-italic">M. bovis</span> infection was evaluated using the CCK8 assay. A two-tailed unpaired <span class="html-italic">t</span>-test and a two-way ANOVA were employed to evaluate the statistical significance between groups, utilizing data from three independent replicates. Statistical significance is denoted as follows: *<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, and ns for no significant difference.</p>
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<p>The impact of <span class="html-italic">NMRAL2</span> knockout or knockdown on intracellular <span class="html-italic">M. bovis</span> viability. (<b>a</b>,<b>b</b>) Assessing the impact of NMRAL2 on <span class="html-italic">M. bovis</span> invasion and adhesion capabilities. EBL cells, with or without <span class="html-italic">NMRAL2</span> expression, were infected with <span class="html-italic">M. bovis</span> at an MOI of 10. The invasion assay was conducted at 37 °C for 1 h, while the adhesion assay was performed at 4 °C for 30 min. The CFU assay quantified <span class="html-italic">M. bovis</span> numbers. (<b>c</b>–<b>f)</b> The impact of <span class="html-italic">NMRAL2</span> knockout or knockdown on intracellular <span class="html-italic">M. bovis</span> survival. WT, NMRAL2-KO, and si-NMRAL2 EBL cells were infected with <span class="html-italic">M. bovis</span> or BCG. Intracellular survival of <span class="html-italic">M. bovis</span> or BCG was evaluated using DsRed-expressing BCG (<b>c</b>) and plate counting methods (<b>d</b>–<b>f</b>). A two-tailed unpaired <span class="html-italic">t</span>-test and a two-way ANOVA were employed to assess the statistical significance of differences among various groups, utilizing three independent replicates for each analysis. *<span class="html-italic">* p</span> &lt; 0.01 and **<span class="html-italic">* p</span> &lt; 0.001 indicate statistically significant differences, and ns for no significant difference.</p>
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<p>Impact of <span class="html-italic">NMRAL2</span> knockout on host gene expression during <span class="html-italic">M. bovis</span> infection. (<b>a</b>,<b>b</b>) Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis for differentially expressed genes from the RNA-seq data comparing NMRAL2-KO-<span class="html-italic">M. bovis</span> versus WT-<span class="html-italic">M. bovis</span>. BP: biological process, MF: molecular function, and CC: cellular component. Words marked in red indicate the representative pathways that the research focuses on. (<b>c</b>) Quantification of NO concentrations. Intracellular NO level detected using the NO probe DAF-FM DA (Green). EBL cells were uninfected and infected with the indicated BCG (DsRed) at a MOI of 10 for 6 h. Cells were incubated with 1μM DAF-FM DA for an additional 30 min before being analyzed via confocal microscopy observation (Left). Scale bars: 100 μm. Ten random images were captured from each sample, and fluorescence intensities were quantified using ImageJ software. (<b>d</b>) Quantitative analysis of intracellular NO levels by Griess Reagent. (<b>e</b>) Immunoblotting analysis of iNOS expression in WT and NMRAL2-KO EBL cells. Various EBL cells were infected by <span class="html-italic">M. bovis</span> at an MOI of 10 for 48 h, followed by immunoblotting. The statistical significance among the different groups was evaluated utilizing a two-tailed unpaired <span class="html-italic">t</span>-test and a two-way ANOVA, based on three independent replicates. <span class="html-italic">* p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and **<span class="html-italic">* p</span> &lt; 0.001 indicate statistically significant differences, and ns for no significant difference.</p>
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<p>The effect of NMRAL2 on cell inflammation during <span class="html-italic">M. bovis</span> infection. (<b>a</b>–<b>d</b>) Expression of proinflammatory cytokines in EBL cells. EBL cells, both WT and NMRAL2-KO, were infected with <span class="html-italic">M. bovis</span> for 24 h. The mRNA levels of ATP6V1C2 (<b>a</b>), MMP2 (<b>b</b>), IL-6 (<b>c</b>), and TNF-α (<b>d</b>) were assessed via RT-qPCR. The mRNA levels of these cytokines were normalized to those of β-actin mRNA. (<b>e</b>) The knockout of <span class="html-italic">NMRAL2</span> resulted in an enhanced activation of the NF-κB and MAPK signaling pathways in response to <span class="html-italic">M. bovis</span>; EBL cells were exposed to <span class="html-italic">M. bovis</span> at an MOI of 10 for a designated infection period. Subsequently, the cells were lysed on ice for 30 min and analyzed via Western blot to determine the total and phosphorylated levels of the proteins p65, p38, JNK, and ERK1/2. The grayscale values of the phosphorylated to total protein ratios (p-p65/p65, p-p38/p38, p-JNK/JNK, and p-ERK/ERK) were normalized against β-actin. The statistical significance among the different groups was evaluated utilizing a two-tailed unpaired <span class="html-italic">t</span>-test and a two-way ANOVA, based on three independent replicates. Significance levels are denoted as follows: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, and *** <span class="html-italic">p</span> &lt; 0.001, and ns for no significant difference.</p>
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<p>Proposed model of NMRAL2 function during <span class="html-italic">M. bovis</span> infection. This schematic illustrates the proposed role of NMRAL2 in bovine lung epithelial cells during <span class="html-italic">M. bovis</span> infection, focusing on the effects observed following <span class="html-italic">NMRAL2</span> knockout. The absence of NMRAL2 results in reduced intracellular survival of <span class="html-italic">M. bovis</span>, along with enhanced production of NO and increased expression of proinflammatory cytokines. The upregulation of these cytokines is mediated through activation of the NF-κB and MAPK signaling pathways, highlighting NMRAL2 as a negative regulator of the host immune response to <span class="html-italic">M. bovis</span>.</p>
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23 pages, 4629 KiB  
Article
Dynamic Changes in Intestinal Microorganisms and Hematological Indices in Giraffes of Different Ages, and the Effect of Diarrhea on Intestinal Microbiota
by Baisheng Yu, Hangfan Li, Qiong Chen, Chuang Yang, Yongqing Guo and Baoli Sun
Animals 2024, 14(23), 3379; https://doi.org/10.3390/ani14233379 - 24 Nov 2024
Viewed by 264
Abstract
This study employed high-throughput sequencing to explore bacterial diversity and hematological variations across different age groups of giraffes, as well as the impact of diarrhea on their intestinal microbiota. Additionally, the correlation between intestinal flora and hematological indices was examined for the first [...] Read more.
This study employed high-throughput sequencing to explore bacterial diversity and hematological variations across different age groups of giraffes, as well as the impact of diarrhea on their intestinal microbiota. Additionally, the correlation between intestinal flora and hematological indices was examined for the first time. Firmicutes, Bacteroides, and Proteobacteria were the predominant bacterial groups in the giraffe’s intestinal flora. The α-diversity analysis indicated significant variations in microbial diversity among giraffes of varying ages (p < 0.05). Furthermore, giraffes suffering from diarrhea exhibited significant alterations in the abundance of Proteobacteria and Actinobacteriota at the phylum level (p < 0.05). At the genus level, Rikenellaceae_RC9_gut_group, Monoglobus, and Prevotellaceae_UCG-004 had significant differences compared to healthy counterparts (p < 0.05). Hematological parameters such as lymphocyte count (Lym), red blood cell count (RBC), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), platelet count (PLT), and plateletcrit (PCT) varied significantly across different age groups (p < 0.05). A substantial correlation was observed between the intestinal microbiome composition and hematological parameters (p < 0.05). In conclusion, this study highlights significant differences in both the intestinal microbiome composition and hematological indices among giraffes of different ages. Diarrhea was found to significantly alter the abundance and composition of the intestinal microbial community. These insights provide a valuable theoretical foundation for the microbiological and hematological aspects of giraffe health management and breeding practices. Full article
(This article belongs to the Section Mammals)
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<p>Venn diagrams and sample feasibility analysis between both groups. (<b>a</b>,<b>b</b>) A Venn diagram illustrating the distribution of ASVs in the feces of giraffes with different ages, stratified at both the phylum and genus levels. (<b>c</b>,<b>d</b>) A Venn diagram illustrating the distribution of ASVs in the feces of giraffes with diarrhea and healthy giraffes, stratified at both the phylum and genus levels. (<b>e</b>) Rank abundance curve.</p>
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<p>(<b>a</b>,<b>b</b>) Bar diagram of intestinal dominant bacteria abundance in the phylum and genera of different developmental stages for giraffes. (<b>c</b>,<b>d</b>) Bar diagram of intestinal dominant bacteria abundance in the phylum and genera of diarrhea and healthy giraffes.</p>
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<p>(<b>a</b>,<b>b</b>) Significant shifts in gut microbial abundance at the phylum and genus levels in giraffes.</p>
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<p>Alpha diversity (Chao1 and Shannon) indices of intestinal microbiota between different ages of giraffes. (B, C, and D represent cubs, sub-adults, and adults, respectively). (<b>a</b>) Box diagram of shannon index between different groups. (<b>b</b>) Box diagram of Chao1 index between different groups. ** represents <span class="html-italic">p</span> &lt; 0.01; *** represents <span class="html-italic">p</span> &lt; 0.001; ns represents not significant.</p>
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<p>(<b>a</b>,<b>c</b>) PCoA analysis of intestinal bacterial communities in giraffes. (<b>b</b>,<b>d</b>) Box chart of Adonis difference analysis. (B, C, and D represent cubs, sub-adults, and adults, respectively).</p>
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<p>(<b>a</b>–<b>c</b>) LDA scores displaying the distinct bacterial differences between different ages of giraffes. LDA scores &gt; 3.0 were considered statistically significant. (B, C, and D represent cubs, sub-adults, and adults, respectively; cubs: 0–1 years old healthy giraffes; sub-adults: 1–3 years old healthy giraffes; adults: 10–12-year-old healthy giraffes).</p>
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<p>(<b>a</b>) LDA scores displaying the distinct bacterial differences between diarrhea and healthy. LDA scores &gt; 3.0 were considered statistically significant. (<b>b</b>) Cladogram revealing phylogenetic distribution of the gut bacterial community associated with diarrhea and healthy giraffes.</p>
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<p>(<b>a</b>) The correlation heatmap of microbial abundance in giraffe feces and routine blood values. (<b>b</b>) The correlation heatmap of microbial abundance in giraffe feces and serum chemistry. (<b>c</b>) The correlation heatmap of microbial abundance between different ages of giraffes. The magnitude of the correlation coefficient was reflected by color intensity. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>(<b>a</b>) Network analysis revealing potential correlation between different bacterial genera. Sizes of circles represent bacterial genera abundance. Correlative strength between both bacteria genera can be evaluated by thickness of the lines. (<b>b</b>) Predicted functional distribution of the intestinal microbiota in captive giraffes. The magnitude of the correlation coefficient was reflected by color intensity.</p>
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30 pages, 1068 KiB  
Article
Peripheral Blood Leukocyte Subpopulation Changes in Reaction to an Acute Psychosocial Stressor as Compared to an Active Placebo-Stressor in Healthy Young Males: Mediating Effects of Major Stress-Reactive Endocrine Parameters
by Lisa-Marie Walther, Angelina Gideon, Christine Sauter, Marcel Leist and Petra H. Wirtz
Cells 2024, 13(23), 1941; https://doi.org/10.3390/cells13231941 - 22 Nov 2024
Viewed by 299
Abstract
Psychosocial stress has been proposed to induce a redistribution of immune cells, but a comparison with an active placebo-psychosocial stress control condition is lacking so far. We investigated immune cell redistribution due to psychosocial stress compared to that resulting from an active placebo-psychosocial [...] Read more.
Psychosocial stress has been proposed to induce a redistribution of immune cells, but a comparison with an active placebo-psychosocial stress control condition is lacking so far. We investigated immune cell redistribution due to psychosocial stress compared to that resulting from an active placebo-psychosocial stress but otherwise identical control condition. Moreover, we tested for mediating effects of endocrine parameters and blood volume changes. The final study sample comprised 64 healthy young men who underwent either a psychosocial stress condition (Trier Social Stress Test; TSST; n = 38) or an active placebo-psychosocial stress control condition (PlacTSST; n = 26). Immune cell counts and hemoglobin, epinephrine, norepinephrine, ACTH, renin, and aldosterone levels, as well as those of saliva cortisol, were determined before and up to 30 min after the TSST/PlacTSST. The TSST induced greater increases in total leukocyte, monocyte, and lymphocyte levels as compared to the PlacTSST (p’s ≤ 0.001), but in not granulocyte counts. Neutrophil granulocyte counts increased in reaction to both the TSST and PlacTSST (p’s ≤ 0.001), while eosinophil and basophil granulocyte counts did not. The psychosocial stress-induced increases in immune cell counts from baseline to peak (i.e., +1 min after TSST cessation) were independently mediated by parallel increases in epinephrine (ab’s ≤ −0.43; 95% CIs [LLs ≤ −0.66; ULs ≤ −0.09]). Subsequent decreases in immune cell counts from +1 min to +10 min after psychosocial stress cessation were mediated by parallel epinephrine, renin, and blood volume decreases (ab’s ≥ 0.17; 95% CIs [LLs ≥ 0.02; ULs ≥ 0.35]). Our findings indicate that psychosocial stress specifically induces immune cell count increases in most leukocyte subpopulations that are not secondary to the physical or cognitive demands of the stress task. Increases in the number of circulating neutrophil granulocytes, however, are not psychosocial stress-specific and even occur in situations with a low probability of threat or harm. Our findings point to a major role of epinephrine in mediating stress-induced immune cell count increases and of epinephrine, renin, and blood volume changes in mediating subsequent immune cell count decreases from +1 min to +10 min after psychosocial stress cessation. Full article
(This article belongs to the Special Issue Innate Immunity in Health and Disease)
12 pages, 2746 KiB  
Article
The Intensity of BCL2A1 Expression Increases According to the Stage Progression of Acute Histologic Chorioamnionitis in the Extra-Placental Membranes of Spontaneous Preterm Birth
by Chan-Wook Park, Eun-Mi Lee, Seung-Han Shin, Chul Lee and Jae-Kyung Won
Life 2024, 14(12), 1535; https://doi.org/10.3390/life14121535 - 22 Nov 2024
Viewed by 319
Abstract
Our prior findings showed that BCL2A1 in neutrophils is highly expressed in the extra-placental membranes (EPMs) of both the human spontaneous preterm-birth (PTB) (i.e., PTL or preterm PROM) and nonhuman-primate PTB model. However, no data exist on whether the intensity of BCL2A1 expression [...] Read more.
Our prior findings showed that BCL2A1 in neutrophils is highly expressed in the extra-placental membranes (EPMs) of both the human spontaneous preterm-birth (PTB) (i.e., PTL or preterm PROM) and nonhuman-primate PTB model. However, no data exist on whether the intensity of BCL2A1 expression quantitatively increases according to the stage progression of acute histologic chorioamnionitis (acute HCA) in EPM. The objective is to investigate whether the intensity of BCL2A1 expression quantitatively increases according to the stage progression of acute HCA in EPM among spontaneous PTB cases, as measured using QuPath. The study population included 121 singleton PTBs (gestational age [GA] at delivery < 34 weeks) due to either preterm labor or preterm PROM. With digital image analysis, we calculated the percentage of BCL2A1-positive cells in immunohistochemistry according to the stage progression of acute HCA in EPMs as the primary outcome and examined the relationship between the percentage of BCL2A1-positive cells and either the GA at delivery or the amniotic-fluid (AF) WBC count as the secondary outcome. The median percentage of BCL2A1-positive cells progressively increases with the stage progression of acute HCA in EPM (group-1 vs. group-2 vs. group-3 vs. group-4 vs. group-5; 7.62 vs. 5.15 vs. 43.57 vs. 71.07; γ = 0.552, p < 0.000001). The percentage of BCL2A1-positive cells in EPMs and the AFWBC count shows a positive correlation (γ = 0.492, p = 0.000385). Moreover, the percentage of BCL2A1-positive cells in EPMs continuously decreased with increasing GA at delivery (γ = −0.253, p = 0.005148). In conclusion, the intensity of BCL2A1 expression increases according to the stage progression of acute HCA in EPMs and the elevation of AFWBC among spontaneous PTB cases. This finding suggests BCL2A1 in EPMs may be a promising marker and target for acute HCA. Full article
(This article belongs to the Special Issue Clinical Management and Prevention of Adverse Pregnancy Outcomes)
15 pages, 943 KiB  
Article
In Vitro Analysis of LPS-Induced miRNA Differences in Bovine Endometrial Cells and Study of Related Pathways
by Xinmiao Li, Zhihao Zhang, Xiangnan Wang, Ligang Lu, Zijing Zhang, Geyang Zhang, Jia Min, Qiaoting Shi, Shijie Lyu, Qiuxia Chu, Xingshan Qi, Huimin Li, Yongzhen Huang and Eryao Wang
Animals 2024, 14(23), 3367; https://doi.org/10.3390/ani14233367 - 22 Nov 2024
Viewed by 219
Abstract
Lipopolysaccharide (LPS) is one of the main factors inducing endometritis in dairy cows. However, the specific pathogenesis of LPS-induced endometritis in dairy cows is not fully understood. The objective of this study was to establish an in vitro endometritis model using LPS-induced bovine [...] Read more.
Lipopolysaccharide (LPS) is one of the main factors inducing endometritis in dairy cows. However, the specific pathogenesis of LPS-induced endometritis in dairy cows is not fully understood. The objective of this study was to establish an in vitro endometritis model using LPS-induced bovine endometrial epithelial (BEND) cells. BEND cells were treated with LPS of different concentrations and times. The cell-counting kit-8 (CCK-8) was used to detect the cell survival rate after LPS treatment, and quantitative real-time PCR (RT-qPCR) was used to detect the expression of control group and LPS-treated group of inflammatory factors interleukin-1 beta (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α). The results showed that the survival rate of endometrial epithelial cells stimulated by 5 μg/mL LPS for 6 h was 75.13%, and the expression of inflammatory factors was significantly increased. Therefore, 5 μg/mL LPS for 6 h could be selected as a suitable model for the study of inflammation. In addition, miRNA sequencing and target gene prediction was performed on normal and LPS-treated BEND cells. Among twenty-one differentially expressed miRNAs, six miRNAs were selected and their expression levels were detected by RT-qPCR, which were consistent with the sequencing results. Twenty-one differentially expressed miRNAs collectively predicted 17,050 target genes. This study provides a theoretical basis for further investigation of the pathogenesis of endometritis. Full article
(This article belongs to the Section Cattle)
9 pages, 2076 KiB  
Article
Milking Reactivity in Primiparous Saanen Goats During Early Lactation: Effects on Milk Yield, Milk Quality and Plasma Cortisol Concentration
by Mayara Andrioli, Joseph K. Grajales-Cedeño, Aline C. Sant’Anna, João A. Negrão and Mateus J. R. Paranhos da Costa
Animals 2024, 14(23), 3365; https://doi.org/10.3390/ani14233365 - 22 Nov 2024
Viewed by 238
Abstract
The reactivity of primiparous goats during milking can significantly affect animal welfare and performance. The aim of this study was to investigate the progression of milking reactivity during lactation and its effects at early lactation on milk yield, milk quality, and plasma cortisol [...] Read more.
The reactivity of primiparous goats during milking can significantly affect animal welfare and performance. The aim of this study was to investigate the progression of milking reactivity during lactation and its effects at early lactation on milk yield, milk quality, and plasma cortisol concentration in primiparous Saanen goats. The milking reactivity of 31 primiparous Saanen goats was evaluated on days 10, 30, and 60 of the lactation period by scoring step-kick behavior (SK) and head, ear, and tail movements (MOV). Milk yield, milk protein and fat percentages, somatic cell count (SCC), and plasma cortisol concentration were measured on day 10 of lactation. Goats’ milking reactivity was classified as low or high (according to the SK and MOV scores measured on day 10 of lactation). SK and MOV scores decreased over the lactation period, indicating that the goats habituated to the milking procedures. Milking reactivity only affected milk fat percentage, with the less reactive goats having more milk fat than the highly reactive ones. There was no evidence of stress when goats were subjected to milking procedures during early lactation. Further studies are needed to clarify the relationships of milking reactivity with stress response and their effects on milk yield and quality. Full article
(This article belongs to the Section Animal Welfare)
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<p>Distribution of the number of animals (n = 31) according to step-kick (SK) and head, ear, and tail movement (MOV) scores during pre-dipping procedures according to the lactation period (days 10, 30, and 60 of the lactation period).</p>
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<p>Box plots of (<b>a</b>) the step-kick behavior scores (SK) and (<b>b</b>) head, ear, and tail movement (MOV) scores in primiparous Saanen goats (<span class="html-italic">n</span> = 31) according to the lactation period (days 10, 30, and 60). Different letters indicate significant differences between the assessment days (<span class="html-italic">p</span> &lt; 0.05). Bold lines indicate medians, triangles represent means, and circles the outliers.</p>
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<p>Box plots of milk yield, kg (<b>a</b>), milk fat, % (<b>b</b>), milk protein, % (<b>c</b>), and somatic cell score (log cel/mL) (<b>d</b>) in primiparous Saanen goats (<span class="html-italic">n</span> = 31) according to the classes of milking reactivity (low and high reactivity). Different letters indicate significant differences between the milking reactivity classes (<span class="html-italic">p</span> &lt; 0.05). Bold lines indicate medians, triangles represent means, and circles the individuals.</p>
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<p>Box plots of plasma cortisol concentration (ng/mL) in primiparous Saanen goats (<span class="html-italic">n</span> = 31) according to the classes of milking reactivity (low and high). Bold lines indicate medians, triangles represent means, and circles the individuals.</p>
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17 pages, 1817 KiB  
Article
Physiological Stress Response and Oxidative Status in Tambaqui (Colossoma macropomum) Fed Diets Supplemented with Selenium
by Celma Maria Ferreira, Valéria Dornelles Gindri Sinhorin, Márcio Aquio Hoshiba and Janessa Sampaio de Abreu
Biology 2024, 13(12), 959; https://doi.org/10.3390/biology13120959 - 22 Nov 2024
Viewed by 294
Abstract
The present study aimed to evaluate the physiological responses to transport stress in juvenile tambaqui (Colossoma macropomum) fed a diet supplemented with hydroxy-selenomethionine (OH-SeMet; Selisseo®, Adisseo) and determine through stress biomarkers whether selenium supplementation could reduce the impact of [...] Read more.
The present study aimed to evaluate the physiological responses to transport stress in juvenile tambaqui (Colossoma macropomum) fed a diet supplemented with hydroxy-selenomethionine (OH-SeMet; Selisseo®, Adisseo) and determine through stress biomarkers whether selenium supplementation could reduce the impact of transport stress on tambaqui resilience. Juvenile fish (15.71 ± 1.90 g) were randomly distributed into five experimental groups; one fed a basal diet without inclusion of OH-SeMet (0.0 mg Se/kg) and four other groups fed supplemented diets with selenium in the form of OH-SeMet at concentrations of 0.3, 0.6, 0.9, and 1.2 mg Se/kg over 75 days. After this period, the fish were transported for 4 h in plastic bags (60 L), and sampling took place before transport (baseline), immediately after transport, and 24 h later. The effects of dietary OH-SeMet in tambaqui challenged by transport were studied in terms of antioxidative status (superoxide dismutase, catalase, glutathione peroxidase, glutathione s-transferase, and reduced glutathione); stress markers (serum cortisol, blood glucose, ionic and hematological parameters), and immunological status (respiratory burst activity and total protein, A/G ratio). Following transport, we observed an elevation in cortisol and blood glucose concentrations, accompanied by hemoconcentration, as evidenced by elevated hematocrit, erythrocyte count, hemoglobin concentration, and mean corpuscular hemoglobin concentration (MCHC). Additionally, there was a decrease in immune response, indicated by reduced respiratory activity of leukocytes and an osmotic imbalance, which culminated in hypochloremia, increased plasma protein concentration, and a rise in red blood cell volume, all of which were measured 24 h after the stressor was applied. Twenty-four hours after transport, the antioxidant defense system was activated, and no recovery of homeostasis was observed for most of the parameters analyzed. The dietary inclusion of selenium did not alleviate the hormonal, metabolic, hematological, and ionic changes caused by transport, but at a concentration of 0.9 mg Se/kg, it was able to increase the activity of enzymes related to oxidative stress (catalase and superoxide dismutase, mainly) in tambaqui juveniles submitted to this challenge. Full article
(This article belongs to the Special Issue Aquatic Animal Nutrition and Feed)
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<p>Serum chloride from tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Different lowercase letters show significantly different data between treatments and capital letters between times (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Mean corpuscular volume (MCV) of tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Different capital letters show significantly different data between each treatment at different times and lower-case letters between treatments within each time (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Results obtained from lymphocytes, monocytes, and neutrophils of tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Each bar represents the mean ± standard error of all treatments. Different capital letters show significantly different data between sampling times (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). For statistical analysis, the data were log (x) transformed. BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Results obtained from leukocyte respiratory burst activity of tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Different lowercase letters show significantly different data between treatments and capital letters between times (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Albumin from tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Different capital letters show significantly different data between each treatment at different times and lower-case letters between treatments within each time. (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Analysis of enzymatic activities in the liver of tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. Superoxide dismutase—SOD (<b>A</b>), catalase—CAT (<b>B</b>), and glutathione peroxidase—GPx (<b>C</b>). Different capital letters show significantly different data between treatments at different times (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05). For statistical analysis, data were transformed into log x. BT: before transport; AT: after transport; 24 h AT: 24 h after transport.</p>
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<p>Concentration of reduced glutathione—GSH (<b>A</b>) and activity of glutathione s-transferase—GST (<b>B</b>) in the liver of tambaqui (<span class="html-italic">Colossoma macropomum</span>) supplemented with selenium in the diet (hydroxy-selenomethionine; 0.0, 0.3, 0.6, 0.9, and 1.2 mg Se/kg) and submitted to transport stress. For GSH concentration (ANOVA; Tukey test; <span class="html-italic">p</span> &lt; 0.05) and GST activity (Kruskal–Wallis’s Test; Dunn’s test; <span class="html-italic">p</span> &lt; 0.05). For statistical analysis, GSH data were transformed into log x.</p>
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16 pages, 929 KiB  
Systematic Review
Efficacy of Prophylactic Antibiotics in COPD: A Systematic Review
by Anh Tuan Tran, Amr Sayed Ghanem, Marianna Móré, Attila Csaba Nagy and Ágnes Tóth
Antibiotics 2024, 13(12), 1110; https://doi.org/10.3390/antibiotics13121110 - 21 Nov 2024
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Abstract
Background/Objectives: Chronic obstructive pulmonary disease (COPD) is a global health problem and the third leading contributor to mortality worldwide. This systematic review aims to summarize the results of previous studies tackling the question of the efficacy of long-term prophylaxis of antibiotics in [...] Read more.
Background/Objectives: Chronic obstructive pulmonary disease (COPD) is a global health problem and the third leading contributor to mortality worldwide. This systematic review aims to summarize the results of previous studies tackling the question of the efficacy of long-term prophylaxis of antibiotics in COPD patients, with particular regard to exacerbation rate, time to first exacerbation, health status, airway bacterial load, inflammatory markers, cell counts in sputum samples, and potential adverse events. Results: Four studies found significant improvement in the exacerbation rate in patients receiving antibiotic intervention. One study found doxycycline to have negative effects on patients’ exacerbation outcomes. Two studies recorded a reduction in total airway bacterial load using quantitative culture of sputum samples, but the prevalence of antibiotic-resistant bacteria increased in all studies that measured it. No change in inflammatory markers was observed; however, there was a decline in neutrophil cell counts and, subsequently, reductions in neutrophil elastase concentrations. Methods: PubMed and Web of Science databases were searched for English-language studies presenting data on the prophylactic use of antibiotics in COPD management. All included studies are randomized controlled trials (RCTs) and meet the inclusion criteria. Conclusions: Based on current evidence from RCTs, the prophylactic antibiotic approach utilizing macrolides is the most effective in reducing the incidence of COPD exacerbation. However, the emergence of antibiotic-resistant pathogens is notable. Whether the beneficial effects of macrolides on exacerbation are due to their antibacterial or immunomodulant properties is still inconclusive. Future studies are needed to better understand the interactions between antibiotics and the airway microbiome during COPD exacerbation. Full article
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<p>PRISMA flow diagram.</p>
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9 pages, 532 KiB  
Article
COVID-19 Vaccinations, Infections, and Outcomes Among 784 People Living with HIV
by Keren Mahlab-Guri, Irina Komarova, Laliv Kadar, Shay Nemet, Ramon Cohen, Sara Radian-Sade, Achiel Tova, Alex Guri, Shira Rosenberg-Bezalel and Daniel Elbirt
Viruses 2024, 16(12), 1805; https://doi.org/10.3390/v16121805 - 21 Nov 2024
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Abstract
Introduction: Variants of COVID-19 are responsible for 700 million infections and 7 million deaths worldwide. Vaccinations have high efficiency in preventing infection and secondary benefits of reducing COVID-19 hospital admissions, attenuating disease severity and duration of illness. Conflicting reports were published regarding COVID-19 [...] Read more.
Introduction: Variants of COVID-19 are responsible for 700 million infections and 7 million deaths worldwide. Vaccinations have high efficiency in preventing infection and secondary benefits of reducing COVID-19 hospital admissions, attenuating disease severity and duration of illness. Conflicting reports were published regarding COVID-19 among PLWH. Objective: The aim of this study was to evaluate COVID-19 morbidity, hospitalization, and the magnitude of immunological response to sequential BNT 162b2 mRNA vaccines in PLWH regarding demographic and clinical factors. Results: Our retrospective study included 784 PLWH who had at least one anti- SARS-CoV-2 antibody test between March 2021 and October 2021. Half of our patients (392) had CD4 cell counts above 500 cells/µL, 40.2% (315) had 200 < CD4 < 500 cells/µL and only 9.8% (77) had CD4 < 200 cells/µL at their last laboratory workup. The mean age was 50.2 ± 12.2 years. About 90% of our patients were given at least two doses of the BNT 162b2 Pfizer vaccines; about 60% received three doses of the vaccine. About a quarter of our patients (27.6%) had COVID-19 infection. Only six patients required hospital admission. All six patients recovered from COVID-19 infection. Titers of COVID-19 antibodies were lower for patients with CD4 cell counts of less than 200 cells/µL in the first, second, and third serological tests with statistical significance. In a multinomial logistic regression, the influence of other factors such as age, sex, and previous COVID-19 infection on first COVID-19 antibody titers was not significant. Conclusions: PLWH are responsive to COVID-19 vaccines. As was expected, patients with higher CD4 cell counts had higher titers of COVID-19 antibodies and lower hospitalization rate. Age, sex, and previous COVID-19 infection did not significantly affect antibody titers according to our study. Larger prospective studies with control groups are needed to further characterize immunologic response to COVID-19 vaccination among PLWH. Full article
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<p>Antibody titers over time for 46 patients with three measurements. * <span class="html-italic">p</span> &lt; 0.001.</p>
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