Alteration of Plasma Indoles in Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is one of the most common endocrinopathies in reproductive-aged women. The occurrence of PCOS was reported to be associated with the alteration of gut microbiota. Microbiota-derived indoles may possibly play a key role in glycemic control. The purpose of this work is to reveal the alteration of plasma indoles in PCOS patients and to investigate the correlation between indoles levels and glucose metabolism. Sixty-five patients with PCOS and twenty-eight age-matched women were enrolled in this work. The concentrations of plasma indoles, including indoxyl sulfate (IS), indole-3-acetic acid (IAA), indole-3-propionate (IPA), indole (IND), and 3-methylindole (3-MI), were measured by HPLC with the fluorescence detection. The plasma levels of IS, IAA, and IND were significantly elevated in patients with PCOS compared to those in the control group (p < 0.05). Furthermore, the plasma levels of IS, IAA, and IND were positively correlated with fasting glucose, fasting insulin, and the homeostatic model of insulin resistance index (HOMA-IR) (p < 0.05). Besides, the 3-MI level in the plasma was positively correlated with the fasting glucose level, whereas plasma levels of IS, IAA, IND, and 3-MI were negatively correlated with glucagon-like peptide 1 (p < 0.05). Moreover, IS and IND were considered to be risk factors for PCOS after age, BMI, T, LH, and HOMA-IR adjustment. The area under the receiver-operating characteristic curve of the combined index of five indoles was 0.867 for PCOS diagnosis. Additionally, plasma indoles altered in PCOS, which was closely associated with the glucose metabolism.

Efficient benzo(a)pyrene degradation by coal gangue-based catalytic material for peroxymonosulfate activation.

A low cost and green peroxymonosulfate (PMS) activation catalyst (CG-Ca-N) was successfully prepared with coal gangue (CG), calcium chloride, and melamine as activator. Under the optimal conditions, the CG-Ca-N can remove 100 % for benzo(a)pyrene (Bap) in an aqueous solution after 20 min and 72.06 % in soil slurry medium within 60 min, which also display excellent reuse ability toward Bap after three times. The removal of Bap is significantly decreased when the initial pH value was greater than 9 and obviously inhibited in the presence of HCO3- or SO42-. The characterization results indicated that the addition of calcium chloride could stabilize and increase the content of pyridinic N during thermal annealing, resulting in the production of •OH, SO4•- and 1O2. Based on electron paramagnetic resonance (EPR) and active radical scavenging experiments, 1O2 could be identified to be the dominant role in Bap degradation. Overall, this work opened a new perspective for the low cost and green PMS catalysts and offered great promise in the practical remediation of organic pollution of groundwater and soil.

[Functional peptidomics analysis of Saccharomyces pastorianus protein hydrolysates based on different enzyme treatments].

The aim of this study is to explore differences in the peptidomics of Saccharomyces pastorianus protein hydrolysates treated with different enzymes. Briefly, differences in the peptide fingerprints and active peptides of neutral protease/papain-hydrolyzed S. pastorianus were analyzed using ultra-high performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS) combined with PEAKS Online 1.7 analysis software, Peptide Ranker, and the BIOPEP database. Compared to traditional databases, the PEAKS Online uses de novo sequencing for analysis to obtain oligopeptides smaller than pentapeptides. It provides more comprehensive data of the peptide sample. In this study, enzymatic hydrolysates of S. pastorianus protein were prepared under the optimum conditions of neutral protease and papain respectively. In total, 7221 and 7062 polypeptides were identified in the hydrolysates of neutral protease and papain, respectively; among these polypeptides, 980 were common to the two enzymes. The 6241 and 6082 unique peptides found in the hydrolysates of neutral protease and papain, respectively, indicated that the peptide fingerprints of the two hydrolysates are quite different. Peptide Ranker predicted that 3013 (41.73%) and 3095 (43.83%) peptides were potentially bioactive in the hydrolysates of neutral protease and papain, respectively. According to the BIOPEP database, neutral protease and papain contained 295 and 357 active peptides, respectively; these peptides were mainly composed of angiotensin converting enzyme (ACE) inhibitors and dipeptidyl peptidase IV inhibitors and antioxidant peptides. The number of active peptides in the hydrolysate of papain was higher than that in the hydrolysate of neutral protease, but the total ion intensity of active peptides in the former was lower than that in the latter. This study revealed the influence of protease type on the composition of enzymatic hydrolysates from S. pastorianus protein. The above results provide a reference for the development of functional products of S. pastorianus protein peptides and the high-value utilization of yeast resources.

The Relationship between Mastitis and Antimicrobial Peptide S100A7 Expression in Dairy Goats.

S100A7 is an inflammation-related protein and plays an essential role in host defenses, yet there is little research about the relationship between mastitis and S100A7 expression in dairy goats. Here, according to the clinical diagnosis of udders, SCC, and bacteriological culture (BC) of milk, 84 dairy goats were grouped into healthy goats (n = 25), subclinical mastitis goats (n = 36), and clinical mastitis goats (n = 23). The S100A7 concentration in subclinical mastitis goats was significantly upregulated than in healthy dairy goats (p = 0.0056) and had a limited change with clinical mastitis dairy goats (p = 0.8222). The relationship between log10 SCC and S100A7 concentration in milk was positive and R = 0.05249; the regression equation was Y = 0.1446 × X + 12.54. According to the three groups, the log10 SCC and S100A7 were analyzed using the receiver operating characteristics (ROC) curve; in subclinical mastitis goats, the area under the ROC curve (AUC) of log10 SCC was 0.9222 and p < 0.0001, and the AUC of S100A7 concentration was 0.7317 and p = 0.0022, respectively; in clinical mastitis goats, the AUC of log10 SCC was 0.9678 and p < 0.0001, and the AUC of S100A7 concentration was 0.5487 and p = 0.5634, respectively. In healthy goats, S100A7 was expressed weakly in the alveolus of the mammary gland of healthy goats while expressed densely in the collapsed alveolus of mastitis goats. Moreover, S100A7 expression increased significantly in mastitis goats than in healthy dairy goats. In this research, results showed the effects of mastitis on the S100A7 expression in the mammary gland and S100A7 concentration in milk and the limited relationship between SCC and mastitis, which provided a new insight into S100A7's role in the host defenses of dairy goats.

Immunolocalization of antibacterial peptide S100A7 in mastitis goat mammary gland and lipopolysaccharide induces the expression and secretion of S100A7 in goat mammary gland epithelial cells via TLR4/NFκB signal pathway.

The antimicrobial peptide S100A7, with antimicrobial activities for a broad spectrum of bacteria, has attracted more and more attention for the prevention and treatment of mastitis. However, there is little information about the expression and regulation mechanism of S100A7 in mastitis goats. This study revealed that S100A7 was mainly expressed in the stratified squamous epithelium of teat skin and streak canal, and S100A7 was present weakly in the healthy goat alveolus yet densely in the mastitis goat collapsed alveolus. Goat mammary epithelial cells (MECs) were isolated and treated with 2.5, 5, 10 and 20 µg/mL lipopolysaccharide (LPS) respectively for a different time, S100A7 mRNA expression and protein secretion were upregulated significantly with LPS treatment for 3 h, and the secretion level of S100A7 descended after 48 h treatment for all of these four groups. Moreover, after treatment with LPS, the mRNA levels of Toll-like receptor 4 (TLR4) and MyD88 were up-regulated, and the phosphorylation of p65 was up-regulated markedly. However, adding TLR4 inhibitor TAK-242 or/and NF-κB inhibitor QNZ significantly suppressed the phosphorylation of p65, and then inhibited the expression and secretion of S100A7 induced by LPS treatment. In conclusion, LPS induced the expression and secretion of S100A7 in goat MECs via TLR4/NF-κB signaling pathway.

Attitudes and willingness to pay for clean heating by typical households: a case study of rural areas in Yongcheng City, Henan Province, China.

Understanding households' attitudes and willingness to pay (WTP) for clean heating can provide a scientific basis for decision-makers to assess the potential to develop clean heating, choose heating methods, and formulate subsidy standards in the region. In this paper, the double-bounded dichotomous contingent valuation method-modified by the spike model-was used to better estimate the actual WTP of households through a sample survey of 456 households in rural areas of Yongcheng City, China, in 2021. The factors influencing attitudes and WTP were examined to reveal mechanisms of accepting clean heating. The results showed that 94.96% of households were willing to pay for clean heating. The annual WTP was 1071 yuan per household, more than eight times the current heating cost. Factors that affect clean heating attitudes do not necessarily affect the WTP. Specifically, gender, length of time spent living at home, and family income had significant influences on WTP, whereas the educational level, adaptive perceptions in relation to environmental perceptions, and the recognition variables for gender equality in energy consumption had a significant impact on attitudes. It is worth noting that elderly people and females were identified as vulnerable groups in the implementation of clean heating.

Estradiol Regulates the Expression and Secretion of Antimicrobial Peptide S100A7 via the ERK1/2-Signaling Pathway in Goat Mammary Epithelial Cells.

S100A7 has received extensive attention in the prevention and treatment of mastitis across a broad spectrum, yet there is a little information about its mechanism, especially in the immunomodulatory effects of estrogen. In the present study, based on the milk bacteriological culture (BC) of 30 dairy goats, the concentration of both estrogen and S100A7 in the BC-positive samples was not significantly different than in the BC-negative samples; the estrogen abundance in subclinical and clinical mastitis samples also showed only a limited difference; compared with healthy samples, the S100A7 abundance in subclinical mastitis samples differed little, while it was significantly decreased in clinical mastitis samples. Moreover, the relationship between estrogen and S100A7 was positive, and the regression equation was y = 0.3206x + 23.459. The goat mammary epithelial cells (gMECs) were isolated and treated with 1, 10, 100 nM E2 and/or 5 µg/mL lipopolysaccharide (LPS), respectively, for 6 h. Compared with control samples, 5 µg/mL LPS, 10 nM E2 and 100 nM E2 markedly induced S100A7 expression and secretion. More than separated treatment, the cooperation of LPS and E2 also significantly increased S100A7 expression, rather than S100A7 secretion. The p-ERK was up-regulated markedly with 100 nM E2 treatment, while the expression of p-JNK, p-p38 and p-Akt had little effect. The G protein-coupled estrogen receptor 1(GPER1) agonist G1 markedly induced S100A7 expression and secretion in gMECs, and the estrogen nuclear receptor antagonist ICI and GPER1 antagonist G15 significantly repressed this process. In conclusion, E2 binds to nuclear and membrane receptors to regulate the expression and secretion of S100A7 via the ERK1/2-signaling pathway in gMECs.

C-Type Natriuretic Peptide Regulates the Expression and Secretion of Antibacterial Peptide S100A7 in Goat Mammary Gland Through PKG/JNK/c-Jun Signaling Pathway.

During infection, the infected tissue secretes a variety of endogenous peptides to resist further invasion of pathogens. Among these endogenous peptides, the natriuretic peptides and the antimicrobial peptides attracted the most attention. C-type natriuretic peptide (CNP) and its receptor natriuretic peptide receptor B (NPR-B) were members of the natriuretic peptide system. The antimicrobial peptide S100A7 plays an important role to resist infection of bacteria in mastitis. It is reported that the expression of S100A7 is regulated by an activator protein-1 (AP-1)-responsive promoter. As a subunit of AP-1, c-Jun is a downstream target of CNP/NPR-B signaling pathway. Therefore, it is a hypothesis that the CNP/NPR-B signaling pathway induces the expression and secretion of S100A7 in mammary glands to take part in local mammary gland innate immunity. To verify this hypothesis, goat mammary gland and isolated mammary epithelial cells (MECs) were used to explore the expression of CNP/NPR-B and their physiological roles in goat mammary gland. The results showed that goat mammary gland expressed NPR-B, but not CNP. The expression and secretion of S100A7 in goat MECs were obviously induced by CNP/NPR-B signaling pathway. After treatment with CNP, the cyclic guanosine monophosphate (cGMP) level in goat MECs was significantly upregulated. Along with the upregulation of cGMP level, the phosphorylation levels of c-Jun N-terminal kinase (JNK) and its target c-Jun were also increased gradually. KT5823 is a specific inhibitor for protein kinase G (PKG). KT5823 remarkably inhibited the phosphorylation of JNK and c-Jun induced by CNP. Correspondingly, KT5823 evidently inhibited the expression and secretion of S100A7 induced by CNP. On the other hand, the expression of NPR-B and S100A7 was upregulated in the mastitis goat mammary gland. But, there was no significant difference in expression of CNP between healthy and mastitis goat mammary gland tissues. The goat mastitis model was established in vitro using goat MECs treated by lipopolysaccharide (LPS). LPS treatment also could increase the expression of NPR-B and S100A7. In conclusion, goat mammary gland expressed NPR-B, indicating mammary gland was the target organ for natriuretic peptide system. Moreover, CNP, through NPR-B/JNK/c-Jun signaling pathway to regulate the expression and secretion of S100A7 in MECs, played an important role in mammary gland innate immunity.

NF-κB-Dependent Snail Expression Promotes Epithelial-Mesenchymal Transition in Mastitis.

Mastitis is a common and important clinical disease in ruminants. This may be associated with inflammatory fibrosis if not treated promptly. Inflammation-derived fibrosis is usually accompanied by epithelial-mesenchymal transition (EMT) in epithelial cells. However, the precise molecular mechanism underlying mastitis-induced fibrosis remains unclear. Nuclear factor kappa-B (NF-κB) and Snail are key regulators of EMT. In this study, primary goat mammary epithelial cells (GMECs) were treated with 10 µg/mL lipopolysaccharide (LPS) for 14 d to mimic the in vivo mastitis environment. After LPS treatment, the GMECs underwent mesenchymal morphological transformation and expressed mesenchymal cell markers. Snail expression was induced by LPS and was inhibited by suppression of the TLR4/NF-κB signaling pathway. Snail knockdown alleviated LPS-induced EMT and altered the expression of inflammatory cytokines. Finally, we found that the expression of key molecules of the TLR4/NF-κB/Snail signaling pathway was increased in mastitis tissues. These results suggest that Snail plays a vital role in LPS-induced EMT in GMECs and that the mechanism is dependent on the activation of the TLR4/NF-κB signaling pathway.

Inhibition of cell proliferation and promotion of acinus-like structure formation from goat mammary epithelial cells via Wnt/ß-catenin signaling.

Mammary epithelial cells have been suggested to be central to control the expansion and remodeling of mammary gland. Wnt/ß-catenin signaling modulates cell fate in animals throughout their life span, and represents indispensable roles in tissue homeostasis, cell renewal, and regeneration in organs. Here, we utilized the small molecule 6-bromoindirubin-3'-oxime (BIO), an activator of Wnt/ß-catenin signaling, and investigated whether Wnt/ß-catenin signaling regulated the proliferation and acinus-like structure formation of goat mammary epithelial cells (GMECs). We showed that isolated GMECs displayed the typical epithelial cobblestone morphology and expressed specific markers of mammary epithelial cells. BIO inhibited the proliferation of GMECs and decreased the expression of proliferation marker c-myc and cell cycle protein cyclin D1. However, the ability of GMECs to form spheroids was accelerated, and the level of E-cadherin mRNA was upregulated with BIO treatment. E-cadherin showed a bright cytomembrane with DMSO treatment, yet E-cadherin was present in cytomembrane and cytoplasm in GMECs with BIO treatment. Meanwhile, BIO increased the protein level of ß-catenin and enhanced the translocation of ß-catenin into the nucleus in GMECs. Furthermore, the mRNA level of Axin2 was also upregulated. This study suggested that Wnt/ß-catenin signaling may play an important role in the proliferation and the acinus-like formation of GMECs.

Towards improved breast mass detection using dual-view mammogram matching.

Breast cancer screening benefits from the visual analysis of multiple views of routine mammograms. As for clinical practice, computer-aided diagnosis (CAD) systems could be enhanced by integrating multi-view information. In this work, we propose a new multi-tasking framework that combines craniocaudal (CC) and mediolateral-oblique (MLO) mammograms for automatic breast mass detection. Rather than addressing mass recognition only, we exploit multi-tasking properties of deep networks to jointly learn mass matching and classification, towards better detection performance. Specifically, we propose a unified Siamese network that combines patch-level mass/non-mass classification and dual-view mass matching to take full advantage of multi-view information. This model is exploited in a full image detection pipeline based on You-Only-Look-Once (YOLO) region proposals. We carry out exhaustive experiments to highlight the contribution of dual-view matching for both patch-level classification and examination-level detection scenarios. Results demonstrate that mass matching highly improves the full-pipeline detection performance by outperforming conventional single-task schemes with 94.78% as Area Under the Curve (AUC) score and a classification accuracy of 0.8791. Interestingly, mass classification also improves the performance of mass matching, which proves the complementarity of both tasks. Our method further guides clinicians by providing accurate dual-view mass correspondences, which suggests that it could act as a relevant second opinion for mammogram interpretation and breast cancer diagnosis.

Harnessing Photocatalytic and Photothermal Effects of C-Doped Graphitic Carbon Nitride for Efficient Bacterial Disinfection.

In this work, the photocatalytic and photothermal effects of carbon-ring-doped graphitic carbon nitride materials against bacteria were systematically studied in a dispersed solution and on a membrane. C-doped graphitic carbon nitride materials C-CN 0.15, 1.5, and 7.5 were synthesized by mixing urea precursor with 0.15, 1.5, and 7.5 wt % glucose. With the increase in the doping level, the photothermal effect was clearly enhanced while the generation of reactive oxygen species (ROS) was slightly inhibited. With exposure to irradiation under a 100 mW cm-2 Xeon lamp with a cutoff filter (λ ≥ 420 nm), the ROS concentration of C-CN 1.5 increased 30% in the dispersed solution and its temperature increased about 10 °C in the dispersed solution and on the membrane compared to that of pristine carbon nitride. As a result, the bactericidal activity of C-CN 1.5 was improved by an order of magnitude in the dispersed solution and more than 2 orders of magnitude on the membrane immersed in a solution at 40 °C. To investigate the fundamental light absorption process on the membrane, an optical model using the finite-difference time-domain method was developed based on the topography of the membrane. The simulation results may explain that although C-CN produces more ROS in a solution; however, with a larger extinction coefficient, the power absorption is lower near the surface of the membrane. The ROS production is therefore inhibited and the bactericidal activity is dominated by the photothermal effect. Our experimental and simulation results provide a basis for designing high-performance photoactive disinfection materials and surfaces.