Identification and function analysis of BCL2 in immune response of Pteria penguin.

B-cell lymphoma/leukemia-2 (BCL2), an anti-apoptotic factor in the mitochondrial regulatory pathway of apoptosis, is critically important in immune defenses. In this study, a novel BCL2 gene was characterized from Pteria penguin (P. penguin). The PpBCL2 was 1482 bp long, containing an open reading frame (ORF) of 588 bp encoding 195 amino acids. Four highly conserved BCL-2 homology (BH) domains were found in PpBCL2. Amino acid alignment and phylogenetic tree showed that PpBCL2 had the highest similarity with BCL2 of Crassostrea gigas at 65.24 %. Tissue expression analysis showed that PpBCL2 had high constitutive expression in gill, digestive diverticulum and mantle, and was significantly increased 72 h of Vibrio parahaemolyticus (V. parahaemolyticus) challenge in these immune tissues. Furthermore, PpBCL2 silencing significantly inhibited antimicrobial activity of hemolymph supernatant by 1.4-fold, and significantly reduced the survival rate by 51.7 % at 72 h post infection in P. penguin. These data indicated that PpBCL2 played an important role in immune response of P. penguin against V. parahaemolyticus infection.

Clinical outcomes of self-glazed zirconia veneers produced by 3D gel deposition: a retrospective study.

BACKGROUND: Self-glazed zirconia (SZ) restorations are made by a novel additive three-dimensional gel deposition approach, which are suitable for a straightforward completely digital workflow. SZ has recently been used as minimally invasive veneer, but its clinical outcomes have not been clarified yet. This study aimed to evaluate the preliminary clinical outcomes of SZ veneers compared with the widely used lithium disilicate glass-ceramic veneers made by either pressing (PG) or milling (MG) process. METHODS: Fifty-six patients treated with SZ, PG, and MG veneers by 2 specialists between June 2018 and October 2022 were identified. Patients were recalled for follow-up at least 1 year after restoration. Clinical outcomes were assessed by 2 independent evaluators according to the modified United States Public Health Service (USPHS) criteria. Overall patient satisfaction was assessed using visual analogue scale (VAS), and analyzed by one-way ANOVA. Chi-square test was applied to compare the difference in the success and survival rates among the 3 groups. RESULTS: A total of 51 patients restored with 45 SZ, 40 PG, and 41 MG veneers completed the study, with a patient dropout rate of 8.9%. Mean and standard deviation of follow-up period was 35.0 ± 14.7 months. All restorations performed well at baseline, except for 2 SZ veneers with mismatched color (rated Bravo). During follow-up, marginal discrepancy (rated Bravo) was found in 4 MG veneers and 1 PG veneer, and partially fractured (rated Charlie) was found in another 2 PG veneers. The survival rate of SZ, PG, and MG veneers was 100%, 95%, and 100%, with a success rate of 95.56%, 92.50%, and 90.24%, respectively, none of which were significantly different (p = 0.099 and 0.628, respectively). The mean VAS score of SZ, PG, and MG was 95.00 ± 1.57, 93.93 ± 2.40, and 94.89 ± 2.00 respectively, without significant difference (p > 0.05). CONCLUSION: SZ veneers exhibited comparable preliminary clinical outcomes to PG and MG veneers, which could be considered as a feasible option for minimally invasive restorative treatment.

Insights into a functional model of key deubiquitinases UBP12/13 in plants.

Understanding the complexities of protein ubiquitination is crucial, as it plays a multifaceted role in controlling protein stability, activity, subcellular localization, and interaction, which are central to diverse biological processes. Deubiquitinases (DUBs) serve to reverse ubiquitination, but research progress in plant DUBs is noticeably limited. Among existing studies, UBIQUITIN-SPECIFIC PROTEASE 12 (UBP12) and UBP13 have garnered attention for their extensive role in diverse biological processes in plants. This review systematically summarizes the recent advancements in UBP12/13 studies, emphasizing their function, and their substrate specificity, their relationship with E3 ubiquitin ligases, and the similarities and differences with their mammalian orthologue, USP7. By unraveling the molecular mechanisms of UBP12/13, this review offers in-depth insights into the ubiquitin-proteasome system (UPS) in plants and aims to catalyze further explorations and comprehensive understanding in this field.

Evaluation of choroidal thickness and retinal nerve fiber layer thickness in Chinese pregnant women and healthy non-pregnant women.

Purpose: To evaluate choroidal thickness and retinal nerve fiber layer (RNFL) thickness in different trimesters using enhanced depth imaging optical coherence tomography (EDI-OCT). Methods: A prospective comparative study included 45 healthy pregnant women in the first trimester, 45 women in the second, 45 women in the third and 45 healthy non-pregnant women as the control group. Macular choroidal thickness was measured at three locations: The subfoveal, 1 â€‹mm temporal, and 1 â€‹mm nasal from the fovea with EDI-OCT. Peripapillary choroidal thickness (PPCT) and RNFL thickness parameters were automatically calculated by the Spectralis OCT. Results: The subfoveal, temporal and nasal macular choroidal thickness were all significantly thicker in the second trimester, compared with those parameters in the first, the third trimesters and the control group (all P â€‹< â€‹0.05). The PPCT was significantly increased in the second trimeter compared with the control group at global, temporal, temporal inferior, nasal and nasal inferior positions (all P â€‹< â€‹0.05). The RNFL thickness was also significantly increased in pregnant women at nasal superior and nasal inferior quadrants (all P â€‹< â€‹0.05). Conclusions: The choroidal thickness in pregnant women was found to be thicker than the control group, regardless of macular or optic disc location. Findings of RNFL thickening might indicate subclinical involvement of the central nervous system.

ER-associated ubiquitin-conjugating enzyme: a key regulator of grain yield and stress resistance in crops.

Recent research reveals the critical roles of endoplasmic reticulum (ER)-associated protein degradation (ERAD)-related ubiquitin-conjugating enzyme AtUBC32 orthologs and their partnering E3 ligases, which play dual roles in enhancing both crop yield and stress resistance. These findings open avenues for breeding high-yield, stress-tolerant crops and inspire further exploration of the ERAD pathway in agricultural innovation.

A smartphone-based fluorescent sensor for rapid detection of multiple pathogenic bacteria.

In this study, we developed a fluorescent sensor for the sensitive detection of multiple pathogenic bacteria based on magnetic separation, fluorescent probes, and smartphone image processing. A microchannel device was assembled using high-transparency resin and 3D printing technology. This device was combined with a smartphone and an external lens to develop a fluorescent sensor for autonomous detection of multiple pathogenic bacteria. Three fluorescence probes with different fluorescence were synthesized from highly specific aptamers and tetraphenylethylene derivatives. These fluorescent probes can make Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa emit different colors of fluorescence. Using the enrichment performance of molecularly imprinted materials, separation and detection of bacteria can be achieved simultaneously. Finally, with the Red-Green-Blue (RGB) analysis functionality of a smartphone, real-time field detection was realized with a sensitivity of 102 CFU/mL and a detection time of 40 min. This work provides a simple, inexpensive, and real-time sensor for the detection of multiple pathogens in medical diagnostics, food testing, and environmental analyses.

Structure and activation mechanism of the rice Salt Overly Sensitive 1 (SOS1) Na+/H+ antiporter.

Salinity is one of the most severe abiotic stresses that adversely affect plant growth and agricultural productivity. The plant Na+/H+ antiporter Salt Overly Sensitive 1 (SOS1) located in the plasma membrane extrudes excess Na+ out of cells in response to salt stress and confers salt tolerance. However, the molecular mechanism underlying SOS1 activation remains largely elusive. Here we elucidate two cryo-electron microscopy structures of rice (Oryza sativa) SOS1, a full-length protein in an auto-inhibited state and a truncated version in an active state. The SOS1 forms a dimeric architecture, with an NhaA-folded transmembrane domain portion in the membrane and an elongated cytosolic portion of multiple regulatory domains in the cytoplasm. The structural comparison shows that SOS1 adopts an elevator transport mechanism accompanied by a conformational transition of the highly conserved Pro148 in the unwound transmembrane helix 5 (TM5), switching from an occluded conformation in the auto-inhibited state to a conducting conformation in the active state. These findings allow us to propose an inhibition-release mechanism for SOS1 activation and elucidate how SOS1 controls Na+ homeostasis in response to salt stress.

Patient satisfaction with humanistic nursing in Chinese secondary and tertiary public hospitals: a cross-sectional survey.

Background: Humanistic care pertains to the abilities, attitudes, and behaviors central to patient-centered care, contributing to patients' sense of safety and wellbeing. This study aimed to assess the satisfaction of patients with humanistic nursing care in Chinese secondary and tertiary public hospitals. Methods: A national cross-sectional survey was conducted across 30 provinces and 83 hospitals in China. Patient satisfaction with humanistic care was assessed using the Methodist Health Care System Nurse Caring Instrument (NCI), which encompasses 20 items across 12 dimensions. Each item was rated on a 7-point Likert scale, yielding a total score of 140. Multiple linear regression analysis was employed to identify factors associated with patients' satisfaction. Results: Moderate satisfaction (mean score 91.26 ± 13.14) with humanistic nursing care was observed among the 17,593 participants. Factors significantly associated with patient satisfaction included age, hospital type, presence of children, educational attainment, place of residence, family monthly income, and medical insurance type. Conclusion: The study findings highlight the importance of tailored interventions, evidence-based practice guidelines, and patient-centered care in improving patients' satisfaction with humanistic nursing care. Continuous emphasis on nursing education and professional development is crucial for enhancing humanistic care and patient satisfaction.

The enhancement in toxic potency of oxidized functionalized polyethylene-microplastics in mice gut and Caco-2 cells.

Microplastics (MPs) are inevitably oxidized in the environment, however, to date, no studies have discussed the biological toxicity of oxidized polyethylene (Ox-PE) MPs. In this study, oxidized low-density polyethylene (Ox-LDPE), a representative Ox-PE, was prepared using a selective oxidation method. The difference in toxicity between LDPE-MPs and Ox-LDPE-MPs were evaluated in C57BL/6 mice and Caco-2 cells. The proton nuclear magnetic resonance (1H NMR) and Fourier transform infrared (FTIR) spectroscopy analyses revealed that some hydrocarbon-containing groups were transformed into carboxyl and ketone groups during selective oxidation. In vivo experiment results showed that LDPE-MPs and Ox-LDPE-MPs exists in the intestinal (duodenum and colon) of mice, and Ox-LDPE-MPs caused more severe intestinal histological changes, oxidative stress, and inflammatory response. The gut microbiota data showed that the relative abundance of Lactobacillus decreased significantly in the LDPE-MP- and Ox-LDPE-MP-exposed groups (P < 0.05). The predicted Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway suggested that exposure to LDPE-MPs or Ox-LDPE-MPs inhibited glycan biosynthesis and metabolism in the flora (P < 0.05). In vitro experiment results showed that selective oxidation to LDPE promoted its uptake by cells and aggravated adverse effects on cells, including reduced cell viability, damaged cell membrane, oxidative stress, and mitochondrial depolarization. The major mechanism of the increased toxicity of Ox-LDPE-MPs may be its easier accumulation and the ionic effect of oxygen-containing functional groups. Overall, these findings provide insights on the differences in toxicity between LDPE-MPs and Ox-LDPE-MPs. They also provide new perspectives for understanding the biohazards of MPs, which are necessary to accurately assess the potential environmental and health risks of these plastic pollutants.

The intestinal microbiota and metabolic profiles of Strauchbufo raddei underwent adaptive changes during hibernation.

The intestinal microbiota help regulate hibernation in vertebrates. However, it needs to be established how hibernation modulates the gut microbiome and intestinal metabolism. In the present study, we used an artificial hibernation model to examine the responses of the gut microbiota of the Strauchbufo raddei to the environmental changes associated with this behavior. Hibernation significantly lowered the diversity of the microbiota and altered the microbial community of the gut. Proteobacteria, Firmicutes, and Bacteroidota were the major bacterial phyla in the intestines of S. raddei. However, Firmicutes and Proteobacteria predominated in the gut of active and hibernating S. raddei, respectively. Certain bacterial genera such as Pseudomonas, Vibrio, Ralstonia, and Rhodococcus could serve as biomarkers distinguishing hibernating and non-hibernating S. raddei. The gut microbiota was more resistant to environmental stress in hibernating than active S. raddei. Moreover, metabolomics revealed that metabolites implicated in fatty acid biosynthesis were highly upregulated in the intestines of hibernating S. raddei. The metabolites that were enriched during hibernation enabled S. raddei to adapt to the low temperatures and the lack of exogenous food that are characteristic of hibernation. A correlation analysis of the intestinal microbiota and their metabolites revealed that the gut microbiota might participate in the metabolic regulation of hibernating S. raddei. The present study clarified the modifications that occur in the intestinal bacteria and their symbiotic relationship with their host during hibernation. These findings are indicative of the adaptive changes in the metabolism of amphibians under different environmental conditions.

Artificial intelligence in the diagnosis of dental diseases on panoramic radiographs: a preliminary study.

BACKGROUND: Artificial intelligence (AI) has been introduced to interpret the panoramic radiographs (PRs). The aim of this study was to develop an AI framework to diagnose multiple dental diseases on PRs, and to initially evaluate its performance. METHODS: The AI framework was developed based on 2 deep convolutional neural networks (CNNs), BDU-Net and nnU-Net. 1996 PRs were used for training. Diagnostic evaluation was performed on a separate evaluation dataset including 282 PRs. Sensitivity, specificity, Youden's index, the area under the curve (AUC), and diagnostic time were calculated. Dentists with 3 different levels of seniority (H: high, M: medium, L: low) diagnosed the same evaluation dataset independently. Mann-Whitney U test and Delong test were conducted for statistical analysis (ɑ=0.05). RESULTS: Sensitivity, specificity, and Youden's index of the framework for diagnosing 5 diseases were 0.964, 0.996, 0.960 (impacted teeth), 0.953, 0.998, 0.951 (full crowns), 0.871, 0.999, 0.870 (residual roots), 0.885, 0.994, 0.879 (missing teeth), and 0.554, 0.990, 0.544 (caries), respectively. AUC of the framework for the diseases were 0.980 (95%CI: 0.976-0.983, impacted teeth), 0.975 (95%CI: 0.972-0.978, full crowns), and 0.935 (95%CI: 0.929-0.940, residual roots), 0.939 (95%CI: 0.934-0.944, missing teeth), and 0.772 (95%CI: 0.764-0.781, caries), respectively. AUC of the AI framework was comparable to that of all dentists in diagnosing residual roots (p > 0.05), and its AUC values were similar to (p > 0.05) or better than (p < 0.05) that of M-level dentists for diagnosing 5 diseases. But AUC of the framework was statistically lower than some of H-level dentists for diagnosing impacted teeth, missing teeth, and caries (p < 0.05). The mean diagnostic time of the framework was significantly shorter than that of all dentists (p < 0.001). CONCLUSIONS: The AI framework based on BDU-Net and nnU-Net demonstrated high specificity on diagnosing impacted teeth, full crowns, missing teeth, residual roots, and caries with high efficiency. The clinical feasibility of AI framework was preliminary verified since its performance was similar to or even better than the dentists with 3-10 years of experience. However, the AI framework for caries diagnosis should be improved.

IDO Regulates Macrophage Functions by Inhibiting the CCL2/CCR2 Signaling Pathway in Fungal Keratitis.

PURPOSE: The aim of this study was to investigate the effects of indoleamine 2,3-dioxygenase (IDO) on macrophage polarization, phagocytosis, and killing through regulation of the CCL2/CCR2 signaling pathway in Aspergillus fumigatus keratitis. METHODS: In vivo and in vitro experiments were conducted in mice and mouse peritoneal macrophages after infection with A. fumigatus . Clinical scoring, reverse transcription-polymerase chain reaction, and immunofluorescence staining were used to evaluate the fungal keratitis lesions, macrophage-related cytokines, and macrophage recruitment. The expression of CCL2 and CCR2 was detected by reverse transcription-polymerase chain reaction and western blot after pretreatment with or without an IDO inhibitor (1-MT). After pretreatment with 1-MT, a CCR2 antagonist, a CCL2 neutralizing antibody, an IDO agonist (IFNG), and recombinant CCL2 protein (CCL2), the flow cytometry and colony-forming unit counts were used to detect the polarization, phagocytosis, and killing function. RESULTS: Compared with the control group, the infected eyes showed increased clinical scores, macrophage-related cytokine expression, and macrophage recruitment. 1-MT pretreatment increased the expression of CCL2 and CCR2 and the proportion of CD206+/CD86+ macrophages; macrophages polarized toward the M2 type, with enhanced killing function. CCR2 antagonists and CCL2 neutralizing antibodies reversed the effects of 1-MT. Compared with the infected group, IFNG pretreatment decreased the proportion of CD206+/CD86+ macrophages, and macrophages polarized toward the M1 type, with decreased phagocytosis and impaired killing function. CCL2 reversed the effect of IFNG. CONCLUSIONS: IDO can promote the polarization of macrophages to the M1 type by blocking the CCL2/CCR2 signaling pathway, inhibiting the phagocytosis and killing function of macrophages, and mediating the protective immune role of A. fumigatus .

Creb2 involved in innate immunity by activating PpMitf-mediated melanogenesis in Pteria penguin.

cAMP response element binding protein 2 (CREB2) acts as an intracellular transcriptional factor and regulates many physiological processes, including melanogenesis and melanocyte differentiation. In our previous research, the Creb2 gene has been characterized from Pteria penguin (P. penguin), but its role and regulatory mechanism in P. penguin are still unclear. In this study, first, the function of PpCreb2 in melanogenesis and innate immunity were identified. PpCreb2 silencing significantly decreased the tyrosinase activity and melanin content, indicating PpCreb2 played an important role in melanogenesis. Meanwhile, PpCreb2 silencing visibly suppressed the antibacterial activity of hemolymph supernatant, indicating that PpCreb2 was involved in innate immunity of P. penguin. Second, the PpCreb2 was confirmed to perform immune function by regulating the melanogenesis. The decreased melanin oxidation product due to PpCreb2 silencing triggered the declining of antibacterial activity of hemolymph supernatant, which then could be rescued by adding exogenous melanin oxidation products. Third, the regulation pathway of PpCreb2 involved in innate immunity was analyzed. The promoter sequence analysis of PpMitf discovered 5 conserved cAMP response element (CRE), which were specifically recognized by basic Leucine zipper domain (bZIP) of upstream activation transcription factor. The luciferase activities analysis showed that PpCreb2 could activate the CRE in PpMitf promoter via highly conserved bZIP domain and regulate the expression of PpMitf, which further regulated the PpTyr expression. Therefore, the results collectively demonstrated that PpCreb2 participated in innate immunity by activating PpMitf-mediated melanogenesis in P. penguin.

Exposure to nitrate induced growth, intestinal histology and microbiota alterations of Bufo raddei Strauch tadpoles.

Nitrate (NO3-) is one of the ubiquitous environmental chemicals which multiplies negative impacts on aquatic life such as amphibian larvae. However, the data involving the dynamics of amphibians in response to NO3-N are scarce. This study investigated the effects of NO3-N on locomotor ability, growth performance, oxidative stress parameters, intestinal histology, and intestinal microbiota of Bufo raddei Strauch tadpoles. The tadpoles were chronically exposed to different concentrations of NO3-N (10, 50, 100, and 200 mg/L) from Gosner stage 26 to 38. Our results revealed that NO3-N exposure caused significantly reduced body weight and length, impaired locomotor activity, and severe oxidative damage to liver tissue. Moreover, the high NO3-N (50, 100, and 200 mg/L) exposure caused irregular arrangement and indistinct cell borders of mucosal epithelial cells in the tadpoles intestine. The NO3-N exposure significantly changed the structure of the intestinal microbiota. The phylum Cyanobacteria occupy the main niche of intestinal microbes and have a certain negative correlation with the growth and motility of tadpoles. In addition, the functional prediction revealed that NO3-N exposure obviously downregulated the metabolism of enzyme families in tadpoles. Our comprehensive research shows the toxicity of NO3-N exposure in B. raddei Strauch, explores the potential links between development and intestinal microbiota of tadpole, and provides a new framework for the potential health risk of nitrate in amphibians.

A Gγ protein regulates alkaline sensitivity in crops.

The use of alkaline salt lands for crop production is hindered by a scarcity of knowledge and breeding efforts for plant alkaline tolerance. Through genome association analysis of sorghum, a naturally high-alkaline-tolerant crop, we detected a major locus, Alkaline Tolerance 1 (AT1), specifically related to alkaline-salinity sensitivity. An at1 allele with a carboxyl-terminal truncation increased sensitivity, whereas knockout of AT1 increased tolerance to alkalinity in sorghum, millet, rice, and maize. AT1 encodes an atypical G protein γ subunit that affects the phosphorylation of aquaporins to modulate the distribution of hydrogen peroxide (H2O2). These processes appear to protect plants against oxidative stress by alkali. Designing knockouts of AT1 homologs or selecting its natural nonfunctional alleles could improve crop productivity in sodic lands.

TLR4 involved in immune response against Vibrio Parahaemolyticus by MyD88-dependent pathway in Crassostrea hongkongensis.

Vibrio parahaemolyticus (V. parahaemolyticus) is a salt-loving gram-negative bacterium, and is the leading cause of mortality in cultured shellfish in recent years. Toll-like Receptor 4 (TLR4) is a classical pattern recognition receptor (PRRs) that recognizes pathogen-associated molecular patterns (PAMPs) of pathogenic microorganism and activates the immune response. However, the function and signal pathway of TLR4 in oyster are still unknown. In this study, a new TLR4 gene was identified from the Crassostrea hongkongensis (C. hongkongensis). The ChTLR4 contained an open reading frame of 2643 bp, encoding 880 amino acids with seven leucine-rich repeat (LRR) domains and a Toll/IL-1R (TIR) domain. The ChTLR4 shared the highest sequence identity (83.0%) with TLR4 of Crassostrea gigas. Tissue expression analysis revealed that ChTLR4 showed the highest constitutive expression in the gill and hepatopancreas, and was significantly upregulated in immune tissues post V. parahaemolyticus infection, especially in gill and hemocytes. Moreover, TLR4 silencing significantly inhibited the immune-enzyme activities, including SOD, CAT, ACP, AKP in gill and LZM in hemolymph supernatant, and increased MDA content in hemolymph supernatant. Meanwhile, the antimicrobial activities of the hemolymph supernatant were also significantly inhibited by TLR4 silencing. These data demonstrated that the ChTLR4 involved in innate immune response of C. hongkongensis against V. parahaemolyticus challenge. Finally, qRT-PCR analysis showed that ChTLR4 silencing clearly inhibited the expression of genes in TLR4-MyD88 pathway, indicating that MyD88-dependent pathway played a crucial role in ChTLR4-mediated immune response against V. parahaemolyticus.