The Dual Role of the NFATc2/galectin-9 Axis in Modulating Tumor-Initiating Cell Phenotypes and Immune Suppression in Lung Adenocarcinoma.

Tumor-initiating cells (TICs) resilience and an immunosuppressive microenvironment are aggressive oncogenic phenotypes that contribute to unsatisfactory long-term outcomes in lung adenocarcinoma (LUAD) patients. The molecular mechanisms mediating the interaction between TICs and immune tolerance have not been elucidated. The role of Galectin-9 in oncogenesis and immunosuppressive microenvironment is still unknown. This study explored the potential role of galectin-9 in TIC regulation and immune modulation in LUAD. The results show that galectin-9 supports TIC properties in LUAD. Co-culture of patient-derived organoids and matched peripheral blood mononuclear cells showed that tumor-secreted galectin-9 suppressed T cell cytotoxicity and induced regulatory T cells (Tregs). Clinically, galectin-9 is upregulated in human LUAD. High expression of galectin-9 predicted poor recurrence-free survival and correlated with high levels of Treg infiltration. LGALS9, the gene encoding galectin-9, is found to be transcriptionally regulated by the nuclear factor of activated T cells 2 (NFATc2), a previously reported TIC regulator, via in silico prediction and luciferase reporter assays. Overall, the results suggest that the NFATc2/galectin-9 axis plays a dual role in TIC regulation and immune suppression.

Thermally and Photothermally Triggered Cytocompatible Triple-Shape-Memory Polymer Based on a Graphene Oxide-Containing Poly(ε-caprolactone) and Acrylate Composite.

Triple-shape-memory polymers (triple-SMPs) are a class of polymers capable of fixing two temporary shapes and recovering sequentially from the first temporary shape to the second temporary shape and, last, to the permanent shape. To accomplish a sequential shape change, a triple-SMP must have two separate shape-fixing mechanisms triggerable by distinct stimuli. Despite the biomedical potential of triple-SMPs, a triple-SMP that with cells present can undergo two different shape changes via two distinct cytocompatible triggers has not previously been demonstrated. Here, we report the design and characterization of a cytocompatible triple-SMP material that responds separately to thermal and light triggers to undergo two distinct shape changes under cytocompatible conditions. Tandem triggering was achieved via a photothermally triggered component, comprising poly(ε-caprolactone) (PCL) fibers with graphene oxide (GO) particles physically attached, embedded in a thermally triggered component, comprising a tert-butyl acrylate-butyl acrylate (tBA-BA) matrix. The material was characterized in terms of thermal properties, surface morphology, shape-memory performance, and cytocompatibility during shape change. Collectively, the results demonstrate cytocompatible triple-shape behavior with a relatively larger thermal shape change (an average of 20.4 ± 4.2% strain recovered for all PCL-containing groups) followed by a smaller photothermal shape change (an average of 3.5 ± 0.8% strain recovered for all PCL-GO-containing groups; samples without GO showed no recovery) with greater than 95% cell viability on the triple-SMP materials, establishing the feasibility of triple-shape memory to be incorporated into biomedical devices and strategies.

Exploring the Potential of ChatGPT-4 in Responding to Common Questions About Abdominoplasty: An AI-Based Case Study of a Plastic Surgery Consultation.

BACKGROUND: With the increasing integration of artificial intelligence (AI) in health care, AI chatbots like ChatGPT-4 are being used to deliver health information. OBJECTIVES: This study aimed to assess the capability of ChatGPT-4 in answering common questions related to abdominoplasty, evaluating its potential as an adjunctive tool in patient education and preoperative consultation. METHODS: A variety of common questions about abdominoplasty were submitted to ChatGPT-4. These questions were sourced from a question list provided by the American Society of Plastic Surgery to ensure their relevance and comprehensiveness. An experienced plastic surgeon meticulously evaluated the responses generated by ChatGPT-4 in terms of informational depth, response articulation, and competency to determine the proficiency of the AI in providing patient-centered information. RESULTS: The study showed that ChatGPT-4 can give clear answers, making it useful for answering common queries. However, it struggled with personalized advice and sometimes provided incorrect or outdated references. Overall, ChatGPT-4 can effectively share abdominoplasty information, which may help patients better understand the procedure. Despite these positive findings, the AI needs more refinement, especially in providing personalized and accurate information, to fully meet patient education needs in plastic surgery. CONCLUSIONS: Although ChatGPT-4 shows promise as a resource for patient education, continuous improvements and rigorous checks are essential for its beneficial integration into healthcare settings. The study emphasizes the need for further research, particularly focused on improving the personalization and accuracy of AI responses. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

CFTR Modulates Hypothalamic Neuron Excitability to Maintain Female Cycle.

Cystic fibrosis transmembrane conductance regulator (CFTR), known as an epithelial Cl- channel, is increasingly noted to be expressed in the nervous system, although whether and how it plays a role in neuronal excitability is unclear. Given the association of CFTR with fertility, we tested here possible involvement of CFTR in regulating hypothalamic neuron excitability. Patch-clamp and Ca2+ imaging showed that pharmacological inhibition of CFTR evoked electrical pulses and Ca2+ spikes in primary rat hypothalamic neurons, which was dependent on extracellular Cl-. Hypothalamic neurons in brain-slice preparations from adult female mice with CFTR mutation (DF508) exhibited significantly reduced electrical pulses as compared to the wild-type controls. Removal of extracellular Cl- eliminated hypothalamic electrical pulses in the wild-type brain slices, which was reversible by subsequent addition of Cl-. In adult female mice, Ca2+ indicator (GCaMP6s)-based fiber-photometry showed that hypothalamic Ca2+ activities in vivo were enhanced at the proestrus/estrus phase as compared to the diestrus phase of the female cycle. Such estrus-associated hypothalamic activities were largely diminished in DF508 female mice, together with delayed puberty and disturbed female cycles. Therefore, these findings suggest a critical role of CFTR in modulating hypothalamic neuron excitability, which may account for the disturbed female cycles and reduced female fertility associated with CFTR mutations.

Metagenomics insights into high-rate nitrogen removal from municipal wastewater by integrated nitrification, partial denitrification and Anammox at an extremely short hydraulic retention time.

To achieve high-rate nitrogen removal in municipal wastewater treatment through anaerobic ammonia oxidation (Anammox), the nitrification, partial denitrification, and Anammox processes were integrated by a step-feed strategy. An exceptional nitrogen removal load of 0.224 kg N/(m3·d) was achieved by gradient-reducing the hydraulic retention time (HRT) to 5 h. Metagenomic analysis demonstrated that Nitrosospira could express all genes encoding ammonia oxidation under low nitrogen and dissolved oxygen conditions (less than 0.5 mg/L), enabling complete nitrification. With the short of HRT, the relative abundance of Thauera increased from 2.8 % to 6.4 %. Frequent substrate exchanges at such extremely short HRT facilitated enhanced synergistic interactions among Nitrosospira, Thauera, and Candidatus Brocadia. These findings provide a comprehensive understanding of the utilization of Anammox combined processes for high-speed nitrogen removal in municipal wastewater treatment and the microbial interactions involved.

Efficient nitrogen removal from municipal wastewater by an autotrophic-heterotrophic coupled anammox system: The up-regulation of key functional genes.

The metabolic pathways based on key functional genes were innovatively revealed in the autotrophic-heterotrophic coupled anammox system for real municipal wastewater treatment. The nitrogen removal performance of the system was stabilized at 88.40 ± 3.39 % during the treatment of real municipal wastewater. The relative abundances of the nitrification functional genes ammonia oxidase (amoA/B/C), hydroxylamine oxidoreductase (hao), and nitrite oxidoreductases (nxrA/B) were increased by 1.2-2.4 times, and these three nitrification functional genes were mostly contributed by Nitrospira that dominated the efficient nitrification of the system. The relative abundance of anammox bacteria Candidatus Brocadia augmented from 0.35 % to 0.75 %, accompanied with the increased expression of hydrazine synthase (hzs) and hydrazine dehydrogenase (hdh), resulting in the major role of anammox (81.24 %) for nitrogen removal. The expression enhancement of the functional genes nitrite reductase (narG/H, napA/B) that promoted partial denitrification (PD) of the system weakened the adverse effects of the sharp decline in the population of PD microbe Thauera (from 5.7 % to 2.2 %). The metabolic module analysis indicated that the carbon metabolism pathways of the system mainly included CO2 fixation and organic carbon metabolism, and the stable enrichment of autotrophic bacteria ensured stable CO2 fixation. Furthermore, the enhanced expression of the glucokinases (glk, GCK, HK, ppgk) and the abundant pyruvate kinase (PK) achieved stable hydrolysis ability of organic carbon metabolism function of the system. This study offers research basics to practical application of the mainstream anammox process.

A critical review of improving mainstream anammox systems: Based on macroscopic process regulation and microscopic enhancement mechanisms.

Full-scale anaerobic ammonium oxidation (anammox) engineering applications are vastly limited by the sensitivity of anammox bacteria to the complex mainstream ambience factors. Therefore, it is of great necessity to comprehensively summarize and overcome performance-related challenges in mainstream anammox process at the macro/micro level, including the macroscopic process variable regulation and microscopic biological metabolic enhancement. This article systematically reviewed the recent important advances in the enrichment and retention of anammox bacteria and main factors affecting metabolic regulation under mainstream conditions, and proposed key strategies for the related performance optimization. The characteristics and behavior mechanism of anammox consortia in response to mainstream environment were then discussed in details, and we revealed that the synergistic nitrogen metabolism of multi-functional bacterial genera based on anammox microbiome was conducive to mainstream anammox nitrogen removal processes. Finally, the critical outcomes of anammox extracellular electron transfer (EET) at the micro level were well presented, carbon-based conductive materials or exogenous electron shuttles can stimulate and mediate anammox EET in mainstream environments to optimize system performance from a micro perspective. Overall, this review advances the extensive implementation of mainstream anammox practice in future as well as shedding new light on the related EET and microbial mechanisms.

Targeting GSTP1 as Therapeutic Strategy against Lung Adenocarcinoma Stemness and Resistance to Tyrosine Kinase Inhibitors.

Glutathione S-transferase pi (GSTP1), a phase II detoxification enzyme, is known to be overexpressed and mediates chemotherapeutic resistance in lung cancer. However, whether GSTP1 supports cancer stem cells (CSCs) and the underlying mechanisms in lung adenocarcinoma (LUAD) remain largely unknown. This study unveiled that GSTP1 is upregulated in lung CSCs and supports tumor self-renewal, metastasis, and resistance to targeted tyrosine kinase inhibitors of LUAD both in vitro and in vivo. Mechanistically, CaMK2A (calcium/calmodulin-dependent protein kinase 2 isoform A)/NRF2 (nuclear factor erythroid 2-related factor 2)/GSTP1 is uncovered as a regulatory axis under hypoxia. CaMK2A increased GSTP1 expression through phosphorylating the Sersine558 residue of NRF2 and promoting its nuclear translocation, a novel mechanism for NRF2 activation apart from conventional oxidization-dependent activation. Upregulation of GSTP1 in turn suppressed reactive oxygen species levels and supported CSC phenotypes. Clinically, GSTP1 analyzed by immunohistochemistry is upregulated in a proportion of LUAD and serves as a prognostic marker for survival. Using patient-derived organoids from an ALK-translocated LUAD, the therapeutic potential of a specific GSTP1 inhibitor ezatiostat in combination treatment with the ALK inhibitor crizotinib is demonstrated. This study demonstrates GSTP1 to be a promising therapeutic target for long-term control of LUAD through targeting CSCs.

The associated factors of earphone usage on hearing impairment among students aged 14 to 28 / 中国学校卫生

Objective@#To investigate the relationship between the use of earphone and hearing impairment and its influencing factors among students aged 14-28, so as to provide a reference for appropriate earphone usage and hearing impairment prevention.@*Methods@#A cross sectional survey was conducted through the questionnaire star platform, and 983 students aged 14 to 28 were recruited across China by snowball sampling during April 3 to May 1, 2022. The χ 2 test was used to identify indicators affecting hearing, the Logistic regression model was used to further selection.@*Results@#There were 366 students with hearing impairment, accounting for 37.23%. Univariate analysis showed significant differences in hearing impairment by gender, earphone usage duration and volume, wearing during sleep, and replacement frequency ( χ 2=6.03, 6.86, 14.87, 12.22, 11.15, P <0.05). The Logistic regression model analysis showed that girls ( OR=1.43, 95%CI =1.10-1.88), maximum earphone volume ( OR=3.08, 95%CI = 1.56- 6.08), earphone usage for >1.5-3 h each time ( OR=1.44, 95%CI =1.04-1.99), sleep with headphone ( OR= 1.53 , 95%CI = 1.11- 2.11) were positively associated with hearing impairment ( P <0.05), earphone replacement every 4-<6 months ( OR= 0.38, 95%CI =0.17-0.86) and earphone replacement every six months or longer ( OR=0.39, 95%CI =0.18-0.85) were negatively associated with hearing impairment ( P < 0.05 ).@*Conclusion@#Students aged 14 to 28 earphone usage shows adverse impact on hearing. When using earphone, it is recommended to limit time spent on earphone usage, low the volume of earphone, avoid sleeping with earphone and replace earphone frequently.

Potassium channel-related genes are a novel prognostic signature for the tumor microenvironment of renal clear cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) accounts for 80% of renal cell carcinomas (RCCs), and its morbidity and prognosis are unfavorable. Surgical resection is the first-line treatment for ccRCC, but the oncogenesis of ccRCC is very complex. With the development of high-throughput sequencing technology, it is necessary to analyze the transcriptome to determine more effective treatment methods. The tumor microenvironment (TME) is composed of tumor cells, various immune-infiltrating cells, fibroblasts, many cytokines, and catalysts. It is a complex system with a dynamic balance that plays an essential role in tumor growth, invasion, and metastasis. Previous studies have confirmed that potassium channels can affect the immune system, especially T lymphocytes that require potassium channel activation. However, the effect of potassium channels on the TME of ccRCC remains to be studied. Therefore, this study aims to construct a prognostic signature for ccRCC patients based on potassium ion channel-related genes (PCRGs), assess patient risk scores, and divide patients into high- and low-risk groups based on the cutoff value. In addition, we investigated whether there were differences in immune cell infiltration, immune activator expression, somatic mutations, and chemotherapeutic responses between the high- and low-risk groups. Our results demonstrate that the PCRG signature can accurately assess patient prognosis and the tumor microenvironment and predict chemotherapeutic responses. In summary, the PCRG signature could serve as an auxiliary tool for the precision treatment of ccRCC.

Biphasic regulation of CFTR expression by ENaC in epithelial cells: The involvement of Ca2+-modulated cAMP production.

The regulatory interaction between two typical epithelial ion channels, cystic fibrosis transmembrane conductance regulator (CFTR) and the epithelial sodium channel (ENaC), for epithelial homeostasis has been noted, although the underlying mechanisms remain unclear. Here, we report that in a human endometrial epithelial cell line (ISK), shRNA-based stable knockdown of ENaC produced a biphasic effect: a low (∼23%) degree of ENaC knockdown resulted in significant increases in CFTR mRNA and protein levels, CFTR-mediated Cl- transport activity as well as intracellular cAMP concentration, while a higher degree (∼50%) of ENaC knockdown did not further increase but restored CFTR expression and cAMP levels. The basal intracellular Ca2+ level of ISK cells was lowered by ENaC knockdown or inhibition in a degree-dependent manner. BAPTA-AM, an intracellular Ca2+ chelator that lowers free Ca2+ concentration, elevated cAMP level and CFTR mRNA expression at a low (5 µM) but not a high (50 µM) dose, mimicking the biphasic effect of ENaC knockdown. Moreover, KH-7, a selective inhibitor of soluble adenylyl cyclase (sAC), abolished the CFTR upregulation induced by low-degree ENaC knockdown or Ca2+ chelation, suggesting the involvement of sAC-driven cAMP production in the positive regulation. A luciferase reporter to indicate CFTR transcription revealed that all tested degrees of ENaC knockdown/inhibition stimulated CFTR transcription in ISK cells, suggesting that the negative regulation on CFTR expression by the high-degree ENaC deficiency might occur at post-transcription stages. Additionally, similar biphasic effect of ENaC knockdown on CFTR expression was observed in a human bronchial epithelial cell line. Taken together, these results have revealed a previously unidentified biphasic regulatory role of ENaC in tuning CFTR expression involving Ca2+-modulated cAMP production, which may provide an efficient mechanism for dynamics and plasticity of the epithelial tissues in various physiological or pathological contexts.

Cell-Responsive Shape Memory Polymers.

Recent decades have seen substantial interest in the development and application of biocompatible shape memory polymers (SMPs), a class of "smart materials" that can respond to external stimuli. Although many studies have used SMP platforms triggered by thermal or photothermal events to study cell mechanobiology, SMPs triggered by cell activity have not yet been demonstrated. In a previous work, we developed an SMP that can respond directly to enzymatic activity. Here, our goal was to build on that work by demonstrating enzymatic triggering of an SMP in response to the presence of enzyme-secreting human cells. To achieve this phenomenon, poly(ε-caprolactone) (PCL) and Pellethane were dual electrospun to form a fiber mat, where PCL acted as a shape-fixing component that is labile to lipase, an enzyme secreted by multiple cell types including HepG2 (human hepatic cancer) cells, and Pellethane acted as a shape memory component that is enzymatically stable. Cell-responsive shape memory performance and cytocompatibility were quantitatively and qualitatively analyzed by thermal analysis (thermal gravimetric analysis and differential scanning calorimetry), surface morphology analysis (scanning electron microscopy), and by incubation with HepG2 cells in the presence or absence of heparin (an anticoagulant drug present in the human liver that increases the secretion of hepatic lipase). The results characterize the shape-memory functionality of the material and demonstrate successful cell-responsive shape recovery with greater than 90% cell viability. Collectively, the results provide the first demonstration of a cytocompatible SMP responding to a trigger that is cellular in origin.

Effective utilization of refractory dissolved organic matters in domestic sewage allows to enhanced nitrogen removal by integrated fermentation, nitrification, denitratation and anammox process.

To take full advantage of refractory dissolved organic matters (rDOMs) and generate sufficient nitrate for domestic sewage treatment, this study presented a novel integrated fermentation, nitrification, denitratation and anammox (IFNDA) process in a combined ABR-CSTR reactor. The results showed that an advanced total nitrogen (TN) removal efficiency of 94.1% was obtained after over 190 days operation, resulting in effluent TN concentration as low as 3.6 mg/L. The system nitrogen removal was dominated by anammox with a high proportion of 88.6%. The high conversion rate of acetic acid (54.0%) and volatile fatty acids (64.5%) from rDOMs in domestic sewage by in-situ fermentation drove efficient denitratation. Microflora analysis indicated that the enriched Commamonas (3.5%) and Longilinea (3.3%) dominated hydrolysis and acidogenesis of organics, and Methanosaeta (9.0%) obligated acetoclastic methanogenesis in two-stage fermentation process. Thauera (8.4%) and Candidatus Brocadia (2.5%) were the core bacteria for nitrogen metabolism in the IFNDA system.

Direction of plant-soil feedback determines plant responses to drought.

Droughts exarcerbate Plant-soil feedbacks (PSFs) making positive PSFs more positive and negative PSFs more negative. Alterations in PSFs that droughts induce could relate to the rooting depth of the tested plants. We present some rare evidence on how a driver of global change will alter a biotic interaction.

Photonic Nanojet-Mediated Optogenetics.

Optogenetics has become a widely used technique in neuroscience research, capable of controlling neuronal activity with high spatiotemporal precision and cell-type specificity. Expressing exogenous opsins in the selected cells can induce neuronal activation upon light irradiation, and the activation depends on the power of incident light. However, high optical power can also lead to off-target neuronal activation or even cell damage. Limiting the incident power, but enhancing power distribution to the targeted neurons, can improve optogenetic efficiency and reduce off-target effects. Here, the use of optical lenses made of polystyrene microspheres is demonstrated to achieve effective focusing of the incident light of relatively low power to neighboring neurons via photonic jets. The presence of microspheres significantly localizes and enhances the power density to the target neurons both in vitro and ex vivo, resulting in increased inward current and evoked action potentials. In vivo results show optogenetic stimulation with microspheres that can evoke significantly more motor behavior and neuronal activation at lowered power density. In all, a proof-of-concept of a strategy is demonstrated to increase the efficacy of optogenetic neuromodulation using pulses of reduced optical power.

Deep Neural Network for Automatic Classification of Pathological Voice Signals.

OBJECTIVES: Computer-aided pathological voice detection is efficient for initial screening of pathological voice, and has received high academic and clinical attention. This paper proposes an automatic diagnosis method of pathological voice based on deep neural network (DNN). Other two classification models (support vector machines and random forests) were used to verify the effectiveness of DNN. METHODS: In this paper, we extracted 12 Mel frequency cepstral coefficients of each voice sample as row features. The constructed DNN consists a two-layer stacked sparse autoencoders network and a softmax layer. The stacked sparse autoencoders layer can learn high-level features from raw Mel frequency cepstral coefficients features. Then, the softmax layer can diagnose pathological voice according to high-level features. The DNN and the other two comparison models used the same train set and test set for the experiment. RESULTS: Experimental results reveal that the value of sensitivity, specificity, precision, accuracy, and F1 score of the DNN can reach 97.8%, 99.4%, 99.4%, 98.6%, and 98.4%, respectively. The five indexes of DNN classification results are at least 6.2%, 5%, 5.6%, 5.7%, and 6.2% higher than the comparison models (support vector machine and random forest). CONCLUSIONS: The proposed DNN can learn advanced features from raw acoustic features, and distinguish pathological voice from healthy voice. To the extent of this preliminary study, future studies can further explore the application of DNN in other experiments and clinical practice.

Magnesium implantation or supplementation ameliorates bone disorder in CFTR-mutant mice through an ATF4-dependent Wnt/ß-catenin signaling.

Magnesium metal and its alloys are being developed as effective orthopedic implants; however, the mechanisms underlying the actions of magnesium on bones remain unclear. Cystic fibrosis, the most common genetic disease in Caucasians caused by the mutation of CFTR, has shown bone disorder as a key clinical manifestation, which currently lacks effective therapeutic options. Here we report that implantation of magnesium-containing implant stimulates bone formation and improves bone fracture healing in CFTR-mutant mice. Wnt/ß-catenin signaling in the bone is enhanced by the magnesium implant, and inhibition of Wnt/ß-catenin by iCRT14 blocks the magnesium implant to improve fracture healing in CFTR-mutant mice. We further demonstrate that magnesium ion enters osteocytes, increases intracellular cAMP level and activates ATF4, a key transcription factor known to regulate Wnt/ß-catenin signaling. In vivo knockdown of ATF4 abolishes the magnesium implant-activated ß-catenin in bones and reverses the improved-fracture healing in CFTR-mutant mice. In addition, oral supplementation of magnesium activates ATF4 and ß-catenin as well as enhances bone volume and density in CFTR-mutant mice. Together, these results show that magnesium implantation or supplementation may serve as a potential anabolic therapy for cystic fibrosis-related bone disease. Activation of ATF4-dependent Wnt/ß-catenin signaling in osteocytes is identified as a previously undefined mechanism underlying the beneficial effect of magnesium on bone formation.