Homeostasis Research Articles

Trophic homogeneity due to seasonal variation in nitrogen in shallow subtropical lakes

Lakes play a crucial role in the nitrogen (N) cycle, and eutrophication disrupts the balance of the nitrogen cycle within lakes, including both the N removal process and the N supplement process. However, the mechanisms by which different nutrient levels affect seasonal nitrogen variations in the water columns are not clear, especially for long-term and large- scale studies. In this study, we used 206 independent spatial samples from a total of 108 subtropical shallow lakes from four surveys in the middle and lower reaches of the Yangtze River, as well as time-case study data from Lake Taihu and Lake Donghu of up to 23 and 14 years, respectively, to analyze the changes in summer TN compared to spring (delta TN). Delta TN was significantly negatively correlated with initial spring TN concentrations, with similar trends observed in both space and time. Furthermore, the slopes of spring TN vs. delta TN varied little across lakes in both time and space, suggesting a consistent relationship between initial spring TN and summer TN changes. When initial TN or TN: TP ratio was low, N fixation by algae played a significant role in compensating for summer N removal, thus mitigating summer N reductions; when TN was high or TN: TP ratio was high, ammonia stress reduced the compensatory effect of algae and denitrification played a significant role in summer N removal, thus increasing summer N reductions. Our study suggested that no matter what the initial conditions are, lakes tend to evolve towards a common nutrient status through biological regulation.

Impact of singular versus combinatorial environmental stress on RONS generation in Drosophila melanogaster larvae.

We investigated environmentally correlated abiotic stressor desiccation (D), heat (H), and starvation (S) in the generation of reactive oxygen and nitrogen species (RONS) using Drosophila melanogaster larvae as an experimental model, subjected to either individual stressors or exposed to a combinatorial form of stressors (D + H, H + S, and D + S). The study was also extended to find synergistic endpoints where the impacts of all three stressors (D + H + S) were exerted simultaneously. We estimated the lethal time (LT20) at specific doses using regression and probit analyses based on the larval survival. LT20 values were used as the base-level parameter for further oxidative stress experimental analysis work. First, all stressors led to the activation of a typical common oxidative stress-mediated response irrespective of the mode of exposure. As envisaged, D. melanogaster larvae exhibited a homeostatic stress tolerance mechanism, triggering an antioxidant defense mechanism, indicated by an elevated level of total antioxidant capacity and enhanced activities of superoxide dismutase, catalase, glutathione reductase, and glutathione peroxidase. In all types of stress-exposed regimes, we found a negative impact of stressors on the activity of mitochondrial enzyme aconitase. Elevated levels of other oxidative stress markers, viz., lipid peroxidation, protein carbonyl content, and advanced oxidative protein products, were obvious although the increment was treatment-specific. Desiccation stress proved to be the most dominant stressor compared to heat and starvation. Among the combination of stressors, rather than a single stressor, D + H impacted more than other binary stress exposures. Focusing on the impact of singular versus combinatorial stress exposure on RONS generation, we observed an increase in the RONS level in both singular and combinatorial forms of stress exposure although the magnitude of the increment varied with the nature of stressors and their combinations. The present study indicated an "additive" effect when all three stressors (D + H + S) operate simultaneously, rather than a "synergistic" effect.

Research Advances on Natural Killer Cells in Infections

Natural killer (NK) cells play a crucial role in the human immune system, swiftly recognizing and eliminating infected cells through non-specific immune mechanisms. This paper reviews the mechanisms by which NK cells act in various infections, including those caused by viruses, bacteria, parasites, and fungi, and explores their biological characteristics and functional regulation in the context of infection. By thoroughly analyzing the regulatory mechanisms of cytokines, receptor-ligand interactions, and the microenvironment on NK cell function, this paper further elucidates the response mechanisms of NK cells under different infection scenarios. Finally, it discusses the clinical application prospects of NK cells in treating infectious diseases and outlines future research directions, providing new ideas and possibilities for NK cell-based immunotherapy strategies.

Moesin controls cell-cell fusion and osteoclast function.

Cell-cell fusion is an evolutionarily conserved process that is essential for many functions, including fertilisation and the formation of placenta, muscle and osteoclasts, multinucleated cells that are unique in their ability to resorb bone. The mechanisms of osteoclast multinucleation involve dynamic interactions between the actin cytoskeleton and the plasma membrane that are still poorly characterized. Here, we found that moesin, a cytoskeletal linker protein member of the Ezrin/Radixin/Moesin (ERM) protein family, is activated during osteoclast maturation and plays an instrumental role in both osteoclast fusion and function. In mouse and human osteoclast precursors, moesin inhibition favors their ability to fuse into multinucleated osteoclasts. Accordingly, we demonstrated that moesin depletion decreases membrane-to-cortex attachment and enhances the formation of tunneling nanotubes (TNTs), F-actin-based intercellular bridges that we reveal here to trigger cell-cell fusion. Moesin also controls HIV-1- and inflammation-induced cell fusion. In addition, moesin regulates the formation of the sealing zone, the adhesive structure determining osteoclast bone resorption area, and thus controls bone degradation, via a β3-integrin/RhoA/SLK pathway. Supporting our results, moesin - deficient mice present a reduced density of trabecular bones and increased osteoclast abundance and activity. These findings provide a better understanding of the regulation of cell-cell fusion and osteoclast biology, opening new opportunities to specifically target osteoclast activity in bone disease therapy.

Association of urinary glyphosate levels with iron homeostasis among a representative sample of US adults: NHANES 2013–2018

PurposeGlyphosate and glyphosate-based herbicides (GBH), widely used globally, were initially considered harmless to humans. Experimental studies have suggested that these substances can disrupt iron homeostasis by interfering with iron uptake or triggering inflammatory responses. However, their potential impact on human iron homeostasis remains underexplored. Approach and resultsWe analyzed data from 5812 participants aged three and older from the 2013 to 2018 NHANES. We investigated the relationships between urinary glyphosate levels, oral iron intake, and markers of iron homeostasis, including serum iron, unsaturated iron-binding capacity (UIBC), total iron-binding capacity (TIBC), transferrin saturation, ferritin, and transferrin receptor. Higher urinary glyphosate levels were positively associated with oral iron intake (β = 1.310, S.E. = 0.382, P = 0.001). A one-unit increase in the natural logarithm (ln)-glyphosate was associated with lower serum iron (β = − 4.236, 95 % CI = − 6.432 to − 2.039, P < 0.001) and ferritin (β = − 9.994, 95 % CI = − 17.342 to − 2.647, P = 0.009), and higher UIBC (β = 5.431, 95 % CI = 1.061–9.800, P = 0.018) and transferrin receptor levels (β = 0.139, 95 % CI = 0.015–0.263, P = 0.029). Increasing glyphosate exposure was associated with significant decreases in serum iron and ferritin across exposure quintiles (trend P-values = 0.003 and 0.018, respectively). ConclusionsHigher glyphosate exposure is associated with reduced iron availability, suggesting potential disruptions in iron absorption. These findings underscore the need for further research into the health implications of glyphosate exposure on iron homeostasis.

Age Specifics of Homeostasis Response to Cold Stress in Wistar Rats: Endurance Improvement or Disorganization?

Cold exposure, especially in combination with exercise, is a common procedure to fortify the body. However, its effects in old age are poorly understood. Using the homeostasis stability coefficient as an indicator, the body response to a 20-min swim in cold water was for the first time studied comprehensively in old and young Wistar rats. Effects on the hormonal, hematological, and morphofunctional systems of the liver and thymus were assessed. Unfavorable age-related changes in the stability of homeostasis were observed in old rats. The changes complicated the recovery after cold stress and required the involvement of a greater number of homeostatic mechanisms than in young rats. The liver was found to be the most vulnerable to cold stress. It was concluded that fortifying the body by cold exposure is possible to use in old age, but with due regard to age-related restrictions.

Beyond monopole electrostatics in regulating conformations of intrinsically disordered proteins.

Conformations and dynamics of an intrinsically disordered protein (IDP) depend on its composition of charged and uncharged amino acids, and their specific placement in the protein sequence. In general, the charge (positive or negative) on an amino acid residue in the protein is not a fixed quantity. Each of the ionizable groups can exist in an equilibrated distribution of fully ionized state (monopole) and an ion-pair (dipole) state formed between the ionizing group and its counterion from the background electrolyte solution. The dipole formation (counterion condensation) depends on the protein conformation, which in turn depends on the distribution of charges and dipoles on the molecule. Consequently, effective charges of ionizable groups in the IDP backbone may differ from their chemical charges in isolation-a phenomenon termed charge-regulation. Accounting for the inevitable dipolar interactions, that have so far been ignored, and using a self-consistent procedure, we present a theory of charge-regulation as a function of sequence, temperature, and ionic strength. The theory quantitatively agrees with both charge reduction and salt-dependent conformation data of Prothymosin-alpha and makes several testable predictions. We predict charged groups are less ionized in sequences where opposite charges are well mixed compared to sequences where they are strongly segregated. Emergence of dipolar interactions from charge-regulation allows spontaneous coexistence of two phases having different conformations and charge states, sensitively depending on the charge patterning. These findings highlight sequence dependent charge-regulation and its potential exploitation by biological regulators such as phosphorylation and mutations in controlling protein conformation and function.

Responses of invertebrate traits to litter chemistry accelerate decomposition under nitrogen enrichment

Nitrogen (N) enrichment shapes litter chemistry, subsequently influencing soil invertebrates and litter decomposition. However, there has been a lack of attention to how soil invertebrates respond to changes in litter chemistry and then drive litter decomposition under N enrichment. Here, trait-based approaches were adopted to explore functional responses of Collembola, a crucial and functional group of invertebrates. We conducted reciprocal transplantation of plant litter between ambient N levels (0 kg N ha−1 yr−1) and N enrichment (90 kg N ha−1 yr−1) plots in a field experiment, quantifying Collembola traits and litter mass loss during litter decomposition process. Results showed that N enrichment-derived litter recruited Collembola with long antenna, legs, and strong mandibles in enrichment environment, while Collembola with same traits were recruited by ambient-derived litter in ambient environment. Collembola traits, including antenna to body ratio, leg to body ratio, and mandible width to length ratio, coincided with high litter decomposition rates under N enrichment. Overall, the results provide evidence that Collembola with strong resource acquisition abilities responded to changes in litter chemistry, and such shifts further accelerate litter decomposition under N enrichment. Our findings demonstrate that adopting trait-based approaches to link litter and invertebrates would advance the understanding of ecosystem processes governed by biological regulation under global change.

Fabrication and Temporal Dependency Osteogenic Regulation of Dual-Scale Hierarchical Microstructures on Medical Metal Surface.

The structural characteristics at the interface of bone implants can guide biological regulation. In this study, a dual-scale hierarchical microstructure is proposed and customized using hybrid machining to achieve temporal dependency osteogenic regulation. It is observed that osteoblasts induced by dual-scale hierarchical structure exhibit adequate protrusion development and rapid cell attachment through the modulation of mechanical forces in the cell growth environment, and further promot the upregulation of the cell membrane receptor PDGFR-α, which is related to cell proliferation. Afterward, transcriptomic analysis reveals that during the differentiation stage, the DSH structure regulates cellular signaling cascades primarily through integrin adhesion mechanisms and then accelerates osteogenic differentiation by activating the TGF-β pathway and cAMP signaling pathway. Furthermore, the calcium nodules are preferentially deposited within the lower honeycomb-like channels, thereby endowing the proposed dual-scale hierarchical structure with the potential to induce oriented deposition and improve the long-term stability of the implant.

Quantitative proteomic analysis of PK-15 cells infected with porcine circovirus type 3 using 4D-DIA approach.

Porcine circovirus type 3 (PCV3) infection is clinically related to various diseases, including porcine dermatitis and nephrotic syndrome (PDNS)-like disease, respiratory disease, reproductive disorders, and gastrointestinal and neurological diseases. Since PCV3 infection was discovered in 2016, it has developed rapidly and has attracted much attention worldwide. However, specific preventive and therapeutic interventions are currently lacking. In this study, four-dimensional (4D) data-independent acquisition (DIA)-based quantitative proteomics detection combined with bioinformatics analysis were employed to quantitatively identify the differentially expressed proteins in PK-15 cells from the PCV3-infected group compared with those from the uninfected control group. A total of 194 cellular proteins were significantly altered in response to PCV3 infection, including 58 upregulated proteins and 136 downregulated proteins. In our Gene Ontology (GO) enrichment analysis, these differentially expressed proteins were mostly associated with cellular anatomical entities, binding, cellular processes, biological regulation, catalytic activity, metabolic processes, developmental processes, protein-containing complexes and responses to stimuli. Our Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the DEPs were predominantly involved in metabolic pathways, the cAMP signaling pathway, protein processing in the endoplasmic reticulum, the PI3K-Akt signaling pathway, and the calcium signaling pathway. For the experiments, Western blotting (WB) was used to confirm the changes in important molecules. The differentially expressed proteins identified should contribute to a greater understanding of the mechanism of PCV3 replication and pathogenesis, as well as the host response.

Tumour lysis syndrome.

Tumour lysis syndrome (TLS) represents a critical oncological emergency characterized by extensive tumour cell breakdown, leading to the swift release of intracellular contents into the systemic circulation, outpacing homeostatic mechanisms. This process results in hyperuricaemia (a by-product of intracellular DNA release), hyperkalaemia, hyperphosphataemia, hypocalcaemia and the accumulation of xanthine. These electrolyte and metabolic imbalances pose a significant risk of acute kidney injury, cardiac arrhythmias, seizures, multiorgan failure and, rarely, death. While TLS can occur spontaneously, it usually arises shortly after the initiation of effective treatment, particularly in patients with a large cancer cell mass (defined as ≥500 g or ≥300 g/m2 of body surface area in children). To prevent TLS, close monitoring and hydration to improve renal perfusion and urine output and to minimize uric acid or calcium phosphate precipitation in renal tubules are essential. Intervention is based on the risk of a patient of having TLS and can include rasburicase and allopurinol. Xanthine, typically enzymatically converted to uric acid, can accumulate when xanthine oxidases, such as allopurinol, are administered during TLS management. Whether measurement of xanthine is clinically useful to optimize the use of allopurinol or rasburicase remains to be determined.

Time-of-day effects of cancer drugs revealed by high-throughput deep phenotyping

The circadian clock, a fundamental biological regulator, governs essential cellular processes in health and disease. Circadian-based therapeutic strategies are increasingly gaining recognition as promising avenues. Aligning drug administration with the circadian rhythm can enhance treatment efficacy and minimize side effects. Yet, uncovering the optimal treatment timings remains challenging, limiting their widespread adoption. In this work, we introduce a high-throughput approach integrating live-imaging and data analysis techniques to deep-phenotype cancer cell models, evaluating their circadian rhythms, growth, and drug responses. We devise a streamlined process for profiling drug sensitivities across different times of the day, identifying optimal treatment windows and responsive cell types and drug combinations. Finally, we implement multiple computational tools to uncover cellular and genetic factors shaping time-of-day drug sensitivity. Our versatile approach is adaptable to various biological models, facilitating its broad application and relevance. Ultimately, this research leverages circadian rhythms to optimize anti-cancer drug treatments, promising improved outcomes and transformative treatment strategies.

A new perspective on microRNA-guided gene regulation specificity, and its potential generalization to transcription factors and RNA-binding proteins.

Our conception of gene regulation specificity has undergone profound changes over the last 20years. Previously, regulators were considered to control few genes, recognized with exquisite specificity by a 'lock and key' mechanism. However, recentlygenome-wide exploration of regulator binding site occupancy (whether on DNA or RNA targets) revealed extensive lists of molecular targets for every studied regulator. Such poor biochemical specificity suggested that each regulator controls many genes, collectively contributing to biological phenotypes. Here, I propose a third model, whereby regulators' biological specificity is only partially due to 'lock and key' biochemistry. Rather, regulators affect many genes at the microscopic scale, but biological consequences for most interactions are attenuated at the mesoscopic scale: only a few regulatory events propagate from microscopic to macroscopic scale;others are made inconsequential by homeostatic mechanisms. This model is well supported by the microRNA literature, and data suggest that it extends to other regulators. It reconciles contradicting observations from biochemistry and comparative genomics on one hand and in vivo genetics on the other hand, but this conceptual unification is obscured by common misconceptions and counter-intuitive modes of graphical display. Profound understanding of gene regulation requires conceptual clarification, and better suited statistical analyses and graphical representation.

Lysozyme 1 Inflamed CCR2+ Macrophages Promote Obesity-Induced Cardiac Dysfunction.

Macrophages are key players in obesity-associated cardiovascular diseases, which are marked by inflammatory and immune alterations. However, the pathophysiological mechanisms underlying macrophage's role in obesity-induced cardiac inflammation are incompletely understood. Our study aimed to identify the key macrophage population involved in obesity-induced cardiac dysfunction and investigate the molecular mechanism that contributes to the inflammatory response. In this study, we used single-cell RNA-sequencing analysis of Cd45+CD11b+F4/80+ cardiac macrophages to explore the heterogeneity of cardiac macrophages. The CCR2+ (C-C chemokine receptor 2) macrophages were specifically removed by a dual recombinase approach, and the macrophage CCR2 was deleted to investigate their functions. We also performed cleavage under target and tagmentation analysis, chromatin immunoprecipitation-polymerase chain reaction, luciferase assay, and macrophage-specific lentivirus transfection to define the impact of lysozyme C in macrophages on obesity-induced inflammation. We find that the Ccr2 cluster undergoes a functional transition from homeostatic maintenance to proinflammation. Our data highlight specific changes in macrophage behavior during cardiac dysfunction under metabolic challenge. Consistently, inducible ablation of CCR2+CX3CR1+ macrophages or selective deletion of macrophage CCR2 prevents obesity-induced cardiac dysfunction. At the mechanistic level, we demonstrate that the obesity-induced functional shift of CCR2-expressing macrophages is mediated by the CCR2/activating transcription factor 3/lysozyme 1/NF-κB (nuclear factor kappa B) signaling. Finally, we uncover a noncanonical role for lysozyme 1 as a transcription activator, binding to the RelA promoter, driving NF-κB signaling, and strongly promoting inflammation and cardiac dysfunction in obesity. Our findings suggest that lysozyme 1 may represent a potential target for the diagnosis of obesity-induced inflammation and the treatment of obesity-induced heart disease.

Intracellular pH differentially regulates transcription of metabolic and signaling pathways in normal epithelial cells

Intracellular pH (pHi) dynamics regulate normal cell function, and dysregulated pHi dynamics is an emerging hallmark of cancer (constitutively increased pHi) and neurodegeneration (constitutively decreased pHi). However, the molecular mechanisms by which pHi dynamics regulate cell biology are poorly understood. Here, we discovered that altering pHi in normal human breast epithelial cells triggers global transcriptional changes. We identified 176 genes differentially regulated by pHi, with pHi-dependent genes clustering in signaling and glycolytic pathways. Using various normal epithelial cell models, we showed pH-dependent Notch homolog 1 protein expression, with increased protein abundance at high pHi. This resulted in pH-dependent downstream signaling, with increased Notch homolog 1 signaling at high pHi. We also found that high pHi increased the expression of glycolytic enzymes and regulators of pyruvate fate, including lactate dehydrogenase and pyruvate dehydrogenase kinase. These transcriptional changes were sufficient to alter lactate production, with high pHi shifting these normal epithelial cells toward a glycolytic metabolism and increasing lactate production. Thus, pHi dynamics transcriptionally regulate signaling and metabolic pathways in normal epithelial cells. Our data reveal new molecular regulators of pHi-dependent biology and a role for increased pHi in driving the acquisition of cancer-associated signaling and metabolic changes in normal human epithelial cells.

C-reactive protein across pregnancy in individuals exposed to childhood maltreatment: The role of psychological and physical sequelae of maltreatment

BackgroundChildhood maltreatment (CM) has long-term consequences for the regulation of stress biology which are particularly pronounced when mental and physical health sequelae have manifested. C-reactive protein (CRP) has been shown to be elevated in the non-pregnant state in association with CM as well as in the setting of CM-associated mental and physical health sequelae. In pregnancy, however, the association between CM and CRP is less clear. We sought to examine this association and consider the moderating role of four common health sequelae of CM (maternal depressive symptoms, overweight/obesity, smoking, and hypertensive disorders during pregnancy). MethodsA prospective, longitudinal study of 744 healthy pregnant participants was conducted, with analyses focusing on a sample of 643 participants. CM was assessed with the Childhood Trauma Questionnaire (CTQ) and categorized by whether no vs. one or more moderate to severe CM experiences were reported. Blood serum concentrations of CRP, maternal depression severity (continuous scores of the Center for Epidemiologic Studies Depression Scale, CES-D) and smoking during pregnancy were assessed in early (16.52 ± 2.50 weeks gestation) and late (33.65 ± 1.18 weeks gestation) pregnancy. Pre-pregnancy body mass index (BMI) was obtained at the first study visit and hypertensive disorders diagnosed during pregnancy were obtained from the medical record. Linear mixed effects models were employed to assess main effects of CM as well as interactive effects of CM and four common CM-associated sequelae as well as a sum score of these sequelae on repeatedly measured CRP concentration. In secondary analyses, we conducted latent class analyses to classify participants based on their specific experiences of childhood abuse and/or neglect and to assess the association of these CM subgroups with CM sequelae and CRP. All analyses were adjusted for potential confounders (maternal race and ethnicity and education/income). ResultsCRP concentration decreased from early to late pregnancy (B = −0.06, SE = 0.01, p < 0.001). While there was no main effect of CM on CRP (p = 0.49), the interaction of CM and depressive symptoms was associated with CRP concentration (B = 0.08, SE = 0.04, p < 0.05), indicating higher CRP across pregnancy with increasing levels of depressive symptoms during pregnancy in participants with CM experience. This interaction was mainly driven by participants with co-occurring physical and emotional maltreatment. For none of the other CM-associated sequelae a statistically significant interaction with CM on CRP concentration was observed. ConclusionsThese results add to the growing empirical evidence suggesting higher inflammation during pregnancy in participants exposed to CM who experience depressive symptoms and highlight the detrimental effects of multiple co-occurring experiences of maltreatment. Given the negative consequences of chronic inflammatory state for the mother and the developing fetus, monitoring and treating psychiatric sequelae during pregnancy among participants exposed to CM is potentially an important opportunity to dampen long-term detrimental effects of CM, serving at least two generations.

Cadmium Exposure in Male Rats Results in Ovarian Granulosa Cell Apoptosis in Female Offspring and Paternal Genetic Effects.

The aim of this study was to investigate whether the damage to male offspring induced by cadmium (Cd) exposure during embryonic period leads to the apoptosis of ovarian granulosa cells (OGCs) in the next generation of female offspring, and whether this apoptosis in the offspring was due to paternal genetic effects. Pregnant Sprague-Dawley (SD) rats were exposed to CdCl2 (0, 0.5, 2.0, or 8.0 mg/kg) by gavage daily for 20 days to produce the filial 1 (F1) generation. F1 males were mated with newly purchased females to produce the F2 generation, and the F3 generation was generated in the same way. No apoptotic bodies were observed in the OGCs of either the F2 or F3 generation as shown by electron microscopy, and a reduced OGC apoptosis rate (detected by flow cytometry) was observed in F2 OGCs from the Cd-exposed group. Moreover, the mRNA (qRT-PCR) levels of Bax and Bcl-2 and the protein (western blotting) level of pro-caspase-8 increased in the F2 generation (p < 0.05). The expression of apoptosis-related miRNAs (qRT-PCR) and methylation of apoptosis-related genes (determined via bisulfite-sequencing PCR) in OGCs were further determined. Compared with those of the controls, the expression patterns of microRNAs (miRNAs) in the F2 offspring were different in the Cd-exposed group. The miR-92a-2-5p expression levels were decreased in both the F2 and F3 generations (p < 0.05), while the average methylation level of apoptosis-related genes did not change significantly (except for individual loci). In summary, this study showed that the paternal genetic intergenerational effect of male Cd exposure during embryonic period induced apoptosis of OGCs in the offspring was weakened, and the transgenerational effect disappeared; nevertheless, intergenerational and transgenerational changes in apoptosis-related genes, epigenetic modifications, DNA methylation, and miRNAs were observed, and may be important for understanding the homeostatic mechanisms of the body to alleviate the intergenerational transmission of Cd-induced damage.

Proteomics and Bioinformatics Identify Drug-Resistant-Related Genes with Prognostic Potential in Cholangiocarcinoma.

Drug resistance is a major challenge in the treatment of advanced cholangiocarcinoma (CCA). Understanding the mechanisms of drug resistance can aid in identifying novel prognostic biomarkers and therapeutic targets to improve treatment efficacy. This study established 5-fluorouracil- (5-FU) and gemcitabine-resistant CCA cell lines, KKU-213FR and KKU-213GR, and utilized comparative proteomics to identify differentially expressed proteins in drug-resistant cells compared to parental cells. Additionally, bioinformatics analyses were conducted to explore the biological and clinical significance of key proteins. The drug-resistant phenotypes of KKU-213FR and KKU-213GR cell lines were confirmed. In addition, these cells demonstrated increased migration and invasion abilities. Proteomics analysis identified 81 differentially expressed proteins in drug-resistant cells, primarily related to binding functions, biological regulation, and metabolic processes. Protein-protein interaction analysis revealed a highly interconnected network involving MET, LAMB1, ITGA3, NOTCH2, CDH2, and NDRG1. siRNA-mediated knockdown of these genes in drug-resistant cell lines attenuated cell migration and cell invasion abilities and increased sensitivity to 5-FU and gemcitabine. The mRNA expression of these genes is upregulated in CCA patient samples and is associated with poor prognosis in gastrointestinal cancers. Furthermore, the functions of these proteins are closely related to the epithelial-mesenchymal transition (EMT) pathway. These findings elucidate the potential molecular mechanisms underlying drug resistance and tumor progression in CCA, providing insights into potential therapeutic targets.

Role of Autophagy and AMPK in Cancer Stem Cells: Therapeutic Opportunities and Obstacles in Cancer.

Cancer stem cells represent a resilient subset within the tumor microenvironment capable of differentiation, regeneration, and resistance to chemotherapeutic agents, often using dormancy as a shield. Their unique properties, including drug resistance and metastatic potential, pose challenges for effective targeting. These cells exploit certain metabolic processes for their maintenance and survival. One of these processes is autophagy, which generally helps in energy homeostasis but when hijacked by CSCs can help maintain their stemness. Thus, it is often referred as an Achilles heel in CSCs, as certain cancers tend to depend on autophagy for survival. Autophagy, while crucial for maintaining stemness in cancer stem cells (CSCs), can also serve as a vulnerability in certain contexts, making it a complex target for therapy. Regulators of autophagy like AMPK (5' adenosine monophosphate-activated protein kinase) also play a crucial role in maintaining CSCs stemness by helping CSCs in metabolic reprogramming in harsh environments. The purpose of this review is to elucidate the interplay between autophagy and AMPK in CSCs, highlighting the challenges in targeting autophagy and discussing therapeutic strategies to overcome these limitations. This review focuses on previous research on autophagy and its regulators in cancer biology, particularly in CSCs, addresses the remaining unanswered questions, and potential targets for therapy are also brought to attention.

Blockade of GluN2B-Containing NMDA Receptors Prevents Potentiation and Depression of Responses during Ocular Dominance Plasticity.

Monocular deprivation (MD) causes an initial decrease in synaptic responses to the deprived eye in juvenile mouse primary visual cortex (V1) through Hebbian long-term depression (LTD). This is followed by a homeostatic increase, which has been attributed either to synaptic scaling or to a slide threshold for Hebbian long-term potentiation (LTP) rather than scaling. We therefore asked in mice of all sexes whether the homeostatic increase during MD requires GluN2B-containing NMDA receptor activity, which is required to slide the plasticity threshold but not for synaptic scaling. Selective GluN2B blockade from 2-6 d after monocular lid suture prevented the homeostatic increase in miniature excitatory postsynaptic current (mEPSC) amplitude in monocular V1 of acute slices and prevented the increase in visually evoked responses in binocular V1 in vivo. The decrease in mEPSC amplitude and visually evoked responses during the first 2 d of MD also required GluN2B activity. Together, these results support the idea that GluN2B-containing NMDA receptors first play a role in LTD immediately following eye closure and then promote homeostasis during prolonged MD by sliding the plasticity threshold in favor of LTP.