Pretreatment Serum Lactate Dehydrogenase and Metastases Numbers as Potential Determinants of Anti-PD-1 Therapy Outcome in Nasopharyngeal Carcinoma.

BACKGROUND: We aimed to investigate the determinant factors of anti-PD-1 therapy outcome in nasopharyngeal carcinoma (NPC). METHODS: In this retrospective study, we included 64 patients with recurrent/metastatic NPC. The association of patients' characteristics, C-reactive protein (CRP), neutrophil to lymphocyte ratio (NLR), and lactate dehydrogenase (LDH) with survival benefit of anti-PD-1 therapy were analyzed using Cox regression models and Kaplan-Meier analyses. Patients were divided based on the median value of CRP, NLR or LDH into different subgroups. RESULTS: At a median follow-up time of 11.4 months (range: 1-28 months), median progression-free survival (PFS) and overall survival (OS) were 1.9 months (95% CI, .18-3.6) and 15 months (95% CI, 10.9-19.1) months, respectively. Pretreatment metastases numbers was significant predictor of PFS (HR = 1.99; 95% CI 1.10-3.63; P = .024) and OS (HR = 2.77; 95% CI 1.36-5.61; P = .005). Baseline LDH level was independent predictor of OS (HR = 7.01; 95% CI 3.09-15.88; P < .001). Patients with LDH level >435 U/L at the baseline had significantly shorter PFS and OS compared to patients with LDH level ≤435 U/L (median PFS: 1.7 vs 3.5 months, P = .040; median OS: 3.7 vs 18.5 months, P < .001). Patients with non-durable clinical benefit (NDB) had significantly higher LDH level at the baseline compared to patients who achieved durable clinical benefit (DCB) (P = .025). Post-treatment levels of CRP, LDH, and NLR were decreased compared to baseline in patients with DCB (P = .030, P = .088, and P = .066, respectively), whereas, there was a significant increase in post-treatment level of LDH compared with baseline in patients with NDB (P = .024). CONCLUSIONS: LDH level at the baseline was an independent predictor of OS and pretreatment metastases numbers was a significant predictor of PFS and OS.

Biochar assisted cellulose anaerobic digestion under the inhibition of dodecyl dimethyl benzyl ammonium chloride: Dose-response kinetic assays, performance variation, potential promotion mechanisms.

This study evaluated the inhibitory effect and mitigation strategy of dodecyl dimethyl benzyl ammonium chloride (DDBAC) suppression on anaerobic digestion. With the 12 h-suppression, only 16.64% of anaerobes were alive, and acetotrophic methanogens were significantly inhibited. As for batch test, DDBAC suppression significantly prolonged the start-up of systems and decreased the biogas production. In cellulose semi-continuous digestion process, the DDBAC suppression induced volatile fatty acids accumulation and pH decrease. However, the biochar amended reactor effectively mitigated the DDBAC suppression and achieved 370.5 mL/d·g-chemical-oxygen-demand biogas production. Moreover, 17.8% more protein in extracellular polymeric substances was secreted as the bio-barrier to defense the DDBAC suppression. Furthermore, microbial analysis showed that biochar addition selectively enriched directed interspecies electron transfer (DIET) participant bacteria (Anaerolineaceae and Syntrophomonas) and methanogens (Methanosaeta and Methanobacterium). Meanwhile, the potential metabolic pathway analysis showed that the abundance of amino acids and energy metabolism were increased 28% and 8%, respectively. The abundance of encoding enzyme related to hydrogenotrophic and acetotrophic methanogenesis enriched 1.88 times and 1.48 times, respectively. These results showed the performance and mechanisms involved in DIET establishment with ethanol stimulation biochar addition.

Endonuclease EEPD1 Is a Gatekeeper for Repair of Stressed Replication Forks.

Replication is not as continuous as once thought, with DNA damage frequently stalling replication forks. Aberrant repair of stressed replication forks can result in cell death or genome instability and resulting transformation to malignancy. Stressed replication forks are most commonly repaired via homologous recombination (HR), which begins with 5' end resection, mediated by exonuclease complexes, one of which contains Exo1. However, Exo1 requires free 5'-DNA ends upon which to act, and these are not commonly present in non-reversed stalled replication forks. To generate a free 5' end, stalled replication forks must therefore be cleaved. Although several candidate endonucleases have been implicated in cleavage of stalled replication forks to permit end resection, the identity of such an endonuclease remains elusive. Here we show that the 5'-endonuclease EEPD1 cleaves replication forks at the junction between the lagging parental strand and the unreplicated DNA parental double strands. This cleavage creates the structure that Exo1 requires for 5' end resection and HR initiation. We observed that EEPD1 and Exo1 interact constitutively, and Exo1 repairs stalled replication forks poorly without EEPD1. Thus, EEPD1 performs a gatekeeper function for replication fork repair by mediating the fork cleavage that permits initiation of HR-mediated repair and restart of stressed forks.

EEPD1 Rescues Stressed Replication Forks and Maintains Genome Stability by Promoting End Resection and Homologous Recombination Repair.

Replication fork stalling and collapse is a major source of genome instability leading to neoplastic transformation or cell death. Such stressed replication forks can be conservatively repaired and restarted using homologous recombination (HR) or non-conservatively repaired using micro-homology mediated end joining (MMEJ). HR repair of stressed forks is initiated by 5' end resection near the fork junction, which permits 3' single strand invasion of a homologous template for fork restart. This 5' end resection also prevents classical non-homologous end-joining (cNHEJ), a competing pathway for DNA double-strand break (DSB) repair. Unopposed NHEJ can cause genome instability during replication stress by abnormally fusing free double strand ends that occur as unstable replication fork repair intermediates. We show here that the previously uncharacterized Exonuclease/Endonuclease/Phosphatase Domain-1 (EEPD1) protein is required for initiating repair and restart of stalled forks. EEPD1 is recruited to stalled forks, enhances 5' DNA end resection, and promotes restart of stalled forks. Interestingly, EEPD1 directs DSB repair away from cNHEJ, and also away from MMEJ, which requires limited end resection for initiation. EEPD1 is also required for proper ATR and CHK1 phosphorylation, and formation of gamma-H2AX, RAD51 and phospho-RPA32 foci. Consistent with a direct role in stalled replication fork cleavage, EEPD1 is a 5' overhang nuclease in an obligate complex with the end resection nuclease Exo1 and BLM. EEPD1 depletion causes nuclear and cytogenetic defects, which are made worse by replication stress. Depleting 53BP1, which slows cNHEJ, fully rescues the nuclear and cytogenetic abnormalities seen with EEPD1 depletion. These data demonstrate that genome stability during replication stress is maintained by EEPD1, which initiates HR and inhibits cNHEJ and MMEJ.

PARP1 is required for chromosomal translocations.

Chromosomal translocations are common contributors to malignancy, yet little is known about the precise molecular mechanisms by which they are generated. Sequencing translocation junctions in acute leukemias revealed that the translocations were likely mediated by a DNA double-strand break repair pathway termed nonhomologous end-joining (NHEJ). There are major 2 types of NHEJ: (1) the classical pathway initiated by the Ku complex, and (2) the alternative pathway initiated by poly ADP-ribose polymerase 1 (PARP1). Recent reports suggest that classical NHEJ repair components repress translocations, whereas alternative NHEJ components were required for translocations. The rate-limiting step for initiation of alternative NHEJ is the displacement of the Ku complex by PARP1. Therefore, we asked whether PARP1 inhibition could prevent chromosomal translocations in 3 translocation reporter systems. We found that 2 PARP1 inhibitors or repression of PARP1 protein expression strongly repressed chromosomal translocations, implying that PARP1 is essential for this process. Finally, PARP1 inhibition also reduced both ionizing radiation-generated and VP16-generated translocations in 2 cell lines. These data define PARP1 as a critical mediator of chromosomal translocations and raise the possibility that oncogenic translocations occurring after high-dose chemotherapy or radiation could be prevented by treatment with a clinically available PARP1 inhibitor.

Involvement of glutamate-cystine/glutamate transporter system in aspirin-induced acute gastric mucosa injury.

Large-dose or long-term use of aspirin tends to cause gastric mucosa injury, which is recognized as the major side effect of aspirin. It has been demonstrated that glutamate exerts a protective effect on stomach, and the level of glutamate is critically controlled by cystine/glutamate transporter (Xc(-)). In the present study, we investigated the role of glutamate-cystine/glutamate transporter system in aspirin-induced acute gastric mucosa injury in vitro and in vivo. Results showed that in human gastric epithelial cells, aspirin incubation increased the activity of LDH and the number of apoptotic cells, meanwhile down-regulated the mRNA expression of Xc(-) accompanied with decreased glutamate release. Similar results were seen in a rat model. In addition, exogenous l-glutamate attenuated the gastric mucosa injury and cell damage induced by aspirin both in vitro and in vivo. Taken together, our results demonstrated that acute gastric mucosa injury induced by aspirin is related to reduction of glutamate-cystine/glutamate transporter system activity.

Improvement of Stress Resistance of Microencapsulated Lactobacillus plantarum by Emulsion Electrospinning.

Probiotics have become increasingly recognized for their potential health-promoting properties; however, the viability of probiotics can be affected by storage and transportation processes as well as the stressful environment of the human digestive tract, preventing them from achieving effective concentration (107 CFU/mL). In this regard, the embedding technology of probiotics provides an effective protection method. Dextran-based water in water (W/W) emulsion loaded with Lactobacillus plantarum was used as spinning solution to prepare Lactobacillus plantarum-loaded electrospun fibers. The structure of the W/W emulsion and the electrospun fibers was charactered. Lactobacillus plantarum were uniformly embedded in the internal phase of the W/W emulsion and the loading efficiency was 9.70 ± 0.40 log CFU/g. After 240 min digestion in the gastrointestinal tract, and temperature treatment in 65 °C and 72 °C, the loaded probiotics maintained high activity. Even after 5 days of storage in room temperature and 4 °C, the loaded probiotic activity levels remained high, with counts >8 log CFU/g. These results suggest that probiotics encapsulated by emulsion electrospinning could be potentially delivered in a novel food delivery system used in the future food industry.

Induction of CX3CL1 expression by LPS and its impact on invasion and migration in oral squamous cell carcinoma.

Purpose: This study aimed to explore the expression of CX3CL1 induced by lipopolysaccharide (LPS) in oral squamous cell carcinoma (OSCC) and its impact on biological characteristics such as invasion and migration, taking the foundation for new targets for the treatment and prognosis of OSCC. Methods: This study utilized a variety of techniques, including bioinformatics, molecular biology, and cell experiments, to investigate the expression of CX3CL1 and its receptor CX3CR1 in OSCC patients' cancer tissues or OSCC cell lines. Extracting, organizing, and analyzing the TCGA database on the expression of CX3CL1 and its receptor CX3CR1 in cancer tissues and corresponding paracancerous normal tissues of OSCC patients by bioinformatics methods. The expression of CX3CL1 in cancerous and normal tissues of OSCC patients was verified by IHC, and the changes in mRNA and protein expression of CX3CL1 and its receptor CX3CR1 in OSCC cell lines were detected before and after lipopolysaccharide LPS stimulation by RT-PCR, ELISA, and WB. Changes in cell biological behavior by overexpression of CX3CL1 in OSCC cell lines were detected by CCK-8, Transwell, scratch healing assay, and cloning assay. The effects of overexpressing cell lines on the AKT pathway and Epithelial-mesenchymal Transition (EMT)-related protein expression before and after LPS stimulation were detected by Western Blot. Results: (1) CX3CL1 and its receptor CX3CR1 were found to be downregulated in OSCC tissues of patients or OSCC cell lines. (2) After LPS stimulation, CX3CL1 gene expression increased in both OSCC cell lines, while CX3CR1 expression remained unchanged. (3) OSCC cell lines overexpressing CX3CL1 showed changes in cell biological characteristics, including decreased proliferation, invasion, migration, and stemness, which were more pronounced after LPS stimulation. (4) Overexpression of CX3CL1 in OSCC cell lines decreased EMT-related protein expression and AKT phosphorylation. On the contrary were promoted by LPS stimulation. Conclusion: CX3CL1 and CX3CR1 are downregulated in OSCC cancer tissues and cell lines compared to adjacent normal tissues and cells. LPS stimulation increases CX3CL1 expression in OSCC cell lines, suggesting that inflammation may induce CX3CL1 expression and that the CX3CL1 gene may play an important role in OSCC progression. Overexpression of CX3CL1 inhibits OSCC cell proliferation, migration, invasion, and stemness, suggesting that CX3CL1 plays a critical role in suppressing OSCC development. CX3CL1 suppresses OSCC invasion and migration by affecting EMT progression and AKT phosphorylation, and partially reverse the process that LPS causes and affects the development of OSCC.

A new ChatGPT-empowered, easy-to-use machine learning paradigm for environmental science.

The quantity and complexity of environmental data show exponential growth in recent years. High-quality big data analysis is critical for performing a sophisticated characterization of the complex network of environmental pollution. Machine learning (ML) has been employed as a powerful tool for decoupling the complexities of environmental big data based on its remarkable fitting ability. Yet, due to the knowledge gap across different subjects, ML concepts and algorithms have not been well-popularized among researchers in environmental sustainability. In this context, we introduce a new research paradigm-"ChatGPT + ML + Environment", providing an unprecedented chance for environmental researchers to reduce the difficulty of using ML models. For instance, each step involved in applying ML models to environmental sustainability, including data preparation, model selection and construction, model training and evaluation, and hyper-parameter optimization, can be easily performed with guidance from ChatGPT. We also discuss the challenges and limitations of using this research paradigm in the field of environmental sustainability. Furthermore, we highlight the importance of "secondary training" for future application of "ChatGPT + ML + Environment".

pH Effect on the Structure, Rheology, and Electrospinning of Maize Zein.

As a simple and convenient technology to fabricate micron-to-nanoscale fibers with controllable structure, electrostatic spinning has produced fiber films with many natural advantages, including a large specific surface area and high porosity. Maize zein, as a major storage protein in corn, showed high hydrophobicity and has been successfully applied as a promising carrier for encapsulation and controlled release in the pharmaceutical and food areas. Proteins exhibit different physical and chemical properties at different pH values, and it is worth investigating whether this change in physical and chemical properties affects the properties of electrospun fiber films. We studied the pH effects on zein solution rheology, fiber morphology, and film properties. Rotational rheometers were used to test the rheology of the solutions and establish a correlation between solution concentration and fiber morphology. The critical concentrations calculated by the cross-equation fitting model were 17.6%, 20.1%, 20.1%, 17.1%, and 19.5% (w/v) for pH 4, 5, 6, 7, and 8, respectively. The secondary structure of zein changed with the variation in solution pH. Furthermore, we analyzed the physical properties of the zein films. The contact angles of the fiber membranes prepared with different pH spinning solutions were all above 100, while zein films formed by solvent evaporation showed hydrophilic properties. The results indicated that the rheological properties of zein solutions and the surface properties of the film were affected by the pH value. This study showed that zein solutions can be stabilized to form electrospun fibers at a variety of pH levels and offered new opportunities to further enhance the encapsulation activity of zein films for bioactive materials.

Formation and application of edible oleogels prepared by dispersing soy fiber particles in oil phase.

Oleogels containing less saturated and trans-fats were considered as an ideal option to replace the solid fats in foods. In this research, oleogel was fabricated by dispersing soy fiber particles (SFP) in soy oil, and further it was used in bread preparation. Effect of the particle size, particle content and the second fluid content on the formation of oleogels were evaluated, based on the appearance and rheological properties. Results showed that the suspension of SFP in soy oil (24%, w/w) could be transformed into gel-like state, upon the addition of the second fluid. The SFP based networks were dominated by the capillary force which was originated from the second fluid. The rheological properties and yield stress of the oleogels could be modulated by particle size and particle content of SFP in oil phase, as well as the second fluid content in the system. When the oleogels were applicated in bread preparation, a layered structure could be formed in the bread, indicating the possibility of replacing the solid fats in bakery products by our oleogels. Our results offered a feasibility approach for oil structuring with natural raw materials, and developed a new approach to replace the solid fats in foods.

Fabrication, characterization and emulsifying properties of agarose microgel.

Agarose microgels were prepared with bottom-up approach, and emulsifying properties of agarose microgels were investigated. Physical properties of microgels are varied with agarose concentration, and further affect the emulsifying performance of microgels. Enhanced surface hydrophobicity index and decreased particle size of microgels were recorded with the increasing of agarose concentration, which were conducive to emulsifying properties of microgels. Improved interfacial adsorption of microgels was evidenced by Dynamic surface tension and SEM. However, microscopic morphology of microgel at O/W interface indicated that increasing agarose concentration could weaken the deformability of microgels. The influence of external conditions (pH and NaCl) on the physical properties of microgels were investigated, and their effects on emulsion stability were evaluated. Compared with acidification, NaCl was appeared to be more destructive to emulsion stability. Results indicated acidification and NaCl could decrease surface hydrophobicity index of microgels, but there was differentiation in the variation of particle size. It was inferred that deformability of microgels could make contribution to the stability of emulsion. This study verified that microgelation was a feasible scheme to improve the interfacial properties of agarose, and the influence of agarose concentration, pH, and NaCl on the emulsifying performance of microgels was investigated.

Modulating in vitro fecal fermentation behavior of sodium alginate by Ca2+ cross-linking.

Slow fermentable dietary fibers can be utilized by human gut microbiota in the distal region of the colon and thus exert a sufficient short-chain fatty acids (SCFAs) supplement in the distal region of the human colon. Alginate (Alg) based microgels are widely fabricated and used to control their digestion by digestive enzymes releasing active substances site-specifically. Herein, sodium alginate microgels with gradient calcium-ion (Ca2+) cross-linking densities were developed, restricting their degradation by gut microbiota. Alg microgels were prepared using high-speed shearing after Alg was cross-linked with 10, 40, and 60 mmol/L Ca2+, respectively (named 10-Alg, 40-Alg, and 60-Alg). The fluorescence and atomic force microscopic results showed that the 40-Alg particle has the densest structure among the three cross-linked Alg. In vitro human fecal fermentation results revealed that the Ca2+ cross-linking exerted more restricting effects than delaying effects on the fermentation of Alg, and the 40-Alg exhibited the slowest fermentation rate and the least fermentation extent, by characterizing the residual total carbohydrate content, residual monosaccharide content, pH, and total short-chain fatty acids. The 16S rRNA gene sequencing results indicated that cross-linking structures shaped a high specifical Bacteroides-type microbial community and that OTU205 (Bacteroides_xylanisolvens) highly correlated to the cross-linking density (R = 0.65, p = 0.047). In sum, Ca2+ cross-linking generated a dense and compact structure of sodium alginate that facilitated a more restricted fermentation property and specificity-targeting microbial community structure in comparison to the original sodium alginate.

Influence of interfacial properties/structure on oxygen diffusion in oil-in-water emulsions.

Oxygen diffusion played an important role in the lipid oxidation of food emulsions. In this study, a simple method was developed to quantitatively observe the oxygen diffusion in the oil-water biphasic system, and it was further applied to investigate the relationship between the oxygen diffusion and lipid oxidation in O/W emulsions. Various factors that related to the emulsion oxidation were considered, from their influence on the oxygen diffusion and lipid oxidation in the emulsions. Results showed that there was obvious correlation between the oxygen diffusion and lipid oxidation in O/W emulsions, which reveals the inhibition of oxygen diffusion could apparently slow down the lipid oxidation. Moreover, the changes of oil phase, water phase and interfacial layer of the emulsions, which were related to the oxygen diffusion, could improve the oxidative stability of the emulsions effectively. Our findings are helpful for deep understanding the mechanisms of the lipid oxidation in food emulsions.

RCN1 deficiency inhibits oral squamous cell carcinoma progression and THP-1 macrophage M2 polarization.

Reticulocalbin 1 (RCN1), a calcium-binding protein located in the endoplasmic reticulum (ER) lumen, contains six conserved regions. Its main functions include maintaining intracellular homeostasis and regulating cell proliferation and apoptosis, and it plays an important role in the development of various tumours. However, the exact function of RCN1 in oral squamous cell carcinoma (OSCC) is not fully understood. Therefore, the aim of this study was to investigate the effects of RCN1 on the biological behaviour of OSCC and the regulation of tumour-associated macrophage (TAM) polarization. The expression of RCN1 in OSCC and normal oral mucosa was evaluated through bioinformatics analysis and immunohistochemical staining. The growth, migration, and invasion of OSCC cells were observed after knockdown of RCN1 using CCK-8 and Transwell assays. Apoptosis was detected by flow cytometry. The effect of tumour cell-derived RCN1 on the polarization of THP-1 macrophages was investigated by establishing a coculture model of THP-1 macrophages and OSCC cells. Additionally, changes in the expression levels of relevant proteins were detected using Western blotting. The upregulation of RCN1 in tumour tissues compared to normal oral mucosal tissues is associated with a poor prognosis and can be utilized as a prognostic indicator for OSCC. Knockdown of RCN1 inhibited the proliferation, migration, and invasion of OSCC cells. Additionally, knockdown of RCN1 in Cal-27 and SCC-25 cells resulted in inhibition of the M2 polarization of THP-1 macrophages. RCN1 knockdown inhibits OSCC progression and M2 macrophage polarization. Targeting RCN1 may be a promising approach for OSCC treatment.

Emulsifying performance of the hexadecyltrimethylammonium bromide (CTAB) complexed alginate microgels: Effects from their deformability on oil-water interface.

Hexadecyltrimethylammonium bromide complexed alginate-Ca2+ microgels (C/AMGs) were developed as emulsifiers, which shown remarkably improved emulsifying performance than non-complexed alginate-Ca2+ microgels (AMGs) in previous study. This work focus on the impact of deformability on the emulsifying performance of C/AMGs. By regulating alginate concentration (1.0-4.0 wt%), microgels with different deformability were prepared. Deformability was proved to have great influence on the emulsifying performance of C/AMGs, which was evaluated by Langmuir trough measurements, emulsion appearance, centrifugation stability, digestive behavior, and oxidative stability. Particle size and SEM images indicated microgels prepared with lower alginate concentration are more deformable. C/AMGs (2.0 wt%) exhibits the best emulsifying performance, which could be ascribed to the appreciated deformability and mechanical strength. Digestive behavior and oxidative stability of alginate-Ca2+ microgel (2.0 wt%) stabilized emulsions were further investigated. Compared with alginate-Ca2+ microgel (2.0 wt%) stabilized emulsions, C/AMGs (2.0 wt%) stabilized emulsions shown delayed lipid digestion and lower POV. Results of this work supporting that Mickering mechanism have potential in fabricating functional emulsions based on natural polysaccharides.

Developmental downregulation of Xenopus cyclin E is phosphorylation and nuclear import dependent and is mediated by ubiquitination.

Cyclins are regulatory subunits that bind to and activate catalytic Cdks. Cyclin E associates with Cdk2 to mediate the G1/S transition of the cell cycle. Cyclin E is overexpressed in breast, lung, skin, gastrointestinal, cervical, and ovarian cancers. Its overexpression correlates with poor patient prognosis and is involved in the etiology of breast cancer. We have been studying how cyclin E is normally downregulated during development in order to determine if disruption of similar mechanisms could either contribute to its overexpression in cancer, or be exploited to decrease its expression. In Xenopus laevis embryos, cyclin E protein level is high and constant until its abrupt destabilization by an undefined mechanism after the 12th cell cycle, which corresponds to the midblastula transition (MBT) and remodeling of the embryonic to the adult cell cycle. Since degradation of mammalian cyclin E is regulated by the ubiquitin proteasome system and is phosphorylation dependent, we examined the role of phosphorylation in Xenopus cyclin E turnover. We show that similarly to human cyclin E, phosphorylation of serine 398 and threonine 394 plays a role in cyclin E turnover at the MBT. Immunofluorescence analysis shows that cyclin E relocalizes from the cytoplasm to the nucleus preceding its degradation. When nuclear import is inhibited, cyclin E stability is markedly increased after the MBT. To investigate whether degradation of Xenopus cyclin E is mediated by the proteasomal pathway, we used proteasome inhibitors and observed a progressive accumulation of cyclin E in the cytoplasm after the MBT. Ubiquitination of cyclin E precedes its proteasomal degradation at the MBT. These results show that cyclin E destruction at the MBT requires both phosphorylation and nuclear import, as well as proteasomal activity.

MG53 E3 Ligase-Dead Mutant Protects Diabetic Hearts From Acute Ischemic/Reperfusion Injury and Ameliorates Diet-Induced Cardiometabolic Damage.

Cardiometabolic diseases, including diabetes and its cardiovascular complications, are the global leading causes of death, highlighting a major unmet medical need. Over the past decade, mitsugumin 53 (MG53), also called TRIM72, has emerged as a powerful agent for myocardial membrane repair and cardioprotection, but its therapeutic value is complicated by its E3 ligase activity, which mediates metabolic disorders. Here, we show that an E3 ligase-dead mutant, MG53-C14A, retains its cardioprotective function without causing metabolic adverse effects. When administered in normal animals, both the recombinant human wild-type MG53 protein (rhMG53-WT) and its E3 ligase-dead mutant (rhMG53-C14A) protected the heart equally from myocardial infarction and ischemia/reperfusion (I/R) injury. However, in diabetic db/db mice, rhMG53-WT treatment markedly aggravated hyperglycemia, cardiac I/R injury, and mortality, whereas acute and chronic treatment with rhMG53-C14A still effectively ameliorated I/R-induced myocardial injury and mortality or diabetic cardiomyopathy, respectively, without metabolic adverse effects. Furthermore, knock-in of MG53-C14A protected the mice from high-fat diet-induced metabolic disorders and cardiac damage. Thus, the E3 ligase-dead mutant MG53-C14A not only protects the heart from acute myocardial injury but also counteracts metabolic stress, providing a potentially important therapy for the treatment of acute myocardial injury in metabolic disorders, including diabetes and obesity.

Paclitaxel exposure-toxicity analysis reveals a pharmacokinetic determinant for dose-limiting neutropenia in East-Asian solid tumor patients: results from two prospective, phase II studies.

PURPOSE: The time of a paclitaxel (PTX) concentration remains above 0.05 µM (Tc > 0.05) has been associated with PTX-induced adverse effects in Caucasians, while limited studies were reported in Asians. This study was aimed to explore the characteristics of Tc > 0.05 and the relationship between PTX exposure and toxicity in East-Asian patients. METHODS: This study was based on two prospective phase II clinical trials and patients with advanced nasopharyngeal cancer (NPC) and non-small cell lung cancer (NSCLC) who were naïve to PTX were included independently. Eligible patients receive PTX (175 mg/m2) and carboplatin (AUC = 5) treatment every 3 weeks. PTX pharmacokinetic analysis was accessed. The relationship between PTX exposure and toxicities after first cycle as well as clinical efficacy was evaluated. RESULTS: A total of 93 NPC and 40 NSCLC patients were enrolled. PTX exposure was consistent in two trials with average Tc > 0.05 duration of 38.8 h and 38.4 h, respectively. Average Tc > 0.05 in patients with grade 3/4 neutropenia was significantly higher than those without severe neutropenia in NPC patients (P = 0.003) and NSCLC patients (P = 0.007). Cut-off value of Tc > 0.05 were identified from the NPC cohort and then verified in the NSCLC cohort, dividing patients into high exposure Tc > 0.05 group (> 39 h) and low exposure group (≤ 39 h). Incidence of grade 3/4 neutropenia were significantly higher in the high exposure group in NPC cohort (43.3% vs 10.0%, P < 0.001) and NSCLC cohort (42.1% vs 9.5%, P = 0.028). No significant relationship between Tc > 0.05 and efficacy were observed. CONCLUSION: Patients with PTX Tc > 0.05 duration above 39 h experience more severe neutropenia than those under 39 h. Prospective studies are needed to verify this threshold.

Three-dimensional collagen represses cyclin E1 via ß1 integrin in invasive breast cancer cells.

The behavior of breast epithelial cells is influenced by their microenvironment which includes stromal cells and extracellular matrix (ECM). During cancer progression, the tissue microenvironment fails to control proliferation and differentiation, resulting in uncontrolled growth and invasion. Upon invasion, the ECM encountered by breast cancer cells changes from primarily laminin and collagen IV to primarily collagen I. We show here that culturing invasive breast cancer cells in 3-dimensional (3D) collagen I inhibits proliferation through direct regulation of cyclin E1, a G(1)/S regulator that is overexpressed in breast cancer. When the breast cancer cell line MDA-MB-231 was cultured within 3D collagen I gels, the G(1)/S transition was inhibited as compared to cells cultured on conventional 2D collagen or plastic dishes. Cells in 3D collagen downregulated cyclin E1 protein and mRNA, with no change in cyclin D1 level. Cyclin D1 was primarily cytoplasmic in 3D cultures, and this was accompanied by decreased phosphorylation of Rb, a nuclear target for both cyclin E1- and cyclin D1-associated kinases. Positive regulators of cyclin E1 expression, the transcription factor c-Myc and cold-inducible RNA binding protein (CIRP), were decreased in 3D collagen cultures, while the collagen I receptor ß1 integrin was greatly increased. Inhibition of ß1 integrin function rescued proliferation and cyclin E1 expression as well as c-Myc expression and Rb phosphorylation, but cyclin D1 remained cytoplasmic. We conclude that cyclin E1 is repressed independent of effects on cyclin D1 in a 3D collagen environment and dependent on ß1 integrin interaction with collagen I, reducing proliferation of invasive breast cancer cells.