Intratumoral Fc-optimized agonistic CD40 antibody induces tumor rejection and systemic antitumor immunity in patients with metastatic cancer.

While CD40 agonism is an attractive approach for activating antigen-presenting cells and initiating antitumor responses, previous attempts have encountered limited clinical efficacy coupled with toxicity. We previously demonstrated that interactions between the antibody Fc domain and the inhibitory receptor FcγRIIB are critical for enhanced antitumor activity. Here, we present the results of a phase 1 study on intratumoral administration of an anti-CD40 agonistic antibody (2141-V11) Fc-engineered to enhance FcγRIIB binding. Primary endpoints included safety, maximum tolerated dose (MTD), and recommended phase 2 dose. Secondary objectives included preliminary clinical activity and correlative studies from biospecimens. 2141-V11 was well-tolerated without dose-limiting toxicities and MTD was not reached. In ten evaluable patients with metastatic cancer, the overall response rate was 20%, with complete responses in two patients (melanoma and breast carcinoma) and stable disease in six patients. 2141-V11 induced tumor regression in injected and non-injected lesions, with increased leukocyte infiltration and tertiary lymphoid structures (TLS) formation in post-treatment biopsies. In a humanized mouse model for CD40 and FcγRs, 2141-V11 induced TLS formation in mice bearing orthotopic breast carcinoma, correlating with local and abscopal antitumor effects, systemic immune activation, and immune memory. These findings support the safety and efficacy of 2141-V11, warranting phase 2 studies and suggesting a unique mechanism of action for this Fc-enhanced immunotherapy (NCT04059588).

Trade-offs inside the black box of neoantigen prediction.

Success of precision neoantigen-based immunotherapies hinges on the selection of immunogenic neoantigens, yet currently neither large-scale datasets nor streamlined methods are available to achieve this goal. Müller et al. present a large experimental dataset resource along with machine learning-based models to classify immunogenic neoantigens.

Severe and fatal adverse events of immune checkpoint inhibitor combination therapy in patients with metastatic renal cell carcinoma: a systematic review and meta-analysis.

Introduction: Immune checkpoint inhibitor (ICI) combination therapy has changed the treatment landscape for metastatic renal cell carcinoma (mRCC). However, little evidence exists on the treatment-related severe adverse events (SAEs) and fatal adverse events (FAEs) of ICI combination therapy in mRCC. Method: We searched PubMed, Embase, and Cochrane Library databases to evaluate randomized controlled trials (RCTs) of ICI combination therapy versus conventional tyrosine kinase inhibitor (TKI)-targeted therapy in mRCC. Data on SAEs and FAEs were analyzed using revman5.4 software. Results: Eight RCTs (n=5380) were identified. The analysis showed no differences in SAEs (60.5% vs. 64.5%) and FAEs (1.2% vs. 0.8%) between the ICI and TKI groups (odds ratio [OR], 0.83; 95%CI 0.58-1.19, p=0.300 and OR, 1.54; 95%CI 0.89-2.69, p=0.120, respectively). ICI-combination therapy was associated with less risk of hematotoxicities, including anemia (OR, 0.24, 95%CI 0.15-0.38, p<0.001), neutropenia (OR, 0.07, 95%CI 0.03-0.14, p<0.001), and thrombocytopenia (OR, 0.05, 95%CI 0.02-0.12, p<0.001), but with increased risks of hepatotoxicities (ALT increase [OR, 3.39, 95%CI 2.39-4.81, p<0.001] and AST increase [OR, 2.71, 95%CI 1.81-4.07, p<0.001]), gastrointestinal toxicities (amylase level increase [OR, 2.32, 95%CI 1.33-4.05, p=0.003] and decreased appetite [OR, 1.77, 95%CI 1.08-2.92, p=0.020]), endocrine toxicity (adrenal insufficiency [OR, 11.27, 95%CI 1.55-81.87, p=0.020]) and nephrotoxicity of proteinuria (OR, 2.21, 95%CI 1.06-4.61, p=0.030). Conclusions: Compared with TKI, ICI combination therapy has less hematotoxicity in mRCC but more specific hepatotoxicity, gastrointestinal toxicity, endocrine toxicity, and nephrotoxicity, with a similar severe toxicity profile. Systematic review registration: https://www.crd.york.ac.uk/prospero/, identifier CRD42023412669.

ATF4 renders human T-cell acute lymphoblastic leukemia cell resistance to FGFR1 inhibitors through amino acid metabolic reprogramming.

Abnormalities of FGFR1 have been reported in multiple malignancies, suggesting FGFR1 as a potential target for precision treatment, but drug resistance remains a formidable obstacle. In this study, we explored whether FGFR1 acted a therapeutic target in human T-cell acute lymphoblastic leukemia (T-ALL) and the molecular mechanisms underlying T-ALL cell resistance to FGFR1 inhibitors. We showed that FGFR1 was significantly upregulated in human T-ALL and inversely correlated with the prognosis of patients. Knockdown of FGFR1 suppressed T-ALL growth and progression both in vitro and in vivo. However, the T-ALL cells were resistant to FGFR1 inhibitors AZD4547 and PD-166866 even though FGFR1 signaling was specifically inhibited in the early stage. Mechanistically, we found that FGFR1 inhibitors markedly increased the expression of ATF4, which was a major initiator for T-ALL resistance to FGFR1 inhibitors. We further revealed that FGFR1 inhibitors induced expression of ATF4 through enhancing chromatin accessibility combined with translational activation via the GCN2-eIF2α pathway. Subsequently, ATF4 remodeled the amino acid metabolism by stimulating the expression of multiple metabolic genes ASNS, ASS1, PHGDH and SLC1A5, maintaining the activation of mTORC1, which contributed to the drug resistance in T-ALL cells. Targeting FGFR1 and mTOR exhibited synergistically anti-leukemic efficacy. These results reveal that FGFR1 is a potential therapeutic target in human T-ALL, and ATF4-mediated amino acid metabolic reprogramming contributes to the FGFR1 inhibitor resistance. Synergistically inhibiting FGFR1 and mTOR can overcome this obstacle in T-ALL therapy.

DUSP4 promotes esophageal squamous cell carcinoma progression by dephosphorylating HSP90ß.

The molecular and pathogenic mechanisms of esophageal squamous cell carcinoma (ESCC) development are still unclear, which hinders the development of effective treatments. In this study, we report that DUSP4 is highly expressed in human ESCC and is negatively correlated with patient prognosis. Knockdown of DUSP4 suppresses cell proliferation and patient-derived xenograft (PDX)-derived organoid (PDXO) growth and inhibits cell-derived xenograft (CDX) development. Mechanistically, DUSP4 directly binds to heat shock protein isoform ß (HSP90ß) and promotes the ATPase activity of HSP90ß by dephosphorylating HSP90ß on T214 and Y216. These dephosphorylation sites are critical for the stability of JAK1/2-STAT3 signaling and p-STAT3 (Y705) nucleus translocation. In vivo, Dusp4 knockout in mice significantly inhibits 4-nitrochinoline-oxide-induced esophageal tumorigenesis. Moreover, DUSP4 lentivirus or treatment with HSP90ß inhibitor (NVP-BEP800) significantly impedes PDX tumor growth and inactivates the JAK1/2-STAT3 signaling pathway. These data provide insight into the role of the DUSP4-HSP90ß-JAK1/2-STAT3 axis in ESCC progression and describe a strategy for ESCC treatment.

Discovery of potential biomarkers for lung cancer classification based on human proteome microarrays using Stochastic Gradient Boosting approach.

PURPOSE: Early identification of lung cancer (LC) will considerably facilitate the intervention and prevention of LC. The human proteome micro-arrays approach can be used as a "liquid biopsy" to diagnose LC to complement conventional diagnosis, which needs advanced bioinformatics methods such as feature selection (FS) and refined machine learning models. METHODS: A two-stage FS methodology by infusing Pearson's Correlation (PC) with a univariate filter (SBF) or recursive feature elimination (RFE) was used to reduce the redundancy of the original dataset. The Stochastic Gradient Boosting (SGB), Random Forest (RF), and Support Vector Machine (SVM) techniques were applied to build ensemble classifiers based on four subsets. The synthetic minority oversampling technique (SMOTE) was used in the preprocessing of imbalanced data. RESULTS: FS approach with SBF and RFE extracted 25 and 55 features, respectively, with 14 overlapped ones. All three ensemble models demonstrate superior accuracy (ranging from 0.867 to 0.967) and sensitivity (0.917 to 1.00) in the test datasets with SGB of SBF subset outperforming others. The SMOTE technique has improved the model performance in the training process. Three of the top selected candidate biomarkers (LGR4, CDC34, and GHRHR) were highly suggested to play a role in lung tumorigenesis. CONCLUSION: A novel hybrid FS method with classical ensemble machine learning algorithms was first used in the classification of protein microarray data. The parsimony model constructed by the SGB algorithm with the appropriate FS and SMOTE approach performs well in the classification task with higher sensitivity and specificity. Standardization and innovation of bioinformatics approach for protein microarray analysis need further exploration and validation.

Chromosome-scale genomics, metabolomics, and transcriptomics provide insight into the synthesis and regulation of phenols in Vitis adenoclada grapes.

Vitis adenoclada is a wild grape unique to China. It exhibits well resistance to heat, humidity, fungal disease, drought, and soil infertility. Here, we report the high-quality, chromosome-level genome assembly of GH6 (V. adenoclada). The 498.27 Mb genome contained 221.78 Mb of transposable elements, 28,660 protein-coding genes, and 481.44 Mb of sequences associated with 19 chromosomes. GH6 shares a common ancestor with PN40024 (Vitis vinifera) from approximately 4.26-9.01 million years ago, whose divergence occurred later than Vitis rotundifolia and Vitis riparia. Widely-targeted metabolome and transcriptome analysis revealed that the profiles and metabolism of phenolic compounds in V. adenoclada varieties significantly were differed from other grape varieties. Specifically, V. adenoclada varieties were rich in phenolic acids and flavonols, whereas the flavan-3-ol and anthocyanin content was lower compared with other varieties that have V. vinifera consanguinity in this study. In addition, ferulic acid and stilbenes content were associated with higher expressions of COMT and STSs in V. adenoclada varieties. Furthermore, MYB2, MYB73-1, and MYB73-2 were presumably responsible for the high expression level of COMT in V. adenoclada berries. MYB12 (MYBF1) was positively correlated with PAL, CHS, FLS and UFGT.Meanwhile, MYB4 and MYBC2-L1 may inhibit the synthesis of flavan-3-ols and anthocyanins in two V. adenoclada varieties (YN2 and GH6). The publication of the V. adenoclada grape genome provides a molecular foundation for further revealing its flavor and quality characteristics, is also important for identifying favorable genes of the East Asian species for future breeding.

Efficient degradation of N-nitrosopyrrolidine using CoFe-LDH/AC particle electrode via heterogeneous Fenton-like reaction.

With the rapid development of drinking water disinfection technology, extensive attentions are paid to the nitrogenous disinfection by-products (N-DBPs) that has strong carcinogenicity, thus their degradation becomes important for the health of human beings. In this work, for the first time, CoFe-LDH material used as particle electrode is proposed to treat trace N-nitrosopyrrolidine (NPYR) in a three-dimensional aeration electrocatalysis reactor (3DAER). The factors on the degradation efficiency and energy consumption of NPYR are systematically investigated, and the results of radical quenching experiments show that the degradation of NPYR is completed by combining with ·OH, ·O2and direct oxidation together. CoFe-LDH particle electrode plays a vital role in generating ·OH via heterogeneous ‾Fenton-like reaction. Moreover, the adsorbed saturated CoFe-LDH particle electrode can be regenerated by electrochemical action to induce further recycle adsorption and form in-situ electrocatalysis. This work pave a way for the removal of NPYR with high efficiency, low energy conservation and environmental protection.

Impacts of crossfit training on the tennis athlete's performance / Impactos do treinamento de crossfit sobre o desempenho dos atletas de tênis / Impacto del entrenamiento de crossfit en el rendimiento de atletas de tenis

ABSTRACT Introduction: The competitive level of tennis has increased in recent years, challenging researchers to promote a higher level of endurance and performance of its practitioners. Objective: Analyze the impacts of CrossFit training on the performance of tennis athletes. Methods: This work conducted a four-week experiment with 50 professional tennis players, introducing a protocol based on CrossFit training to the experimental group. Levels of sports endurance and competition performance were statistically analyzed before and after the experiment. It was also analyzed whether CrossFit training could effectively improve tennis players' sports endurance and performance level. Results: After the intervention, the time required for the tennis players to take off and hit the ball 20 times in three steps after the recoil was reduced from 71.12 seconds before the experiment to 60.04 seconds in the experimental group. The forehand strike's linear and diagonal speeds increased by 11.00% and 6.57%, respectively. And the number of effective and accurate balls in the recoil increased by 5.87% and 5.58%. Conclusion: CrossFit training can improve tennis players' sporting endurance, playing a positive role in improving the players' level. Level of evidence II; Therapeutic studies - investigation of treatment outcomes.

RESUMO Introdução: O nível competitivo do tênis tem se elevado nos últimos anos, desafiando os pesquisadores na promoção de um maior nível de resistência e desempenho dos seus praticantes. Objetivo: Analisar os impactos do treinamento de CrossFit sobre o desempenho dos atletas de tênis. Métodos: Este trabalho conduziu uma experiência de quatro semanas com 50 tenistas profissionais, introduzindo um protocolo baseado no treinamento de CrossFit ao grupo experimental. Níveis de resistência esportiva e o desempenho na competição foram analisados estatisticamente antes e depois do experimento, também foi analisado se o treinamento CrossFit poderia efetivamente melhorar a resistência esportiva e o nível de desempenho dos tenistas. Resultados: Após a intervenção, o tempo necessários para os tenistas decolarem e baterem na bola por 20 vezes em três etapas após o recuo foi reduzido de 71,12 segundos antes do experimento para 60,04 segundos, no grupo experimental. A velocidade linear e a velocidade diagonal da batida dianteira também aumentaram em 11,00% e 6,57%, respectivamente. Sendo que o número de bolas efetivas e de bolas precisas no recuo aumentaram em 5,87% e 5,58%. Conclusão: O treinamento de CrossFit pode melhorar a resistência esportiva dos tenistas, desempenhando um papel positivo na melhora do nível dos jogadores. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción: El nivel competitivo del tenis ha aumentado en los últimos años, desafiando a los investigadores en la promoción de un mayor nivel de resistencia y rendimiento de sus practicantes. Objetivo: Analizar los impactos del entrenamiento de Crossfit en el rendimiento de atletas de tenis. Métodos: Este trabajo realizó un experimento de cuatro semanas con 50 tenistas profesionales, introduciendo en el grupo experimental un protocolo basado en el entrenamiento CrossFit. Se analizaron estadísticamente los niveles de resistencia deportiva y rendimiento en competición antes y después del experimento, también se analizó si el entrenamiento de Crossfit podía mejorar eficazmente la resistencia deportiva y el nivel de rendimiento de los tenistas. Resultados: Después de la intervención, el tiempo requerido por los tenistas para despegar y golpear la pelota durante 20 veces en tres pasos después del retroceso se redujo de 71,12 segundos antes del experimento a 60,04 segundos en el grupo experimental. La velocidad lineal y la velocidad diagonal del golpe de derecha también aumentaron un 11,00% y un 6,57%, respectivamente. Siendo que el número de pelotas efectivas y de pelotas precisas en el retiro aumentaron en 5,87% y 5,58%. Conclusión: El entrenamiento de Crossfit puede mejorar la resistencia deportiva de los tenistas, desempeñando un papel positivo en la mejora del nivel de los jugadores. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Effect of abdominal adipose content on spine phantom bone mineral density measured by rapid kilovoltage-switching dual-energy CT and quantitative CT.

Background: The purpose of this study was to investigate the effect of abdominal adiposity, as measured by abdominal total adipose tissue (TAT), on the accuracy of rapid kilovoltage-switching dual-energy computed tomography (DECT) and quantitative computed tomography (QCT) measurements of bone mineral density (BMD) in a spine phantom model. Methods: Fresh porcine fat was wrapped around the European Spine Phantom (ESP) and divided into four groups according to the TAT cross-sectional areas, S=0, 100, 200, and 350 cm2, to simulate different TAT contents. The hydroxyapatite (HAP) (water) values of each vertebra were measured by DECT, and the BMD values by QCT. A one-sample t-test was used to analyze the differences between the measurements and the true values of the ESP. One-way analysis of variance (ANOVA) was applied to compare the differences between measurements under different TAT conditions, and the root-mean-square errors (RMSEs) of the BMD measurements were calculated and compared. Moreover, Pearson's correlation analysis was performed for the RMSE and TAT. Linear regression analysis was conducted on the measurements, the true values, and the TAT to obtain the correction equations for the BMD and to compare the RMSE before and after correction. Results: At higher TAT content, the measurements of both scanning methods were more affected, and the measurements of the TAT =350 cm2 group were significantly different from the remaining groups (P<0.05). There was a positive correlation between the RMSE and TAT (r>0, P<0.05), with the RMSE of the L1 vertebrae the largest under the same TAT content. The corrected equations for BMD were derived, and the RMSE of BMD was significantly reduced after correction. Conclusions: The measurements of ESP BMD for both rapid kilovoltage-switching DECT and QCT changed with TAT content. Along with the increase of TAT, the RMSE of measurements increased and the accuracy decreased; moreover, the lower the value of BMD, the more significant the RMSE. The linear regression analysis allowed the corrected BMD measurements to be very close to true values.

PHLDA1 promotes glioblastoma cell growth via sustaining the activation state of Ras.

Activation of the Ras signaling pathway promotes the growth of malignant human glioblastoma multiforme (GBM). Mutations in Ras are rare in GBM, elevated levels of activated Ras are prevalently observed in GBM. However, the potential mechanism of how Ras is activated in GBM remains unclear. In this study, we screened a new interacted protein of Ras, PHLDA1. Our findings confirmed that PHLDA1 acted as an oncogene and promoted glioma progression and recurrence. We demonstrated that PHLDA1 was upregulated in GBM tissues and cells. PHLDA1 overexpression promoted cell proliferation and tumor growth. In terms of mechanism, PHLDA1 promoted cell proliferation by regulating Ras/Raf/Mek/Erk signaling pathway. Moreover, Src promotes GTPase activity of Ras via tyrosine 32 phosphorylation. PHLDA1 and Src competed for binding with Ras, inhibiting Ras phosphorylation by Src and rescuing Ras activity. This study may provide a new idea of the molecular mechanism underlying glioma progression and a novel potential therapeutic target for comprehensive glioblastoma treatment.

Distinct cellular dynamics associated with response to CAR-T therapy for refractory B cell lymphoma.

Chimeric antigen receptor (CAR)-T cell therapy has revolutionized the treatment of hematologic malignancies. Approximately half of patients with refractory large B cell lymphomas achieve durable responses from CD19-targeting CAR-T treatment; however, failure mechanisms are identified in only a fraction of cases. To gain new insights into the basis of clinical response, we performed single-cell transcriptome sequencing of 105 pretreatment and post-treatment peripheral blood mononuclear cell samples, and infusion products collected from 32 individuals with large B cell lymphoma treated with either of two CD19 CAR-T products: axicabtagene ciloleucel (axi-cel) or tisagenlecleucel (tisa-cel). Expansion of proliferative memory-like CD8 clones was a hallmark of tisa-cel response, whereas axi-cel responders displayed more heterogeneous populations. Elevations in CAR-T regulatory cells among nonresponders to axi-cel were detected, and these populations were capable of suppressing conventional CAR-T cell expansion and driving late relapses in an in vivo model. Our analyses reveal the temporal dynamics of effective responses to CAR-T therapy, the distinct molecular phenotypes of CAR-T cells with differing designs, and the capacity for even small increases in CAR-T regulatory cells to drive relapse.

CDK15 promotes colorectal cancer progression via phosphorylating PAK4 and regulating ß-catenin/ MEK-ERK signaling pathway.

Colorectal cancer (CRC) is the third most diagnosed cancer and the second leading cause of cancer-related deaths. However, there are few effective therapeutic targets for CRC patients. Here, we found that CDK15 was highly expressed in human CRC and negatively correlated with patient prognosis and overall survival in tissue microarray. Knockdown of CDK15 suppressed cell proliferation and anchorage-independent growth of CRC cells and inhibited tumor growth in cell line-derived xenograft (CDX) model. Importantly, knockout of CDK15 in mice retarded AOM/DSS-induced tumorigenesis and CDK15 silencing by lentivirus significantly suppressed tumor progression in patient-derived xenograft (PDX) model. Mechanistically, CDK15 could bind PAK4 and phosphorylate PAK4 at S291 site. Phosphorylation of PAK4 at the S291 residue promoted cell proliferation and anchorage-independent growth through ß-catenin/c-Myc, MEK/ERK signaling pathway in CRC. Moreover, inhibition of PAK4 reversed the tumorigenic function of CDK15 in CRC cells and pharmacological targeting PAK4 suppressed tumor growth in PDX models. Thus, our data reveal the pivotal role of CDK15 in CRC progression and demonstrate CDK15 promotes CRC tumorigenesis by phosphorylating PAK4. Hence, the CDK15-PAK4 axis may serve as a novel therapeutic target for CRC.

Investigation on the Explosion Characteristics of an Aluminum Dust-Diethyl Ether-Air Mixture.

In this work, the explosion characteristics of an aluminum (Al)-diethyl ether (DEE)-air mixture were investigated in a 20 L spherical vessel. The effect factors of the explosion characteristics considered were fuel concentration, component proportion, and ignition energy. With the increasing concentration of the mixed fuel (Al/DEE = 1:1), the maximum pressure (P max), the maximum rate of pressure rise ((dP/dt)max), and the flame propagation speed (νF) exhibit an inversely "U-shaped" curve. The maximum P max, (dP/dt)max, and νF values are 901.2 kPa, 148.3 MPa/s, and 15.3 m/s, respectively, corresponding to an optimum concentration of 600 g/m3. The P max, the (dP/dt)max, and the νF increase with the addition of DEE when the proportion of DEE is below 55% but have a decrease tendency when the proportion of DEE is over 55%. As the explosions of Al and DEE were mutually promoting, the studied explosion characteristics of the Al-DEE-air mixture are obviously higher than those of pure Al or DEE in air. The minimum ignition energy (MIE) of the Al-DEE-air mixture is 1.9 mJ, between the MIE of Al and DEE. With the increase of ignition energy, P max, (dP/dt)max, and νF all increase, while the minimum explosion concentration presents a linear decreasing trend. This work could provide significant scientific evidence for evaluating the explosion risk of the Al-DEE-air mixture.

SDCBP/MDA-9/syntenin phosphorylation by AURKA promotes esophageal squamous cell carcinoma progression through the EGFR-PI3K-Akt signaling pathway.

SDCBP is an adapter protein containing two tandem PDZ domains mediating cell adhesion. The role and underlying molecular mechanism of SDCBP in ESCC remain obscure. Here, we report that SDCBP is frequently overexpressed in ESCC tissues and cells compared to normal controls and that its overexpression is correlated with late clinical stage and predicts poor prognosis in ESCC patients. Functionally, high expression of SDCBP is positively related to ESCC progression both in vitro and in vivo. Furthermore, mechanistic studies show that SDCBP activates the EGFR-PI3K-Akt signaling pathway by binding to EGFR and preventing EGFR internalization. Moreover, we provide evidence that AURKA binds to SDCBP and phosphorylates it at the Ser131 and Thr200 sites to inhibit ubiquitination-mediated SDCBP degradation. More importantly, the sites at which AURKA phosphorylates SDCBP are crucial for the EGFR signaling-mediated oncogenic function of SDCBP. Taken together, we propose that SDCBP phosphorylation by AURKA prevents SDCBP degradation and promotes ESCC tumor growth through the EGFR-PI3K-Akt signaling pathway. Our findings unveil a new AURKA-SDCBP-EGFR axis that is involved in ESCC progression and provide a promising therapeutic target for ESCC treatment in the clinic.

Hydrophilic Glyceryl Ferulates Preparation Catalyzed by Free Lipase B from Candida antartica.

Ferulic acid (FA), 4-hydroxyl-3-methoxy-2-benzylacrylic acid, has antioxidant, anticancer and ultraviolet absorption activities. However, the low hydrophilicity of FA has limited its application. Glyceryl ferulate (FG), which is an all-natural hydrophilic derivative of FA, can be used as an antioxidant and UV filter in food and cosmetic formulations. However, the applications of FG in these fields are limited due to its low content in nature. In this work, free liquid lipase was firstly used as a catalyst for FG preparation. Several different free liquid lipases (Candida antartica lipase-B, Candida antartica lipase-A, Thermomyces lanuginosus (Lipozyme TL 100L)) were screened and compared. The effects of the transesterification parameters (time, temperature, enzyme load and substrate ratio) were optimized and evaluated by response surface methodology. A reaction thermodynamic investigation was also performed. The results showed that, among the tested free lipases, the maximum FG yield (84.8±1.5%) was achieved using free Candida antartica lipase-B. Under the optimized conditions (an atmospheric system, an enzyme load of 11.1% and a 20:1 molar ratio of glycerol to EF at 70°C for 39.5 h), the FG yield and EF conversion were 84.8±1.5% and 95.7±1.2%, respectively. The activation energies of FG formation and EF conversion were 56.4 and 58.0kJ/mol, respectively.

Assessment of candidate biomarkers in paired saliva and plasma samples from oral cancer patients by targeted mass spectrometry.

For oral cancer, numerous saliva- and plasma-derived protein biomarker candidates have been discovered and/or verified; however, it is unclear about the behavior of these candidates as saliva or plasma biomarkers. In this study, we developed two targeted assays, MRM and SISCAPA-MRM, to quantify 30 potential biomarkers in both plasma and saliva samples collected from 30 healthy controls and 30 oral cancer patients. Single point measurements were used for target quantification while response curves for assay metric determination. In comparison with MRM assay, SISCAPA-MRM effectively improved (>1.5 fold) the detection sensitivity of 11 and 21 targets in measurement of saliva and plasma samples, respectively. The integrated results revealed that the salivary levels of these 30 selected biomarkers weakly correlated (r < 0.2) to their plasma levels. Five candidate biomarkers (MMP1, PADI1, TNC, CSTA and MMP3) exhibited significant alterations and disease-discriminating powers (AUC = 0.914, 0.827, 0.813, 0.77, and 0.753) in saliva sample; nevertheless, no such targets could be found in plasma samples. Our data support the notion that saliva may be more suitable for the protein biomarker-based detection of oral cancer, and the newly developed SISCAPA-MRM assay could be applied to verify multiple oral cancer biomarker candidates in saliva samples. SIGNIFICANCE: In this work we systematically determined the abundance of 30 selected targets in the paired saliva and plasma samples to evaluate the utility of saliva and plasma samples for protein biomarker-based detection of oral cancer. Our study provides significant evidence to support the use of saliva, but not blood samples, offer more opportunity to achieve the success of protein biomarker discovery for oral cancer detection.

IL17RB expression might predict prognosis and benefit from gemcitabine in patients with resectable pancreatic cancer.

BACKGROUND: (Interleukin 17 Receptor Beta) IL17RB has been implicated in several malignancies. However, its role in the progression of and chemosensitivity in pancreatic cancer remains unknown. We aimed to determine the clinical significance of IL17RB expression in the prognosis of resectable pancreatic cancer and in the benefits from gemcitabine treatment. MATERIALS AND METHODS: We used microarray and immunohistochemical staining techniques to evaluate IL17RB expression in 91 resectable pancreatic cancer tissues and their respective matched adjacent non-cancerous tissues. Quantitative real-time PCR and Western blotting were used to evaluate IL17RB in human pancreatic cancer cell lines after gemcitabine treatment. The correlation between IL17RB expression and overall survival and disease-free survival times were analyzed. RESULTS: IL17RB expression correlated with lymph node metastasis and (Vascular endothelial growth factor) VEGF expression. Cox proportional model showed that high IL17RB expression is a significant negative prognostic factor for both (overall survival) OS and (disease-free survival) DFS. Kaplan-Meier survival curves confirmed significantly reduced median overall and DFS time in high IL17RB patients as compared with low IL17RB patients. Furthermore, Cox proportional model confirmed a correlation between adjuvant treatment with gemcitabine-based chemotherapy and IL17RB expression. Kaplan-Meier survival curves showed that low IL17RB expression was associated with longer OS and DFS in patients than high IL17RB expression and gemcitabine-based adjuvant chemotherapy. In human pancreatic cancer cell lines, the messenger RNA and protein levels of IL17RB were significantly enhanced after gemcitabine treatment. CONCLUSIONS: IL17RB predicts the prognosis and benefit from gemcitabine in patients with resectable pancreatic cancer.