Enhancing vulnerability assessment through spatially explicit modeling of mountain social-ecological systems exposed to multiple environmental hazards.

The evaluation of the vulnerability of coupled socio-ecological systems is critical for addressing and preventing the adverse impacts of various environmental hazards and devising strategies for climate change adaptation. The initial step in vulnerability assessment involves exposure assessment, which entails quantifying and mapping the risks posed by multiple environmental hazards, thereby offering valuable insights for the implementation of vulnerability assessment methodologies. Consequently, this study sought to model the exposure of coupled social-ecological systems in mountainous regions to various environmental hazards. By a set of socio-economic, climatic, geospatial, hydrological, and demographic data, as well as satellite imagery, and examining 11 hazards, including droughts, pests, dust storms, winds, extreme temperatures, evapotranspiration, landslides, floods, wildfires, and social vulnerability, this research employed machine learning (ML) techniques and the fuzzy analytical hierarchy process (FAHP). Expert opinions were utilized to guide hazard weighting and calculate the exposure index (EI). Through the precise spatial mapping of EI variations across the socio-ecological systems in mountainous areas, this investigation provides insights into vulnerability to multiple environmental hazards, thereby laying the groundwork for future endeavors in supporting national-level vulnerability assessments aimed at fostering sustainable environments. The findings reveal that social vulnerability and pests receive the highest weighting, while floods and landslides are ranked lower. All hazards demonstrate significant correlations with the EI, with droughts exhibiting the strongest correlation (r > 0.81). Spatial analysis indicates a north-south gradient in forest exposure, with southern regions showing higher exposure hotspots (EI 29.08) compared to northern areas (EI 10.60). Validation based on Area Under Curve (AUC) and Consistency Rate (CR) in FAHP demonstrates robustness, with AUC values exceeding 0.78 and CR values below 0.1. Considering the anticipated intensification of hazards, management strategies should prioritize reducing social vulnerability, restore degraded areas using drought-resistant species, combat pests, and mitigate desertification. By integrating multidisciplinary data and expert opinions, this research contributes to informed decision-making regarding sustainable forest management and climate resilience in mountain ecosystems.

A point-of-care testing assay for clonorchiasis using a EuNPs-CsTR1 fluorescent probe-based immunoassay.

Clonorchis sinensis is one of the most important fish-borne zoonotic parasitic worms in humans, and is distributed in several countries with more than 15 million people infected globally. However, the lack of a point-of-care testing (POCT) method is still the critical barrier to effectively prevent clonorchiasis. With the application of novel fluorescent nanomaterials, the development of on-site testing methods with high signal enhancement can provide a simple, precise and inexpensive tool for disease detection. In this study, Eu-(III) nanoparticles (EuNPs) were used as indicative probes, combined with C. sinensis tandem repeat sequence 1 (CSTR1) antigen to capture specific antibodies. Afterward, the complex binds to mouse anti-human IgG immobilized on the test line (T-line) producing a fluorescent signal under UV light. The EuNPs-fluorescent immunoassay (EuNPs-FIA) was successfully constructed, allowing sample detection within 10 min. It enabled both qualitative determination with the naked eye under UV light and quantitative detection by scanning the fluorescence intensity on the test line and control line (C-line). A total of 133 clinical human sera (74 negative, 59 clonorchiasis, confirmed by conventional Kato-Katz (KK) methods and PCR via testing fecal samples corresponding to each serum sample) were used in this study. For qualitative analysis, the cut-off value of fluorescence for positive serum was 31.57 by testing 74 known negative human samples. The assay had no cross-reaction with other 9 parasite-infected sera, and could recognize the mixed infection sera of C. sinensis and other parasites. The sensitivity and specificity of EuNPs-FIA were both 100% compared with KK smear method. Taking advantage of its high precision and user-friendly procedure, the established EuNPs-FIA provides a powerful tool for the diagnosis and epidemiological survey of clonorchiasis.

Predefined-time position tracking optimization control with prescribed performance of the induction motor based on observers.

To solve the position control problem of the induction motor with parameter perturbations, load disturbances, and modeling errors, a predefined-time position tracking optimization control method with prescribed performance is proposed. In practice, the rotor flux linkage of the induction motor can't be measured, and a predefined-time sliding mode observer (PTSMO) is applied to accurately estimate it. Additionally, predefined-time disturbance observers (PTDOs) are employed to identify the uncertainties in the motor system. The position and flux linkage controllers are then designed by integrating the predefined-time control approach with the prescribed performance function method, realizing accurate tracking control of the induction motor within a predefined time. Next, the adaptive genetic algorithm (AGA) and the improved particle swarm optimization (IPSO) technique are combined to optimize the designed controllers, enhancing the convergence rate and steady-state accuracy of the induction motor. Finally, comparative analyses through simulations and dSPACE simulated experiments validate the efficacy of the proposed control method, highlighting its applicability in practical motor systems.

Cancer associated fibroblasts serve as an ovarian cancer stem cell niche through noncanonical Wnt5a signaling.

Frequent relapse and chemoresistance cause poor outcome in ovarian cancer (OC) and cancer stem cells (CSCs) are important contributors. While most studies focus exclusively on CSCs, the role of the microenvironment in providing optimal conditions to maintain their tumor-initiating potential remains poorly understood. Cancer associated fibroblasts (CAFs) are a major constituent of the OC tumor microenvironment and we show that CAFs and CSCs are enriched following chemotherapy in patient tumors. CAFs significantly increase OC cell resistance to carboplatin. Using heterotypic CAF-OC cocultures and in vivo limiting dilution assay, we confirm that the CAFs act by enriching the CSC population. CAFs increase the symmetric division of CSCs as well as the dedifferentiation of bulk OC cells into CSCs. The effect of CAFs is limited to OC cells in their immediate neighborhood, which can be prevented by inhibiting Wnt. Analysis of single cell RNA-seq data from OC patients reveal Wnt5a as the highest expressed Wnt in CAFs and that certain subpopulations of CAFs express higher levels of Wnt5a. Our findings demonstrate that Wnt5a from CAFs activate a noncanonical Wnt signaling pathway involving the ROR2/PKC/CREB1 axis in the neighboring CSCs. While canonical Wnt signaling is found to be predominant in interactions between cancer cells in patients, non-canonical Wnt pathway is activated by the CAF-OC crosstalk. Treatment with a Wnt5a inhibitor sensitizes tumors to carboplatin in vivo. Together, our results demonstrate a novel mechanism of CSC maintenance by signals from the microenvironmental CAFs, which can be targeted to treat OC chemoresistance and relapse.

Qi-Ju-Di-Huang-Pill delays the progression of diabetic retinopathy.

ETHNOPHARMACOLOGICAL RELEVANCE: Qi-Ju-Di-Huang-Pill (QJDH pill) is a Chinese decoction. Although it is commonly used to treat eye conditions, such as diabetic retinopathy (DR), its exact mechanism of action is unknown. AIM OF THE STUDY: To investigate the specific mechanism by which QJDH pill slows the progression of diabetic retinopathy (DR) based on animal and cellular experiments. MATERIAL AND METHODS: The major components of QJDH pill were characterized by ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLCMS/MS). C57BL/6J mice were randomly divided into five groups as follows: normal group (control group), model group (STZ group), low-dosage QJDH pill group (QJDH-L group), medium-dosage QJDH pill group (QJDH-M group) and high-dosage QJDH pill group (QJDH-H group). Changes in water intake, urination, food intake, and body mass were monitored weekly, while changes in blood glucose were monitored monthly. Fluorescein fundus angiography (FFA), optical coherence tomography angiography (OCTA), and optical coherence tomography (OCT) were utilized to analyze the changes in fundus imaging indications. Hematoxylin & eosin (H&E) and transmission electron microscopy (TEM) were employed to examine histopathologic and ultrastructural changes in retina. The levels of interleukin-6 (IL-6), interleukin-17 (IL-17), tumor necrosis factor-α (TNF-α), and vascular endothelial growth factor (VEGF) in peripheral blood were detected using Enzyme-linked immunosorbent assay (ELISA). The mouse retina apoptotic cells were labeled with green fluorescence via terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (Tunel). The protein levels of Bcl-2-Associated X (Bax), B cell lymphoma 2 (Bcl-2), Caspase-3, PI3K, phosphorylated PI3K (p-PI3K), protein kinase B (AKT) and phosphorylated AKT (p-AKT) were quantified by Western blot (WB). The retinal pigment epithelium (RPE) cells were cultured and classified into five groups as follows: normal glucose group (NG group), high glucose group (HG group), high glucose + QJDH pill group (HG + QJDH group), high glucose + inhibitor group (HG + LY294002 group), and high glucose + inhibitor + QJDH pill group (HG + LY294002 + QJDH group). Cell viability and apoptosis were detected via Cell Counting Kit-8 (CCK8) and then analyzed by flow cytometry. RESULTS: In vivo experiments revealed that the QJDH pill effectively reduced blood glucose, symptoms of increased water intake, elevated urination, increased food intake and decreased body mass in DR mice. QJDH pill also slowed the development of a series of fundus imaging signs, such as retinal microangiomas, tortuous dilatation of blood vessels, decreased vascular density, and thinning of retinal thickness, downregulated IL-6, IL-17, TNF-α, and VEGF levels in peripheral blood, and inhibited retinal cell apoptosis by activating the PI3K/AKT signaling pathway. Moreover, in vitro experiments showed that high glucose environment inhibited RPE cell viability and activated RPE cell apoptosis pathway. In contrast, lyophilized powder of QJDH pill increased RPE cell viability, protected RPE cells from high glucose-induced damage, and decreased apoptosis of RPE cells by activating the pi3k pathway. CONCLUSION: QJDH pill induces hypoglycemic, anti-inflammatory effects, anti-VEGF and anti-retinal cell apoptosis by activating PI3K/AKT signaling pathway, and thus can protect the retina and slow the DR progression.

Efficacy of Arthroscopic Discopexy on Condylar Growth in Temporomandibular Joint Anterior Disc Displacement: A Randomized Clinical Trial.

BACKGROUND: Adolescents with temporomandibular joint (TMJ) anterior disc displacement (ADD) frequently develop dentofacial deformities. It is unknown if adjunctive arthroscopic discopexy compared to orthodontic treatment alone increases condylar growth and then improves dentofacial deformity. This study aimed to determine whether arthroscopic discopexy before functional appliance (joint-occlusal treatment) or single functional appliance (occlusal treatment) increases condylar growth and improves dentofacial deformity among adolescents with TMJ ADD. METHODS: A multicenter, randomized, parallel controlled trial was conducted in three centers in China. Adolescents diagnosed with TMJ ADD and dentofacial deformity were enrolled. Eligible participants were randomly assigned to a joint-occlusal group or occlusal group at a ratio of 2:1. MRI, cephalometric radiographs were evaluated at baseline, 8 months and 14 months. The primary outcome was changes in condylar height from 14 months to baseline. Secondary outcomes were changes in skeletal position. RESULTS: A total of 240 patients (14.65±1.88 years of age) were randomized (joint-occlusal group: 160; occlusal group: 80). The overall difference in condylar height between groups was 3.65 mm (95% CI, 3.10 to 4.19; p < 0.001). The between-group differences in condylar height on the left and right sides were 3.60 mm (95% CI, 2.92 to 4.28; p < 0.001) and 3.69 mm (95% CI, 3.06 to 4.32; p < 0.001), respectively. Significant between-group differences were noted in skeletal position (all p < 0.001). CONCLUSIONS: Joint-occlusal treatment can promote condylar growth and improve dentofacial deformity in adolescents after 14 months when compared with single occlusal treatment.

Attention-aware contrastive learning for predicting T cell receptor-antigen binding specificity.

MOTIVATION: It has been proven that only a small fraction of the neoantigens presented by major histocompatibility complex (MHC) class I molecules on the cell surface can elicit T cells. This restriction can be attributed to the binding specificity of T cell receptor (TCR) and peptide-MHC complex (pMHC). Computational prediction of T cells binding to neoantigens is a challenging and unresolved task. RESULTS: In this paper, we proposed an attention-aware contrastive learning model, ATMTCR, to infer the TCR-pMHC binding specificity. For each TCR sequence, we used a transformer encoder to transform it to latent representation, and then masked a percentage of amino acids guided by attention weights to generate its contrastive view. Compared to fully-supervised baseline model, we verified that contrastive learning-based pretraining on large-scale TCR sequences significantly improved the prediction performance of downstream tasks. Interestingly, masking a percentage of amino acids with low attention weights yielded best performance compared to other masking strategies. Comparison experiments on two independent datasets demonstrated our method achieved better performance than other existing algorithms. Moreover, we identified important amino acids and their positional preference through attention weights, which indicated the potential interpretability of our proposed model.

Birth defects data from hospital-based birth defect surveillance in Guilin, China, 2018-2020.

Objectives: Birth defects (BDs) are a major contributor to perinatal and infant mortality, morbidity and lifelong disability worldwide. A hospital-based study on birth defects was designed in Guilin city in the Guangxi province of Southwestern China aiming to determine the prevalence of BDs in the studied region, and the classify the BDs based on clinical presentation and causation. Methods: The study involved BDs among all pregnancy outcomes (live births, stillbirths, death within 7 days, and pregnancy terminations) born in the 42 registered hospitals of Guilin between 2018 and 2020. The epidemiological characteristics of BDs and the etiologic profile of BDs were evaluated in this study. Results: Of the total 147,817 births recorded during the study period, 2,003 infants with BDs were detected, giving a total prevalence rate of 13.55 per 1,000 births. The top five BD types were congenital heart defects, polydactyly, syndactyly, malformations of the external ear, and talipes equinovarus, whereas, neural tube defects, congential esophageal atresia, gastroschisis, extrophy of urinary bladder, were the least common BD types in these 3 years. Only 8.84% of cases were assigned a known etiology, while most cases (91.16%) could not be conclusively assigned a specific cause. Conclusion: This study provides an epidemiological description of BDs in Guilin, which may be helpful for understanding the overall situation in Southwest China of BDs and aid in more comprehensive studies of BDs in future healthcare systems, including funding investment, policy-making, monitor, prevention. Strong prevention strategies should be the priority to reduce BDs and improve the birth quality.

Binocular treatment for individual with amblyopia: A systematic review and meta-analysis.

OBJECTIVE: To assess the efficacy of binocular treatment for individual with amblyopia. METHODS: In this meta-analysis, a comprehensive search of literatures was performed from PubMed, Embase, Cochrane Library and Web of Science databases up to December 21, 2020. Sensitivity analysis was performed for all outcomes. The Begg's test was used to assess the publication bias. Heterogeneity test was conducted for each effect indicator. Indicators were analyzed by random-effects model when the heterogeneity statistic I2 ≥ 50%, on the contrary, indicators were analyzed by fixed-effect model. Standard mean difference (SMD) or weighted mean difference (WMD) was adopted as effect indicators, and the effect amount was expressed as 95% confidence intervals (CIs). RESULTS: A total of 13 literatures including 1146 participants were finally enrolled, with 595 in the intervention group and 551 in the control group. The results indicated that the improvement of amblyopic eye visual acuity [SMD: 0.882, 95%CI: (0.152, 1.613), P = 0.018] in binocular treatment group was better than that in control group. And binocular treatment could improve stereo acuity in individual with amblyopia [WMD: 0.138, 95%CI: (0.068, 0.208), P < 0.001]. CONCLUSION: Binocular treatment may be beneficial to visual acuity, stereo acuity and binocular function improvement for individual with amblyopia. In clinical practice, binocular treatment can be used as one of the treatments for individual with amblyopia.

Use of an ultraviolet light-activated persulfate process to degrade humic substances: effects of wavelength and persulfate dose.

Natural organic matter (NOM), commonly found in surface and ground waters, form disinfection by-products in drinking water. Generally, advanced oxidation processes (AOPs) featuring hydrogen peroxide are used to treat water; however, sulfate radical recently has been used to treat recalcitrant organics, because it is associated with a higher oxidation potential and more effective removal than hydroxyl radicals. Hence, in this research, we evaluated persulfate oxidation efficiency in terms of reductions in humic substance levels and investigated the degradation mechanism. The results showed that ultraviolet-activated persulfate effectively treated humic substances compared with hydrogen peroxide and direct irradiation. Treatment was dose and wavelength dependent; higher persulfate concentrations or shorter UV wavelengths were more effective for treating humic substances as high concentration sulfate radicals were created. The degradation mechanism was similar to that of hydrogen peroxide. Aromatic and chromophore components were more susceptible to degradation than were lower molecular weight components, being initially decomposed into the latter, reducing UV254 absorbance and the SUVA254. Lower molecular weight materials were eventually degraded to end products: NPOC levels fell. And we also treated the inflow of a drinking water treatment plant with persulfate, and humic substances were effectively removed.

Surface modification of titanium with collagen/hyaluronic acid and bone morphogenetic protein 2/7 heterodimer promotes osteoblastic differentiation.

The aim of this study was to evaluate the effects of a collagen/hyaluronic acid coating without or with incorporated heterodimeric bone morphogenetic protein 2/7 (BMP2/7) on in-vitro osteoblastic differentiation on titanium discs. The multilayer collagen/hyaluronic acid coatings without or without incorporated BMP2/7 were deposited on titanium discs via a layer-by-layer technique. The effects of the coatings were evaluated by assessing the alkaline phosphatase (ALP) activity (an early osteoblastic differentiation marker) and the osteocalcin expression (a late osteoblastic differentiation marker). The expression levels of the osteoblastic genes, such as alkaline phosphatase 2 (AKP2) and osteocalcin (OC) were detected using real-time RT-PCR. ALP activity and OC expression were significantly increased when cells were cultured with collagen/hyaluronic acid+BMP2/7 heterodimer (p<0.05). The same result was found in cells with the expression of a BMP2/7 fusion gene, OC and AKP2. These results indicated that collagen/hyaluronic acid+BMP2/7 heterodimer-coated discs might have the potential to greatly enhance osseointegration than a either BMP2 or BMP7 solution or a mixture of BMP2 and BMP7 BMP2/7.

Chemical and flavor profile changes of cocoa beans (Theobroma cacao L.) during primary fermentation.

This survey reports for the first time the changed of quality of fermented cocoa (Theobroma cacao L.) beans. The quality evaluation and simultaneous detection of amino acids, flavor, procyanidin, color, fat, protein, antioxidant activity, and enthalpy were obtained for different fermentation stages of cocoa beans. The results showed that total essential amino acids contents ranged from 2.64 g/100 g to 3.68 g/100 g. A total of 88 compounds identified at the end of the fermentation belonged to alcohols, acids, esters, ketones, pyrazines, aldehydes, and terpenoids. One of the chemical groups that were present in highest abundance in the consummation treatments was acids, representing 56.04% of the total extracted area, followed by alcohols (22.95%) and ketones (9.40%). The colors of the beans in different fermentation stages were different, from deep purple to deep red-brown. Fermented cocoa beans were shown to be 53.45% and 13.51% bean butter and protein content, respectively. The value of denaturation enthalpy (ΔH) ranged from 30.4 (J/g) to 43.38 (J/g). The 3-day fermented sample had the highest ΔH (43.38 J/g). When the fermentation process was complete, the procyanidin concentration of the beans decreased, with the final yield of procyanidin at 6.2%. During fermentation, the antioxidant capacity of beans gradually reduced. The fermenting of cocoa beans had a significant effect on the quality formation. The findings of this study constitute a basis for further investigations on the quality formation of cocoa during fermentation.

Nobiletin inhibits cell growth through restraining aerobic glycolysis via PKA-CREB pathway in oral squamous cell carcinoma.

BACKGROUND/AIM: Nobiletin is a polymethoxylated flavone enriched in Citrus and is used as an important drug in traditional Chinese medicine for various kinds of diseases. Among its multiple functions, it has shown that nobiletin inhibits proliferation of various cancer cells. However, it is unclear whether nobiletin inhibits the growth of oral squamous cell carcinoma (OSCC) cells. MATERIALS AND METHODS: We explored the antitumor effects of nobiletin in TCA-8113 and CAL-27 oral squamous cells. The Cell Counting Kit-8 (CCK8) assay was used to measure cell vitality. Flow cytometry was performed to measure the number of cells in the various phases of the cell cycle. PCR and Western blot were applied to determine mRNA and protein expression, respectively. RESULTS: Nobiletin inhibited proliferation of TCA-8113 and CAL-27 cells via inducing cell cycle arrest at the G1 phase. In addition, the levels of phosphorylated-PKA and phosphorylated-CREB were reduced in nobiletin-treated TCA-8113 and CAL-27 cells. Importantly, our results showed that nobiletin treatment resulted in impaired mitochondrial function and altered glucose consumption, and pyruvate and lactate production. Lastly, nobiletin was found to inhibit the generation of xenografts in vivo. Interestingly, administration of 50 µmol/L Sp-cAMP, a potent PKA activator, rescued all phenotypes caused by nobiletin. CONCLUSIONS: Nobiletin inhibits OSCC cell proliferation in a mitochondria-dependent manner, indicating that it may have a promising role in cancer treatment and attenuation of drug resistance.

Cytocompatible cerium oxide-mediated antioxidative stress in inhibiting ocular inflammation-associated corneal neovascularization.

As oxidative stress is involved with inflammation and neovascularization, blocking oxidative stress may be beneficial for reducing inflammation. To investigate the potential use of cerium oxide nanoparticles (CeNPs) in treating neovascularization-related ophthalmic diseases, various CeNP samples were synthesized, and the sample with the best antioxidant efficacy was used in a rat model of inflammation-associated corneal neovascularization. This synthesized cerium oxide showed good biocompatibility and was capable of mediating a decrease in the expression levels of inflammatory factors via antioxidative stress. Additionally, in vitro tests showed that the Ce3+/Ce4+ ratio of the CeNPs directly affected the antioxidative activity, with higher ratios achieving better efficacy. The anti-inflammatory efficacy of the functional CeNPs was examined both in vitro and in vivo. Slit-lamp biomicroscopy and histological analysis revealed the gradual development of corneal neovascularization, suggesting that inflammation and neovascularization could be controlled by reducing the level of oxidative stress. CeNP-induced antioxidation could serve as a new strategy in the development of long-acting functional agents for treating ophthalmic diseases.

Cancer Associated Fibroblasts: Naughty Neighbors That Drive Ovarian Cancer Progression.

Ovarian cancer is the most lethal gynecologic malignancy, and patient prognosis has not improved significantly over the last several decades. In order to improve therapeutic approaches and patient outcomes, there is a critical need for focused research towards better understanding of the disease. Recent findings have revealed that the tumor microenvironment plays an essential role in promoting cancer progression and metastasis. The tumor microenvironment consists of cancer cells and several different types of normal cells recruited and reprogrammed by the cancer cells to produce factors beneficial to tumor growth and spread. These normal cells present within the tumor, along with the various extracellular matrix proteins and secreted factors, constitute the tumor stroma and can compose 10⁻60% of the tumor volume. Cancer associated fibroblasts (CAFs) are a major constituent of the tumor microenvironment, and play a critical role in promoting many aspects of tumor function. This review will describe the various hypotheses about the origin of CAFs, their major functions in the tumor microenvironment in ovarian cancer, and will discuss the potential of targeting CAFs as a possible therapeutic approach.

Nontargeted Metabolomics for Phenolic and Polyhydroxy Compounds Profile of Pepper (Piper nigrum L.) Products Based on LC-MS/MS Analysis.

In the present study, nontargeted metabolomics was used to screen the phenolic and polyhydroxy compounds in pepper products. A total of 186 phenolic and polyhydroxy compounds, including anthocyanins, proanthocyanidins, catechin derivatives, flavanones, flavones, flavonols, isoflavones and 3-O-p-coumaroyl quinic acid O-hexoside, quinic acid (polyhydroxy compounds), etc. For the selected 50 types of phenolic compound, except malvidin 3,5-diglucoside (malvin), l-epicatechin and 4'-hydroxy-5,7-dimethoxyflavanone, other compound contents were present in high contents in freeze-dried pepper berries, and pinocembrin was relatively abundant in two kinds of pepper products. The score plots of principal component analysis indicated that the pepper samples can be classified into four groups on the basis of the type pepper processing. This study provided a comprehensive profile of the phenolic and polyhydroxy compounds of different pepper products and partly clarified the factors responsible for different metabolite profiles in ongoing studies and the changes of phenolic compounds for the browning mechanism of black pepper.

Curcumin analog EF24 induces apoptosis and downregulates the mitogen activated protein kinase/extracellular signal-regulated signaling pathway in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common malignancies worldwide. Diphenyldifluoroketone (EF24) is a curcumin analog that has been demonstrated to improve anticancer activity; however, its therapeutic potential and mechanisms in oral cancer remain unknown. In the present study, the effect of EF24 on apoptosis induction and its potential underlying mechanism in the CAL­27 human OSCC cell line was investigated. To achieve this, various concentrations of cisplatin or EF24 were administrated to CAL­27 cells for 24 h, and cell viability, apoptotic DNA fragmentation, and cleaved caspase 3 and 9 levels were evaluated. To investigate the potential underlying mechanism, the levels of mitogen­activated protein kinase kinase 1 (MEK1) and extracellular signal­regulated kinase (ERK), two key proteins in the mitogen­activated protein kinase/ERK signaling pathway, were additionally examined. The results indicated that EF24 and cisplatin treatment decreased cell viability. EF24 treatment increased the levels of activated caspase 3 and 9, and decreased the phosphorylated forms of MEK1 and ERK. Sequential treatments of EF24 and 12­phorbol­13­myristate acetate, a MAPK/ERK activator, resulted in a significant increase of activated MEK1 and ERK, and reversed cell viability. These results suggested that EF24 has potent anti­tumor activity in OSCC via deactivation of the MAPK/ERK signaling pathway. Further analyses using animal models are required to confirm these findings in vivo.

Comparative metabolomics in vanilla pod and vanilla bean revealing the biosynthesis of vanillin during the curing process of vanilla.

High-performance liquid chromatography-mass spectrometry (LC-MS) was used for comprehensive metabolomic fingerprinting of vanilla fruits prepared from the curing process. In this study, the metabolic changes of vanilla pods and vanilla beans were characterized using MS-based metabolomics to elucidate the biosynthesis of vanillin. The vanilla pods were significantly different from vanilla beans. Seven pathways of vanillin biosynthesis were constructed, namely, glucovanillin, glucose, cresol, capsaicin, vanillyl alcohol, tyrosine, and phenylalanine pathways. Investigations demonstrated that glucose, cresol, capsaicin, and vanillyl alcohol pathway were detected in a wide range of distribution in microbial metabolism. Thus, microorganisms might have participated in vanillin biosynthesis during vanilla curing. Furthermore, the ion strength of glucovanillin was stable, which indicated that glucovanillin only participated in the vanillin biosynthesis during the curing of vanilla.

A Cisplatin Derivative Tetra-Pt(bpy) as an Oncotherapeutic Agent for Targeting ALT Cancer.

Background: In approximately 15% of human cancers, telomere length is maintained independently of telomerase by the homologous recombination (HR)-mediated alternative lengthening of telomeres (ALT) pathway. Whether the ALT pathway can be exploited for therapeutic treatment remains unknown. The purpose of this study is to develop oncotherapeutic agent to target ALT cancers. Methods: Surface plasmon resonance assay, antibody to G-quadruplex, and fluorescence in situ hybridization (FISH) were used to discover Tetra-Pt(bpy), a cisplatin derivative that specifically targets telomeric G-quadruplex. We used immunofluorescence, FISH, C-circle assay, and chromosome orientation FISH to evaluate the inhibitory effect of Tetra-Pt(bpy) on ALT activity in human ALT cancers. The shortening of telomere length induced by Tetra-Pt(bpy) was determined by telomere restriction fragment or Q-FISH. Cell destination after Tetra-Pt(bpy) treatment was determined by ß-gal staining or apoptosis assay. Nude mice (n = 4 per group) were injected with U2OS cells to evaluate the effects of Tetra-Pt(bpy) on tumor growth. All statistical tests were two-sided. Results: Tetra-Pt(bpy) inhibits the strand invasion/annealing step of telomeric homologous recombination by selectively converting telomeric ssDNA to a G-quadruplex. ALT-cells treated with Tetra-Pt(bpy) show fewer ALT-associated promyelocytic leukemia bodies (untreated: mean±SD = 5.9±0.2 vs treated: mean±SD = 3.1±0.1, P < .001), fewer extrachromosomal C-circles (untreated: mean±SD = 100.5±1.6 vs treated: mean±SD = 18.0±1.7, P < .001), and reduced telomere sister chromatin exchanges (untreated: mean±SD = 25.2%±1.5% vs treated: mean±SD = 13.1%±1.9%, P < .001). Consequently, critically short telomeres accumulate after multiple population doublings (untreated: mean±SD = 18.9%±1.7% vs treated: mean±SD = 57.4%±2.2%, P < .001), resulting in cell death by apoptosis or senescence. In vivo, Tetra-Pt(bpy) severely inhibits the growth of ALT-cell xenograft tumors in mice (untreated: mean±SD = 57.1±3.7 mm 3 vs treated: mean±SD = 19.0±3.2 mm 3 , P < .001). Importantly, Tetra-Pt(bpy) exhibits no adverse effects on proliferation, gene expression, or telomere metabolism in normal cells. Conclusions: These results reveal the potential of Tetra-Pt(bpy) as a novel oncotherapeutic agent for targeting ALT cancer cells.