A multidimensional atlas of human glioblastoma-like organoids reveals highly coordinated molecular networks and effective drugs.

Recent advances in the genomics of glioblastoma (GBM) led to the introduction of molecular neuropathology but failed to translate into treatment improvement. This is largely attributed to the genetic and phenotypic heterogeneity of GBM, which are considered the major obstacle to GBM therapy. Here, we use advanced human GBM-like organoid (LEGO: Laboratory Engineered Glioblastoma-like Organoid) models and provide an unprecedented comprehensive characterization of LEGO models using single-cell transcriptome, DNA methylome, metabolome, lipidome, proteome, and phospho-proteome analysis. We discovered that genetic heterogeneity dictates functional heterogeneity across molecular layers and demonstrates that NF1 mutation drives mesenchymal signature. Most importantly, we found that glycerol lipid reprogramming is a hallmark of GBM, and several targets and drugs were discovered along this line. We also provide a genotype-based drug reference map using LEGO-based drug screen. This study provides new human GBM models and a research path toward effective GBM therapy.

Optical Temperature Sensing Based on Linear Change of Luminescence Intensity Ratio in Y2O3: Tm3+, Eu3+ Phosphors.

In this work, Y2O3: Tm3+, Eu3+ phosphors were made by homogeneous precipitation with urea as precipitator. The emission spectra varying with temperature of Y2O3: Tm3+, Eu3+ phosphors were measured and analyzed. Analysis show that the luminescence of Eu3+ represents a normal thermal quenching change, while that of Tm3+ exhibits slow thermal enhancement phenomenon. In the temperature range of 303-503 K, the luminescence of Tm3+ showed a trend of first strengthening and then weakening. The reason for this phenomenon of Tm3+ is that there is energy transfer from Eu3+ to Tm3+, and the energy transfer efficiency increases gradually with temperature. Meanwhile, the luminescence of Tm3+ also have thermal quenching effect. Under the combined influence of thermal quenching and energy transfer, the luminescence of Tm3+ first becomes stronger and then then becomes weaker. According to the calculation, the luminescence intensity ratio (LIR) of Tm3+ and Eu3+ conforms to the linear empirical formula with increasing temperature. The relative sensitivity of phosphors decreases with Eu3+ concentration increased, and the maximum Sr reaches 0.460% K-1 (1% Tm3+, 0.3% Eu3+, at 303 K). Moreover, the temperature cycle test present that the LIR of phosphors has good repeatability.

A high-precision thermometry strategy by replacing the infrared with visible light for detection.

We have developed a high-precision thermometry strategy based on 4I9/2-4I15/2 (I800 nm) and 4S3/2-4I15/2 (I551 nm) transitions of Er3+, after replacing the measurement of the 4I11/2-4I15/2 (I1000 nm) transition with the 4S3/2-4I15/2 transition, i.e., using visible light for detection instead of infrared. Through rate equation model analysis, (I1000 nm)2 and I551 nm can be substituted for each other under certain conditions. Further, because the 4I9/2 and 4I11/2 levels of Er3+ are thermally coupled, a new idea of ratiometric thermometry is proposed based on the ratio of (I800 nm)2 and I551 nm, which has the advantages of anti-interference of excitation light source fluctuation and background-free detection. The feasibility of the idea was verified by researching the power-dependent emission spectra at different temperatures and temperature-dependent emission spectra of a CaWO4:Er3+,Yb3+ sample under 980-nm laser excitation. The maximum relative sensitivity for the new ratiometric thermometry reaches up to 7.4% K-1 and the optimal temperature uncertainty calculated is 0.03 K at 303 K. This study provides guidance for solving the problem of a weak response of an infrared detector.

Reaction of the anammox granules to various antibiotics and operating the anammox coupled denitrifying reactor for oxytetracycline wastetwater treatment.

The anaerobic ammonium oxidation (anammox) basedtechnology has been considered as an economic and efficient way to remove nitrogen. However, the anammox bacteria could be strongly inhibited by antibiotics. In present research, inhibiting properties of oxytetracycline, penicillin and polymyxin sulfate upon the anammox activity were investigated through batch experiment. The results implied that anammox activity was significantly inhibited by oxytetracycline and polymyxin sulfate. The non-competitive inhibiting model showed that the inhibiting constants (Ki) of oxytetracycline and polymyxin sulfate were 188.5 and 17.7 mg/L, respectively. Meanwhile, the anammox process was not suppressed while the concentration of penicillin reached 3000 mg/L. In long-run experiment, the influent oxytetracycline concentration of the anammox coupled denitrifying reactor was operated at 20 mg/L. It was observed that the anammox performance completely deteriorated, while the NO2--N removing efficiency reached 15.8%. The obtained findings could provide important instruction for the treatment of antibiotic contaminated wastewater.

Ratiometric optical thermometry based on upconversion luminescence with different multi-photon processes in CaWO4:Tm3+/Yb3+ phosphor.

Ratiometric optical thermometry based on upconversion (UC) luminescence with different multi-photon processes in CaWO4:Tm3+,Yb3+ phosphor was developed. A new fluorescence intensity ratio (FIR) thermometry, utilizing the ratio of the cube of 3F2,3 emission to the square of 1G4 emission of Tm3+ and retaining the feature of anti-interference of excitation light source fluctuations, is proposed. Under the hypotheses of the UC terms being neglected in the rate equations and the ratio of the cube of 3H4 emission to the square of 1G4 emission of Tm3+ being a constant in a relatively narrow temperature range, the new FIR thermometry is valid. The correctness of all hypotheses was confirmed by testing and analyzing the power-dependent emission spectra at different temperatures and the temperature-dependent emission spectra of CaWO4:Tm3+,Yb3+ phosphor. The results prove that the new ratiometric thermometry based on UC luminescence with different multi-photon processes is feasible through optical signal processing, and maximum relative sensitivity of the thermometry is 6.61% K-1 at 303 K. This study provides guidance in selecting UC luminescence with different multi-photon processes to construct ratiometric optical thermometers with anti-interference of excitation light source fluctuation.

Assessment of Normal Tissue Radiosensitivity by Evaluating DNA Damage and Repair Kinetics in Human Brain Organoids.

DNA-double strand break (DSB), detected by immunostaining of key proteins orchestrating repair, like γH2AX and 53BP1, is well established as a surrogate for tissue radiosensitivity. We hypothesized that the generation of normal brain 3D organoids ("mini-brains") from human induced pluripotent stem cells (hiPSC) combined with detection of DNA damage repair (DDR) may hold the promise towards developing personalized models for the determination of normal tissue radiosensitivity. In this study, cerebral organoids, an in vitro model that stands in its complexity between 2D cellular system and an organ, have been used. To quantify radiation-induced response, immunofluorescent staining with γH2AX and 53BP1 were applied at early (30 min, initial damage), and late time points (18 and 72 h, residual damage), following clinical standard 2 Gy irradiation. Based on our findings, assessment of DDR kinetics as a surrogate for radiosensitivity in hiPSC derived cerebral organoids is feasible. Further development of mini-brains recapitulating mature adult neuronal tissue and implementation of additional signaling and toxicity surrogates may pave the way towards development of next-generation personalized assessment of radiosensitivity in healthy neuronal tissue.

Rapid cultivation of the aerobic granules for simultaneous phenol degradation and ammonium oxidation in a sequencing batch reactor.

The rapid cultivation strategy of aerobic granular sludge (AGS) for simultaneous phenol degradation and ammonium oxidation was studied in a sequence batch reactor (SBR). The short-term inhibitory kinetics of phenol was studied through batch experiments. For the sodium acetate fed AGS, the phenol inhibition constants (Ki) of specific oxygen utilization rate by heterotrophic bacteria (SOURh) was 508.6 mg/L. The Ki of specific ammonium utilization rate (SAUR) was 232.3 mg/L. After 28 days' acclimatization, the phenol and NH4+-N removal rates of the SBR reached 94.0% and 96.4% when the influent phenol and NH4+-N concentrations were 1000 and 33.5 mg/L, respectively. The phenol removal loading rate was 1.69 kg/(m3·d). For the mature phenol&ammonium degrading AGS, the polysaccharide (PS) and protein (PN) concentrations were 247.4 ±â€¯10.3 and 68.6 ±â€¯6.5 mg/g VSS, respectively. The functional groups analysis showed that the amount of OH, NH, CO and CC groups remained unchanged in the mature phenol&ammonium degrading AGS.

Structural Insight into African Swine Fever Virus dUTPase Reveals a Novel Folding Pattern in the dUTPase Family.

The African swine fever virus (ASFV) is the deadly pathogen of African swine fever (ASF) that induces high mortality, approaching 100% in domestic pigs, causes enormous losses to the global pig industry, and threatens food security. Currently, there is no effective treatment or preventive countermeasure. dUTPases (deoxyuridine 5'-triphosphate pyrophosphatases) are ubiquitous enzymes that are essential for the hydrolysis of dUTP and prevent the misincorporation of dUTP into newly synthesized DNA. Here, we present the crystal structures of the ASFV dUTPase in complex with the product dUMP and cofactor Mg2+ at a resolution of 2.2 Å. We observed that a unique "turning point" at G125 plays an unexpected critical role in the swapping region of the C-terminal segment, which is further stabilized by the interactions of the last C-terminal ß strand with the ß1 and ß2 strands, thereby positioning the catalytic motif 5 into the active site of its own subunit instead of into a third subunit. Therefore, the ASFV dUTPase employs a novel two-subunit active site that is different than the classic trimeric dUTPase active site, which is composed of all three subunits. Meanwhile, further results confirmed that the configuration of motifs 1 to 5 has high structural homology with and a catalytic mechanism similar to that of the known trimeric dUTPases. In general, our study expands the information not only on the structural diversity of the conserved dUTPase family but also on the details needed to utilize this dUTPase as a novel target in the treatment of ASF.IMPORTANCE African swine fever virus (AFSV), a large enveloped double-stranded DNA virus, causes a deadly infection in domestic pigs. In addition to Africa, Europe, and South America, countries in Asia, such as China, Vietnam, and Mongolia, have suffered the hazards posed by ASFV outbreaks in recent years. Until now, there has been no vaccine for protection from ASFV infection or effective treatments to cure ASF. Here, we solved the crystal structure of the ASFV dUTPase-dUMP-Mg2+ complex. The ASFV dUTPase displays a noncanonical folding pattern that differs from that of the classic homotrimeric dUTPase, in which the active site is composed of two subunits. In addition, several nonconserved residues within the 3-fold axis channel play a vital role in ASFV dUTPase homotrimer stability. Our finding on these unique structural features of the ASFV dUTPase could be explored for the design of potential specific inhibitors that target this unique enzyme.

Construction of prognostic microRNA signature for human invasive breast cancer by integrated analysis.

BACKGROUND: Despite the advances in early detection and treatment methods, breast cancer still has a high mortality rate, even in those patients predicted to have a good prognosis. The purpose of this study is to identify a microRNA signature that could better predict prognosis in breast cancer and add new insights to the current classification criteria. MATERIALS AND METHODS: We downloaded microRNA sequencing data along with corresponding clinicopathological data from The Cancer Genome Atlas (TCGA). Of 1,098 breast cancer patients identified, 253 patients with fully characterized microRNA profiles were selected for analysis. A three-microRNA signature was generated in the training set. Subsequently, the performance of the signature was confirmed in a validation set. After construction of the signature, we conducted additional experiments, including flow cytometry and the Cell Counting Kit-8 assay, to illustrate the correlation of this microRNA signature with breast cancer cell cycle, apoptosis, and proliferation. RESULTS: Three microRNAs (hsa-mir-31, hsa-mir-16-2, and hsa-mir-484) were identified to be significantly and independently correlated with patient prognosis, and performed with good stability. Our results suggest that higher expression of hsa-mir-484 indicated worse prognosis, while higher expression of hsa-mir-31 and hsa-mir-16-2 indicated better prognosis. Moreover, additional experiments confirmed that this microRNA signature was related to breast cancer cell cycle and proliferation. CONCLUSION: Our results indicate a three-microRNA signature that can accurately predict the prognosis of breast cancer, especially in basal-like and hormone receptor-positive breast cancer subtypes. We recommend more aggressive therapy and more frequent follow-up for high-risk groups.

Combination of SIRT1 and Src overexpression suggests poor prognosis in luminal breast cancer.

OBJECTIVES: 1) Analyze the correlation of SIRT1 and Src with human breast cancer (BC) prognosis; 2) explore the roles of SIRT1 and Src in BC cell proliferation, tumor invasion, and metastasis; and 3) analyze the correlation and interaction between SIRT1 and Src. MATERIALS AND METHODS: 1) Tissue microarray was used to analyze the expression of SIRT1 and Src in human BC tissues and the correlation between protein expression and cancer prognosis; 2) CCK8 assay was used to determine the influence of SIRT1 and Src inhibitors on BC cell proliferation; 3) Transwell migration assay and wound healing assay were used to determine the effect of SIRT1 and Src inhibitors on BC cell migration and invasion; and 4) Western blotting was used to analyze the correlation and interaction between SIRT1 and Src. RESULTS: 1) Combination of SIRT1 and/or Src positivity is a prognosis factor in BC, especially in luminal type; 2) MCF-7 cell proliferation is suppressed by SIRT1 inhibitor Ex527, and cell migration and invasion were inhibited by Src inhibitor bosutinib; 3) combined with Ex527, bosutinib has a significantly increased effect on MCF-7 cell migration suppression; and 4) there is a positive association between SIRT1 and Src both in BC tissues and in MCF-7 cells. CONCLUSION: 1) SIRT1 and Src overexpression are both correlated with poor prognosis in human BC; 2) SIRT1 + Src (SIRT1 and/or Src positivity) is a fine prognosis model for luminal-type BC; 3) SIRT1 is a copromotor of Src in BC migration and invasion, but not in cell proliferation; and 4) our results suggest a potential interaction or a common regulation pathway between SIRT1 and Src expression and activity.

The prognostic role of Sirt1 expression in solid malignancies: a meta-analysis.

Although many studies have discussed the association of abnormally expressed silent information regulator 1 (Sirt1) with the prognosis of patients with a variety of solid carcinomas, they failed to agree on whether excessive Sirt1 indicates a good or poor overall survival for the patients. We conducted the current meta-analysis to illustrate the prognostic value of Sirt1 in solid malignancies. Articles published before December 2016 were searched using Pubmed and Web of Science. The studies were selected for the meta-analysis based on certain criteria. A total of 7,369 cases from 37 studies were included, in which 48.6% of the patients overexpressed Sirt1. The overall survival (OS) and clinical features, such as age and TNM stage, were analyzed using RevMan 5.3 software. Sirt1 overexpression was significantly correlated with the OS (HR: 1.52, 95% CI: [1.23, 1.88], P = 0.0001), especially in liver cancer (HR: 1.78, 95% CI: [1.46, 2.18], P < 0.00001) and lung cancer (HR: 1.80, 95% CI: [1.06, 3.05], P = 0.03), which suggested that the overexpression of Sirt1 indicates poor prognosis of patients with solid cancers.

Dephosphorylated cofilin expression is associated with poor prognosis in cases of human breast cancer: a tissue microarray analysis.

BACKGROUND: Proteins in the cofilin pathway regulate actin dynamics and may be involved in cancer cell migration and invasion. However, there are no direct data that suggest that dephosphorylated cofilin can affect breast cancer prognosis. METHODS: We assessed the expressions of cofilin and phosphorylated cofilin (P-cofilin) in breast cancer tissue microarrays (290 patients, mean follow-up: 95.7±2.49 months) to evaluate dephosphorylated cofilin and its relationship with breast cancer prognosis. The associations of pathological characteristics with cumulative survival were evaluated using Kaplan-Meier analysis. RESULTS: Univariate analyses revealed that overall survival was associated with cofilin levels, N category, TNM stage, estrogen receptor status, progesterone receptor status, and molecular subtypes. Cofilin status and TNM stage independently affected overall survival, although P-cofilin expression was not associated with patient survival. In the P-cofilin-negative subgroup, cofilin expression was significantly associated with patient survival, although cofilin expression was not significantly associated with patient survival in the P-cofilin-positive subgroup. We further analyzed the P-cofilin-negative cases and found that Ki-67 expression was significantly elevated in the subgroup that was strongly positive for cofilin (P=0.002). CONCLUSION: Among P-cofilin-negative patients with breast cancer, cofilin expression defines a population of patients with lower overall survival, which suggests that dephosphorylated cofilin expression might predict the prognosis in cases of P-cofilin-negative breast cancer. Furthermore, our results suggest that inhibitors of dephosphorylated cofilin expression may provide therapeutic benefits in patients with breast cancer.

The Size Confinement Effect for Eu3+ Concentration Quenching and Energy Transfer in YVO4 Nanocrystal.

YVO4: Eu3+ nanocrystal powders (~30 nm) with different doping concentrations were prepared using a precipitation method. Bulky powders (~500 nm) were obtained by annealing the nanopowders at high temperature. The concentration quenching of luminescent centers and energy transfer in YVO4: Eu3+ powders were investigated. It was found that quenching concentration for Eu3+ 5D0--> 7F2 transition emission in nanopowders is distinctly higher than that in bulk powders. The type of energy transfer that caused concentration quenching was identified to be electric dipole-dipole interaction in bulk powders and exchange interaction in nanopowders. The electric dipole-dipole interaction is a long-range interaction (operating range of several nanometers). The size confinement effect of boundary in nanoparticles has obvious inhibitory effect on electric dipole-dipole interaction, and hardly affect the exchange interaction which is a short-range interaction (operating range several angstroms). The electric dipole-dipole interaction is restrained by particle boundary in nanopowders. So energy transfer of Eu3+ ions in nanomaterials is dominated by exchange interaction, and quenching concentration of nanomaterials is higher than in bulky materials.

Effects of fibroblast growth factor 21 on cell damage in vitro and atherosclerosis in vivo.

Fibroblast growth factor 21 (FGF-21), which is a modulator of glucose and lipid homeostasis, acts as a novel therapeutic reagent for many metabolic perturbations. However, its potential as a treatment for cardiovascular disease, especially atherosclerosis (AS) has not been fully explored. Here, we report that recombinant FGF-21 improves resistance to cell damage from oxidative stress in vitro, and from atherosclerosis in vivo. Human umbilical vein endothelial cells (HUVECs) were induced with H2O2, followed by treatment with high purity recombinant FGF-21. The results indicated that FGF-21 significantly enhanced cell viability and decreased the degree of DNA fragmentation in HUVECs, as caused by H2O2 stress induction. Further studies revealed that FGF-21 inhibited H2O2-induced cell apoptosis by preventing the activation of mitogen-activated protein kinase (MAPK) signaling pathways. In an established rat model, FGF-21 dramatically improved the condition of atherosclerotic rats by decreasing serum levels of total triglyceride (TG), low density lipoprotein cholesterol (LDL-C), and total cholesterol (TC), and by increasing the serum levels of high density lipoprotein cholesterol (HDL-C). FGF-21 also has antioxidant effects in the atherosclerotic rat, such that increased levels of superoxide dismutase, reduced glutathione, and reduced malondialdehyde were observed. These data provide novel insight into the potential use of FGF-21 in the prevention and treatment of human cardiovascular diseases.

The size confinement effect for Ln3+ (Ln = Tm or Eu) concentration quenching and energy transfer in Y2O3 nanocrystals.

Y2O3:Ln (Ln = Tm or Eu) nano-powders with different particle sizes and various doping concentrations were prepared by using a combustion method. The bulk powders doped with the same concentrations were obtained by annealing the nano-powders at high temperatures. Emission spectra of the phosphors were measured. The crystal structure and morphology of the phosphors were characterized by XRD (X-ray diffraction) and FE-SEM (field emission scanning electron microscopy), respectively. The concentration quenching of luminescent centers and energy transfer between luminescent centers in Y2O3:Ln nanocrystal powders were investigated. It is found that the behavior of luminescent concentration quenching for Eu3+ 5D0 --> 7F2 in nano-powders is similar to that in bulk powders. On the contrary, the quenching concentration for Tm3+ 1D2 --> 3H4 is distinctly higher than that in bulk powders. This owes to the size confinement effect which will restrain the electric dipole-dipole interaction as a long-rang interaction (e.g., energy transfer between Tm3+ ions), and will hardly affect the exchange interaction which is a short-rang interaction (e.g., energy transfer between Eu3+ ions).

Wetting and reaction promoted by ultrasound between sapphire and liquid Al-12Si alloy.

Ultrasonic-assisted wetting between sapphire bulks and liquid Al-12Si alloy in an atmospheric environment at 620 °C is carried out in this study. Complete, rather than partial, wetting and joining can be achieved with the aid of ultrasound. Growth of epitaxial alumina on sapphire bulks is promoted dramatically during ultrasonic-assisted wetting comparing to that during hot-dipping without ultrasound. XRD results show that the epitaxial alumina is non-crystalline. This indicates that the temperature on the surface of the sapphire substrate is not more than 1200 °C even though the collapse of acoustic cavitation bubbles could theoretically produce extremely high temperature. The bonding force at the interface between the Al-Si alloy and sapphire is strengthened because of the epitaxial alumina. The interfacial shear strength of sapphire/Al-Si alloy can reach as high as 60-65 MPa. The fracture morphology shows that cracks initiated at the interface between Si grains and the epitaxial alumina on sapphire. This result is especially useful for the joining of metals and ceramics.

Specific PCR detection of tiger, leopard, and lion ingredients from test samples.

A PCR method was developed for specific detection of tiger, leopard, and lion DNA from test specimens for inspection and quarantine or for law-enforced animal protection. Three pairs of specific primers were designed based on the mitochondrial cytochrome b gene of tiger, leopard, and lion and used in the PCR testing. To mimic the effect of food processing on the sensitivity of the test, the tiger muscle and bovine bonemeal powder samples were treated at 133 degrees C for 30 min. At this processing condition, the method was sensitive enough to detect as low as 0.05% of tiger-derived ingredients from the mixed bonemeal powders. The data demonstrate that our PCR method is convenient and economic, with high sensitivity and repeatability, and can be used to detect and identify tiger, leopard, and lion ingredients from various test samples.

Neuroprotective effects of chlorogenic acid against apoptosis of PC12 cells induced by methylmercury.

Chlorogenic acid (CGA) widely exists in edible and medicinal plants. We aimed to evaluate the effect of CGA on the protection from apoptosis by methylmercury (MeHg) in PC12 cells. Cell viability was evaluated by MTT assay. Apoptosis was assayed by flow cytometry detection. Caspase-3 activity was measured by confocal microscopy. Intracellular GSH levels were determined by bicinchoninic acid protein assay. Intracellular reactive oxygen species (ROS) was assessed by means of chloromethyl-dihydrodichlorofluorescein diacetate. Glutathione peroxidase (GPx) activity was determined by UV. In order to elucidate the action of CGA, the protective effects of CGA were compared to Vit.E. CGA was effective at protecting PC12 cells against MeHg-induced damage in dose-dependent manner. CGA not only suppressed the generation of ROS, the decrease of activity in GPx and the decrease of GSH, but also attenuated caspase-3 activation in PC12 cells by MeHg. CGA eventually protected PC12 cells against MeHg-induced apoptosis. The results highlighted that CGA may exert neuroprotective effects through its antioxidant actions.