Suppression of Hypoxia-Inducible Factor 1α (HIF-1α) by Tirapazamine Is Dependent on eIF2α Phosphorylation Rather Than the mTORC1/4E-BP1 Pathway

Hypoxia-inducible factor 1 (HIF-1), a heterodimeric transcription factor that mediates the adaptation of tumor cells and tissues to the hypoxic microenvironment, has attracted considerable interest as a potential therapeutic target. Tirapazamine (TPZ), a well-characterized bioreductive anticancer agent, is currently in Phase II and III clinical trials. A major aspect of the anticancer activity of TPZ is its identity as a tumor-specific topoisomerase IIα inhibitor. In the study, for the first time, we found that TPZ acts in a novel manner to inhibit HIF-1α accumulation driven by hypoxia or growth factors in human cancer cells and in HepG2 cell-derived tumors in athymic nude mice. We investigated the mechanism of TPZ on HIF-1α in HeLa human cervical cancer cells by western blot analysis, reverse transcription-PCR assay, luciferase reporter assay and small interfering RNA (siRNA) assay. Mechanistic studies demonstrated that neither HIF-1α mRNA levels nor HIF-1α protein degradation are affected by TPZ. However, TPZ was found to be involved in HIF-1α translational regulation. Further studies revealed that the inhibitory effect of TPZ on HIF-1α protein synthesis is dependent on the phosphorylation of translation initiation factor 2α (eIF2α) rather than the mTOR complex 1/eukaryotic initiation factor 4E-binding protein-1 (mTORC1/4E-BP1) pathway. Immunofluorescence analysis of tumor sections provide the in vivo evidences to support our hypothesis. Additionally, siRNA specifically targeting topoisomerase IIα did not reverse the ability of TPZ to inhibit HIF-1α expression, suggesting that the HIF-1α inhibitory activity of TPZ is independent of its topoisomerase IIα inhibition. In conclusion, our findings suggest that TPZ is a potent regulator of HIF-1α and provide new insight into the potential molecular mechanism whereby TPZ serves to reduce HIF-1α expression

Utjecaj lantana na mikrostrukturu i svojstva čelika ASTM A216

In order to satisfy the rudder horn casting standards of the International Association of Classification Societies, the properties of ASTM A216 steel should be improved. Therefore, in this article the rudder horn casting and accompanying specimens were cast moulded by arc furnace smelting, external refining, and modification treatment of the molten steel by lanthanum. The samples were first underwent normalizing treatment at 900 °C for 10 hours, then air cooled, followed by tempering treatment at 600 °C for 7 hours and samples were air cooled again. The mechanical properties and microstructures of the samples were measured. The crystallography relationships between lanthanum compounds formed in the molten steel and primary δ-Fe were analysed. The nucleation effect of lanthanum compounds as a heterogeneous nucleation core of primary δ-Fe were calculated and discussed based on two-dimensional mismatch theory. The results indicated that the strip MnS inclusions in ASTM A216 steel became granular rare earth compound inclusions due to La. The refined microstructures were obtained by a synergistic effect of the enhanced condensate depression and the nucleation rate of melt and La compounds as the heterogeneous nucleation caused by La.Da bi se zadovoljili standardi Međunarodnog udruženja klasifikacijskih zavoda (IACS) za lijevanje krmenog roga, potrebno je poboljšati svojstva čelika ASTM A216. Odljevci krmenog roda i prateći uzorci, nakon taljenja čelika u elektrolučnoj peći, rafiniranja i dodatka lantana, izrađeni su lijevanjem čelika u kalupe. Uzorci su prvo normalizirani zagrijavanjem 10 sati na 900 °C, ohlađeni na zraku, potom podvrgnuti popuštanju 7 sati na 600 °C i ponovno ohlađeni na zraku. Određena su mehanička svojstva uzoraka i proučena mikrostruktura. Analizirana je sličnost kristalne građe spojeva lantana nastalih u rastaljenom čeliku i δ-željeza. Izračunata je učinkovitost lantanovih spojeva kao podloge za heterogenu nukleaciju δ-željeza na temelju dvodimenzijskog nepodudaranja struktura. Dodatkom lantana vrpčasti uklopci MnS postaju zrnasti uklopci spoja rijetke zemlje. Finija mikrostruktura dobivena je sinergijskim djelovanjem povećane depresije kondenzata i brže nukleacije uz lantan

Effects of Changes in the Levels of Damage-Associated Molecular Patterns Following Continuous Veno–Venous Hemofiltration Therapy on Outcomes in Acute Kidney Injury Patients With Sepsis

Background: We investigated the association of damage-associated molecular pattern (DAMP) removal with mortality in sepsis patients undergoing continuous veno–venous hemofiltration (CVVH).Methods: Circulating levels of DAMPs [mitochondrial DNA (mtDNA); nuclear DNA (nDNA); heat shock protein 70 (HSP70); and high mobility group box 1 (HMGB1)] and cytokines were measured at baseline, 6 and 12 h after initiation of CVVH. Urinary DNA levels were analyzed at baseline and end of CVVH. The expression of human leukocyte antigen (HLA)-DR was assayed at 0, 3, and 7 days after initiation of CVVH. Moreover, the effects of HSP70 and HMGB1 clearance on survival were analyzed.Results: We evaluated 43 patients with acute kidney injury (AKI) (33 sepsis patients). Twenty-two sepsis patients (67%) and three non-sepsis patients (30%) expired (P = 0.046). Significant reductions in the levels of circulating interleukin-6 (P = 0.046) and tumor necrosis factor-α (P = 0.008) were found in the sepsis group. The levels of mtDNA were increased (ND2, P = 0.035; D-loop, P = 0.003), whereas that of HSP70 was reduced (P = 0.000) in all patients during the first 12 h. The levels of DAMPs in the plasma were markedly increased after blood passage from the inlet through the dialyzer in survivor sepsis patients. The clearance rates of HSP70 and HMGB1 were good predictors of mortality [area under the curve (AUC) = 0.937, P = 0.000; AUC = 0.90, P = 0.001, respectively]. The level of HLA-DR was increased in response to higher HSP70 clearance (P = 0.006). Survival was significantly worse in groups with higher clearance rates of HSP70 and HMGB1 than the cut-off value (log-rank test: P = 0.000 for both). Higher HSP70 clearance was a significant independent predictor of mortality (odds ratio = 1.025, 95% confidence interval [CI]: 1.012–1.039, P = 0.000). The urinary nDNA (β-globin) level before CVVH was an independent risk factor for the duration of CVVH in patients with sepsis (sRE = 0.460, 95% CI: 1.720–8.857, P = 0.005).Conclusion: CVVH removes inflammatory factors, reduces urinary DAMPs, and removes plasma DAMPs. However, survival decreases in response to higher HSP70 clearance

Dual regulatory switch through interactions of Tcf7l2/Tcf4 with stage-specific partners propels oligodendroglial maturation

Constitutive activation of Wnt/β-catenin inhibits oligodendrocyte myelination. Tcf7l2/Tcf4, a β-catenin transcriptional partner, is required for oligodendrocyte differentiation. How Tcf7l2 modifies β-catenin signalling and controls myelination remains elusive. Here we define a stage-specific Tcf7l2-regulated transcriptional circuitry in initiating and sustaining oligodendrocyte differentiation. Multistage genome occupancy analyses reveal that Tcf7l2 serially cooperates with distinct co-regulators to control oligodendrocyte lineage progression. At the differentiation onset, Tcf7l2 interacts with a transcriptional co-repressor Kaiso/Zbtb33 to block β-catenin signalling. During oligodendrocyte maturation, Tcf7l2 recruits and cooperates with Sox10 to promote myelination. In that context, Tcf7l2 directly activates cholesterol biosynthesis genes and cholesterol supplementation partially rescues oligodendrocyte differentiation defects in Tcf712 mutants. Together, we identify stage-specific co-regulators Kaiso and Sox10 that sequentially interact with Tcf7l2 to coordinate the switch at the transitions of differentiation initiation and maturation during oligodendrocyte development, and point to a previously unrecognized role of Tcf7l2 in control of cholesterol biosynthesis for CNS myelinogenesis

5-FU-hydrogel inhibits colorectal peritoneal carcinomatosis and tumor growth in mice

<p>Abstract</p> <p>Background</p> <p>Colorectal peritoneal carcinomatosis (CRPC) is a common form of systemic metastasis of intra-abdominal cancers. Intraperitoneal chemotherapy is a preferable option for colorectal cancer. Here we reported that a new system, 5-FU-loaded hydrogel system, can improve the therapeutic effects of intraperitoneal chemotherapy.</p> <p>Methods</p> <p>A biodegradable PEG-PCL-PEG (PECE) triblock copolymer was successfully synthesized. The biodegradable and temperature sensitive hydrogel was developed to load 5-FU. Methylene blue-loaded hydrogel were also developed for visible observation of the drug release. The effects and toxicity of the 5-FU-hydrogel system were evaluated in a murine CRPC model.</p> <p>Results</p> <p>The hydrogel system is an injectable flowing solution at ambient temperature and forms a non-flowing gel depot at physiological temperature. 5-FU-hydrogel was subsequently injected into abdominal cavity in mice with CT26 cancer cells peritoneal dissemination. The results showed that the hydrogel delivery system prolonged the release of methylene blue; the 5-FU-hydrogel significantly inhibited the peritoneal dissemination and growth of CT26 cells. Furthermore, intraperitoneal administration of the 5-FU-hydrogel was well tolerated and showed less hematologic toxicity.</p> <p>Conclusions</p> <p>Our data indicate that the 5-FU-hydrogel system can be considered as a new strategy for peritoneal carcinomatosis, and the hydrogel may provide a potential delivery system to load different chemotherapeutic drugs for peritoneal carcinomatosis of cancers.</p

Spatio-Temporal Analysis of Ecological Vulnerability and Driving Factor Analysis in the Dongjiang River Basin, China, in the Recent 20 Years

The global ecological environment faces many challenges. Landsat thematic mapper time-series, digital elevation models, meteorology, soil types, net primary production data, socio-economic data, and auxiliary data were collected in order to construct a comprehensive evaluation system for ecological vulnerability (EV) using multi-source remote sensing data. EV was divided into five vulnerability levels: potential I, slight II, mild III, moderate IV, and severe V. Then, we analyzed and explored the spatio-temporal patterns and driving mechanisms of EV in the region over the past 20 years. Our research results showed that, from 2001 to 2019, the DRB was generally characterized as being in the severe vulnerability class, with higher upstream and downstream EV classes and a certain amount of reduction in the midstream EV classes. Moreover, EV in the DRB continues to decrease. The spatio-temporal EV patterns in the DRB were significantly influenced by the relative humidity, average annual temperature, and vegetation cover over the past 20 years. Our work can provide a basis for decision-making and technical support for ecosystem protection, ecological restoration, and ecological management in the DRB