Peptide-Driven Proton Sponge Nano-Assembly for Imaging and Triggering Lysosome-Regulated Immunogenic Cancer Cell Death.

Triggering lysosome-regulated immunogenic cell death (ICD, e.g., pyroptosis and necroptosis) with nanomedicines is an emerging approach for turning an "immune-cold" tumor "hot"-a key challenge faced by cancer immunotherapies. Proton sponge such as high-molecular-weight branched polyethylenimine (PEI) is excellent at rupturing lysosomes, but its therapeutic application is hindered by uncontrollable toxicity due to fixed charge density and poor understanding of resulted cell death mechanism. Here, a series of proton sponge nano-assemblies (PSNAs) with self-assembly controllable surface charge density and cell cytotoxicity are created. Such PSNAs are constructed via low-molecular-weight branched PEI covalently bound to self-assembling peptides carrying tetraphenylethene pyridinium (PyTPE, an aggregation-induced emission-based luminogen). Assembly of PEI assisted by the self-assembling peptide-PyTPE leads to enhanced surface positive charges and cell cytotoxicity of PSNA. The self-assembly tendency of PSNAs is further optimized by tuning hydrophilic and hydrophobic components within the peptide, thus resulting in the PSNA with the highest fluorescence, positive surface charge density, cell uptake, and cancer cell cytotoxicity. Systematic cell death mechanistic studies reveal that the lysosome rupturing-regulated pyroptosis and necroptosis are at least two causes of cell death. Tumor cells undergoing PSNA-triggered ICD activate immune cells, suggesting the great potential of PSNAs to trigger anticancer immunity.

A novel severe cerebral venous thrombosis rat model based on semi-ligation combined with ferric chloride and thrombin.

AIMS: An applicable cerebral venous sinus thrombosis (CVST) model is imperative for exploring its pathophysiology. We established a novel severe CVST model using semi-ligation, ferric chloride, and thrombin. METHODS: A total of 138 male Sprague-Dawley rats were randomly divided into semi-ligation (n = 75) and non-semi-ligation (n = 63) groups. A sham group (n = 46) was also included. We compared short-term and long-term neurological and cognitive dysfunction, mortality rates, thrombus load, venous infarction volume, the blood-brain barrier permeability, brain water content, and microglia activation among the three groups. RESULTS: Thrombi involving multiple venous sinuses appeared in all semi-ligation rats within 2 days postoperatively. Compared with the non-semi-ligation group, short-term and long-term neurological dysfunction were more severe (p < 0.05), and thrombus weight, venous infarction volumes, and microglia activation were more significant (p < 0.05) in the semi-ligation group. Further, the cognitive function of the semi-ligation group significantly decreased (p < 0.05) on postoperative day 21. Cumulative mortality rates between the semi-ligation and non-semi-ligation groups did not differ significantly. CONCLUSION: Semi-ligation combined with ferric chloride and thrombin can produce a severe CVST model with multiple venous sinus involvement, which is suitable for short- and long-term neurological and cognitive dysfunction assessment.

Glioma Imaging by O-(2-18F-Fluoroethyl)-L-Tyrosine PET and Diffusion-Weighted MRI and Correlation With Molecular Phenotypes, Validated by PET/MR-Guided Biopsies.

Gliomas exhibit high intra-tumoral histological and molecular heterogeneity. Introducing stereotactic biopsy, we achieved a superior molecular analysis of glioma using O-(2-18F-fluoroethyl)-L-tyrosine (FET)-positron emission tomography (PET) and diffusion-weighted magnetic resonance imaging (DWI). Patients underwent simultaneous DWI and FET-PET scans. Correlations between biopsy-derived tumor tissue values, such as the tumor-to-background ratio (TBR) and apparent diffusion coefficient (ADC)/exponential ADC (eADC) and histopathological diagnoses and those between relevant genes and TBR and ADC values were determined. Tumor regions with human telomerase reverse transcriptase (hTERT) mutation had higher TBR and lower ADC values. Tumor protein P53 mutation correlated with lower TBR and higher ADC values. α-thalassemia/mental-retardation-syndrome-X-linked gene (ATRX) correlated with higher ADC values. 1p/19q codeletion and epidermal growth factor receptor (EGFR) mutations correlated with lower ADC values. Isocitrate dehydrogenase 1 (IDH1) mutations correlated with higher TBRmean values. No correlation existed between TBRmax/TBRmean/ADC/eADC values and phosphatase and tensin homolog mutations (PTEN) or O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation. Furthermore, TBR/ADC combination had a higher diagnostic accuracy than each single imaging method for high-grade and IDH1-, hTERT-, and EGFR-mutated gliomas. This is the first study establishing the accurate diagnostic criteria for glioma based on FET-PET and DWI.

RELA Fusion in Supratentorial Extraventricular Ependymomas: A Morphologic, Immunohistochemical, and Molecular Study of 43 Cases.

Supratentorial extraventricular ependymomas (STEEs) are relatively rare ependymomas, and their pathologic and genetic characteristics are still poorly understood. The aim of this study was to determine the histologic, immunohistochemical, and RELA fusion features, as well as to clarify in more detail the clinical courses of STEEs. Data from a total of 43 patients with STEEs was analyzed retrospectively. The status of RELA fusion was evaluated using fluorescence in situ hybridization. The expression levels of L1CAM, p65, cyclin D1, and p53 were assessed using immunohistochemistry. Progression-free survival and overall survival were calculated via Kaplan-Meier estimation using the log-rank test. Among all 43 STEEs, 65.1% (28/43) are positive for RELA fusion. Interestingly, almost half of the patients with RELA fusion-positive ependymomas are adults (13/28), and 89.3% (25/28) cases are anaplastic ependymomas, which suggests that RELA fusion testing is necessary in adults with STEEs. We investigated the immunohistochemical status of p65, L1CAM and CCND1 protein expression for their ability to predict RELA fusion status. RELA fusion-positive STEEs are frequently associated with expression of p65 (85.2%), L1CAM (85.2%), and CCND1 (81.5%). The accuracy of predicting RELA fusion status was much higher when the expression of p65 and L1CAM was combined, that is, when both were immunopositive. The status of RELA fusion, p53 overexpression, and extent of tumor resection are significantly associated with prognosis.

H3 K27M-mutant diffuse midline gliomas in different anatomical locations.

The histone H3 K27M mutation has been frequently reported in most diffuse midline gliomas. However, the relationship between the H3 K27M mutation and clinical outcomes of gliomas from different anatomical locations is still not fully understood. A total of 120 patients with diffuse midline gliomas were selected for this retrospective observational study. The status of H3 K27M, ATRX, TP53, and IDH was evaluated using immunohistochemistry and Sanger sequencing. Of the 120 patients aged from 4 to 76 years (median, 27 years), 61 (50.8%) were harboring the H3 K27M mutation. Tumors were mainly located in the brainstem, ATRX thalamus, and spinal cord, but also in the cerebellum, corpus callosum, and the lateral ventricle. Patients with H3 K27M-mutant diffuse midline gliomas had a significantly shorter overall survival than H3 wild-type counterparts (P = .001). However, the H3 K27M mutation was mainly associated with a poorer prognosis in infratentorial gliomas compared with the corresponding H3 wild-type gliomas (P < .0001), but not in supratentorial gliomas (P = .603). Moreover, patients with H3 K27M-mutant gliomas in unusual anatomical locations had a better prognosis than did those with corresponding tumors in the brainstem. This study may provide guidance for better treatment stratification decisions for diffuse gliomas bearing the H3 K27M mutation, which arise in different locations of the brainstem, thalamus, spinal cord, or other sites.

NHC Pd(II) and Ag(I) Complexes: Synthesis, Structure, and Catalytic Activity in Three Types of C-C Coupling Reactions.

Four bis-benzimidazolium salts, 1,4-bis[1'-(N-R-benzimidazoliumyl)methyl]-2,3,5,6-tetramethylbenzene 2X- (L 1 H 2 ·(PF 6 ) 2 : R = ethyl, X = PF6; L 2 H 2 ·Br 2 : R = picolyl, X = Br; L 3 H 2 ·Br 2 : R = benzyl, X = Br; and L 4 H 2 ·Br 2 : R = allyl, X = Br), and their three N-heterocyclic carbene (NHC) Pd(II) and Ag(I) complexes, L 1 Pd 2 Cl 4 (1), L 2 Ag 2 Br 2 (2), and L 4 (AgBr) 2 (3), as well as one anionic complex L 3 H 2 ·(Ag 4 Br 8 ) 0.5 (4), have been synthesized and characterized. Complex 1 adopts a funnel-like type of structure, complex 2 adopts a cyclic structure, and complex 3 is an open structure. In the crystal packing of 1-4, one-dimensional polymeric chains and two-dimensional supramolecular layers are formed via intermolecular weak interactions, including hydrogen bonds, π-π interactions, and C-H···π contacts. The catalytic activities of NHC Pd(II) complex 1 in three types of C-C coupling reactions (Suzuki-Miyaura, Heck-Mizoroki, and Sonogashira reactions) were studied. The results show that this catalytic system is efficient for these C-C coupling reactions.

Comparative assessment of three methods to analyze MGMT methylation status in a series of 350 gliomas and gangliogliomas.

MGMT promoter methylation is considered as a prognostic and predictive biomarker indicating response to chemotherapy and radiotherapy in glioblastoma. A number of different methods and platforms including pyrosequencing (PSQ), quantitative methylation-specific PCR (qMSP) and immunohistochemistry (IHC), methylation-sensitive high resolution melting (MS-HRM) and NGS (Next Generation Sequencing) have been used to detect MGMT promoter methylation in gliomas. However, controversy remains about the most appropriate method to use for analyzing MGMT status. The MGMT promoter methylation status of a total of 350 gliomas and gangliogliomas was examined using PSQ, qMSP and IHC in parallel. Using PSQ as a recommended standard method, the sensitivity, specificity, positive/negative predictive value and correlation with the other assays were calculated. Among 350 glioma and ganglioglioma cases, the MGMT promoter tested positive for methylation in 53.1%, 55.4%, and 70.3% of the cases by PSQ, qMSP and IHC, respectively. The sensitivity and specificity of qMSP were 97.8% and 92.7%, respectively. Twelve cases that tested positive for methylation using qMSP were negative according to PSQ, and four cases that were negative according to qMSP tested positive according to PSQ. The concordance rate between PSQ and qMSP was 90.8%. The sensitivity and specificity of IHC for the detection of MGMT at the protein level were 84.4% and 45.7%, respectively. The concordance rate between PSQ and IHC was 30.8%. This study demonstrated that qMSP is an effective and rapid detection method for routine use in pathology laboratories for the identification of MGMT promoter methylation. A combination of IHC and qMSP assays can provide high sensitivity and specificity for the prediction of MGMT status. A few cases that tested negative with PSQ did harbor MGMT promoter methylation, as confirmed by qMSP and sequencing, and this subgroup of patients may benefit from temozolomide.

Synthesis and structural studies of N-heterocyclic carbene Ag(I) and Hg(II) complexes and recognition of dihydrogen phosphate anion.

Bis-benzimidazolium salt (S)-2,2'-bis[2″-(N-Et-benzimidazoliumyl)ethoxy]-1,1'-binaphthyl hexafluorophosphate [(S)-L1H2]·(PF6)2 and bis-imidazolium salts (S)-2,2'-bis[2″-(N-R-imidazoliumyl)ethoxy]-1,1'-binaphthyl hexafluorophosphate [(S)-L2H2]·(PF6)2 and [(S)-L3H2]·(PF6)2 (R = ethyl or benzyl), as well as their five N-heterocyclic carbene Hg(II) and Ag(I) complexes such as [(S)-L1Hg(HgBr4)] (1), [(S)-L2Hg(HgBr4)] (2), [(S)-L2Hg(HgI4)] (3), {[(S)-L2Ag](PF6)}n (4) and [(S)-L3Ag](PF6) (5) have been prepared and characterized. Each of complexes 1-3 consists of two rings (one 6-membered ring and one 11-membered ring), in which the oxygen atom in the ligand participates in coordination with Hg(II) ion. In complex 4, 1D helical polymeric chain is formed via biscarbene ligand (S)-L2 and Ag(I) ion. A 15-membered macrometallocycle is constructed through a ligand (S)-L3 and a Ag(I) ion in complex 5. Additionally, the selective recognition of H2PO4- using complex 5 as a receptor was investigated on the basis of fluorescence and UV/vis spectroscopic titrations. The results indicate that complex 5 can distinguish effectively H2PO4- from other anions.

[Correlation of chromosome 1p and 19q status and expression of R132H mutant IDH1 protein in oligodendroglial tumors].

OBJECTIVE: To correlate the presence of chromosome 1p/19q deletion with the expression of R132H mutant IDH1 status in oligodendroglial tumors, and to explore molecular markers for predicting chemosensitivity of oligodendroglial tumors. METHODS: The study included 75 oligodendroglial tumors (38 oligodendrogliomas and 37 oligoastrocytomas). Immunohistochemistry was used to detect the expression of R132H mutant IDH1 protein, and fluorescence in situ hybridization (FISH) was employed to detect 1p/19q deletion. RESULTS: Deletion of chromosome 1p and/or 19q was detected in 37 cases (37/75, 49.3%), among which co-deletion of 1p and 19q was seen in 34 cases (closely correlated, P < 0.01). Oligodendrogliomas WHOIIhad a slightly higher deletion rate than oligodendrogliomas WHO III, although without statistical significance. Oligodendrogliomas WHO IIand WHO III had a significantly higher deletion rate of chromosome 1p/19q than oligoastrocytomas WHO II and WHO III (P < 0.05). While combined loss of 1p/19q was always detected in oligodendrogliomas when FISH was positive, isolated 1p or 19q deletion was only found in oligoastrocytomas. The expression of R132H mutant IDH1 was detected in 51 of 75 cases (68.0%), in which oligodendrogliomas had a higher positive rate than oligoastrocytomas. Statistical analysis demonstrated a significant correlation between the expression of R132H mutant IDH1 protein and the presence of combined 1p/19q deletion in oligodendrogliomas (P < 0.05). CONCLUSIONS: A significant correlation was observed between the expression of R132H mutant protein and 1p/19q LOH.Expression of 132H mutant IDH1 protein is the potential biomarker for predicating the presence of 1p/19q deletion in oligodendrogliomas.