Targeting KRAS Mutant Cancers with a Covalent G12C-Specific Inhibitor.

KRASG12C was recently identified to be potentially druggable by allele-specific covalent targeting of Cys-12 in vicinity to an inducible allosteric switch II pocket (S-IIP). Success of this approach requires active cycling of KRASG12C between its active-GTP and inactive-GDP conformations as accessibility of the S-IIP is restricted only to the GDP-bound state. This strategy proved feasible for inhibiting mutant KRAS in vitro; however, it is uncertain whether this approach would translate to in vivo. Here, we describe structure-based design and identification of ARS-1620, a covalent compound with high potency and selectivity for KRASG12C. ARS-1620 achieves rapid and sustained in vivo target occupancy to induce tumor regression. We use ARS-1620 to dissect oncogenic KRAS dependency and demonstrate that monolayer culture formats significantly underestimate KRAS dependency in vivo. This study provides in vivo evidence that mutant KRAS can be selectively targeted and reveals ARS-1620 as representing a new generation of KRASG12C-specific inhibitors with promising therapeutic potential.

Contribution of structural accessibility to the cooperative relationship of TF-lncRNA in myopia.

Transcriptional regulation is associated with complicated mechanisms including multiple molecular interactions and collaborative drive. Long noncoding RNAs (lncRNAs) have highly structured characteristics and play vital roles in the regulation of transcription in organisms. However, the specific contributions of conformation feature and underlying molecular mechanisms are still unclear. In the present paper, a hypothesis regarding molecular structure effect is presented, which proposes that lncRNAs fold into a complex spatial architecture and act as a skeleton to recruit transcription factors (TF) targeted binding, and which is involved in cooperative regulation. A candidate set of TF-lncRNA coregulation was constructed, and it was found that structural accessibility affected molecular binding force. In addition, transcription factor binding site (TFBS) regions of myopia-related lncRNA transcripts were disturbed, and it was discovered that base mutations affected the occurrence of significant molecular allosteric changes in important elements and variable splicing regions, mediating the onset and development of myopia. The results originated from structureomics and interactionomics and created conditions for systematic research on the mechanisms of structure-mediated TF-lncRNA coregulation in transcriptional regulation. Finally, these findings will help further the understanding of key regulatory roles of molecular allostery in cell physiological and pathological processes.

A direct interaction between CENTLEIN and RABIN8 is required for primary cilium formation.

Primary cilia are formed in nearly all growth-arrested cells and are essential for mammalian development and tissue homeostasis. Defects in primary cilia result in a range of disorders in humans, named ciliopathies. The spatiotemporal localization of RABIN8 on the pericentrosome is an early step in ciliogenesis. Here, we show that CENTLEIN depletion causes the persistent accumulation of RABIN8 on the pericentrosome and primary cilium loss in hTERT-immortalized retinal pigment epithelial cells and murine embryonic fibroblasts. CENTLEIN interacts with RABIN8 directly. A stretch of a 31-amino acid sequence located in the 200‒230 region of the RABIN8 GEF domain is responsible for its physical interaction with CENTLEIN, while expression of the full-length but not the internal deletion lacking the RABIN8-binding site of CENTLEIN largely rescues the ciliogenesis defect provoked by CENTLEIN depletion. Expression of activated RAB8A partially reverses cilium loss in CENTLEIN-null RPE1 cells, so the functional importance of the CENTLEIN-RABIN8 interaction is defined.

Effect of aqueous extract of seed of broccoli on inflammatory cytokines and Helicobacter pylori infection: a randomized, double-blind, controlled trial in patients without atrophic gastritis.

The purpose of this study was to investigate the anti-inflammatory effect of an aqueous extract of seed of broccoli (AESB) in Helicobacter pylori (HP)-infected patients without atrophic gastritis. This was a double-centre, randomized, double-blind, controlled study. A total of 110 HP-infected subjects were randomized to receive either AESB or placebo for 2 months. Inflammatory cytokine (IL-8, IFN-γ, TNF-α, CRP, IL-17A, IL-1ß, IL-18), pepsinogen I, II (PG I, PG II), and gastrin-17 (G-17) measurements and 13C-urea breath tests were performed at baseline and at 60 days. At 60 days, there was no significant difference in any of the inflammatory cytokines, pepsinogen or gastrin between the two groups. However, IL-8, IFN-γ, PG I, PG I/PG II ratio (PGR), and G-17 were reduced by 9.02 pg/mL, 5.08 pg/mL, 24.56 ng/mL, 1.75 and 0.3 pmol/L, respectively, in the AESB group compared with baseline (all P < 0.05). The HP eradication rates in the AESB group and placebo group were 11.11 and 3.70% at 60 days, respectively (P > 0.05). No treatment-related adverse events were reported. Thus, AESB may reduce the risk of gastric mucosal lesions and decrease the risk of gastric cancer by relieving inflammatory cytokines. The safety profile of AESB was satisfactory. This study is registered with the Chinese Clinical Trials Registry (Registration No. ChiCTR2100054249).

Selective inhibition of phosphoinositide 3-kinase p110α preserves lymphocyte function.

Class IA phosphoinositide 3-kinase (PI3K) is essential for clonal expansion, differentiation, and effector function of B and T lymphocytes. The p110δ catalytic isoform of PI3K is highly expressed in lymphocytes and plays a prominent role in B and T cell responses. Another class IA PI3K catalytic isoform, p110α, is a promising drug target in cancer but little is known about its function in lymphocytes. Here we used highly selective inhibitors to probe the function of p110α in lymphocyte responses in vitro and in vivo. p110α inhibition partially reduced B cell receptor (BCR)-dependent AKT activation and proliferation, and diminished survival supported by the cytokines BAFF and IL-4. Selective p110δ inhibition suppressed B cell responses much more strongly, yet maximal suppression was achieved by targeting multiple PI3K isoforms. In mouse and human T cells, inhibition of single class IA isoforms had little effect on proliferation, whereas pan-class I inhibition did suppress T cell expansion. In mice, selective p110α inhibition using the investigational agent MLN1117 (previously known as INK1117) did not disrupt the marginal zone B cell compartment and did not block T cell-dependent germinal center formation. In contrast, the selective p110δ inhibitor IC87114 strongly suppressed germinal center formation and reduced marginal zone B cell numbers, similar to a pan-class I inhibitor. These findings show that although acute p110α inhibition partially diminishes AKT activation, selective p110α inhibitors are likely to be less immunosuppressive in vivo compared with p110δ or pan-class I inhibitors.

Sex Differences in the Relationship Between Emotional Support and Self-rated Health among Chinese Elderly.

PURPOSE: This study aimed to explore sex differences in the association between emotional support and self-rated health among the elderly. DESIGN: This was a cross-sectional survey based on the sub-project of China's National Basic Public Health Service Project-Health Management Services for the Elderly. SETTING: Participants were recruited from ten rural townships in Jingyuan County, Gansu Province, Northwestern China. SUBJECTS: 1405 subjects aged 60 or above. METHODS: Emotional support (consisting of 5 items) and self-rated health (evaluated by EQ-VAS) were investigated in this study. Multiple linear regression was conducted to consider the potential relationship. RESULTS: The frequency of children visit and the number of providers of emotional support were positively associated with self-rated health among older women (ß = 1.13, 95%CI = 0.25-2.02; ß = 1.80, 95%CI = 1.01-2.58), whereas the number of close friends had a positive association with self-rated health among older men (ß = 1.11, 95%CI = 0.20-2.01). The number of close relatives and the frequency of seeking emotional support were not found to be associated with self-rated health among both older men and older women. CONCLUSION: The study has found that the relationship between emotional support and self-rated health was differed by sex, calling attention to the need for sex-specific interventions.

Trace Sc3+-electrolyte additive enabling stable Zn metal anodes for aqueous zinc-ion batteries.

Trace Sc3+ additive (1.0 mol%) is shown to greatly improve the Coulombic efficiency and cycling stability of Zn metal anodes in aqueous ZnSO4 electrolyte due to the decreased nucleation overpotential and increased kinetics for Zn plating/stripping. Both Zn‖Zn and Zn‖V2O5 cells show enhanced cycling stability and rate capability in the Sc3+-modified electrolyte.

A Thin Composite Polymer Electrolyte Functionalized by a Novel Antihydrolysis Additive to Enable All-Solid-State Lithium Battery with Excellent Rate and Cycle Performance.

Composite solid-state electrolyte (CSE) incorporated with fluorine-containing functional additives usually endows the assembled cell with improved electrochemical performance by forming stable electrode/electrolyte interfaces. However, most of fluorine-containing additives are prone to hydrolysis, which is not suitable for the large-scale preparation of CSEs. In this work, an antihydrolysis and fluorine-containing additive of magnesium 2,3,4,5,6-pentafluorophenylacetate (MgPFPAA) is successfully synthesized and then used to regulate the properties of the electrode/electrolyte interfaces of the all-solid-state lithium batteries (ASSLBs). The antihydrolysis property of MgPFPAA facilitates the large-scale preparation of the ultrathin CSEs in atmospheric environment. Both theoretical calculations and experimental results indicate that MgPFPAA can effectively improve the composition and structure of the generated solid electrolyte interface film by providing rich F sources and Mg2+ , thus leading to a stable CSE/Li interface. Furthermore, an ultrathin PEO/PVDF-based CSE (≈30 µm) functionalized by this novel MgPFPAA additive enables the assembled LiFePO4 -based ASSLB with greatly enhanced electrochemical performances, with high discharge specific capacity of 93.7 mAh g-1 at 10 C and a high capacity retention of 74.9% after 1500 cycles at 5.0 C. Also, this MgPFPAA functionalized CSE can be compatible with the high-areal-capacity LiFePO4 and the high-voltage LiNi0.8 Co0.1 Mn0.1 O2 cathodes.

Detector-grade perovskite single-crystal wafers via stress-free gel-confined solution growth targeting high-resolution ionizing radiation detection.

Solution-processed organic‒inorganic halide perovskite (OIHP) single crystals (SCs) have demonstrated great potential in ionizing radiation detection due to their outstanding charge transport properties and low-cost preparation. However, the energy resolution (ER) and stability of OIHP detectors still lag far behind those of melt-grown inorganic perovskite and commercial CdZnTe counterparts due to the absence of detector-grade high-quality OIHP SCs. Here, we reveal that the crystallinity and uniformity of OIHP SCs are drastically improved by relieving interfacial stress with a facial gel-confined solution growth strategy, thus enabling the direct preparation of large-area detector-grade SC wafers up to 4 cm with drastically suppressed electronic and ionic defects. The resultant radiation detectors show both a small dark current below 1 nA and excellent baseline stability of 4.0 × 10-8 nA cm-1 s-1 V-1, which are rarely realized in OIHP detectors. Consequently, a record high ER of 4.9% at 59.5 keV is achieved under a standard 241Am gamma-ray source with an ultralow operating bias of 5 V, representing the best gamma-ray spectroscopy performance among all solution-processed semiconductor radiation detectors ever reported.

Simulations to Assess the Performance of Multifactor Risk Scores for Predicting Myopia Prevalence in Children and Adolescents in China.

Background: Myopia is the most common visual impairment among Chinese children and adolescents. The purpose of this study is to explore key interventions for myopia prevalence, especially for early-onset myopia and high myopia. Methods: Univariate and multivariate analyses were conducted to evaluate potential associations between risk factor exposure and myopia. LASSO was performed to prioritize the risk features, and the selected leading factors were used to establish the assembled simulation model. Finally, two forecasting models were constructed to predict the risk of myopia and high myopia. Results: Children and adolescents with persistently incorrect posture had a high risk of myopia (OR 7.205, 95% CI 5.999-8.652), which was 2.8 times higher than that in students who always maintained correct posture. In the cohort with high myopia, sleep time of less than 7 h per day (OR 9.789, 95% CI 6.865-13.958), incorrect sitting posture (OR 8.975, 95% CI 5.339-15.086), and siblings with spherical equivalent <-6.00 D (OR 8.439, 95% CI 5.420-13.142) were the top three risk factors. The AUCs of integrated simulation models for myopia and high myopia were 0.8716 and 0.8191, respectively. Conclusion: The findings illustrate that keeping incorrect posture is the leading risk factor for myopia onset, while the onset age of myopia is the primary factor affecting high myopia progression. The age between 8 and 12 years is the crucial stage for clinical intervention, especially for children with parental myopia.

[Impact of family members and health care providers on the use of folic acid in pregnant women].

OBJECTIVE: To investigate the impact of family members and health care providers on the use of folic acid supplements in pregnant women, and to provide basic data for improving the effectiveness of folic acid intervention. METHODS: A cross-sectional study was conducted in hospitals and households from June to September in 2009. Face-to-face anonymous questionnaires were distributed to 2094 women, who were pregnant at least three months or postpartum in one year, in two counties of Gansu Province. RESULTS: The awareness rate of folic acid was in 62.2% of 2094 pregnant women, and 25.4% of them have taken folic acid. Higher knowledge about folic acid of family members (OR = 0.268, 95% CI 0.208 - 0.346), agreed with taking folic acid by family members (OR = 0.103, 95% CI 0.031 -0.338), and urging pregnant women to take folic acid by family members (OR = 0.147, 95% CI 0.115 - 0.190) were significant predictors for having folic acid taken by pregnant women. Propagating knowledge related to folic acid (OR = 0.252, 95% CI 0.197 - 0.323) and directing pregnant women to use folic acid (OR = 0.168, 95% CI 0.096 - 0.296) by health care providers were also the important predictors for folic acid intake. CONCLUSION: Family members and health care providers play an important role in affecting the use of folic acid among pregnant women. In order to improve the effectiveness of intervention with folic acid, family members of pregnant women and health care providers should be included into the target population to receive an intensive propaganda campaign on folic acid education to improve the use of folic acid in pregnant women extensively.

Cyclodextrin-Integrated PEO-Based Composite Solid Electrolytes for High-Rate and Ultrastable All-Solid-State Lithium Batteries.

Poly(ethylene oxide) (PEO)-based composite solid electrolytes (CSEs) are considered as one of the most promising candidates for all-solid-state lithium batteries (ASSLBs). However, a key challenge for their further development is to solve the main issues of low ionic conductivity and poor mechanical strength, which can lead to insufficient capacity and stability. Herein, ß-cyclodextrin (ß-CD) is first demonstrated as a multifunctional filler that can form a continuous hydrogen bond network with the ether oxygen unit from the PEO matrix, thus improving the comprehensive performances of the PEO-based CSE. By relevant characterizations, it is demonstrated that ß-CD is uniformly dispersed into the PEO substrate, inducing adequate dissociation of lithium salt and enhancing mechanical strength through hydrogen bond interactions. In a Li/Li symmetric battery, the ß-CD-integrated PEO-based (PEO-LiTFSI-15% ß-CD) CSE works well at a critical current density up to 1.0 mA cm-2 and retains stable lithium plating/stripping for more than 1000 h. Such reliable properties also enable its superior performance in LiFePO4-based ASSLBs, with specific capacities of 123.6 and 114.0 mA h g-1 as well as about 100 and 81.8% capacity retention over 300 and 700 cycles at 1 and 2 C (1 C = 170 mA g-1), respectively.

The missing linker between SUN5 and PMFBP1 in sperm head-tail coupling apparatus.

The sperm head-to-tail coupling apparatus (HTCA) ensures sperm head-tail integrity while defective HTCA causes acephalic spermatozoa, rendering males infertile. Here, we show that CENTLEIN is indispensable for HTCA integrity and function, and that inactivation of CENTLEIN in mice leads to sperm decapitation and male sterility. We demonstrate that CENTLEIN directly interacts with both SUN5 and PMFBP1, two proteins localized in the HTCA and related with acephalic spermatozoa syndrome. We find that the absence of Centlein sets SUN5 and PMFBP1 apart, the former close to the sperm head and the latter in the decapitated tail. We show that lack of Sun5 results in CENTLEIN and PMFBP1 left in the decapitated tail, while disruption of Pmfbp1 results in SUN5 and CENTLEIN left on the detached sperm head. These results demonstrate that CENTLEIN cooperating with SUN5 and PMFBP1 participates in the HTCA assembly and integration of sperm head to the tail, indicating that impairments of CENTLEIN might be associated with acephalic spermatozoa syndrome in humans.

[Factors affecting the use of folic acid supplements in pregnant women in Gansu Province].

OBJECTIVE: To study the factors of folic acid intake among pregnant women, and to provide the basic dates for improving the effectiveness of folic acid intervention. METHODS: A cross-section study was conducted both in hospital and household. Anonymous questionnaires were distributed to 2094 women in two counties of Gansu province, who were pregnant at least three months or postpartum within one year. RESULTS: Only 25.4% of 2094 subjects have taken folic acid. Lack of knowledge on folic acid, the age of pregnant women, the history of birth defects, had a check in hospital before pregnant, the degree of education, planned pregnancy and the history of miscarriages were the significant influential factors for whether or not taking folic acid. CONCLUSION: The history of childbearing and the knowledge on folic are the important factors affecting the use of folic acid supplements. Widely initiating health education on the knowledge of folic acid and advocating family planning in childbearing aged population should be taken as the most important measures. But finding a way of effectively increasing the rate of folic acid intake before pregnancy is a problem still needs to be solved.

Menin inhibitor MI-3454 induces remission in MLL1-rearranged and NPM1-mutated models of leukemia.

The protein-protein interaction between menin and mixed lineage leukemia 1 (MLL1) plays a critical role in acute leukemias with translocations of the MLL1 gene or with mutations in the nucleophosmin 1 (NPM1) gene. As a step toward clinical translation of menin-MLL1 inhibitors, we report development of MI-3454, a highly potent and orally bioavailable inhibitor of the menin-MLL1 interaction. MI-3454 profoundly inhibited proliferation and induced differentiation in acute leukemia cells and primary patient samples with MLL1 translocations or NPM1 mutations. When applied as a single agent, MI-3454 induced complete remission or regression of leukemia in mouse models of MLL1-rearranged or NPM1-mutated leukemia, including patient-derived xenograft models, through downregulation of key genes involved in leukemogenesis. We also identified MEIS1 as a potential pharmacodynamic biomarker of treatment response with MI-3454 in leukemia, and demonstrated that this compound is well tolerated and did not impair normal hematopoiesis in mice. Overall, this study demonstrates, for the first time to our knowledge, profound activity of the menin-MLL1 inhibitor as a single agent in clinically relevant PDX models of leukemia. These data provide a strong rationale for clinical translation of MI-3454 or its analogs for leukemia patients with MLL1 rearrangements or NPM1 mutations.

5,5'- and 6,6'-dialkyl-5,6-dihydro-1H-pyridin-2-ones as potent inhibitors of HCV NS5B polymerase.

The discovery of 5,5'- and 6,6'-dialkyl-5,6-dihydro-1H-pyridin-2-ones as potent inhibitors of the HCV RNA-dependent RNA polymerase (NS5B) is described. Several of these agents also display potent antiviral activity in cell culture experiments (EC50 <0.10 microM). In vitro DMPK data for selected compounds as well as crystal structures of representative inhibitors complexed with the NS5B protein are also disclosed.

5,6-Dihydro-1H-pyridin-2-ones as potent inhibitors of HCV NS5B polymerase.

5,6-Dihydro-1H-pyridin-2-one analogs were discovered as a novel class of inhibitors of genotype 1 HCV NS5B polymerase. Among these, compound 4ad displayed potent inhibitory activities in biochemical and replicon assays (IC(50) (1b)<10nM; IC(50) (1a)<25nM, EC(50) (1b)=16nM), good in vitro DMPK properties, as well as moderate oral bioavailability in monkeys (F=24%).

Discovery of tricyclic 5,6-dihydro-1H-pyridin-2-ones as novel, potent, and orally bioavailable inhibitors of HCV NS5B polymerase.

A novel series of non-nucleoside small molecules containing a tricyclic dihydropyridinone structural motif was identified as potent HCV NS5B polymerase inhibitors. Driven by structure-based design and building on our previous efforts in related series of molecules, we undertook extensive SAR studies, in which we identified a number of metabolically stable and very potent compounds in genotype 1a and 1b replicon assays. This work culminated in the discovery of several inhibitors, which combined potent in vitro antiviral activity against both 1a and 1b genotypes, metabolic stability, good oral bioavailability, and high C(12) (PO)/EC(50) ratios.

Studies toward the duocarmycin prodrugs for the antibody prodrug therapy approach.

A tricyclic precursor for the synthesis of the prodrugs of pro-1,2,9,9a-tetrahydrocyclopropa[c]benz-[e]indole-4-one tetramethoxyindolecarboxamide (CBI-TMI) was prepared using the ring-closing metathesis approach. The tricyclic intermediate was converted to an advanced precursor of a CBI-TMI prodrug equipped with a linker presumably suitable for activation using the aldolase catalytic antibody 38C2. An attempted 38C2-catalyzed two-step activation of the hydroxy-pro-CBI intermediate involving retro-aldol and the ß-elimination reactions was also examined.